Bjarne Bogen

• Lion, Aharon; Israel, Arieh Y.; Merzon, Eugene; Oster, Howard; Weizman, Abraham & Luboshits, Galia [Vis alle 9 forfattere av denne artikkelen] (2026). Simvastatin as an immunomodulator and anti-myeloma agent - epidemiological, in vitro and murine model studies. Frontiers in Immunology. 17, s. 1–11. doi: 10.3389/fimmu.2026.1882291. Fulltekst i vitenarkiv
• Laursen, Marlene Fyrstenberg; Combes, Francis; Gudjonsson, Arnar; Bjerkan, Louise; Parot, Jérémie & Tesfaye, Demo Yemane [Vis alle 14 forfattere av denne artikkelen] (2026). Targeting vaccine fusion proteins to APCs increases immunogenicity of adenoviral and mRNA-LNP vaccines. Molecular Therapy. ISSN 1525-0016. s. 1–18. doi: 10.1016/j.ymthe.2026.06.034. Fulltekst i vitenarkiv Vis sammendrag

  mRNA-lipid nanoparticle (LNP) and adenoviral delivery of vaccines have proven effective during the COVID-19 pandemic. Herein, we explored whether antigen-presenting cell (APC) targeting of antigens could further enhance the immunogenicity of these delivery formats. Experiments were performed in mouse models for malaria (Plasmodium falciparum reticulocyte-binding protein homolog 5 [PfRH5] antigen) and influenza (hemagglutinin [HA] antigen). We used genetic constructs encoding bivalent fusion proteins that target antigens to MHC class II (MHCII) molecules on professional APCs. We demonstrate that MHCII-targeted fusion proteins bound to professional APCs and that such APC-targeting increased antibody and T cell responses as well as protection against the influenza virus. The results suggest that the injected mRNA-LNP and adenoviral vectors resulted in the secretion of fusion proteins that targeted APCs. Employing the APC-targeting principle could enhance the efficiency of adenoviral and mRNA-LNP vaccines against a variety of diseases.

• Werninghaus, Ina Charlotta; Hinke, Daniëla Maria; Katta, Kirankumar; Bogen, Bjarne & Braathen, Ranveig (2026). A combinatorial DNA vaccine encoding APC-targeted, bivalent HA and NA antigens results in an improved influenza vaccine. Journal of Immunology. ISSN 0022-1767. 215(2). doi: 10.1093/jimmun/vkaf378. Fulltekst i vitenarkiv
• Jong, Rineke de; Zhou, Fan; Andersen, Tor Kristian; Stockhofe-Zurwieden, Norbert; Cox, Rebecca Jane & Bogen, Bjarne [Vis alle 7 forfattere av denne artikkelen] (2025). An HLAII-targeted DNA vaccine against influenza H7N9 protected mice and ferrets from a virus challenge. npj Vaccines. 11. doi: 10.1038/s41541-025-01341-4. Fulltekst i vitenarkiv Vis sammendrag

  Due to antigenic drift and shift, influenza A viruses may cause new future pandemics. Currently used seasonal influenza vaccines are of little use against novel viruses with pandemic potential. Genetic vaccines can be rapidly produced and could therefore mitigate pandemic outbreaks. Here, we present preclinical proof of protective efficacy of a DNA vaccine encoding a vaccine protein that targets influenza hemagglutinin (HA) to human leukocyte antigen class II (HLAII) molecules on antigen presenting cells (APC). Vaccination of mice raised robust levels of neutralizing antibodies, and protection against a lethal challenge with influenza H7N1 virus. In ferrets, we observed induction of neutralizing antibodies and T-cell responses after a single vaccination, with levels increasing after a second dose. Protection of vaccinated ferrets against a viral challenge with influenza H7N9 was dose dependent, with ferrets receiving the highest vaccine dose being completely protected from clinical disease. In sum, these results warrant progression to a human clinical Phase I trial.

• Anderson, Ane Marie; Tjärnhage, Elias; Hinke, Daniëla Maria; Braathen, Ranveig; Grødeland, Gunnveig & Bogen, Bjarne (2025). DNA vaccines targeting hemagglutinin from 18 subtypes of influenza A virus to antigen-presenting cells confer broad protection. Molecular Therapy Nucleic Acids. 37(1). doi: 10.1016/j.omtn.2025.102814. Fulltekst i vitenarkiv Vis sammendrag

  Novel vaccines that confer broad protection against influenza A viruses (IAVs) are urgently needed. Hemagglutinin (HA) is the major influenza antigen targeted by protective immune responses. We have here developed a DNA vaccine that simultaneously presents HA from 18 subtypes of IAV to the immune system. The vaccine consists of a DNA plasmid mixture that encodes a variety of dimeric vaccine proteins. Each dimer expresses two different HAs, as well as a targeting moiety directing the vaccine protein to antigen-presenting cells (APCs). When the vaccine proteins were targeted toward chemokine receptors 1, 3, and 5 (CCR1/3/5) on APC by means of macrophage inflammatory protein 1-alpha (MIP1α) (CCL3), vaccinated mice were broadly protected against infection with H1N1, H3N2, H5N1, and H7N1 influenza viruses. Furthermore, antibody-mediated protection against H1N1 was maintained when the H1 antigen was removed from the plasmid mixture, indicating that the diversity of HAs in the mixture promoted formation of antibodies specific for shared, conservative epitopes. The results may guide the development of a broadly protective influenza A vaccine for humans

• Hinke, Daniëla Maria; Anderson, Ane Marie; Katta, Kirankumar; Laursen, Marlene Fyrstenberg; Tesfaye, Demo Yemane & Werninghaus, Ina Charlotta [Vis alle 10 forfattere av denne artikkelen] (2024). Applying valency-based immuno-selection to generate broadly cross-reactive antibodies against influenza hemagglutinins. Nature Communications. 15:850(1), s. 1–17. doi: 10.1038/s41467-024-44889-w. Fulltekst i vitenarkiv Vis sammendrag

  Conserved epitopes shared between virus subtypes are often subdominant, making it difficult to induce broadly reactive antibodies by immunization. Here, we generate a plasmid DNA mix vaccine that encodes protein heterodimers with sixteen different influenza A virus hemagglutinins (HA) representing all HA subtypes except H1 (group 1) and H7 (group 2). Each single heterodimer expresses two different HA subtypes and is targeted to MHC class II on antigen presenting cells (APC). Female mice immunized with the plasmid mix produce antibodies not only against the 16 HA subtypes, but also against non-included H1 and H7. We demonstrate that individual antibody molecules cross-react between different HAs. Furthermore, the mix vaccine induces T cell responses to conserved HA epitopes. Immunized mice are partially protected against H1 viruses. The results show that application of valency-based immuno-selection to diversified antigens can be used to direct antibody responses towards conserved (subdominant) epitopes on viral antigens.

• Ng, Brandon D.; Rajagopalan, Adhithi; Kousa, Anastasia I.; Fischman, Jacob S.; Chen, Sophia & Massa, Alyssa [Vis alle 16 forfattere av denne artikkelen] (2024). IL-18-secreting multiantigen targeting CAR T cells eliminate antigen-low myeloma in an immunocompetent mouse model. Blood. ISSN 0006-4971. 144(2), s. 171–186. doi: 10.1182/blood.2023022293. Fulltekst i vitenarkiv
• Gopalakrishnan, Ramakrishna Prabhu; Østrøm, Marius Sigurdsson; Skjeldal, Frode Miltzow; Bakke, Oddmund; Bogen, Bjarne & Huszthy, Csaba Peter (2024). B Cells With Complementary B Cell Receptors Can Kill Each Other. European Journal of Immunology. ISSN 0014-2980. 55(1), s. 1–13. doi: 10.1002/eji.202350890. Fulltekst i vitenarkiv
• Métivier, Cassandra; Saëc, Patricia Le; Gaschet, Joëlle; Chauvet, Catherine; Marionneau-Lambot, Séverine & Hofgaard, Peter Olaf [Vis alle 16 forfattere av denne artikkelen] (2023). Preclinical Evaluation of a ⁶⁴Cu-Based Theranostic Approach in a Murine Model of Multiple Myeloma. Pharmaceutics. 15:1817(7), s. 1–24. doi: 10.3390/pharmaceutics15071817. Fulltekst i vitenarkiv
• Baert, L.; Manfroi, B.; Quintero, M.; Chavarria, O.; Barbon, P.V. & Clement, E. [Vis alle 14 forfattere av denne artikkelen] (2023). 3-O sulfation of syndecan-1 mediated by the sulfotransferase HS3ST3a1 enhances myeloma aggressiveness. Matrix Biology. ISSN 0945-053X. 120, s. 60–75. doi: 10.1016/j.matbio.2023.05.005. Fulltekst i vitenarkiv Vis sammendrag

  Multiple myeloma is a hematological neoplasm derived from plasma cells invariably developing in the bone marrow (BM). The persisting clinical challenge in MM resides in its high ability to resist drugs as shown by the frequent relapses observed in patients regardless of the treatment applied. In a mouse model of MM, we identified a subpopulation of cells harboring increased resistance to current MM drugs. These cells bound a proliferation inducing ligand (APRIL), a key MM promoting/survival factor. APRIL binding involved the heparan sulfate (HS) chain present on syndecan-1 (SDC-1), and correlated with reactivity to the anti-HS antibody 10e4. 10e4+cells had a high proliferation activity, and were able to form colonies in 3-D cultures. 10e4+ cells were the only cells able to develop in BM after intravenous injection. They also resisted drugs in vivo, since their number increased after treatment in BM. Notably, 10e4+ cells differentiated into 10e4− cells upon in vitro and in vivo expansion. Expression of one sulfotransferase, HS3ST3a1, allowed modification of syndecan-1 to confer reactivity to 10e4 and binding to APRIL. HS3ST3a1 deletion inhibited tumorigenesis in BM. Notably, the two populations coexisted at a variable frequency in the BM of MM patients at diagnosis. In total, our results indicate that 3-O-sulfation on SDC-1 carried out by HS3ST3a1 defines aggressive MM cells, and that targeting of this enzyme could possibly be used to better control drug resistance.

• Westhrin, Marita; Blazevski, Jana; Textor, Ana; Abdollahi, Pegah; Gopalakrishnan, Ramakrishna Prabhu & Ngo, Linda Thuy [Vis alle 13 forfattere av denne artikkelen] (2023). Id-neoantigen vaccine induces therapeutic CD8⁺ T cells against multiple myeloma: H chain-loss escapees cause FLC MM. Journal for ImmunoTherapy of Cancer (JITC). 11:e006944(8), s. 1–13. doi: 10.1136/jitc-2023-006944. Fulltekst i vitenarkiv Vis sammendrag

  Background Multiple myeloma (MM) cancers originate from plasma cells that have passed through the germinal center reaction where somatic hypermutation of Ig V regions takes place. Myeloma protein V regions often express many mutations and are thus a rich source of neoantigens (traditionally called idiotopes (Id)). Therefore, these are highly tumor-specific and excellent targets for immunotherapy. Methods We have developed a DNA Id vaccine which as translated protein targets conventional dendritic cells (cDC) for CCL3-mediated delivery of myeloma protein V regions in a single-chain fragment variable (scFv) format. Vaccine efficacy was studied in the mouse MM model, mineral oil-induced plasmacytoma 315.BM. Results The Id vaccine protected mice against a challenge with MM cells. Moreover, the vaccine had a therapeutic effect. However, in some of the vaccinated mice, MM cells not producing H chains escaped rejection, resulting in free light chain (FLC) MM. Depletion of CD8 + T cells abrogated vaccine efficacy, and protection was observed to be dependent on cDC1s, using Batf3 -/- mice. Modifications of scFv in the vaccine demonstrated that CD8 + T cells were specific for two mutated V H sequences. Conclusions V H neoantigen-specific CD8 + T cells elicited by CCL3-containing Id vaccines had a therapeutic effect against MM in a mouse model. MM cells could escape rejection by losing expression of the H chain, thus giving rise to FLC MM.

• Tjärnhage, Elias; Brown, Diamond; Bogen, Bjarne; Andersen, Tor Kristian & Grødeland, Gunnveig (2023). Trimeric, APC-Targeted Subunit Vaccines Protect Mice against Seasonal and Pandemic Influenza. Journal of Virology. ISSN 0022-538X. 97:e01694-22(2), s. 1–16. doi: 10.1128/jvi.01694-22. Fulltekst i vitenarkiv Vis sammendrag

  Subunit vaccines present only selected viral proteins to the immune system and allow for safe and easy production. Here, we have developed a novel vaccine where influenza hemagglutinin is presented in the natural trimeric form and then steered toward antigen-presenting cells for increased immunogenicity. We demonstrate efficient induction of antibodies and T-cell responses, and demonstrate that the vaccine format can protect mice against influenza subtypes H1N1, H5N1, and H7N1.

• Werninghaus, Ina Charlotta; Hinke, Daniëla Maria; Fossum, Even; Bogen, Bjarne & Braathen, Ranveig (2023). Neuraminidase delivered as an APC-targeted DNA vaccine induces protective antibodies against influenza. Molecular Therapy. ISSN 1525-0016. 31(7), s. 2188–2205. doi: 10.1016/j.ymthe.2023.03.012. Fulltekst i vitenarkiv Vis sammendrag

  Conventional influenza vaccines focus on hemagglutinin (HA). However, antibody responses to neuraminidase (NA) have been established as an independent correlate of protection. Here, we introduced the ectodomain of NA into DNA vaccines that, as translated dimeric vaccine proteins, target antigen-presenting cells (APCs). The targeting was mediated by an single-chain variable fragment specific for major histocompatibility complex (MHC) class II, which is genetically linked to NA via a dimerization motif. A single immunization of BALB/c mice elicited strong and long-lasting NA-specific antibodies that inhibited NA enzymatic activity and reduced viral replication. Vaccine-induced NA immunity completely protected against a homologous influenza virus and out-competed NA immunity induced by a conventional inactivated virus vaccine. The protection was mainly mediated by antibodies, although NA-specific T cells also contributed. APC-targeting and antigen bivalency were crucial for vaccine efficacy. The APC-targeted vaccine was potent at low doses of DNA, indicating a dose-sparing effect. Similar results were obtained with NA vaccines that targeted different surface molecules on dendritic cells. Interestingly, the protective efficacy of NA as antigen compared favorably with HA and therefore ought to receive more attention in influenza vaccine research.

