A Comparison of the Humoral and Cellular Immune Responses at Different Immunological Sites after Split Influenza Virus Vaccination of Mice

ABSTRACTSeasonal influenza is a vaccine-preventable disease that remains a major health problem worldwide, especially in immunocompromised populations. The impact of influenza disease is even greater when strains drift, and influenza pandemics can result when animal-derived influenza virus strains combine with seasonal strains. In this study, we used the SAM technology and characterized the immunogenicity and efficacy of a self-amplifying mRNA expressing influenza virus hemagglutinin (HA) antigen [SAM(HA)] formulated with a novel oil-in-water cationic nanoemulsion. We demonstrated that SAM(HA) was immunogenic in ferrets and facilitated containment of viral replication in the upper respiratory tract of influenza virus-infected animals. In mice, SAM(HA) induced potent functional neutralizing antibody and cellular immune responses, characterized by HA-specific CD4 T helper 1 and CD8 cytotoxic T cells. Furthermore, mice immunized with SAM(HA) derived from the influenza A virus A/California/7/2009 (H1N1) strain (Cal) were protected from a lethal challenge with the heterologous mouse-adapted A/PR/8/1934 (H1N1) virus strain (PR8). Sera derived from SAM(H1-Cal)-immunized animals were not cross-reactive with the PR8 virus, whereas cross-reactivity was observed for HA-specific CD4 and CD8 T cells. Finally, depletion of T cells demonstrated that T-cell responses were essential in mediating heterologous protection. If the SAM vaccine platform proves safe, well tolerated, and effective in humans, the fully synthetic SAM vaccine technology could provide a rapid response platform to control pandemic influenza.IMPORTANCEIn this study, we describe protective immune responses in mice and ferrets after vaccination with a novel HA-based influenza vaccine. This novel type of vaccine elicits both humoral and cellular immune responses. Although vaccine-specific antibodies are the key players in mediating protection from homologous influenza virus infections, vaccine-specific T cells contribute to the control of heterologous infections. The rapid production capacity and the synthetic origin of the vaccine antigen make the SAM platform particularly exploitable in case of influenza pandemic.

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H5N1 influenza virus-like particles produced by transient expression in mammalian cells induce humoral and cellular immune responses in mice

Canadian Journal of Microbiology ◽

10.1139/w2012-006 ◽

2012 ◽

Vol 58 (4) ◽

pp. 391-401 ◽

Cited By ~ 4

Author(s):

Ling Tao ◽

Jianjun Chen ◽

Zhenhua Zheng ◽

Jin Meng ◽

Zhenfeng Zhang ◽

...

Keyword(s):

Influenza Virus ◽

Immune Responses ◽

Cleavage Site ◽

Mammalian Cells ◽

Influenza Virus Infection ◽

Influenza Viruses ◽

Virus Like Particles ◽

Cellular Immune Responses ◽

H5n1 Influenza ◽

Cellular Immune

Vaccination is an effective way to protect from influenza virus infection. Among the new candidates of influenza vaccines, influenza virus-like particles (VLPs) seem to be promising. Here, we generated 2 types of H5N1 influenza VLPs by co-expressing influenza virus Env (envelope protein) and murine leukemia virus (MLV) Gag–Pol. VLPs generated by co-transfection of pHCMV-wtH5 or pHCMV-mtH5 with pSV-Mo-MLVgagpol and pHCMV-N1 were named as wtH5N1 VLPs or mtH5N1 VLPs. The plasmid of pHCMV-wtH5 encoded the wild-type hemagglutinin (HA) (wtH5) from A/swine/Anhui/ca/2004 (H5N1) with a multibasic cleavage site, while pHCMV-mtH5 encoded the modified mutant-type (mtH5) with a monobasic cleavage site. Influenza virus HA VLPs were characterized and equal amounts of them were used to immunize mice subcutaneously, intraperitoneally, or intramuscularly. The levels of HA-specific IgG1, IFN-γ, and neutralization antibodies were significantly induced in mice immunized with wtH5N1 VLPs or mtH5N1 VLPs via all 3 routes, while HA-specific IgG2a was barely detectable. IL-4 secretion was detected in mice subcutaneously immunized with wtH5N1 VLPs or mtH5N1 VLPs, or intramuscularly immunized with mtH5N1 VLPs. Our results indicated that both H5N1 influenza VLPs could induce specific humoral and cellular immune responses in immunized mice. In conclusion, our study provides helpful information for designing new candidate vaccines against H5N1 influenza viruses.

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Analyses of Cellular Immune Responses in Ferrets Following Influenza Virus Infection

Methods in Molecular Biology - Influenza Virus ◽

10.1007/978-1-4939-8678-1_24 ◽

2018 ◽

pp. 513-530 ◽

Cited By ~ 3

Author(s):

Anthony T. DiPiazza ◽

Katherine A. Richards ◽

Wen-Chun Liu ◽

Randy A. Albrecht ◽

Andrea J. Sant

Keyword(s):

Influenza Virus ◽

Virus Infection ◽

Immune Responses ◽

Influenza Virus Infection ◽

Cellular Immune Responses ◽

Cellular Immune

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Cellular immune responses of mice to influenza virus infection

Cellular Immunology ◽

10.1016/0008-8749(80)90124-0 ◽

1980 ◽

Vol 56 (2) ◽

pp. 502-509 ◽

Cited By ~ 10

Author(s):

Carol S. Reiss ◽

Jerome L. Schulman

Keyword(s):

Influenza Virus ◽

Virus Infection ◽

Immune Responses ◽

Influenza Virus Infection ◽

Cellular Immune Responses ◽

Cellular Immune

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Distinct cellular immune responses following primary and secondary influenza virus challenge in cotton rats

Cellular Immunology ◽

10.1016/j.cellimm.2006.12.005 ◽

2006 ◽

Vol 243 (2) ◽

pp. 67-74 ◽

Cited By ~ 9

Author(s):

Maryna C. Eichelberger ◽

Samantha Bauchiero ◽

Dana Point ◽

Bettina W.M. Richter ◽

Gregory A. Prince ◽

...

