title: CD40 ligand preferentially modulates immune response and enhances protection against influenza virus.
abstract: CD40L, a key regulator of the immune system, was studied as both a targeting ligand and a molecular adjuvant in nucleoprotein (NP)-based host defense against influenza in mouse models with different genetic backgrounds. Adenoviral vectors secreting NP-CD40L fusion protein (denoted as rAd-SNP40L) afforded full protection of immunocompetent and immunocompromised mice (CD40L(-/-) and CD4(-/-)) against lethal influenza infection. Mechanistically, rAd-SNP40L preferentially induced early and persistent B cell germinal center formation, and accelerated Ig isotype-switching and Th1-skewed, NP-specific Ab response. Moreover, it drastically augmented primary and memory NP-specific CTL activity and polyfunctional CD8(+) T cells. The markedly enhanced nonneutralizing Abs and CTLs significantly reduced viral burdens in the lungs of mice upon lethal virus challenge. Data generated from CD40L(-/-) and CD4(-/-) mice revealed that the protection was indeed CD40L mediated but CD4(+) T cell independent, demonstrating the viability of the fusion Ags in protecting immunodeficient hosts. Notably, a single dose of rAd-SNP40L completely protected mice from lethal viral challenge 4 mo after immunization, representing the first report, to our knowledge, on NP in conjunction with a molecular adjuvant inducing a robust and long-lasting memory immune response against influenza. This platform is characterized by an increased in vivo load of CD40-targeted Ag upon the secretion of the fusion protein from adenovirus-infected cells and may represent a promising strategy to enhance the breadth, durability, and potency of Ag-specific immune responses.
authors.affiliation: Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario K1A 0K9, Canada; Department of Medical Microbiology and Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah 21589, Saudi Arabia; Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia;
authors.forename: Anwar M
authors.initials: AM
authors.lastname: Hashem
authors.affiliation: Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario K1A 0K9, Canada;
authors.forename: Caroline
authors.initials: C
authors.lastname: Gravel
authors.affiliation: Shanghai Institute of Biological Products, Shanghai 200231, China;
authors.forename: Ze
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Shanghai Institute of Biological Products, Shanghai 200231, China;
authors.forename: Yinglei
authors.initials: Y
authors.lastname: Yi
authors.affiliation: Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario K1A 0K9, Canada;
authors.forename: Monika
authors.initials: M
authors.lastname: Tocchi
authors.affiliation: Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario K1A 0K9, Canada;
authors.forename: Bozena
authors.initials: B
authors.lastname: Jaentschke
authors.affiliation: National Institutes for the Control of Food and Drug, Beijing 10050, People's Republic of China;
authors.forename: Xingliang
authors.initials: X
authors.lastname: Fan
authors.affiliation: National Institutes for the Control of Food and Drug, Beijing 10050, People's Republic of China;
authors.forename: Changgui
authors.initials: C
authors.lastname: Li
authors.affiliation: Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario K1A 0K9, Canada;
authors.forename: Michael
authors.initials: M
authors.lastname: Rosu-Myles
authors.affiliation: Division of Human Gene Therapy, Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294; Department of Obstetrics and Gynecology, University of Alabama at Birmingham, Birmingham, AL 35294; Department of Pathology and Surgery, University of Alabama at Birmingham, Birmingham, AL 35294; Gene Therapy Center, University of Alabama at Birmingham, Birmingham, AL 35294;
authors.forename: Alexander
authors.initials: A
authors.lastname: Pereboev
authors.affiliation: National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba R3E 3R2, Canada; and.
authors.forename: Runtao
authors.initials: R
authors.lastname: He
authors.affiliation: National Institutes for the Control of Food and Drug, Beijing 10050, People's Republic of China; Sean.Li@hc-sc.gc.ca wangjz@nicpbp.org.cn.
authors.forename: Junzhi
authors.initials: J
authors.lastname: Wang
authors.affiliation: Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario K1A 0K9, Canada; Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, Ontario K1H 8M5, Canada Sean.Li@hc-sc.gc.ca wangjz@nicpbp.org.cn.
authors.forename: Xuguang
authors.initials: X
authors.lastname: Li
mesh_terms: D056704:Adaptive Immunity / Q000235:genetics / Q000276:immunology*; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D023201:CD40 Ligand / Q000172:deficiency / Q000235:genetics / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D004285:Dogs; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D007114:Immunization; D009980:Influenza A virus / Q000276:immunology* / Q000502:physiology; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D041681:NIH 3T3 Cells; D009698:Nucleoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D009976:Orthomyxoviridae Infections / Q000235:genetics / Q000276:immunology* / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016019:Survival Analysis; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D009698:Nucleoproteins; D011993:Recombinant Fusion Proteins; D023201:CD40 Ligand