• Kikuchi, Jiro; Kodama, Nobuyuki; Takeshita, Masataka; Ikeda, Sho; Kobayashi, Takahiro & Kuroda, Yoshiaki [Vis alle 13 forfattere av denne artikkelen] (2023). EMD originates from hyaluronan-induced homophilic interactions of CD44 variant-expressing MM cells under shear stress. Blood Advances. ISSN 2473-9529. 7(4), s. 508–524. doi: 10.1182/bloodadvances.2022007291. Fulltekst i vitenarkiv Vis sammendrag

  Abstract Extramedullary disease (EMD) is known to be associated with chemoresistance and poor prognosis in multiple myeloma (MM); however, the mechanisms of its development are not fully understood. Elucidating the mechanism of EMD development and its therapeutic targeting would greatly contribute to further improvement of treatment outcome in patients with MM. Here, we show that bone marrow stroma cell–derived hyaluronan (HA) elicits homophilic interactions of MM cells by binding to surface CD44, especially long-stretch variants, under physiological shear stress and generates cell clusters that might develop into EMD. We recapitulated the development of EMD via administration of HA in a syngeneic murine MM model in a CD44-dependent manner. HA-induced MM cell clusters exhibited the specific resistance to proteasome inhibitors (PIs) in vitro and in murine models via γ-secretase–mediated cleavage of the intracellular domains of CD44, which in turn transactivated PI resistance-inducible genes. Treatment of HA-injected mice with anti-CD44 antibody or γ-secretase inhibitors readily suppressed the development of EMD from transplanted MM cells and significantly prolonged the survival of recipients by overcoming PI resistance. The HA-CD44 axis represents a novel pathway to trigger EMD development and could be a target of the prediction, prevention, and treatment of EMD in patients with MM.

• Bekri, Selma; Rodney-Sandy, Reunet; Gruenstein, Diana; Mei, Anna; Bogen, Bjarne & Castle, John [Vis alle 8 forfattere av denne artikkelen] (2022). Neoantigen vaccine-induced CD4 T cells confer protective immunity in a mouse model of multiple myeloma through activation of CD8 T cells against non-vaccine, tumor-associated antigens. Journal for ImmunoTherapy of Cancer (JITC). 10:e003572(2), s. 1–11. doi: 10.1136/jitc-2021-003572. Fulltekst i vitenarkiv Vis sammendrag

  Background Cancer-associated neoantigens (neoAg) derived from tumor genomic sequencing and predictive algorithms for mutated peptides are a promising basis for therapeutic vaccines under investigation. Although these are generally designed to bind major histocompatibility complex class I and induce CD8 cytolytic T lymphocyte (CTL) activity, results from preclinical and clinical studies demonstrate that the majority of neoAg vaccines efficiently induce CD4 T helper (Th) responses but not CTL. Despite this, these vaccines have demonstrated clinical efficacy. Therefore, understanding the mechanisms of CD4 + T cell-mediated tumor protection is critical to optimizing this immunotherapeutic strategy. Methods We investigated this phenomenon in the mineral oil-induced plasmacytoma (MOPC).315.BM (MOPC315) mouse model of multiple myeloma, a malignancy of plasma cells. MOPC315 cells express in their lambda chain a unique tumor-specific neoAg, an idiotypic (Id) peptide. We generated a vaccine formulated with this Id peptide fused to a heat shock protein HSC70 binding (HSB) motif co-delivered with poly (I:C). The immunogenicity of the Id-vaccine was measured in splenocytes by ELISpot. Mice were challenged with MOPC315 cells and antitumor immunity was assessed by co-incubating splenocytes and bone marrow mononuclear cells derived from vaccinated mice and controls, with the Id antigen and irradiated MOPC315 cells. The frequency of activated CD4 and CD8 T cells and their phenotype were characterized by flow cytometry. Results Id-vaccine efficiently induced antigen-specific CD4 Th activity and antitumor immunity, protecting mice from MOPC315 tumor growth. CD4 cytolytic activity was not detected under these conditions. Polyfunctional CD8 T cells homed to the bone marrow microenvironment of protected mice and preferentially expanded only when restimulated ex vivo with both Id peptide and MOPC315 cells. Protective activity was abrogated by depletion of either CD4 or CD8 lymphocytes. Conclusion These results demonstrate that Id-HSB +poly (I:C) vaccine protects against MOPC315 growth by priming Id-specific CD4 Th cells that confer protection against tumor but are not directly cytotoxic. These data indicate that activation of CD8 CTL against MOPC315-associated antigens not present in the vaccine is one of the major mechanisms of tumor immunity.

• Tesfaye, Demo Yemane; Bobic, Sonja; Lysén, Anna Karin Christina; Huszthy, Peter Csaba; Gudjonsson, Arnar & Braathen, Ranveig [Vis alle 8 forfattere av denne artikkelen] (2022). Targeting Xcr1 on dendritic cells rapidly induce Th1-associated immune responses that contribute to protection against influenza infection. Frontiers in Immunology. 13:752714, s. 1–13. doi: 10.3389/fimmu.2022.752714. Fulltekst i vitenarkiv Vis sammendrag

  Targeting antigen to conventional dendritic cells (cDCs) can improve antigen-specific immune responses and additionally be used to influence the polarization of the immune responses. However, the mechanisms by which this is achieved are less clear. To improve our understanding, we here evaluate molecular and cellular requirements for CD4+ T cell and antibody polarization after immunization with Xcl1-fusion vaccines that specifically target cDC1s. Xcl1-fusion vaccines induced an IgG2a/IgG2b-dominated antibody response and rapid polarization of Th1 cells both in vitro and in vivo. For comparison, we included fliC-fusion vaccines that almost exclusively induced IgG1, despite inducing a more mixed polarization of T cells. Th1 polarization and IgG2a induction with Xcl1-fusion vaccines required IL-12 secretion but were nevertheless maintained in BATF3-/- mice which lack IL-12-secreting migratory DCs. Interestingly, induction of IgG2a-dominated responses was highly dependent on the early kinetics of Th1 induction and was important for optimal protection in an influenza infection model. Early Th1 induction was dominant, since a combined Xcl1- and fliC-fusion vaccine induced IgG2a/IgG2b polarized antibody responses similar to Xcl1-fusion vaccines alone. In summary, our results demonstrate that targeting antigen to Xcr1+ cDC1s is an efficient strategy for enhancing IgG2a antibody responses through rapid Th1 induction, which can be utilized for improved vaccine design.

• Lopes, Raquel; Caetano, Joana; Barahona, Filipa; Pestana, Carolina; Ferreira, Bruna Velosa & Lourenço, Diana [Vis alle 17 forfattere av denne artikkelen] (2022). Multiple Myeloma-Derived Extracellular Vesicles Modulate the Bone Marrow Immune Microenvironment. Frontiers in Immunology. 13:909880, s. 1–14. doi: 10.3389/fimmu.2022.909880. Fulltekst i vitenarkiv Vis sammendrag

  Multiple myeloma (MM), the third most frequent hematological cancer worldwide, is characterized by the proliferation of neoplastic plasma cells in the bone marrow (BM). One of the hallmarks of MM is a permissive BM microenvironment. Increasing evidence suggests that cell-to-cell communication between myeloma and immune cells via tumor cell-derived extracellular vesicles (EV) plays a key role in the pathogenesis of MM. Hence, we aimed to explore BM immune alterations induced by MM-derived EV. For this, we inoculated immunocompetent BALB/cByJ mice with a myeloma cell line, MOPC315.BM, inducing a MM phenotype. Upon tumor establishment, characterization of the BM microenvironment revealed the expression of both activation and suppressive markers by lymphocytes, such as granzyme B and PD-1, respectively. In addition, conditioning of the animals with MOPC315.BM-derived EV, before transplantation of the MOPC315.BM tumor cells, did not anticipate the disease phenotype. However, it induced features of suppression in the BM milieu, such as an increase in PD-1 expression by CD4+ T cells. Overall, our findings reveal the involvement of MOPC315.BM-derived EV protein content as promoters of immune niche remodeling, strengthening the importance of assessing the mechanisms by which MM may impact the immune microenvironment.

• Hinke, Daniëla Maria; Andersen, Tor Kristian; Gopalakrishnan, Ramakrishna Prabhu; Skullerud, Lise Madelen; Werninghaus, Ina Charlotta & Grødeland, Gunnveig [Vis alle 9 forfattere av denne artikkelen] (2022). Antigen bivalency of antigen-presenting cell-targeted vaccines increases B cell responses. Cell reports. 39:110901(9), s. 1–27. doi: 10.1016/j.celrep.2022.110901. Fulltekst i vitenarkiv Vis sammendrag

  Antibodies are important for vaccine efficacy. Targeting antigens to antigen-presenting cells (APCs) increases antibody levels. Here, we explore the role of antigen valency in MHC class II (MHCII)-targeted vaccines delivered as DNA. We design heterodimeric proteins that carry either two identical (bivalent vaccines), or two different antigens (monovalent vaccines). Bivalent vaccines with two identical influenza hemagglutinins (HA) elicit higher amounts of anti-HA antibodies in mice than monovalent versions with two different HAs. Bivalent vaccines increase the levels of germinal center (GC) B cells and long-lived plasma cells. Only HA-bivalent vaccines completely protect mice against challenge with homologous influenza virus. Similar results are obtained with other antigens by targeting CD11c and Xcr1 on dendritic cells (DCs) or when administering the vaccine as protein with adjuvant. Bivalency probably increases B cell responses by cross-linking BCRs in readily observable DC-B cell synapses. These results are important for generating potent APC-targeted vaccines.

• Jung, Keunok; Jeon, You-kyoung; Jeong, Dae Hoon; Byun, Jung Mi; Bogen, Bjarne & Choi, Inhak (2022). VSIG4-expressing tumor-associated macrophages impair anti-tumor immunity. Biochemical and Biophysical Research Communications - BBRC. ISSN 0006-291X. 628, s. 18–24. doi: 10.1016/j.bbrc.2022.08.055. Fulltekst i vitenarkiv
• Andersen, Tor Kristian; Bodin, Johanna Eva; Oftung, Fredrik; Bogen, Bjarne; Mjaaland, Siri & Grødeland, Gunnveig (2021). Pandemic Preparedness Against Influenza: DNA Vaccine for Rapid Relief. Frontiers in Immunology. 12. doi: 10.3389/fimmu.2021.747032. Fulltekst i vitenarkiv Vis sammendrag

  The 2009 “swine flu” pandemic outbreak demonstrated the limiting capacity for egg-based vaccines with respect to global vaccine supply within a timely fashion. New vaccine platforms that efficiently can quench pandemic influenza emergences are urgently needed. Since 2009, there has been a profound development of new vaccine platform technologies with respect to prophylactic use in the population, including DNA vaccines. These vaccines are particularly well suited for global pandemic responses as the DNA format is temperature stable and the production process is cheap and rapid. Here, we show that by targeting influenza antigens directly to antigen presenting cells (APC), DNA vaccine efficacy equals that of conventional technologies. A single dose of naked DNA encoding hemagglutinin (HA) from influenza/A/California/2009 (H1N1), linked to a targeting moiety directing the vaccine to major histocompatibility complex class II (MHCII) molecules, raised similar humoral immune responses as the adjuvanted split virion vaccine Pandemrix, widely administered in the 2009 pandemic. Both vaccine formats rapidly induced serum antibodies that could protect mice already 8 days after a single immunization, in contrast to the slower kinetics of a seasonal trivalent inactivated influenza vaccine (TIV). Importantly, the DNA vaccine also elicited cytotoxic T-cell responses that reduced morbidity after vaccination, in contrast to very limited T-cell responses seen after immunization with Pandemrix and TIV. These data demonstrate that DNA vaccines has the potential as a single dose platform vaccine, with rapid protective effects without the need for adjuvant, and confirms the relevance of naked DNA vaccines as candidates for pandemic preparedness.

• Szodoray, Peter; Andersen, Tor Kristian; Heinzelbecker, Julia; Imbery, John Franklin; Huszthy, Peter C. & Stanford, Stephanie M. [Vis alle 12 forfattere av denne artikkelen] (2021). Integration of T helper and BCR signals governs enhanced plasma cell differentiation of memory B cells by regulation of CD45 phosphatase activity. Cell reports. 36:109525(6), s. 1–24. doi: 10.1016/j.celrep.2021.109525. Fulltekst i vitenarkiv Vis sammendrag

  Humoral immunity relies on the efficient differentiation of memory B cells (MBCs) into antibody-secreting cells (ASCs). T helper (Th) signals upregulate B cell receptor (BCR) signaling by potentiating Src family kinases through increasing CD45 phosphatase activity (CD45 PA). In this study, we show that high CD45 PA in MBCs enhances BCR signaling and is essential for their effective ASC differentiation. Mechanistically, Th signals upregulate CD45 PA through intensifying the surface binding of a CD45 ligand, Galectin-1. CD45 PA works as a sensor of T cell help and defines high-affinity germinal center (GC) plasma cell (PC) precursors characterized by IRF4 expression in vivo. Increasing T cell help in vitro results in an incremental CD45 PA increase and enhances ASC differentiation by facilitating effective induction of the transcription factors IRF4 and BLIMP1. This study connects Th signals with BCR signaling through Galectin-1-dependent regulation of CD45 PA and provides a mechanism for efficient ASC differentiation of MBCs.

