BACKGROUND Influenza is an infectious disease that has significant public health impact due to its high prevalence and mortality. In early 2009, a novel influenza A(H1N1) virus emerged in Mexico and the USA, then rapidly spread worldwide and caused the first influenza pandemic of the 21st century. However, it is still controversial that whether seasonal influenza vaccine can provide a cross-protection against influenza A(H1N1) pdm09 infection. Since the 2009 pandemic occurred, numbers of studies focusing on this issue have been published, yet no confirmed conclusion was drawn. Therefore, further quantitative evaluation is needed to provide more reliable evidence. The objective of this study is to assess the cross-protection of seasonal influenza vaccination against 2009 pandemic A(H1N1) influenza illness, and explore the impact of seasonal influenza activities on this association. METHODS I followed the PRISMA statement and searched the PubMed, MEDLINE, Ovid Embase, The Cochrane Library databases, SCOPUS and CNKI. Randomized control trials, cohort studies, case-control studies assess the effect of seasonal influenza vaccine against influenza A (H1N1)pdm09 illness published in English and Chinese from 2009 to July 2013 were identified. The quality of included studies was assessed by the Jadad scale and the Newcastle-Ottawa Scale. I used the I2statistic, and Begg's funnel plot for assessment of heterogeneity and publication bias respectively. The software Review Manager 5.2 was used for generating the pooled effect with corresponding 95% confidence intervals and forest plots. Subgroup analysis was performed based on the study locations and previous circulating influenza viruses. RESULTS 20 studies were included in the meta-analyses. There is a non-significant 19% reduced risk of pandemic influenza illness in the countries combined data based on case-control studies(OR=0.81, 95% CI=0.60 to 1.08). While, for RCTs, a non-significant increase risk in seasonal influenza vaccinees was observed(RR=1.13, 95% CI=0.56 to 2.29). For the subgroup analysis, a significant 35% to 50% cross-protection was observed in South America and Europe, but an opposite result was observed in Canada(OR=1.44, 95% CI=0.83 to 2.50). Besides, the results indicate that there is no association between seasonal influenza vaccination and ILI. No publication bias was detected. CONCLUSIONS The findings partially support the hypothesis that seasonal vaccine may offer moderate cross-protection against laboratory-confirmed pandemic influenza A(H1N1) illness in general. Further immunological research is needed to understand the mechanism behind these findings. / published_or_final_version / Public Health / Master / Master of Public Health

Since there is an urgent need for development of vaccines and antiviral agents to combat influenza pandemics, this study aimed to develop influenza virus-like particles (VLPs) and aptamers targeting the virus particles as vaccine and antiviral agent candidates. Influenza VLPs containing three structural proteins of hemagglutinin (HA), neuraminidase (NA) and matrix 1 (M1) derived from influenza A/Hong Kong/01/2009 (H1N1) virus (HK/01) were constructed using a Bac-to-Bac baculovirus expression system. The expressed VLPs were purified by sucrose density gradient ultracentrifugation and characterized by Western blotting analysis and transmission electron microscopy. The immune responses and protective efficacy induced by VLPs were compared with those elicited by the clinically used Panenza vaccine in BALB/c mouse model. The results showed that two-dose vaccination with both VLP and the Panenza vaccine could confer complete protection. Single-dose vaccination with VLP could also provide 100% protection against lethal virus challenge, whereas single dose of an equal amount (based on HA content) of the Panenza vaccination just provided incomplete protection (67% survival rate) against the lethal virus challenge. Compared to the Panenza vaccination, the VLP vaccination could induce higher and broader antibody responses and higher viral specific T help (Th) cell and cytotoxic T lymphocyte (CTL) responses. Notably, a novel finding in this study is that the VLP vaccination could induce antibodies to inhibit virus release from infected MDCK cells, although the underlined mechanism needed to be further studied. These results indicated that influenza VLP might be a more effective and safe vaccine candidate which could be developed into an alternative vaccine for the control of epidemic and pandemic influenza in the future. To develop aptamers as antiviral agents against influenza, I sought to use influenza VLPs as target for ssDNA aptamer selection. After 11 rounds of selection using the systemic evolution of ligandsby exponential enrichment (SELEX),the recovered DNA molecules were PCR-amplified, gel purified and cloned into pCR-Blunt II TOPO vector for sequencing. The sequencing results showed that one aptamer Va-1 was markedly enriched, which was accounted for 59% (13/22) of the selected aptamers. Compared to the other non-enriched aptamers, the enriched aptamer Va-1 showed the highest binding affinity to the UV inactivated influenza HK/01 virus. It was also shown that the aptamer Va-1 specifically bound to the HK/01 stain while it could not bind other respiratory viruses even the PR8 strain within the H1N1 subtype. It was further demonstrated that the aptamer Va-1 could only bind to NA protein in a dose-dependent manner but not bind to HA and M1 proteins. Unfortunately, the selected aptamer did not show any antiviral effects. However, it may be potentially developed into a diagnostic and analytic agent because its binding activity was comparable with that of the commercial anti-NA antibody. In conclusion, the influenza VLPs may be a promising vaccine candidate for the control of influenza virus infection and the selected aptamer may be potentially developed into an alternative tool for influenza virus detection. / published_or_final_version / Microbiology / Doctoral / Doctor of Philosophy

