Viral Escape by Selection of Cytotoxic T Cell–Resistant Variants in Influenza a Virus Pneumonia

Antigenic variation is a strategy exploited by influenza viruses to promote survival in the face of the host adaptive immune response and constitutes a major obstacle to efficient vaccine development. Thus, variation in the surface glycoproteins hemagglutinin and neuraminidase is reflected by changes in susceptibility to antibody neutralization. This has led to the current view that antibody-mediated selection of influenza A viruses constitutes the basis for annual influenza epidemics and periodic pandemics. However, infection with this virus elicits a vigorous protective CD8+ cytotoxic T lymphocyte (CTL) response, suggesting that CD8+ CTLs might exert selection pressure on the virus. Studies with influenza A virus–infected transgenic mice bearing a T cell receptor (TCR) specific for viral nucleoprotein reveal that virus reemergence and persistence occurs weeks after the acute infection has apparently been controlled. The persisting virus is no longer recognized by CTLs, indicating that amino acid changes in the major viral nucleoprotein CTL epitope can be rapidly accumulated in vivo. These mutations lead to a total or partial loss of recognition by polyclonal CTLs by affecting presentation of viral peptide by class I major histocompatibility complex (MHC) molecules, or by interfering with TCR recognition of the mutant peptide–MHC complex. These data illustrate the distinct features of pulmonary immunity in selection of CTL escape variants. The likelihood of emergence and the biological impact of CTL escape variants on the clinical outcome of influenza pneumonia in an immunocompetent host, which is relevant for the design of preventive vaccines against this and other respiratory viral infections, are discussed.

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Perforin and Fas Cytolytic Pathways Coordinately Shape the Selection and Diversity of CD8+-T-Cell Escape Variants of Influenza Virus

Journal of Virology ◽

10.1128/jvi.79.13.8545-8559.2005 ◽

2005 ◽

Vol 79 (13) ◽

pp. 8545-8559 ◽

Cited By ~ 18

Author(s):

Graeme E. Price ◽

Lei Huang ◽

Rong Ou ◽

Menghua Zhang ◽

Demetrius Moskophidis

Keyword(s):

Influenza Virus ◽

T Cell ◽

Vaccine Development ◽

Influenza Virus Infection ◽

Influenza Viruses ◽

Viral Escape ◽

Viral Control ◽

Ctl Escape ◽

Selection Of

ABSTRACT Antigenic variation is a viral strategy exploited to promote survival in the face of the host immune response and represents a major challenge for efficient vaccine development. Influenza viruses are pathogens with high transmissibility and mutation rates, enabling viral escape from immunity induced by prior infection or vaccination. Intense selection from neutralizing antibody drives antigenic changes in the surface glycoproteins, resulting in emergence of new strains able to reinfect hosts immune to previously circulating viruses. CD8+ cytotoxic T cells (CTLs) also provide protective immunity from influenza virus infection and may contribute to the antigenic evolution of influenza viruses. Utilizing mice transgenic for an influenza virus NP366-374 peptide-specific T-cell receptor, we demonstrated that the respiratory tract is a suitable site for generation of escape variants of influenza virus selected by CTL in vivo. In this report the contributions of the perforin and Fas pathways utilized by influenza virus-specific CTLs in viral clearance and selection of CTL escape variants have been evaluated. While transgenic CTLs deficient in either perforin- or Fas-mediated pathways are efficient in initial pulmonary viral control, variant virus emergence was observed in all the mice studied, although the spectrum of viral CTL escape variants selected varied profoundly. Thus, a less-restricted repertoire of escape variants was observed in mice with an intact perforin cytotoxic pathway compared with a limited variant diversity in perforin pathway-deficient mice, although maximal variant diversity was observed in mice having both Fas and perforin pathways intact. We conclude that selection of viral CTL escape variants reflects coordinate action between the tightly controlled perforin/granzyme pathway and the more promiscuous Fas/FasL pathway.

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Design, synthesis and primary immunologic evaluation of M2e-CRM197 conjugate as a universal influenza vaccine candidate

Current Pharmaceutical Biotechnology ◽

10.2174/1389201021666201104145006 ◽

2020 ◽

Vol 21 ◽

Author(s):

Lu Xu ◽

Chun Zhang ◽

Jing Zhang ◽

Rong Yu ◽

Zhiguo Su

Keyword(s):