• Lysén, Anna Karin Christina; Gudjonsson, Arnar; Tesfaye, Demo Yemane; Bobic, Sonja; Bern, Malin C. & Bogen, Bjarne [Vis alle 7 forfattere av denne artikkelen] (2021). Intranasal delivery of a cDC1 targeted influenza vaccine with poly(I:C) enhances T cell responses and protects against influenza infection. Scandinavian Journal of Immunology. ISSN 0300-9475. 95:e13128(3), s. 1–12. doi: 10.1111/sji.13128. Fulltekst i vitenarkiv Vis sammendrag

  Targeting antigens to dendritic cells represent a promising method for enhancing immune responses against specific antigens. However, many studies have focused on systemic delivery (intravenous or intraperitoneally) of targeted antigen, approaches that are not easily transferable to humans. Here we evaluate the efficacy of an influenza vaccine targeting Xcr1+ cDC1 administered by intranasal immunization. Intranasal delivery of antigen fused to the chemokine Xcl1, the ligand of Xcr1, resulted in specific uptake by lung CD103+ cDC1. Interestingly, intranasal immunization with influenza A/PR/8/34 haemagglutinin (HA) fused to Xcl1, formulated with poly(I:C), resulted in enhanced induction of antigen-specific IFNγ+CD4+ and IFNγ+CD8+ T cell responses in lung compared non-targeted anti-NIP-HA (αNIP-HA). Induction of antibody responses was, however, similar in Xcl1-HA and αNIP-HA immunized mice, but significantly higher than in mice immunized with monomeric HA. Both Xcl1-HA and αNIP-HA vaccines induced full protection when mice were challenged with a lethal dose of influenza PR8 virus, reflecting the strong induction of HA-specific antibodies. Our results demonstrate that i.n. delivery of Xcl1-HA is a promising vaccine strategy for enhancing T cell responses in addition to inducing strong antibody responses.

• Bjerkan, Louise; Visweswaran, Ganesh Ram R.; Gudjonsson, Arnar; Labbé, Genevieve M.; Quinkert, Doris & Pattinson, David J. [Vis alle 10 forfattere av denne artikkelen] (2021). APC-Targeted DNA Vaccination Against Reticulocyte-Binding Protein Homolog 5 Induces Plasmodium falciparum-Specific Neutralizing Antibodies and T Cell Responses. Frontiers in Immunology. 12:720550(18), s. 1–13. doi: 10.3389/fimmu.2021.720550. Fulltekst i vitenarkiv Vis sammendrag

  Targeted delivery of antigen to antigen presenting cells (APCs) is an efficient way to induce robust antigen-specific immune responses. Here, we present a novel DNA vaccine that targets the Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5), a leading blood-stage antigen of the human malaria pathogen, to APCs. The vaccine is designed as bivalent homodimers where each chain is composed of an amino-terminal single chain fragment variable (scFv) targeting unit specific for major histocompatibility complex class II (MHCII) expressed on APCs, and a carboxyl-terminal antigenic unit genetically linked by the dimerization unit. This vaccine format, named “Vaccibody”, has previously been successfully applied for antigens from other infectious diseases including influenza and HIV, as well as for tumor antigens. Recently, the crystal structure and key functional antibody epitopes for the truncated version of PfRH5 (PfRH5ΔNL) were characterized, suggesting PfRH5ΔNL to be a promising candidate for next-generation PfRH5 vaccine design. In this study, we explored the APC-targeting strategy for a PfRH5ΔNL-containing DNA vaccine. BALB/c mice immunized with the targeted vaccine induced higher PfRH5-specific IgG1 antibody responses than those vaccinated with a non-targeted vaccine or antigen alone. The APC-targeted vaccine also efficiently induced rapid IFN-γ and IL-4 T cell responses. Furthermore, the vaccine-induced PfRH5-specific IgG showed inhibition of growth of the P. falciparum 3D7 clone parasite in vitro. Finally, sera obtained after vaccination with this targeted vaccine competed for the same epitopes as PfRH5-specific mAbs from vaccinated humans. Robust humoral responses were also induced by a similar P. vivax Duffy-binding protein (PvDBP)-containing targeted DNA vaccine. Our data highlight a novel targeted vaccine platform for the development of vaccines against blood-stage malaria.

• Høydahl, Lene; Frigstad, Terje; Rasmussen, Ingunn B; Øynebråten, Inger; Schjetne, Karoline W & Andersen, Jan Terje [Vis alle 10 forfattere av denne artikkelen] (2021). Antibody-mediated delivery of T-cell epitopes to antigen-presenting cells induce strong CD4 and CD8 T-cell responses. Vaccine. ISSN 0264-410X. 39(11), s. 1583–1592. doi: 10.1016/j.vaccine.2021.02.012. Fulltekst i vitenarkiv Vis sammendrag

  Targeted delivery of antigen to antigen-presenting cells (APCs) enhances antigen presentation and thus, is a potent strategy for making more efficacious vaccines. This can be achieved by use of antibodies with specificity for endocytic surface molecules expressed on the APC. We aimed to compare two different antibody-antigen fusion modes in their ability to induce T-cell responses; first, exchange of immunoglobulin (Ig) constant domain loops with a T-cell epitope (Troybody), and second, fusion of T-cell epitope or whole antigen to the antibody C-terminus. Although both strategies are well-established, they have not previously been compared using the same system. We found that both antibody-antigen fusion modes led to presentation of the T-cell epitope. The strength of the T-cell responses varied, however, with the most efficient Troybody inducing CD4 T-cell proliferation and cytokine secretion at 10–100-fold lower concentration than the antibodies carrying antigen fused to the C-terminus, both in vitro and after intravenous injection in mice. Furthermore, we exchanged this loop with an MHCI-restricted T-cell epitope, and the resulting antibody enabled efficient cross-presentation to CD8 T cells in vivo. Targeting of antigen to APCs by use of such antibody-antigen fusions is thus an attractive vaccination strategy for increased activation of both CD4 and CD8 peptide-specific T cells.

• He, Le; Valignat, Marie-Pierre; Zhang, Lichen; Gelard, Lena; Zhang, Fanghui & Guen, Valentin Le [Vis alle 17 forfattere av denne artikkelen] (2021). ARHGAP45 controls naïve T- and B- cell entry into lymph nodes and T-cell progenitor thymus seeding. EMBO Reports. ISSN 1469-221X. 22:e52196(4), s. 1–19. doi: 10.15252/embr.202052196. Fulltekst i vitenarkiv Vis sammendrag

  T and B cells continually recirculate between blood and secondary lymphoid organs. To promote their trans-endothelial migration (TEM), chemokine receptors control the activity of RHO family small GTPases in part via GTPase-activating proteins (GAPs). T and B cells express several RHO-GAPs, the function of most of which remains unknown. The ARHGAP45 GAP is predominantly expressed in hematopoietic cells. To define its in vivo function, we describe two mouse models where ARHGAP45 is ablated systemically or selectively in T cells. We combine their analysis with affinity purification coupled to mass spectrometry to determine the ARHGAP45 interactome in T cells and with time-lapse and reflection interference contrast microscopy to assess the role of ARGHAP45 in T-cell polarization and motility. We demonstrate that ARHGAP45 regulates naïve T-cell deformability and motility. Under physiological conditions, ARHGAP45 controls the entry of naïve T and B cells into lymph nodes whereas under competitive repopulation it further regulates hematopoietic progenitor cell engraftment in the bone marrow, and T-cell progenitor thymus seeding. Therefore, the ARGHAP45 GAP controls multiple key steps in the life of T and B cells.

• Ziouti, Fani; Rummler, Maximilian; Steyn, Beatrice; Thiele, Tobias; Seliger, Anne & Duda, Georg N. [Vis alle 9 forfattere av denne artikkelen] (2021). Prevention of Bone Destruction by Mechanical Loading Is Not Enhanced by the Bruton’s Tyrosine Kinase Inhibitor CC-292 in Myeloma Bone Disease. International Journal of Molecular Sciences. ISSN 1661-6596. 22:3840(8), s. 1–14. doi: 10.3390/ijms22083840. Fulltekst i vitenarkiv Vis sammendrag

  Limiting bone resorption and regenerating bone tissue are treatment goals in myeloma bone disease (MMBD). Physical stimuli such as mechanical loading prevent bone destruction and enhance bone mass in the MOPC315.BM.Luc model of MMBD. It is unknown whether treatment with the Bruton’s tyrosine kinase inhibitor CC-292 (spebrutinib), which regulates osteoclast differentiation and function, augments the anabolic effect of mechanical loading. CC-292 was administered alone and in combination with axial compressive tibial loading in the MOPC315.BM.Luc model for three weeks. However, neither CC-292 alone nor its use in combination with mechanical loading was more effective in reducing osteolytic bone disease or rescuing bone mass than mechanical stimuli alone, as evidenced by microcomputed tomography (microCT) and histomorphometric analysis. Further studies are needed to investigate novel anti-myeloma and anti-resorptive strategies in combination with physical stimuli to improve treatment of MMBD.

• Grødeland, Gunnveig; Baranowska-Hustad, Marta Natalia; Abadejos, Justin; Blane, Tanja R.; Teijaro, John & Nemazee, David [Vis alle 7 forfattere av denne artikkelen] (2020). Induction of Cross-Reactive and Protective Antibody Responses After DNA Vaccination With MHCII-Targeted Stem Domain From Influenza Hemagglutinin. Frontiers in Immunology. 11:431, s. 1–13. doi: 10.3389/fimmu.2020.00431. Fulltekst i vitenarkiv Vis sammendrag

  Novel and more broadly protective vaccines against influenza are needed to efficiently meet antigenic drift and shift. Relevant to this end, the stem domain of hemagglutinin (HA) is highly conserved, and antibodies specific for epitopes located to the stem have been demonstrated to be able to confer broad protection against various influenza subtypes. However, a remaining challenge is to induce antibodies against the poorly immunogenic stem by vaccination strategies that can be scaled up for prophylactic vaccination of the general population. Here, we have developed DNA vaccines where the conserved stem domain of HA from influenza A/PR/8/34 (H1N1) and A/Shanghai/2/2013 (H7N9) was targeted toward MHC class II molecules on antigen-presenting cells (APC) for increased immunogenicity. Each of these vaccines induced antibodies that cross-reacted with other subtypes in the corresponding phylogenetic influenza groups. Importantly, when mixing the MHCII-targeted stem domains from H1N1 and H7N9 influenza viruses into one vaccine bolus, we observed broad protection against candidate stains from both phylogenetic groups 1 and 2.

• Rummler, Maximilian; Ziouti, Fani; Bouchard, Alice L.; Brandl, Andreas; Duda, Georg N. & Bogen, Bjarne [Vis alle 10 forfattere av denne artikkelen] (2020). Mechanical loading prevents bone destruction and exerts anti-tumor effects in the MOPC315.BM.Luc model of myeloma bone disease. Acta Biomaterialia. ISSN 1742-7061. 119, s. 247–258. doi: 10.1016/j.actbio.2020.10.041. Fulltekst i vitenarkiv
• Ziouti, Fani; Soares, Ana Prates; Moreno-Jiménez, Inés; Rack, Alexander; Bogen, Bjarne & Cipitria, Amaia [Vis alle 8 forfattere av denne artikkelen] (2020). An Early Myeloma Bone Disease Model in Skeletally Mature Mice as a Platform for Biomaterial Characterization of the Extracellular Matrix. Journal of Oncology. ISSN 1687-8450. 2020:3985315, s. 1–12. doi: 10.1155/2020/3985315. Fulltekst i vitenarkiv Vis sammendrag

  Multiple myeloma (MM) bone disease is characterized by osteolytic bone tissue destruction resulting in bone pain, fractures, vertebral collapse, and spinal cord compression in patients. Upon initial diagnosis of MM, almost 80% of patients suffer from bone disease. Earlier diagnosis and intervention in MM bone disease would potentially improve treatment outcome and patient survival. New preclinical models are needed for developing novel diagnostic markers of bone structural changes as early as possible in the disease course. Here, we report a proof-of-concept, syngeneic, intrafemoral MOPC315.BM MM murine model in skeletally mature BALB/c mice for detection and characterization of very early changes in the extracellular matrix (ECM) of MM-injected animals. Bioluminescence imaging (BLI) in vivo confirmed myeloma engraftment in 100% of the animals with high osteoclast activity within 21 days after tumor cell inoculation. Early signs of aggressive bone turnover were observed on the outer bone surfaces by high-resolution microcomputed tomography (microCT). Synchrotron phase contrast-enhanced microcomputer tomography (PCE-CT) revealed very local microarchitecture differences highlighting numerous active sites of erosion and new bone at the micrometer scale. Correlative backscattered electron imaging (BSE) and confocal laser scanning microscopy allowed direct comparison of mineralized and nonmineralized matrix changes in the cortical bone. The osteocyte lacunar-canalicular network (OLCN) architecture was disorganized, and irregular-shaped osteocyte lacunae were observed in MM-injected bones after 21 days. Our model provides a potential platform to further evaluate pathological MM bone lesion development at the micro- and ultrastructural levels. These promising results make it possible to combine material science and pharmacological investigations that may improve early detection and treatment of MM bone disease.