Development of effective vaccines against mutable viruses (i.e HIV-1 and influenza) remains a big challenge. Antibody-dependent cell-mediated cytotoxicity (ADCC) has been found to be a key component of immune protection against viral infections in vivo. Therefore, vaccine immunogens that elicit broadly neutralizing antibodies with high ADCC are desired for vaccine development. This study is to identify primary immunogens that can initiate somatic maturation of germline antibodies of known broadly neutralizing HIV-1 antibodies (bnAbs) for HIV vaccine development and to localize dominant ADCC epitopes on hemagglutinin (HA) of pandemic H1N1 influenza virus for development of a flu vaccine. Based on the observations that known HIV-1 bnAbs have extensive somatic mutation compared to their germline versions and that HIV-1 envelope (Env) glycoprotein lacks measurable binding to putative germline antibodies of known bnAbs, we hypothesized that non-HIV-derived (poly)peptides may serve as primary immunogens to trigger somatic maturation of germline antibodies of bnAbs, leading to elicitation of intermediate antibodies (iAbs) that can further mature to HIV-1 bnAbs upon Env vaccination or HIV-1 infection. Using b12 as a model bnAb, we identified five non-HIV-derived (poly)peptides that bound to putative b12 germline and iAbs, and immunized rabbits with the (poly)peptide priming followed by Env boosting. Rabbit immunization with (poly)peptides alone induced high titers of antibodies that were cross-reactive with gp140SF162 trimer and resurfaced Env RSC3, and the serum IgGs neutralized SF162 and JRFL. These results suggest that the (poly)peptides might structurally mimic CD4bs of Env. Priming rabbits with (poly)peptides followed by boosts with gp140SF162 and RSC3 resulted in antibodies capable of competing with b12 for binding to gp140SF162 trimer and neutralizing cross-clade isolates, while control rabbits without priming produced antibodies that were unable to compete with b12 for gp140SF162 trimer binding, and the serum IgGs neutralized only 3 clade B isolates. Our results provide proof of concept that non-HIV-derived (poly)peptides may serve as primary vaccine immunogens to initiate guided immune responses towards bnAbs. HA protein has high level of immunogenicity and considered the most important target for immune protection against influenza virus infection. Several potent HA-specific bnAbs have been reported with their conserved neutralizing epitopes revealed, but there has been no report so far about ADCC epitopes on HA. Using yeast display and flow cytometry assisted cell sorting, we mapped the epitope of convalescent plasma IgGs with different ADCC activity, we identified two dominant ADCC epitopes, designated HA-E1 [AA92-117] and HA-E2 (AA 124-159), on HA of 2009 pandemic H1N1 influenza virus. E1 and E2 overlapped with the immunodominant epitopes of HA. Depletion of purified patient plasma IgGs with yeast cells expressing E1 or E2 peptides decreased ADCC activity of the IgGs. E1 and E2 sequences are highly conserved in H1N1 strains, but less so in other subtypes of influenza A viruses. Our study may aid in designing immunogens that can elicit antibodies with high ADCC activity. Vaccine immunogens designed to include the structural determinants of potent bnAbs and ADCC epitopes may confer a comprehensive immune protection against influenza virus infection. / published_or_final_version / Microbiology / Doctoral / Doctor of Philosophy

Thesis (M.S.)--Baylor University, 2007. / Includes bibliographical references (p. 53-64).

Thesis (M.S.)--Worcester Polytechnic Institute. / Keywords: DNA; Vaccine; Immunology; Virology; Influenza; Antibody; VIrus. Includes bibliographical references (leaves 51-54).

Thesis (M. P. H.)--University of Hong Kong, 2005. / Also available in print.

Thesis (M. P. H.)--University of Hong Kong, 2008. / Includes bibliographical references (leaf 55-58) Also available in print.

Thesis (M.P.H.)--University of Hong Kong, 2008. / Includes bibliographical references (p. 36-38).

Thesis (M. Med. Sc.)--University of Hong Kong, 2004. / Also available in print.

Thesis (M. Med. Sc.)--University of Hong Kong, 2005.