Keyword(s):

Influenza Virus ◽

Immune Responses ◽

Virus Challenge ◽

Cellular Immune Responses ◽

Cotton Rats ◽

Cellular Immune

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Complement C3 Plays a Key Role in Inducing Humoral and Cellular Immune Responses to Influenza Virus Strain-Specific Hemagglutinin-Based or Cross-Protective M2 Extracellular Domain-Based Vaccination

Journal of Virology ◽

10.1128/jvi.00969-18 ◽

2018 ◽

Vol 92 (20) ◽

Cited By ~ 6

Author(s):

Yu-Jin Kim ◽

Ki-Hye Kim ◽

Eun-Ju Ko ◽

Min-Chul Kim ◽

Yu-Na Lee ◽

...

Keyword(s):

Influenza Virus ◽

Immune Responses ◽

Adaptive Immunity ◽

Immune Defense ◽

Innate Immune ◽

Complement C3 ◽

Complement Protein ◽

Cellular Immune Responses ◽

Homologous Virus ◽

Cellular Immune

ABSTRACTThe complement pathway is involved in eliminating antigen immune complexes. However, the role of the C3 complement system remains largely unknown in influenza virus M2 extracellular (M2e) domain or hemagglutinin (HA) vaccine-mediated protection after vaccination. Using a C3 knockout (C3 KO) mouse model, we found that complement protein C3 was required for effective induction of immune responses to vaccination with M2e-based or HA-based vaccines, which include isotype class-switched antibodies and effector CD4 and CD8 T cell responses. C3 KO mice after active immunization with cross-protective nonneutralizing M2e-based vaccine were not protected against influenza virus, although low levels of M2e-specific antibodies were protective after passive coadministration with virus in wild-type mice. In contrast, C3 KO mice that were immunized with strain-specific neutralizing HA-based vaccine were protected against homologous virus challenge despite lower levels of HA antibody responses. C3 KO mice showed impaired maintenance of innate immune cells and a defect in innate immune responses upon exposure to antigens. The findings in this study suggest that C3 is required for effective induction of humoral and cellular adaptive immune responses as well as protective immunity after nonneutralizing influenza M2e vaccination.IMPORTANCEComplement is the well-known innate immune defense system involved in the opsonization and lysis of pathogens but is less studied in establishing adaptive immunity after vaccination. Influenza virus HA-based vaccination confers protection via strain-specific neutralizing antibodies, whereas M2e-based vaccination induces a broad spectrum of protection by immunity against the conserved M2e epitopes. This study revealed the critical roles of C3 complement in inducing humoral and cellular immune responses after immunization with M2e or HA vaccines. C3 was found to be required for protection by M2e-based but not by HA-based active vaccination as well as for maintaining innate antigen-presenting cells. Findings in this study have insight into better understanding the roles of C3 complement in inducing effective innate and adaptive immunity as well as in conferring protection by cross-protective conserved M2e vaccination.

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A DNA Vaccine-Encoded Nucleoprotein of Influenza Virus Fails To Induce Cellular Immune Responses in a Diabetic Mouse Model

Clinical and Vaccine Immunology ◽

10.1128/cvi.00445-09 ◽

2010 ◽

Vol 17 (4) ◽

pp. 683-687 ◽

Cited By ~ 5

Author(s):

Abbas Jamali ◽

Farzaneh Sabahi ◽

Taravat Bamdad ◽

Hamidreza Hashemi ◽

Fereidoun Mahboudi ◽

...

Keyword(s):

Influenza Virus ◽

Virus Infection ◽

Immune Responses ◽

Dna Vaccine ◽

Lymphocyte Proliferation ◽

Influenza Virus Infection ◽

Diabetic Mice ◽

Cellular Immune Responses ◽

Ifn Γ ◽

Cellular Immune

ABSTRACT Influenza virus infections cause yearly epidemics and are a major cause of lower respiratory tract illnesses in humans worldwide. Influenza virus has long been recognized to be associated with higher morbidity and mortality in diabetic patients. Vaccination is an effective tool to prevent influenza virus infection in this group of patients. Vaccines employing recombinant-DNA technologies are an alternative to inactivated virus and live attenuated virus vaccines. Internal highly conserved viral nucleoprotein (NP) can be delivered as a DNA vaccine to provide heterosubtypic immunity, offering resistance against various influenza virus strains. In this study, we investigated the efficacy of an NP DNA vaccine for induction of cell-mediated immune responses and protection against influenza virus infection in a mouse model of diabetes. Healthy and diabetic BALB/c mice were immunized on days 0, 14, and 28 by injection of NP DNA vaccine. Two weeks after the last immunization, the cellular immune response was evaluated by gamma interferon (IFN-γ), lymphocyte proliferation, and cytotoxicity assays. The mice were challenged with influenza virus, and the viral titers in the lungs were measured on day 4. Diabetic mice showed significantly smaller amounts of IFN-γ production, lymphocyte proliferation, and cytotoxicity responses than nondiabetic mice. Furthermore, higher titers of the influenza virus were detected after challenge in the lungs of the diabetic mice. The present data suggest that the NP DNA vaccine with the protocol of immunization described here is not able to induce efficient cellular immune responses against influenza virus infection in diabetic mice.

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