• Haabeth, Ole Audun; Hennig, Kjartan; Fauskanger, Marte; Løset, Geir Åge; Bogen, Bjarne & Tveita, Anders Aune (2020). CD4+ T-cell killing of multiple myeloma cells is mediated by resident bone marrow macrophages. Blood Advances. ISSN 2473-9529. 4(12), s. 2595–2605. doi: 10.1182/bloodadvances.2020001434. Fulltekst i vitenarkiv Vis sammendrag

  Abstract CD4+ T cells may induce potent antitumor immune responses through interaction with antigen-presenting cells within the tumor microenvironment. Using a murine model of multiple myeloma, we demonstrated that adoptive transfer of idiotype-specific CD4+ T cells may elicit curative responses against established multifocal myeloma in bone marrow. This finding indicates that the myeloma bone marrow niche contains antigen-presenting cells that may be rendered tumoricidal. Given the complexity of the bone marrow microenvironment, the mechanistic basis of such immunotherapeutic responses is not known. Through a functional characterization of antitumor CD4+ T-cell responses within the bone marrow microenvironment, we found that killing of myeloma cells is orchestrated by a population of bone marrow–resident CD11b+F4/80+MHC-IIHigh macrophages that have taken up and present secreted myeloma protein. The present results demonstrate the potential of resident macrophages as powerful mediators of tumor killing within the bone marrow and provide a basis for novel therapeutic strategies against multiple myeloma and other malignancies that affect the bone marrow.

• Kwon, Minsuk; Kim, Chang Gon; Lee, Hoyoung; Cho, Hyunsoo; Kim, Youngun & Lee, Eung Chang [Vis alle 17 forfattere av denne artikkelen] (2020). PD-1 Blockade Reinvigorates Bone Marrow CD8+ T Cells from Patients with Multiple Myeloma in the Presence of TGFβ Inhibitors. Clinical Cancer Research. ISSN 1078-0432. 26(7), s. 1644–1655. doi: 10.1158/1078-0432.CCR-19-0267. Fulltekst i vitenarkiv
• Fossum, Even; Tesfaye, Demo Yemane; Bobic, Sonja; Gudjonsson, Arnar; Braathen, Ranveig & Lahoud, Mireille H. [Vis alle 8 forfattere av denne artikkelen] (2020). Targeting Antigens to Different Receptors on Conventional Type 1 Dendritic Cells Impacts the Immune Response. Journal of Immunology. ISSN 0022-1767. 205(3), s. 661–673. doi: 10.4049/jimmunol.1901119. Fulltekst i vitenarkiv
• Braathen, Ranveig; Spång, Heidi Cecilie Larsen; Hinke, Daniëla Maria; Blazevski, Jana; Bobic, Sonja & Fossum, Even [Vis alle 7 forfattere av denne artikkelen] (2020). A DNA Vaccine That Encodes an Antigen-Presenting Cell-Specific Heterodimeric Protein Protects against Cancer and Influenza. Molecular therapy. Methods & clinical development. ISSN 2399-6951. 17, s. P378–P392. doi: 10.1016/j.omtm.2020.01.007. Fulltekst i vitenarkiv Vis sammendrag

  Immunogenicity of DNA vaccines can be increased by constructing the DNA in such a way that it encodes secreted homodimeric fusion proteins that target antigen-presenting cells (APCs). In this study, we have developed novel APC-targeting vaccine molecules with an increased flexibility due to introduction of a heterodimerization motif. The heterodimeric proteins permit four different fusions within a single molecule, thus allowing expression of two different APC-targeting moieties and two different antigens. Two types of heterodimeric fusion proteins were developed that employed either the ACID/BASE or the Barnase/Barstar motifs, respectively. The ACID/BASE heterodimeric vaccines conferred protection against challenges with either influenza virus or tumor cells in separate preclinical models. The ACID/BASE motif was flexible since a large number of different targeting moieties and antigens could be introduced with maintenance of specificity, antigenicity, and secretion. APC-targeting ACID/BASE vaccines expressing two different antigens induced antibody and T cell responses against either of the two antigens. Heterodimeric ACID/BASE DNA vaccines were of approximately the same potency as previously reported homodimeric DNA vaccines. The flexibility and potency of the ACID/BASE format suggest that it could be a useful platform for DNA vaccines that encode APC-targeting fusion proteins.

• Mahalunkar, Sneha; Yadav, Amit Singh; Gorain, Mahadeo; Pawar, Vinay; Braathen, Ranveig & Weiss, Siegfried [Vis alle 9 forfattere av denne artikkelen] (2019). Functional design of pH-responsive folate-targeted polymer-coated gold nanoparticles for drug delivery and in vivo therapy in breast cancer. International Journal of Nanomedicine. ISSN 1176-9114. 14, s. 8285–8302. doi: 10.2147/IJN.S215142. Fulltekst i vitenarkiv Vis sammendrag

  Background: Curcumin has been widely used owing to its various medicinal properties including antitumor effects. However, its clinical application is limited by its instability, poor solubility and low bioavailability. Folic acid (FA)-functionalized nanoformulations may enhance the sustained release of an anticancer drug (curcumin) by tumor-specific targeting to improve therapeutic benefit. This study aims to design a nanoconjugate (NC) comprised of folate–curcumin-loaded gold–polyvinylpyrrolidone nanoparticles (FA–CurAu-PVP NPs) for targeted delivery in breast cancer model systems. Methods: We developed curcumin-loaded FA-functionalized Au-PVP NCs by layer-by-layer assembly. The folic acid–curcumin Au-PVP NCs (FA–CurAu-PVP NCs) were characterized by ultraviolet–visible spectra, Fourier transform infrared spectroscopy, X-ray powder diffraction and thermogravimetric analysis. In vitro anticancer and antimigratory effects of NCs were examined by performing MTT and wound migration assays. The in vivo antitumor efficacy of NCs was investigated using a preclinical breast cancer orthotopic mouse model. Results: Curcumin (40 μg/mL) was loaded along with conjugation of folate onto Au-PVP NPs to form FA–CurAu-PVP NCs. The size and charge of the NCs were increased gradually through layer-by-layer assembly and showed 80% release of curcumin at acidic pH. The NC did not show aggregation when incubated with human serum and mimicked an intrinsic peroxidase-like property in the presence of 3,3ʹ,5,5ʹ-tetramethylbenzidine substrate. The MTT data using these NCs showed efficient anticancer activity at lower doses in estrogen/progesterone receptor (ER/PR)-negative cells compared with ER/PR-positive cells. Furthermore, the NCs did not show cytotoxicity at the investigated concentration in human breast epithelial and mouse fibroblast cell lines. They showed inhibitory effects on cell migration and high antitumor efficacy in in vivo analysis. Conclusion: These results suggest that folate-based tumor targeting using CurAu-PVP NCs is a promising approach for tumor-specific therapy of breast cancer without harming normal cells.

• Gudjonsson, Arnar; Andersen, Tor Kristian; Gjerstad, Vibeke Sundvold; Bogen, Bjarne & Fossum, Even (2019). Endocytosis Deficient Murine Xcl1-Fusion Vaccine Enhances Protective Antibody Responses in Mice. Frontiers in Immunology. 10, s. 1–12. doi: 10.3389/fimmu.2019.01086. Fulltekst i vitenarkiv Vis sammendrag

  Targeting antigen to surface receptors on dendritic cells (DCs) can improve antibody response against subunit vaccines. We have previously observed that human XCL1-fusion vaccines target murine Xcr1+ DCs without actively inducing endocytosis of the antigen, resulting in enhanced antibody responses in mice. However, the use of foreign chemokines for targeting is undesirable when translating this observation to human or veterinary medicine due to potential cross-reactive responses against the endogenous chemokine. Here we have identified a mutant version of murine Xcl1, labeled Xcl1(Δ1) owing to removal of a conserved valine in position 1 of the mature chemokine, that retains specific binding to Xcr1+ DCs without inducing endocytosis of the receptor. DNA immunization with Xcl1(Δ1) conjugated to influenza hemagglutinin (HA) induced improved antibody responses, with higher end point titers of IgG compared to WT Xcl1-HA. The Xcl1(Δ1) fusion vaccine also resulted in an increased number of HA reactive germinal center B cells with higher avidity toward the antigen, and serum transfer experiments show that Xcl1(Δ1)-HA induced antibody responses provided better protection against influenza infection as compared to WT Xcl1-HA. In summary, our observations indicate that targeting antigen to Xcr1+ DCs in an endocytosis deficient manner enhances antibody responses. This effect was obtained by introducing a single mutation to Xcl1, suggesting our strategy may easily be translated to human or veterinary vaccine settings.

• Mooij, Petra; Grødeland, Gunnveig; Koopman, Gerrit; Andersen, Tor Kristian; Mortier, Daniella & Nieuwenhuis, Ivonne G. [Vis alle 10 forfattere av denne artikkelen] (2019). Needle-free delivery of DNA: Targeting of hemagglutinin to MHC class II molecules protects rhesus macaques against H1N1 influenza. Vaccine. ISSN 0264-410X. 37(6), s. 817–826. doi: 10.1016/j.vaccine.2018.12.049. Fulltekst i vitenarkiv Vis sammendrag

  Conventional influenza vaccines are hampered by slow and limited production capabilities, whereas DNA vaccines can be rapidly produced for global coverage in the event of an emerging pandemic. However, a drawback of DNA vaccines is their generally low immunogenicity in non-human primates and humans. We have previously demonstrated that targeting of influenza hemagglutinin to human HLA class II molecules can increase antibody responses in larger animals such as ferrets and pigs. Here, we extend these observations by immunizing non-human primates (rhesus macaques) with a DNA vaccine encoding a bivalent fusion protein that targets influenza virus hemagglutinin (HA) to Mamu class II molecules. Such immunization induced neutralizing antibodies and antigen-specific T cells. The DNA was delivered by pain- and needle-free jet injections intradermally. No adverse effects were observed. Most importantly, the immunized rhesus macaques were protected against a challenge with influenza virus.

• Baghdadi, Muhammad; Ishikawa, Kozo; Nakanishi, Sayaka; Murata, Tomoki; Umeyama, Yui & Kobayashi, Takuto [Vis alle 20 forfattere av denne artikkelen] (2019). A role for IL-34 in osteolytic disease of multiple myeloma. Blood Advances. ISSN 2473-9529. 3(4), s. 541–551. doi: 10.1182/bloodadvances.2018020008. Fulltekst i vitenarkiv Vis sammendrag

  Multiple myeloma (MM) is a hematological malignancy that grows in multiple sites of the axial skeleton and causes debilitating osteolytic disease. Interleukin-34 (IL-34) is a newly discovered cytokine that acts as a ligand of colony-stimulating factor-1 (CSF-1) receptor and can replace CSF-1 for osteoclast differentiation. In this study, we identify IL-34 as an osteoclastogenic cytokine that accelerates osteolytic disease in MM. IL-34 was found to be expressed in the murine MM cell line MOPC315.BM, and the expression of IL-34 was enhanced by stimulation with proinflammatory cytokines or by bone marrow (BM) stromal cells. MM-cell–derived IL-34 promoted osteoclast formation from mouse BM cells in vitro. Targeting Il34 by specific small interfering RNA impaired osteoclast formation in vitro and attenuated osteolytic disease in vivo. In BM aspirates from MM patients, the expression levels of IL-34 in CD138+ populations vary among patients from high to weak to absent. MM cell–derived IL-34 promoted osteoclast formation from human CD14+ monocytes, which was reduced by a neutralizing antibody against IL-34. Taken together, this study describes for the first time the expression of IL-34 in MM cells, indicating that it may enhance osteolysis and suggesting IL-34 as a potential therapeutic target to control pathological osteoclastogenesis in MM patients.

• Lysén, Anna Karin Christina; Braathen, Ranveig; Gudjonsson, Arnar; Tesfaye, Demo Yemane; Bogen, Bjarne & Fossum, Even (2019). Dendritic cell targeted Ccl3- and Xcl1-fusion DNA vaccines differ in induced immune responses and optimal delivery site. Scientific Reports. 9:1820, s. 1–11. doi: 10.1038/s41598-018-38080-7. Fulltekst i vitenarkiv Vis sammendrag

  Fusing antigens to chemokines to target antigen presenting cells (APC) is a promising method for enhancing immunogenicity of DNA vaccines. However, it is unclear how different chemokines compare in terms of immune potentiating effects. Here we compare Ccl3- and Xcl1-fusion vaccines containing hemagglutinin (HA) from influenza A delivered by intramuscular (i.m.) or intradermal (i.d.) DNA vaccination. Xcl1 fusion vaccines target cDC1s, and enhance proliferation of CD4+ and CD8+ T cells in vitro. In contrast, Ccl3 target both cDC1 and cDC2, but only enhance CD4+ T cell proliferation in combination with cDC2. When Ccl3- or Xcl1-HA fusion vaccines were administered by i.m. DNA immunization, both vaccines induced Th1-polarized immune responses with antibodies of the IgG2a/IgG2b subclass and IFNγ-secreting T cells. After i.d. DNA vaccination, however, only Xcl1-HA maintained a Th1 polarized response and induced even higher numbers of IFNγ-secreting T cells. Consequently, Xcl1-HA induced superior protection against influenza infection compared to Ccl3-HA after i.d. immunization. Interestingly, i.m. immunization with Ccl3-HA induced the strongest overall in vivo cytotoxicity, despite not inducing OT-I proliferation in vitro. In summary, our results highlight important differences between Ccl3- and Xcl1- targeted DNA vaccines suggesting that chemokine fusion vaccines can be tailor-made for different diseases.

• Høglund, Rune Alexander; Torsetnes, Silje Bøen; Lossius, Peter Andreas Vold; Bogen, Bjarne; Homan, E. Jane & Bremel, Robert [Vis alle 7 forfattere av denne artikkelen] (2019). Human cysteine cathepsins degrade immunoglobulin G in vitro in a predictable manner. International Journal of Molecular Sciences. ISSN 1661-6596. 20:4843(19), s. 1–19. doi: 10.3390/ijms20194843. Fulltekst i vitenarkiv Vis sammendrag

  Cysteine cathepsins are critical components of the adaptive immune system involved in the generation of epitopes for presentation on human leukocyte antigen (HLA) molecules and have been implicated in degradation of autoantigens. Immunoglobulin variable regions with somatic mutations and random complementarity region 3 amino acid composition are inherently immunogenic. T cell reactivity towards immunoglobulin variable regions has been investigated in relation to specific diseases, as well as reactivity to therapeutic monoclonal antibodies. Yet, how the immunoglobulins, or the B cell receptors, are processed in endolysosomal compartments of professional antigen presenting cells has not been described in detail. Here we present in silico and in vitro experimental evidence suggesting that cysteine cathepsins S, L and B may have important roles in generating peptides fitting HLA class II molecules, capable of being presented to T cells, from monoclonal antibodies as well as from central nervous system proteins including a well described autoantigen. By combining neural net models with in vitro proteomics experiments, we further suggest how such degradation can be predicted, how it fits with available cellular models, and that it is immunoglobulin heavy chain variable family dependent. These findings are relevant for biotherapeutic drug design as well as to understand disease development. We also suggest how these tools can be improved, including improved machine learning methodology.