Immune Responses ◽

Influenza Vaccine ◽

Influenza A Virus ◽

Influenza A ◽

Vaccine Development ◽

Acute Infection ◽

Acid Hydrazide ◽

Influenza Viruses ◽

Carrier Protein ◽

Universal Influenza Vaccine

Background: Influenza is a contagious respiratory illness caused by acute infection of influenza viruses, among which influenza A virus causes epidemic seasonal infection nearly every year. Along with unpredictability of evolving influenza A virus and time-consuming vaccine development cycles, novel universal influenza vaccine designed to induce broadly cross-reactive immune responses against frequently mutant influenza A virus strains are greatly urgent. Objective: The aim of this study was to synthesize a novel vaccine through the dual-site specific conjugation of the constant epitope of 23 amino acids (M2e) of influenza A virus with highly immunogenic carrier protein of cross-reacting material (CRM197) under denaturation, and evaluate its primary immunogenicity in mice. Methods: The antigen (M2e) and the carrier protein (CRM197) were linked with different type of hetero-functionalized linkers, α-maleimide-ε-hydrazide polyethylene glycol 2k (MAL-PEG-HZ) and N-β-maleimidopropionic acid hydrazide (BMPH) separately. The immunogenicity of the M2e-CRM197 conjugates with different type of linkers was evaluated in mice, and the M2e-specific total IgG and IgG-isotypes were determined by ELSIA. Results: Immunogenicity study revealed that anti-M2e antibody could be induced by the conjugate products, M2e-PEGCRM197 and M2e-BMPH-CRM197, were approximately 30 and 90-fold higher than that of M2e group. In addition, the antiM2e antibody level induced by M2e-PEG-CRM197 conjugate was three times higher than that of M2e-BMPH-CRM197 conjugate, and the former could simultaneously activate both cellar and humoral immune responses. Conclusions: The M2e-CRM197 conjugated vaccines we synthesized in this study are highly immunogenic compared with M2e alone. Besides, evidences were presented here indicated that the hydrophilic, non-immunogenic and biocompatible chain of the cross-linker might be a better choice for development of conjugate vaccine.

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Broad neutralization of H1 and H3 viruses by adjuvanted influenza HA stem vaccines in nonhuman primates

Science Translational Medicine ◽

10.1126/scitranslmed.abe5449 ◽

2021 ◽

Vol 13 (583) ◽

pp. eabe5449

Author(s):

Nicole Darricarrère ◽

Yu Qiu ◽

Masaru Kanekiyo ◽

Adrian Creanga ◽

Rebecca A. Gillespie ◽

...

Keyword(s):

Influenza A Virus ◽

Influenza A ◽

Nonhuman Primates ◽

Vaccine Development ◽

Protective Efficacy ◽

Neutralizing Antibody ◽

Gene Families ◽

Influenza Viruses ◽

Public Health Importance ◽

Water Emulsion

Seasonal influenza vaccines confer protection against specific viral strains but have restricted breadth that limits their protective efficacy. The H1 and H3 subtypes of influenza A virus cause most of the seasonal epidemics observed in humans and are the major drivers of influenza A virus–associated mortality. The consequences of pandemic spread of COVID-19 underscore the public health importance of prospective vaccine development. Here, we show that headless hemagglutinin (HA) stabilized-stem immunogens presented on ferritin nanoparticles elicit broadly neutralizing antibody (bnAb) responses to diverse H1 and H3 viruses in nonhuman primates (NHPs) when delivered with a squalene-based oil-in-water emulsion adjuvant, AF03. The neutralization potency and breadth of antibodies isolated from NHPs were comparable to human bnAbs and extended to mismatched heterosubtypic influenza viruses. Although NHPs lack the immunoglobulin germline VH1-69 residues associated with the most prevalent human stem-directed bnAbs, other gene families compensated to generate bnAbs. Isolation and structural analyses of vaccine-induced bnAbs revealed extensive interaction with the fusion peptide on the HA stem, which is essential for viral entry. Antibodies elicited by these headless HA stabilized-stem vaccines neutralized diverse H1 and H3 influenza viruses and shared a mode of recognition analogous to human bnAbs, suggesting that these vaccines have the potential to confer broadly protective immunity against diverse viruses responsible for seasonal and pandemic influenza infections in humans.

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Influenza type A virus M protein expression on infected cells is responsible for cross-reactive recognition by cytotoxic thymus-derived lymphocytes

Infection and Immunity ◽

10.1128/iai.29.2.719-723.1980 ◽

1980 ◽

Vol 29 (2) ◽

pp. 719-723 ◽

Cited By ~ 1

Author(s):

C S Reiss ◽

J L Schulman

Keyword(s):

T Cell ◽

Influenza A Virus ◽

Influenza A ◽

Rabbit Antiserum ◽

Type A ◽

M Protein ◽

Cytotoxic T Cell ◽

Influenza A Viruses ◽

Infected Cells ◽

Cytotoxic T Cell Responses

M protein of influenza A virus was detected with rabbit antiserum by both indirect immunofluorescence and by antibody plus complement-mediated cytolysis on the cell surfaces of both productively and nonproductively infected cells. In contrast, antiserum to nucleoprotein failed to react with unfixed infected cells, but did bind to fixed infected cells, especially in the perinuclear area. Incorporation of antiserum to M protein in a T-cell-mediated cytotoxicity assay produced almost complete abrogation of lysis of H-2-compatible cells infected with an influenza A virus of a subtype which differed from that used to elicit the cytotoxic T cells. However, the antibody did not significantly block 51Cr release from cells infected with the homotypic type A influenza virus. These observations are in accord with the hypothesis that the cross-reactive cytotoxic T-cell responses seen with cells infected by heterotypic influenza A viruses are due to recognition of a common M protein.