• Grødeland, Gunnveig; Fossum, Even & Bogen, Bjarne (2019). Targeting of HA to chemokine receptors induces strong and cross-reactive T cell responses after DNA vaccination in pigs. Vaccine. ISSN 0264-410X. 38(6), s. 1280–1285. doi: 10.1016/j.vaccine.2019.11.084. Fulltekst i vitenarkiv Vis sammendrag

  Efficient influenza vaccination of pigs can reduce disease burdens for the swine industry, but also represents an important measure for reducing the risk from novel viral reassortments that pose pandemic threats to the human population. Here, we have vaccinated pigs with a DNA vaccine encoding influenza virus hemagglutinin (HA) linked to the chemokine MIP1α that bind chemokine receptors 1, 3, and 5 expressed on antigen presenting cells (APC). Such MIP1α targeting of HA to APC enhanced induction of HA reactive antibodies, particularly IgG2. In addition, the MIP1α- HA vaccine induced strong T cell responses that could cross-react with different influenza subtypes. Thus, the strategy of targeting HA to chemokine receptors could be important for inducing broad protection against antigenically diverse influenza strains in pigs.

• Huszthy, Peter Csaba; Gopalakrishnan, Ramakrishna Prabhu; Jacobsen, Johanne Tracey; Haabeth, Ole Audun; Løset, Geir Åge & Braathen, Ranveig [Vis alle 10 forfattere av denne artikkelen] (2019). B cell receptor ligation induces display of V-region peptides on MHC class II molecules to T cells. Proceedings of the National Academy of Sciences of the United States of America (PNAS). ISSN 0027-8424. 116(51), s. 25850–25859. doi: 10.1073/pnas.1902836116. Fulltekst i vitenarkiv Vis sammendrag

  The B cell receptors (BCRs) for antigen express variable (V) regions that are enormously diverse, thus serving as markers on individual B cells. V region-derived idiotypic (Id) peptides can be displayed as pId:MHCII complexes on B cells for recognition by CD4+ T cells. It is not known if naive B cells spontaneously display pId:MHCII in vivo or if BCR ligation is required for expression, thereby enabling collaboration between Id+ B cells and Id-specific T cells. Here, using a mouse model, we show that naive B cells do not express readily detectable levels of pId:MHCII. However, BCR ligation by Ag dramatically increases physical display of pId:MHCII, leading to activation of Id-specific CD4+ T cells, extrafollicular T-B cell collaboration and some germinal center formation, and production of Id+ IgG. Besides having implications for immune regulation, the results may explain how persistent activation of self-reactive B cells induces the development of autoimmune diseases and B cell lymphomas.

• Bogen, Bjarne; Fauskanger, Marte; Haabeth, Ole Audun & Tveita, Anders Aune (2019). CD4+ T cells indirectly kill tumor cells via induction of cytotoxic macrophages in mouse models. Cancer Immunology and Immunotherapy. ISSN 0340-7004. 68(11), s. 1865–1873. doi: 10.1007/s00262-019-02374-0. Fulltekst i vitenarkiv Vis sammendrag

  It is well recognized that CD4+ T cells may play an important role in immunosurveillance and immunotherapy against cancer. However, the details of how these cells recognize and eliminate the tumor cells remain incompletely understood. For the past 25 years, we have focused on how CD4+ T cells reject multiple myeloma cells in a murine model (MOPC315). In our experimental system, the secreted tumor-specific antigen is taken up by tumor-infiltrating macrophages that process it and present a neoepitope [a V region-derived idiotypic (Id) peptide] on MHC class II molecules to Th1 cells. Stimulated Th1 cells produce IFNγ, which activates macrophages in a manner that elicits an M1-like, tumoricidal phenotype. Through an inducible nitric oxide synthetase (iNOS)-dependent mechanism, the M1 macrophages secrete nitric oxide (NO) that diffuses into neighboring tumor cells. Inside the tumor cells, NO-derived reactive nitrogen species, including peroxynitrite, causes nitrosylation of proteins and triggers apoptosis by the intrinsic apoptotic pathway. This mode of indirect tumor recognition by CD4+ T cells operates independently of MHC class II expression on cancer cells. However, secretion of the tumor-specific antigen, and uptake and MHCII presentation on macrophages, is required for rejection. Similar mechanisms can also be observed in a B-lymphoma model and in the unrelated B16 melanoma model. Our findings reveal a novel mechanism by which CD4+ T cells kill tumor cells indirectly via induction of intratumoral cytotoxic macrophages. The data suggest that induction of M1 polarization of tumor-infiltrating macrophages, by CD4+ T cells or through other means, could serve as an immunotherapeutic strategy.

• Andersen, Tor Kristian; Huszthy, Peter Csaba; Gopalakrishnan, Ramakrishna Prabhu; Jacobsen, Johanne Tracey; Fauskanger, Marte & Tveita, Anders Aune [Vis alle 8 forfattere av denne artikkelen] (2019). Enhanced germinal center reaction by targeting vaccine antigen to major histocompatibility complex class II molecules. npj Vaccines. 4:9, s. 1–13. doi: 10.1038/s41541-019-0101-0. Fulltekst i vitenarkiv Vis sammendrag

  Enhancing the germinal center (GC) reaction is a prime objective in vaccine development. Targeting of antigen to MHCII on APCs has previously been shown to increase antibody responses, but the underlying mechanism has been unclear. We have here investigated the GC reaction after targeting antigen to MHCII in (i) a defined model with T and B cells of known specificity using adjuvant-free vaccine proteins, and (ii) an infectious disease model using a DNA vaccine. MHCII-targeting enhanced presentation of peptide: MHCII on APCs, and increased the numbers of GC B cells, TFH, and plasma cells. Antibodies appeared earlier and levels were increased. BCR of GC B cells and serum antibodies had increased avidity for antigen. The improved responses required cross-linking of BCR and MHCII in either cis or trans. The enhanced GC reaction induced by MHCII-targeting of antigen has clear implications for design of more efficient subunit vaccines.

• Tesfaye, Demo Yemane; Gudjonsson, Arnar; Bogen, Bjarne & Fossum, Even (2019). Targeting conventional dendritic cells to fine-tune antibody responses. Frontiers in Immunology. 10:1529(JULY), s. 1–8. doi: 10.3389/fimmu.2019.01529. Fulltekst i vitenarkiv Vis sammendrag

  Dendritic cells (DCs) facilitate cross talk between the innate and adaptive immune system. They sense and phagocytose invading pathogens, and are not only capable of activating naïve T cells, but can also determine the polarization of T cell responses into different effector subtypes. Polarized T cells in turn have a crucial role in antibody class switching and affinity maturation, and consequently the quality of the resulting humoral immunity. Targeting vaccines to DCs thus provides a great deal of opportunities for influencing the humoral immune responses, by fine-tuning the T cell response as well as regulating antigen availability for B cells. In this review we aim to outline how different DC targeted vaccination strategies can be utilized to induce a desired humoral immune response. A range of factors, including route of vaccine administration, use of adjuvants, choice of DC subset and surface receptor to target have been reported to influence the resulting immune response and will be reviewed herein. Finally, we will discuss opportunities for designing improved vaccines and challenges with translating this knowledge into clinical or veterinary medicine.

• Abrahamsen, Greger; Gjerstad, Vibeke Sundvold; Mekonnen, Meseret; Bogen, Bjarne & Spurkland, Anne (2018). Polarity of CD4+ T cells towards the antigen presenting cell is regulated by the Lck adapter TSAd. Scientific Reports. 8:13319, s. 1–13. doi: 10.1038/s41598-018-31510-6. Fulltekst i vitenarkiv Vis sammendrag

  Polarization of T cells towards the antigen presenting cell (APC) is critically important for appropriate activation and differentiation of the naïve T cell. Here we used imaging flow cytometry (IFC) and show that the activation induced Lck and Itk adapter T cell specific adapter protein (TSAd), encoded by SH2D2A, modulates polarization of T cells towards the APC. Upon exposure to APC presenting the cognate antigen Id, Sh2d2a−/− CD4+ T cells expressing Id-specific transgenic T cell receptor (TCR), displayed impaired polarization of F-actin and TCR to the immunological synapse (IS). Sh2d2a−/− T-cells that did polarize F-actin and TCR still displayed impaired polarization of PKCξ, PAR3 and the microtubule-organizing center (MTOC). In vitro differentiation of activated Sh2d2a−/− T cells was skewed towards an effector memory (Tem) rather than a central memory (Tcm) phenotype. A similar trend was observed for Id-specific TCR Sh2d2a−/− T cells stimulated with APC and cognate antigen. Taken together our data suggest that TSAd modulates differentiation of experienced T cells possibly through polarization of CD4+ T cells towards the APC.

• Haabeth, Ole Audun; Fauskanger, Marte; Manzke, Melanie; Lundin, Katrin; Corthay, Alexandre & Bogen, Bjarne [Vis alle 7 forfattere av denne artikkelen] (2018). CD4+ T-cell–Mediated Rejection of MHC Class II–Positive Tumor Cells Is Dependent on Antigen Secretion and Indirect Presentation on Host APCs. Cancer Research. ISSN 0008-5472. 78(16), s. 4573–4585. doi: 10.1158/0008-5472.CAN-17-2426. Fulltekst i vitenarkiv
• Braathen, Ranveig; Spång, Heidi Cecilie Larsen; Lindeberg, Mona Mari; Fossum, Even; Grødeland, Gunnveig & Fredriksen, Agnete [Vis alle 7 forfattere av denne artikkelen] (2018). The Magnitude and IgG Subclass of Antibodies Elicited by Targeted DNA Vaccine Are Influenced by Specificity for APC Surface Molecules. ImmunoHorizons. 2(1), s. 38–53. doi: 10.4049/immunohorizons.1700038. Fulltekst i vitenarkiv Vis sammendrag

  Upon APC-targeted DNA vaccination, transfected cells secrete fusion proteins with targeting units specific for surface molecules on APC. In this study, we have tested several different targeting units for their ability to influence the magnitude and subclass of Ab responses to hemagglutinin from influenza A virus. The experiments employed bivalent homodimeric Ig-based molecules (vaccibodies). The overall efficiency in BALB/c mice depended on the targeting units in the following order: aMHC class II . aCD11c . aCD40 . Xcl-1 = MIP-1a . FliC . GM-CSF . Flt-3L . aDEC205. GM-CSF induced mainly IgG1, whereas Xcl1, MIP-1a, aCD40, and aDEC205 induced predominantly IgG2a. A more balanced mixture of IgG1 and IgG2a was observed with aCD11c, aMHC class II, Flt3L, and FliC. Similar results of IgG subclass–skewing were obtained in Th1-prone C57BL/6 mice with a more limited panel of vaccines. IgG1 responses in BALB/c occurred early after immunization but declined relatively rapidly over time. IgG2a responses appeared later but lasted longer (.252 d) than IgG1 responses. The most efficient targeting units elicited short- and long-term protection against PR8 influenza (H1N1) virus in BALB/c mice. The results suggest that targeting of Xcr1+ conventional type 1 dendritic cells preferentially induces IgG2a responses, whereas simultaneous targeting of several dendritic cell subtypes also induces IgG1 responses. The induction of distinct subclass profiles by different surface molecules supports the APC–B cell synapse hypothesis. The results may contribute to generation of more potent DNA vaccines that elicit high levels of Abs with desired biologic effector functions.

• Anderson, Ane Marie; Baranowska-Hustad, Marta Natalia; Braathen, Ranveig; Grødeland, Gunnveig & Bogen, Bjarne (2018). Simultaneous targeting of multiple hemagglutinins to APCs for induction of broad immunity against influenza. Journal of Immunology. ISSN 0022-1767. 200(6), s. 2057–2066. doi: 10.4049/jimmunol.1701088. Fulltekst i vitenarkiv
• Fauskanger, Marte; Haabeth, Ole Audun; Skjeldal, Frode Miltzow; Bogen, Bjarne & Tveita, Anders (2018). Tumor killing by CD4+ T cells is mediated via induction of inducible nitric oxide synthase-dependent macrophage cytotoxicity. Frontiers in Immunology. 9:1684, s. 1–13. doi: 10.3389/fimmu.2018.01684. Fulltekst i vitenarkiv Vis sammendrag

  CD4+ T cells can induce potent anti-tumor immune responses. Due to the lack of MHC class II expression in most cancer cells, antigen recognition occurs indirectly via uptake and presentation on tumor-infiltrating antigen-presenting cells (APCs). Activation of the APCs can induce tumor rejection, but the mechanisms underlying tumor killing by such cells have not been established. To elucidate the molecular basis of CD4+ T-cell-mediated tumor rejection, we utilized a murine model of multiple myeloma, in which the T cells recognize a secreted tumor neoantigen. Our findings demonstrate that T cell recognition triggers inducible nitric oxide synthase activity within tumor-infiltrating macrophages. Diffusion of nitric oxide into surrounding tumor cells results in intracellular accumulation of toxic secondary oxidants, notably peroxynitrite. This results in tumor cell apoptosis through activation of the mitochondrial pathway. We find that this mode of cytotoxicity has strict spatial limitations, and is restricted to the immediate surroundings of the activated macrophage, thus limiting bystander killing. These findings provide a molecular basis for macrophage-mediated anti-tumor immune responses orchestrated by CD4+ T cells. Since macrophages are abundant in most solid tumors, evoking the secretion of nitric oxide by such cells may represent a potent therapeutic strategy.

• Park, Bo Mi; Kim, Eun Jin; Nam, Hye Yeon; Zhang, Dongdong; Bae, Chu Hyun & Kang, Myeongmo [Vis alle 10 forfattere av denne artikkelen] (2017). Cyclized oligopeptide targeting LRP5/6-DKK1 interaction reduces the growth of tumor burden in a multiple myeloma mouse model. Yonsei medical journal. ISSN 0513-5796. 58(3), s. 505–513. doi: 10.3349/ymj.2017.58.3.505. Fulltekst i vitenarkiv
• Wang, Dong; Fløisand, Yngvar; Myklebust, Camilla Victoria; Bürgler, Simone; Ribes, Anna Maria Parente & Hofgaard, Peter Olaf [Vis alle 13 forfattere av denne artikkelen] (2017). Autologous bone marrow Th cells can support multiple myeloma cell proliferation in vitro and in xenografted mice. Leukemia. ISSN 0887-6924. 31(10), s. 2114–2121. doi: 10.1038/leu.2017.69. Fulltekst i vitenarkiv Vis sammendrag

  Multiple myeloma (MM) is a plasma cell malignancy where MM cell growth is supported by the bone marrow (BM) microenvironment with poorly defined cellular and molecular mechanisms. MM cells express CD40, a receptor known to activate autocrine secretion of cytokines and elicit proliferation. Activated T helper (Th) cells express CD40 ligand (CD40L) and BM Th cells are significantly increased in MM patients. We hypothesized that activated BM Th cells could support MM cell growth. We here found that activated autologous BM Th cells supported MM cell growth in a contact- and CD40L-dependent manner in vitro. MM cells had retained the ability to activate Th cells that reciprocated and stimulated MM cell proliferation. Autologous BM Th cells supported MM cell growth in xenografted mice and were found in close contact with MM cells. MM cells secreted chemokines that attracted Th cells, secretion was augmented by CD40-stimulation. Within 14 days of culture of whole BM aspirates in autologous serum, MM cells and Th cells mutually stimulated each other, and MM cells required Th cells for further expansion in vitro and in mice. The results suggest that Th cells may support the expansion of MM cells in patients.

• Andersen, Tor Kristian; Zhou, Fan; Cox, Rebecca Jane; Bogen, Bjarne & Grødeland, Gunnveig (2017). A DNA vaccine that targets hemagglutinin to antigen-presenting cells protects mice against H7 influenza. Journal of Virology. ISSN 0022-538X. 91:e01340-17(23), s. 1–13. doi: 10.1128/JVI.01340-17. Fulltekst i vitenarkiv Vis sammendrag

  Zoonotic influenza H7 viral infections have a case fatality rate of about 40%. Currently, no or limited human to human spread has occurred, but we may be facing a severe pandemic threat if the virus acquires the ability to transmit between humans. Novel vaccines that can be rapidly produced for global distribution are urgently needed, and DNA vaccines may be the only type of vaccine that allows for the speed necessary to quench an emerging pandemic. Here, we constructed DNA vaccines encoding the hemagglutinin (HA) from influenza A/chicken/Italy/13474/99 (H7N1). In order to increase the efficacy of DNA vaccination, HA was targeted to either major histocompatibility complex class II molecules or chemokine receptors 1, 3, and 5 (CCR1/3/5) that are expressed on antigen-presenting cells (APC). A single DNA vaccination with APC-targeted HA significantly increased antibody levels in sera compared to nontargeted control vaccines. The antibodies were confirmed neutralizing in an H7 pseudotype-based neutralization assay. Furthermore, the APC-targeted vaccines increased the levels of antigen-specific cytotoxic T cells, and a single DNA vaccination could confer protection against a lethal challenge with influenza A/turkey/Italy/3889/1999 (H7N1) in mice. In conclusion, we have developed a vaccine that rapidly could contribute protection against a pandemic threat from avian influenza. IMPORTANCE Highly pathogenic avian influenza H7 constitute a pandemic threat that can cause severe illness and death in infected individuals. Vaccination is the main method of prophylaxis against influenza, but current vaccine strategies fall short in a pandemic situation due to a prolonged production time and insufficient production capabilities. In contrast, a DNA vaccine can be rapidly produced and deployed to prevent the potential escalation of a highly pathogenic influenza pandemic. We here demonstrate that a single DNA delivery of hemagglutinin from an H7 influenza could mediate full protection against a lethal challenge with H7N1 influenza in mice. Vaccine efficacy was contingent on targeting of the secreted vaccine protein to antigen-presenting cells.

• Høglund, Rune Alexander; Lossius, Peter Andreas Vold; Johansen, Jorunn; Homan, Jane; Benth, Jurate Saltyte & Robins, Harlan [Vis alle 9 forfattere av denne artikkelen] (2017). In silico prediction analysis of idiotope-driven T-B cell collaboration in multiple sclerosis. Frontiers in Immunology. 8. doi: 10.3389/fimmu.2017.01255. Fulltekst i vitenarkiv Vis sammendrag

  Memory B cells acting as antigen-presenting cells are believed to be important in multiple sclerosis (MS), but the antigen they present remains unknown. We hypothesized that B cells may activate CD4+ T cells in the central nervous system of MS patients by presenting idiotopes from their own immunoglobulin variable regions on human leukocyte antigen (HLA) class II molecules. Here, we use bioinformatics prediction analysis of B cell immunoglobulin variable regions from 11 MS patients and 6 controls with other inflammatory neurological disorders (OINDs), to assess whether the prerequisites for such idiotope-driven T–B cell collaboration are present. Our findings indicate that idiotopes from the complementarity determining region (CDR) 3 of MS patients on average have high predicted affinities for disease associated HLA-DRB1*15:01 molecules and are predicted to be endosomally processed by cathepsin S and L in positions that allows such HLA binding to occur. Additionally, complementarity determining region 3 sequences from cerebrospinal fluid (CSF) B cells from MS patients contain on average more rare T cell-exposed motifs that could potentially escape tolerance and stimulate CD4+ T cells than CSF B cells from OIND patients. Many of these features were associated with preferential use of the IGHV4 gene family by CSF B cells from MS patients. This is the first study to combine high-throughput sequencing of patient immune repertoires with large-scale prediction analysis and provides key indicators for future in vitro and in vivo analyses.

• Deloizy, Charlotte; Fossum, Even; Barnier-Quer, Christophe; Urien, Céline; Chrun, Tiphany & Duval, Audrey [Vis alle 19 forfattere av denne artikkelen] (2017). The anti-influenza M2e antibody response is promoted by XCR1 targeting in pig skin. Scientific Reports. 7:7639, s. 1–12. doi: 10.1038/s41598-017-07372-9. Fulltekst i vitenarkiv
• Gudjonsson, Arnar; Lysén, Anna Karin Christina; Balan, Sreekumar; Gjerstad, Vibeke Sundvold; Arnold-Schrauf, Catharina & Richter, Lisa Maria [Vis alle 10 forfattere av denne artikkelen] (2017). Targeting influenza virus hemagglutinin to Xcr1+ dendritic cells in the absence of receptor-mediated endocytosis enhances protective antibody responses. Journal of Immunology. ISSN 0022-1767. 198(7), s. 2785–2795. doi: 10.4049/jimmunol.1601881. Fulltekst i vitenarkiv
• Lilly, Cameron L; Villa, Nancy Y; Matos, Ana Lemos de; Ali, Haider M; Dhillon, Jess-Karan S & Hofland, Tom [Vis alle 11 forfattere av denne artikkelen] (2017). Ex Vivo Oncolytic Virotherapy with Myxoma Virus Arms Multiple Allogeneic Bone Marrow Transplant Leukocytes to Enhance Graft versus Tumor. Molecular Therapy Oncolytics. 4, s. 31–40. doi: 10.1016/j.omto.2016.12.002. Fulltekst i vitenarkiv
• Kulkarni, Upasana; Karsten, Christian M.; Kohler, Thomas; Hammerschmidt, Sven; Bommert, Kurt & Tiburzy, Benjamin [Vis alle 22 forfattere av denne artikkelen] (2016). IL-10 mediates plasmacytosis-associated immunodeficiency by inhibiting complement-mediated neutrophil migration. Journal of Allergy and Clinical Immunology. ISSN 0091-6749. 137(5), s. 1487–1497e6. doi: 10.1016/j.jaci.2015.10.018. Fulltekst i vitenarkiv
• Lorvik, Kristina Berg; Hammarström, Clara Louise; Fauskanger, Marte; Haabeth, Ole Audun; Zangani, Michael M & Haraldsen, Guttorm [Vis alle 8 forfattere av denne artikkelen] (2016). Adoptive transfer of tumor-specific Th2 cells eradicates tumors by triggering an in situ inflammatory immune response. Cancer Research. ISSN 0008-5472. 76(23), s. 6864–6876. doi: 10.1158/0008-5472.CAN-16-1219. Fulltekst i vitenarkiv
• Bartee, Eric; Bartee, Mee Y.; Bogen, Bjarne & Yu, Xue-Zhong (2016). Systemic therapy with oncolytic myxoma virus cures established residual multiple myeloma in mice. Molecular Therapy Oncolytics. 3:201632. doi: 10.1038/mto.2016.32. Fulltekst i vitenarkiv
• Tveita, Anders; Fauskanger, Marte; Bogen, Bjarne & Haabeth, Ole Audun (2016). Tumor-specific CD4<sup>+</sup> T cells eradicate myeloma cells genetically deficient in MHC class II display. OncoTarget. 7(41), s. 67175–67182. doi: 10.18632/oncotarget.11946. Fulltekst i vitenarkiv
• Ardouin, Laurence; Luche, Hervé; Chelbi, Rabie; Carpentier, Sabrina; Shawket, Alaa & Sanchis, Frédéric Montanana [Vis alle 23 forfattere av denne artikkelen] (2016). Broad and largely concordant molecular changes characterize tolerogenic and immunogenic dendritic cell maturation in thymus and periphery. Immunity. ISSN 1074-7613. 45(2), s. 305–318. doi: 10.1016/j.immuni.2016.07.019. Fulltekst i vitenarkiv
• Lambert, Laura; Kinnear, Ekaterina; McDonald, Jacqueline U.; Grødeland, Gunnveig; Bogen, Bjarne & Stubsrud, Elisabeth [Vis alle 9 forfattere av denne artikkelen] (2016). DNA vaccines encoding antigen targeted to MHC class II induce influenza-specific CD8+ T cell responses, enabling faster resolution of influenza disease. Frontiers in Immunology. 7:321. doi: 10.3389/fimmu.2016.00321. Fulltekst i vitenarkiv
• Andersen, Aram Nikolai; Landsverk, Ole J. B.; Simonsen, Anne; Bogen, Bjarne; Corthay, Alexandre & Øynebråten, Inger (2016). Coupling of HIV-1 antigen to the selective autophagy receptor SQSTM1/p62 promotes T-cell-mediated immunity. Frontiers in Immunology. 7. doi: 10.3389/fimmu.2016.00167. Fulltekst i vitenarkiv Vis sammendrag

  Vaccines aiming to promote T-cell-mediated immune responses have so far showed limited efficacy, and there is a need for novel strategies. Studies indicate that autophagy plays an inherent role in antigen processing and presentation for CD4+ and CD8+ T cells. Here, we report a novel vaccine strategy based on fusion of antigen to the selective autophagy receptor sequestosome 1 (SQSTM1)/p62. We hypothesized that redirection of vaccine antigen from proteasomal degradation into the autophagy pathway would increase the generation of antigen-specific T cells. A hybrid vaccine construct was designed in which the antigen is fused to the C-terminus of p62, a signaling hub, and a receptor that naturally delivers ubiquitinated cargo for autophagic degradation. Fusion of the human immunodeficiency virus-1 antigen Gagp24 to p62 resulted in efficient antigen delivery into the autophagy pathway. Intradermal immunization of mice revealed that, in comparison to Gagp24 delivered alone, fusion to p62 enhanced the number of Gagp24-specific interferon-γ-producing T cells, including CD8+ T cells. The strategy may also have the potential to modulate the antigenic peptide repertoire. Because p62 and autophagy are highly conserved between species, we anticipate this strategy to be a candidate for the development of T-cell-based vaccines in humans.