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Cross-protection in mice infected with influenza A virus by the respiratory route is correlated with local IgA antibody rather than serum antibody or cytotoxic T cell reactivity

European Journal of Immunology ◽

10.1002/eji.1830140414 ◽

1984 ◽

Vol 14 (4) ◽

pp. 350-356 ◽

Cited By ~ 212

Author(s):

Foo Y. Liew ◽

Stuart M. Russell ◽

Geoffrey Appleyard ◽

Colin M. Brand ◽

John Beale

Keyword(s):

T Cell ◽

Influenza A Virus ◽

Influenza A ◽

Serum Antibody ◽

Cross Protection ◽

Cytotoxic T Cell ◽

Cell Reactivity ◽

Iga Antibody ◽

T Cell Reactivity

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Rectification of Age-Associated Deficiency in Cytotoxic T Cell Response to Influenza A Virus by Immunization with Immune Complexes

The Journal of Immunology ◽

10.4049/jimmunol.179.9.6153 ◽

2007 ◽

Vol 179 (9) ◽

pp. 6153-6159 ◽

Cited By ~ 31

Author(s):

Biao Zheng ◽

Yongxin Zhang ◽

Hongxia He ◽

Ekaterina Marinova ◽

Kirsten Switzer ◽

...

Keyword(s):

T Cell ◽

Influenza A Virus ◽

Immune Complexes ◽

Cell Response ◽

Influenza A ◽

T Cell Response ◽

Cytotoxic T Cell

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Induction of cytotoxic T-cell responses by gene gun DNA vaccination with minigenes encoding influenza A virus HA and NP CTL-epitopes

Vaccine ◽

10.1016/s0264-410x(99)00279-0 ◽

1999 ◽

Vol 18 (7-8) ◽

pp. 681-691 ◽

Cited By ~ 27

Author(s):

Anders Fomsgaard ◽

Henrik V Nielsen ◽

Nikolai Kirkby ◽

Karin Bryder ◽

Sylvie Corbet ◽

...

Keyword(s):

T Cell ◽

Influenza A Virus ◽

Influenza A ◽

T Cell Responses ◽

Dna Vaccination ◽

Cytotoxic T Cell ◽

Gene Gun ◽

Ctl Epitopes ◽

Cell Responses ◽

Cytotoxic T Cell Responses

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Immunomic Analysis of the Repertoire of T-Cell Specificities for Influenza A Virus in Humans

Journal of Virology ◽

10.1128/jvi.01563-08 ◽

2008 ◽

Vol 82 (24) ◽

pp. 12241-12251 ◽

Cited By ~ 135

Author(s):

Erika Assarsson ◽

Huynh-Hoa Bui ◽

John Sidney ◽

Qing Zhang ◽

Jean Glenn ◽

...

Keyword(s):

Influenza Virus ◽

T Cell ◽

Influenza A Virus ◽

Influenza A ◽

T Cell Responses ◽

Class Ii ◽

Influenza Viruses ◽

Class I ◽

Cell Responses ◽

Virus Strains

ABSTRACT Continuing antigenic drift allows influenza viruses to escape antibody-mediated recognition, and as a consequence, the vaccine currently in use needs to be altered annually. Highly conserved epitopes recognized by effector T cells may represent an alternative approach for the generation of a more universal influenza virus vaccine. Relatively few highly conserved epitopes are currently known in humans, and relatively few epitopes have been identified from proteins other than hemagglutinin and nucleoprotein. This prompted us to perform a study aimed at identifying a set of human T-cell epitopes that would provide broad coverage against different virus strains and subtypes. To provide coverage across different ethnicities, seven different HLA supertypes were considered. More than 4,000 peptides were selected from a panel of 23 influenza A virus strains based on predicted high-affinity binding to HLA class I or class II and high conservancy levels. Peripheral blood mononuclear cells from 44 healthy human blood donors were tested for reactivity against HLA-matched peptides by using gamma interferon enzyme-linked immunospot assays. Interestingly, we found that PB1 was the major target for both CD4+ and CD8+ T-cell responses. The 54 nonredundant epitopes (38 class I and 16 class II) identified herein provided high coverage among different ethnicities, were conserved in the majority of the strains analyzed, and were consistently recognized in multiple individuals. These results enable further functional studies of T-cell responses during influenza virus infection and provide a potential base for the development of a universal influenza vaccine.

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