• Grødeland, Gunnveig; Fredriksen, Agnete; Løset, Geir Åge; Vikse, Elisabeth Lea; Fugger, Lars & Bogen, Bjarne (2016). Antigen targeting to human HLA Class II molecules increases efficacy of DNA vaccination. Journal of Immunology. ISSN 0022-1767. 197(9), s. 3575–3585. doi: 10.4049/jimmunol.1600893. Fulltekst i vitenarkiv
• Haabeth, Ole Audun; Lorvik, Kristina Berg; Yagita, Hideo; Bogen, Bjarne & Corthay, Alexandre (2016). Interleukin-1 is required for cancer eradication mediated by tumor-specific Th1 cells. Oncoimmunology. ISSN 2162-4011. 5(1). doi: 10.1080/2162402X.2015.1039763. Fulltekst i vitenarkiv
• Zhou, Peng; Chionh, Yok Teng; Irac, Sergio Erdal; Ahn, Matae; Ng, Justin Han Jia & Fossum, Even [Vis alle 11 forfattere av denne artikkelen] (2016). Unlocking bat immunology: Establishment of Pteropus alecto bone marrow-derived dendritic cells and macrophages. Scientific Reports. 6. doi: 10.1038/srep38597. Fulltekst i vitenarkiv
• Deloizy, Charlotte; Bouguyon, Edwige; Fossum, Even; Sebo, Peter; Osička, Radim & Bole, Angelique [Vis alle 12 forfattere av denne artikkelen] (2016). Expanding the tools for identifying mononuclear phagocyte subsets in swine: Reagents to porcine CD11c and XCR1. Developmental and Comparative Immunology. ISSN 0145-305X. 65, s. 31–40. doi: 10.1016/j.dci.2016.06.015. Fulltekst i vitenarkiv
• Guilliams, Martin; Dutertre, Charles-Antoine; Scott, Charlotte L.; McGovern, Naomi; Sichien, Dorine & Chakarov, Svetoslav [Vis alle 32 forfattere av denne artikkelen] (2016). Unsupervised high-dimensional analysis aligns dendritic cells across tissues and species. Immunity. ISSN 1074-7613. 45(3), s. 669–684. doi: 10.1016/j.immuni.2016.08.015. Fulltekst i vitenarkiv
• Grødeland, Gunnveig; Fossum, Even & Bogen, Bjarne (2015). Polarizing T and B cell responses by APC-targeted subunit vaccines. Frontiers in Immunology. 6. doi: 10.3389/fimmu.2015.00367. Fulltekst i vitenarkiv
• Tveita, Anders; Schjesvold, Fredrik Hellem; Haabeth, Ole Audun; Fauskanger, Marte Hotvedt & Bogen, Bjarne (2015). Tumors escape CD4+ T-cell-mediated immunosurveillance by impairing the ability of infiltrating macrophages to indirectly present tumor antigens. Cancer Research. ISSN 0008-5472. 75(16), s. 3268–3278. doi: 10.1158/0008-5472.CAN-14-3640. Fulltekst i vitenarkiv Vis sammendrag

  Tumors cells can escape cytotoxic CD8+ T cells by preventing MHC I display of tumor antigens. It is unknown how tumors evade CD4+ T-cell responses, but because many tumor cells lack MHC II expression, novel mechanisms would be required. We have investigated this issue in a model in which MHC IINEG myeloma cells secrete a monoclonal Ig containing a V region L chain (VL) epitope recognized by CD4+ T cells. Infiltrating macrophages process and present the secreted tumor antigen to Th1 cells, resulting in induction of macrophage cytotoxicity and apparent rejection of the tumor. Despite long-term tumor protection in VL-specific T-cell receptor transgenic mice, we here describe that some myeloma cells persisted in a dormant state and, eventually, formed expanding tumors. Escape tumor cells maintained their secretion of complete (H+L) monoclonal Ig with unchanged sequence, while secretion of surplus free L chain was severely diminished. Although free L chains were efficiently processed and presented by tumor-infiltrating macrophages to CD4+ T cells, complete (H+L) monoclonal Ig was not. Forced overexpression of free L chain secretion reinstated tumor rejection. These results show that tumors can escape CD4+ T-cell–mediated rejection by impairing indirect presentation of tumor antigen by infiltrating macrophages. This occurs through a novel mechanism of immunoediting, in which modulation of the quaternary structure of the secreted tumor-specific antigen reduces its immunogenicity.

• Kikuchi, Jiro; Koyama, Daisuke; Wada, Taeko; Izumi, Tohru; Hofgaard, Peter Olaf & Bogen, Bjarne [Vis alle 7 forfattere av denne artikkelen] (2015). Phosphorylation-mediated EZH2 inactivation promotes drug resistance in multiple myeloma. Journal of Clinical Investigation. ISSN 0021-9738. 125(12), s. 4375–4390. doi: 10.1172/JCI80325. Fulltekst i vitenarkiv
• Aas-Hanssen, Kristin; Thompson, Keith M; Bogen, Bjarne & Munthe, Ludvig Andre (2015). Systemic lupus erythematosus: Molecular mimicry between anti-dsDNA CDR3 idiotype, microbial and self peptides-as antigens for Th cells. Frontiers in Immunology. 6:00382. doi: 10.3389/fimmu.2015.00382. Fulltekst i vitenarkiv Vis sammendrag

  Systemic lupus erythematosus (SLE) is marked by a T helper (Th) cell-dependent B cell hyperresponsiveness, with frequent germinal center reactions, and gammaglobulinemia. A feature of SLE is the finding of IgG autoantibodies specific for dsDNA. The specificity of the Th cells that drive the expansion of anti-dsDNA B cells is unresolved. However, anti-microbial, anti-histone, and anti-idiotype Th cell responses have been hypothesized to play a role. It has been entirely unclear if these seemingly disparate Th cell responses and hypotheses could be related or unified. Here, we describe that H chain CDR3 idiotypes from IgG+ B cells of lupus mice have sequence similarities with both microbial and self peptides. Matched sequences were more frequent within the mutated CDR3 repertoire and when sequences were derived from lupus mice with expanded anti-dsDNA B cells. Analyses of histone sequences showed that particular histone peptides were similar to VDJ junctions. Moreover, lupus mice had Th cell responses toward histone peptides similar to anti-dsDNA CDR3 sequences. The results suggest that Th cells in lupus may have multiple cross-reactive specificities linked to the IgVH CDR3 Id-peptide sequences as well as similar DNA-associated protein motifs.

• Terhorst, Dorothea; Fossum, Even; Baranska, Anna; Tamoutounour, Samira; Malosse, Camille & Garbani, Mattia [Vis alle 12 forfattere av denne artikkelen] (2015). Laser-Assisted Intradermal Delivery of Adjuvant-Free Vaccines Targeting XCR1+ Dendritic Cells Induces Potent Antitumoral Responses. Journal of Immunology. ISSN 0022-1767. 194(12), s. 5895–5902. doi: 10.4049/jimmunol.1500564. Fulltekst i vitenarkiv
• Matthes, Thomas W.; Manfroi, B.; Zeller, Andreas; Dunand-Sauthier, Isabelle; Bogen, Bjarne & Huard, Bertrand (2015). Autocrine amplification of immature myeloid cells by IL-6 in multiple myeloma-infiltrated bone marrow. Leukemia. ISSN 0887-6924. 29(9), s. 1882–1890. doi: 10.1038/leu.2015.145. Fulltekst i vitenarkiv
• Grødeland, Gunnveig & Bogen, Bjarne (2015). Efficient vaccine against pandemic influenza: Combining DNA vaccination and targeted delivery to MHC class II molecules. Expert Review of Vaccines. ISSN 1476-0584. 14(6), s. 805–814. doi: 10.1586/14760584.2015.1029919. Fulltekst i vitenarkiv
• Bürgler, Simone; Vives, Aleix Gimeno; Ribes, Anna Maria Parente; Wang, Dong; Os, Audun & Devereux, Stephen [Vis alle 10 forfattere av denne artikkelen] (2015). Chronic lymphocytic leukemia cells express CD38 in response to Th1 cell-derived IFN-γ by a T-bet-dependent mechanism. Journal of Immunology. ISSN 0022-1767. 194(2), s. 827–835. doi: 10.4049/jimmunol.1401350. Fulltekst i vitenarkiv
• Fossum, Even; Grødeland, Gunnveig; Terhorst, Dorothea; Tveita, Anders; Vikse, Elisabeth Lea & Mjaaland, Siri [Vis alle 9 forfattere av denne artikkelen] (2015). Vaccine molecules targeting Xcr1 on cross-presenting DCs induce protective CD8+ T-cell responses against influenza virus. European Journal of Immunology. ISSN 0014-2980. 45(2), s. 624–635. doi: 10.1002/eji.201445080. Fulltekst i vitenarkiv
• Baranowska, Marta Natalia; Hauge, Anna G; Hoornaert, Chloe; Bogen, Bjarne & Grødeland, Gunnveig (2015). Targeting of nucleoprotein to chemokine receptors by DNA vaccination results in increased CD8(+)-mediated cross protection against influenza. Vaccine. ISSN 0264-410X. 33(49), s. 6988–6996. doi: 10.1016/j.vaccine.2015.08.094. Fulltekst i vitenarkiv Vis sammendrag

  Vaccination is at present the most efficient way of preventing influenza infections. Currently used inactivated influenza vaccines can induce virus-neutralizing antibodies that are protective against a particular influenza strain, but hamper the induction of cross-protective T-cell responses to later infections. Thus, influenza vaccines need to be updated annually in order to confer protection against circulating influenza strains. This study aims at developing an efficient vaccine that can induce broader protection against influenza. For this purpose, we have used the highly conserved nucleoprotein (NP) from an influenza A virus subtype H7N7 strain, and inserted it into a vaccine format that targets an antigen directly to relevant antigen presenting cells (APCs). The vaccine format consists of bivalent antigenic and targeting units, linked via an Ig-based dimerization unit. In this study, NP was linked to MIP-1α, a chemokine that targets the linked antigen to chemokine receptors 1, 3 and 5 expressed on various APCs. The vaccine protein was indirectly delivered by DNA. Mice were vaccinated intradermally with plasmids, in combination with electroporation to enhance cellular uptake of DNA. We found that a single DNA vaccination was sufficient for induction of both antibody and T cell responses in BALB/c mice. Targeting of nucleoprotein to chemokine receptors enhanced T cell responses but not antibody responses. Moreover, a single dose of MIP1α-NP conferred protection in BALB/c mice against a lethal challenge with an H1N1 influenza virus. The observed cross-protection was mediated by CD8+ T cells.

• Haabeth, Ole Audun; Tveita, Anders; Fauskanger, Marte Hotvedt; Schjesvold, Fredrik Hellem; Lorvik, Kristina Berg & Hofgaard, Peter Olaf [Vis alle 11 forfattere av denne artikkelen] (2014). How do CD4+ T cells detect and eliminate tumor cells that either lack or express MHC class II molecules? Frontiers in Immunology. 5:174. doi: 10.3389/fimmu.2014.00174. Fulltekst i vitenarkiv Vis sammendrag

  CD4+ T cells contribute to tumor eradication, even in the absence of CD8+ T cells. Cytotoxic CD4+ T cells can directly kill MHC class II positive tumor cells. More surprisingly, CD4+ T cells can indirectly eliminate tumor cells that lack MHC class II expression. Here, we review the mechanisms of direct and indirect CD4+ T cell-mediated elimination of tumor cells. An emphasis is put on T cell receptor (TCR) transgenic models, where anti-tumor responses of naïve CD4+ T cells of defined specificity can be tracked. Some generalizations can tentatively be made. For both MHCIIPOS and MHCIINEG tumors, presentation of tumor-specific antigen by host antigen-presenting cells (APCs) appears to be required for CD4+ T cell priming. This has been extensively studied in a myeloma model (MOPC315), where host APCs in tumor-draining lymph nodes are primed with secreted tumor antigen. Upon antigen recognition, naïve CD4+ T cells differentiate into Th1 cells and migrate to the tumor. At the tumor site, the mechanisms for elimination of MHCIIPOS and MHCIINEG tumor cells differ. In a TCR-transgenic B16 melanoma model, MHCIIPOS melanoma cells are directly killed by cytotoxic CD4+ T cells in a perforin/granzyme B-dependent manner. By contrast, MHCIINEG myeloma cells are killed by IFN-γ stimulated M1-like macrophages. In summary, while the priming phase of CD4+ T cells appears similar for MHCIIPOS and MHCIINEG tumors, the killing mechanisms are different. Unresolved issues and directions for future research are addressed.

• Schwarzer, Rolf; Nickel, N.; Godau, J.; Willie, B.M.; Duda, GN & Schwarzer, R. [Vis alle 12 forfattere av denne artikkelen] (2014). Notch pathway inhibition controls myeloma bone disease in the murine MOPC315.BM model. Blood Cancer Journal. 4(6). doi: 10.1038/bcj.2014.37. Fulltekst i vitenarkiv
• Dutertre, Charles-Antoine; Jourdain, Jean-Pierre; Rancez, Magali; Amraoui, Sonia; Fossum, Even & Bogen, Bjarne [Vis alle 13 forfattere av denne artikkelen] (2014). TLR3-responsive, XCR1<sup>+</sup>, CD141(BDCA-3)<sup>+</sup>/ CD8α<sup>+</sup>- equivalent dendritic cells uncovered in healthy and simian immunodeficiency virus-infected rhesus macaques. Journal of Immunology. ISSN 0022-1767. 192(10), s. 4697–4708. doi: 10.4049/jimmunol.1302448. Fulltekst i vitenarkiv

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• Laursen, Marlene Fyrstenberg; Katta, Kirankumar; Tesfaye, Demo Yemane; Werninghaus, Ina Charlotta; Hinke, Daniëla Maria & Bogen, Bjarne [Vis alle 7 forfattere av denne artikkelen] (2023). An APC-targeted influenza H1-mix DNA vaccine induces cross-protective immunity in mice. Fulltekst i vitenarkiv Vis sammendrag

  Influenza continues to be a threat against health, causing several cases of respiratory deaths and severe illness worldwide. The influenza virus mutates constantly and therefore the influenza vaccines must be annually updated and usually have an efficacy of only 20-60%. Prediction of the next season’s circulating strains are one of the challenges. Novel approaches to generate vaccines with broader protection range are as such of high priority. One approach is to use DNA vaccines, which are rapid and cheap to construct and produce. In a novel DNA vaccine, we have combined multiple HAs from H1N1 virus strains. Four H1 variants were selected from viruses known to circulate in humans. These antigens were expressed as fusion protein to a heterodimerization motif, and a single chain variable fragment (scFv) that targets the vaccine to antigen-presenting cells (APCs) via MHC class II. The antigens were fused to the heterodimerization so that only dimers with two different H1s will be formed in the H1-mix vaccine. This H1-mix vaccine induced antibodies that had high binding to the HA-stem and cross-reacted to several HAs including multiple H1 subtypes and H2 and H5. The cross-reactive antibodies were mainly of IgG2a subtype and activated antibody-dependent cellular cytotoxicity (ADCC). In addition, we found high amounts of H1-specific IFN-γ producing cells in the spleen of H1-mix vaccinated mice. Finally, in heterologous H1N1 challenge studies we saw a significant reduction in viral load in lungs of H1-mix vaccinated mice and the mice were completely protected.

• Nylenna, Magne & Bogen, Bjarne (2022). Terapeutiske vaksiner. Tidsskrift for Den norske legeforening (Tidsskriftet). ISSN 0029-2001. 142. doi: 10.4045/tidsskr.21.0717. Fulltekst i vitenarkiv
• Laursen, Marlene Fyrstenberg; Maria, Hinke Daniela; Bogen, Bjarne & Braathen, Ranveig (2021). An MHC class II-targeted H1-mix Influenza Vaccine Induces Cross-protective Antibody Responses in Mice. Fulltekst i vitenarkiv Vis sammendrag

  Influenza infections are a major cause of morbidity and mortality worldwide. The surface glycoprotein hemagglutinin (HA) is the primary target for neutralizing antibodies, protecting against influenza infections. Within the influenza A virus (IAV) family, HA subtypes H1-H18 exist. In humans, circulating IAVs belonging to H1 and H3 subtypes cause seasonal influenza. Applying influenza vaccines can lower the burden of influenza disease. However, minor genetic mutations causing substitutions of single amino acids in the HA sequence lead to antigenic drift and immune escape of the virus. This lowers the efficacy of current vaccines and annual revaccination is needed. To develop novel influenza vaccines which more broadly protect immunized individuals without the need of an annual boost is consequently of high interest. In our studies, we combined HAs from 4 different H1N1 IAVs into an MHC class II-targeting heterodimeric DNA H1-mix vaccine (H1-mix). Vaccinated mice were followed for several weeks and antibody responses analyzed by ELISA. The H1-mix induced antibodies against a variety of H1s as well as against other subtypes not included in the mix. Furthermore, mice vaccinated with the H1-mix showed complete protection against a heterologous viral infection with the H1N1 virus A/Puerto Rico/8/1934 (PR8), which was not included in the mix. Mice vaccinated with the individual H1s used in the mix vaccine, showed no protection against PR8. This strategy enables an easy adaption of the vaccine to better protect against drifted viral strains, probably by inducing cross-reactive antibodies. This is an important step towards generating a universal influenza vaccine.

• Werninghaus, Ina; Hinke, Daniëla Maria; Bogen, Bjarne & Braathen, Ranveig (2021). Neuraminidase in antigen presenting cell-targeted DNA vaccine induces antibody-mediated protection against influenza. Fulltekst i vitenarkiv Vis sammendrag

  Vaccines provide an effective countermeasure against influenza. However, antigenic drift of the virus and varying vaccine efficacy result in a need for novel and flexible vaccine approaches. Conventional influenza vaccines are standardized based on their hemagglutinin (HA) content, however neuraminidase (NA) has also been established as an independent correlate of protection and has great potential as a vaccine antigen. Our lab designed a flexible and rapidly producible DNA vaccine platform, increasing immunogenicity by targeting dimeric vaccine molecules to antigen-presenting cells (APCs). Here, for the first time, NA was included in the APC-targeted DNA vaccine platform encoding MHC class II-targeted heterodimeric antigen-bivalent vaccine proteins. The vaccine with NA was compared to an otherwise identical HA vaccine, which proved successful in previous studies. A single immunization elicited strong and long-lasting NA-specific humoral responses, which had the ability to inhibit NA enzymatic activity. Vaccine-induced NA immunity protected mice completely from challenge with homologous influenza virus. Immunization with NA was overall more effective than the HA-based vaccine and potent at low vaccine doses. Additionally, NA-reactive CD4+ and CD8+ T cells were induced, however protection was mainly antibody-mediated. Furthermore, similarly strong NA-specific immune responses and protection could be obtained with different targeting units as well as dimerization motifs. Overall, our results show that NA is a potent influenza vaccine antigen and can flexibly be used in the APC-targeted DNA vaccine platform. Strategies to combine NA with other influenza antigens like HA to elicit broadened immunity should be explored in future influenza vaccine design.

• Hinke, Daniëla Maria; Andersen, Tor Kristian; Skullerud, Lise Madelen; Werninghaus, Ina; Grødeland, Gunnveig & Fossum, Even [Vis alle 8 forfattere av denne artikkelen] (2021). Antigen bivalency in antigen presenting cell-targeted vaccines enhances humoral responses. Fulltekst i vitenarkiv Vis sammendrag

  Pathogen-specific antibodies can provide protective immunity against infectious diseases. Vaccine-elicited antibody levels can be increased by targeting antigens to antigen presenting cells (APCs). We hypothesize that antigen multivalency can play an additional role to further enhance antibody responses against APC-targeted vaccines. Here, we examine the role of antigen valency in APC-targeted vaccines delivered in the form of DNA. We designed MHC class II-targeted heterodimeric vaccine molecules that carried either two identical antigens (bivalent vaccines) or two different antigens (monovalent vaccines). Besides antigen valency, the vaccine molecules were identical. Bivalent and monovalent vaccines were compared in their efficiency to induce humoral responses in mice. We found that antigen bivalent vaccine molecules with two identical influenza hemagglutinins (HA) induced higher amounts of anti-HA antibodies than monovalent versions. Furthermore, only MHC class II-targeted HA-bivalent vaccines protected mice completely from challenge with homologous influenza virus. The potentiating effect of antigen bivalency was also obtained with other antigens and APC targeting in the DNA vaccine format, and when delivering vaccines as recombinant protein instead of DNA. In vitro data showed that MHC class II-targeted vaccine proteins aid the clustering of APCs and antigen-specific B cells, suggesting the formation of APC-B cell synapses. We hypothesize that antigen bivalency contributes to efficient cross-linking of B cell receptors within the synapse, resulting in increased B cell responses. Overall, our results indicate that combining antigen bivalency with APC-targeting is an attractive strategy to enhance the immunogenicity of vaccines.

• Lysén, Anna Karin Christina; Braathen, Ranveig; Gudjonsson, Arnar; Tesfaye, Demo Yemane; Bogen, Bjarne & Fossum, Even (2020). Author Correction: Dendritic cell targeted Ccl3- and Xcl1-fusion DNA vaccines differ in induced immune responses and optimal delivery site (Scientific Reports, (2019), 9, 1, (1820), 10.1038/s41598-018-38080-7). Scientific Reports. 10(1). doi: 10.1038/s41598-020-62688-3. Fulltekst i vitenarkiv
• Gudjonsson, Arnar; Bjerkan, Louise; Labbe, Genevieve; Draper, Simon; Braathen, Ranveig & Bogen, Bjarne (2019). Targeting PfRH5 to APCs via MHC class II potentiates viral vector immunization against malaria. Fulltekst i vitenarkiv
• Bjerkan, Louise; Gudjonsson, Arnar; Visweswaran, Ganesh Ram; Genevieve, Labbe; Quinkert, Doris & Draper, Simon [Vis alle 8 forfattere av denne artikkelen] (2019). APC-targeted DNA vaccination against RH5 induces Plasmodium falciparum-specific neutralizing antibodies and T cell responses. Fulltekst i vitenarkiv
• Bjerkan, Louise; Braathen, Ranveig; Gudjonsson, Arnar; Grødeland, Gunnveig; Visweswaran, Ganesh Ram & Bogen, Bjarne (2018). Targeted DNA vaccination induce protective immune responses against malaria. Fulltekst i vitenarkiv
• Bjerkan, Louise; Braathen, Ranveig; Gudjonsson, Arnar; Grødeland, Gunnveig; Visweswaran, Ganesh Ram & Bogen, Bjarne (2018). An APC-targeted PfRH5 containing DNA vaccine induce protective immune responses against Plasmodium falciparum. Fulltekst i vitenarkiv
• Hinke, Daniëla Maria; Spång, Heidi; Fossum, Even; Grødeland, Gunnveig; Bogen, Bjarne & Braathen, Ranveig (2018). MHC class II targeted DNA vaccine is most efficient in the induction of protective antibodies against influenza. Fulltekst i vitenarkiv
• Braathen, Ranveig; Spång, Heidi; Hinke, Daniela Maria; Blazevski, Jana & Bogen, Bjarne (2018). Novel heterodimeric DNA vaccine targeting APC protects against MOPC315 myeloma and influenza in BALB/c mice. Fulltekst i vitenarkiv
• Haabeth, Ole Audun; Tveita, Anders; Fauskanger, Marte Hotvedt; Hennig, Kjartan; Hofgaard, Peter Olaf & Bogen, Bjarne (2016). Idiotype-specific CD4+ T cells eradicate disseminated myeloma. Leukemia. ISSN 0887-6924. 30(5), s. 1216–1220. doi: 10.1038/leu.2015.278. Fulltekst i vitenarkiv
• Visweswaran, Ganesh Ram; Bjerkan, Louise; Braathen, Ranveig & Bogen, Bjarne (2016). DEVELOPMENT OF EFFICACIOUS TARGETED DNA VACCINES AGAINST MALARIA. Fulltekst i vitenarkiv Vis sammendrag

  DEVELOPMENT OF EFFICACIOUS TARGETED DNA VACCINES AGAINST MALARIA Ganesh Ram Visweswaran1, Ranveig Braathen1, Louise Bjerkan1, Genevieve Labbe2, Simon J. Draper2 and Bjarne Bogen1 1Institute of Immunology, University of Oslo, Oslo, Norway, 2University of Oxford, Oxford, United Kingdom Objective: To develop novel malaria vaccines (DNA, Protein, and Viral-vectored) that induces long-lasting immune responses and confers protection against malaria. Introduction: Malaria, one of the leading causes of human death (584,000 in 2013) in developing countries, is caused by the protozoan parasite Plasmodium and transmitted by the female anopheles mosquito. To date there are no efficacious malaria vaccines available that would impart protective immune response. In this study, a novel DNA vaccine technology (Vaccibody®) is being used to develop Malaria vaccine using a putative blood-stage antigen P. falciparum Reticulocyte Binding Protein Homologue 5 (PfRH5). Method: Vaccibody is composed of a targeting, dimerization, and an antigenic unit. The first two units are being involved in targeting the MHC-class II molecules on antigen presenting cells and for homodimerization of the vaccibody molecules, respectively. The antigenic unit contains the potential antigen (PfRH5) of interest. Results: All PfRH5 vaccibody proteins were secreted by HEK293E cells and were shown to bind different PfRH5 monoclonal antibodies by ELISA. In vivo experiments revealed that the MHC-class II PfRH5 vaccibodies are better targeted and induced significant antibody titers when compared to the non-targeted and antigen controls. Conclusion: Binding by different PfRH5 monoclonal antibodies confirmed functional PfRH5 vaccibody proteins. The MHC-class II targeted PfRH5 vaccibodies elicited higher immune response in mice than the non-targeted and antigen controls, indicating the targeting potential.

• Bjerkan, Louise; Visweswaran, Ganesh Ram; Braathen, Ranveig & Bogen, Bjarne (2016). Novel malaria vaccines by VaccibodyTM DNA immunization. Fulltekst i vitenarkiv Vis sammendrag

  Novel malaria vaccines by VaccibodyTM DNA immunization L Bjerkan1, GR Visweswaran1, R Braathen1, G Labbe 3, SJ Draper3, B Bogen1,2 1K.G. Jebsen Centre for Research on Influenza Vaccines, Oslo 0450, Norway 2Centre for Immune Regulation Institute of Immunology, University of Oslo and Oslo University Hospital, Oslo 0027, Norway 3 The Jenner Institute, University of Oxford, Oxford, United Kingdom DNA vaccines have a multitude of advantages such as ease of construction, mass production, and safety. In this study, we exploit a new strategy for DNA vaccination where transfected cells secrete bivalent fusion proteins, denoted vaccibodies. These molecules target antigen to antigen presenting cells (APCs) resulting in efficient stimulation of B and T cells. Results from this study show that BALB/c mice immunized with vaccibody homodimers bearing the Plasmodium falciparum blood stage antigen reticulocyte binding homolog RH5, induce high antibody titers and exhibit a targeting effect even after viral boost. This DNA vaccination platform will also be tested in larger animals.

• Haabeth, Ole Audun; Lorvik, Kristina Berg; Hammarström, Clara Louise; Donaldson, Ian M; Haraldsen, Guttorm & Bogen, Bjarne [Vis alle 7 forfattere av denne artikkelen] (2016). Kreft-spesifikke betennelsesreaksjoner kan beskytte mot kreft. Bestpractice Onkologi/Hematologi. ISSN 2245-0440. February. Fulltekst i vitenarkiv
• Ribes, Anna Maria Parente; Skånland, Sigrid Strand; Bürgler, Simone; Os, Audun; Wang, Dong & Bogen, Bjarne [Vis alle 9 forfattere av denne artikkelen] (2016). Spleen tyrosine kinase inhibitors reduce CD40l-induced proliferation of chronic lymphocytic leukemia cells but not normal B cells. Haematologica. ISSN 0390-6078. 101(2), s. e59–e62. doi: 10.3324/haematol.2015.135590. Fulltekst i vitenarkiv
• Osnes, Liv T. N. & Bogen, Bjarne (2015). Primære immundefekter. I Hagve, Tor-Arne & Berg, Jens Petter (Red.), Klinisk biokjemi og fysiologi. Gyldendal Akademisk. ISSN 9788205463677. s. 147–154. Fulltekst i vitenarkiv
• Tjønnfjord, Geir Erland & Bogen, Bjarne (2015). Myeloproliferative og lymfoproliferative sykdommer. I Hagve, Tor-Arne & Berg, Jens Petter (Red.), Klinisk biokjemi og fysiologi. Gyldendal Akademisk. ISSN 9788205463677. s. 140–154. Fulltekst i vitenarkiv

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