7/14/2014 VACCINE-INDUCED ANTI-HA2 ANTIBODIES PROMOTE VIRUS FUSION AND ENHANCE INFLUENZA VIRUS RESPIRATORY DISEASE (VAERD)
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1 7/14/214 VACCINATION & ENHANCED DISEASE VACCINE-INDUCED ANTI-HA2 ANTIBODIES PROMOTE VIRUS FUSION AND ENHANCE INFLUENZA VIRUS RESPIRATORY DISEASE (VAERD) HANA GOLDING & SURENDER KHURANA DIVISION OF VIRAL PRODUCTS, CBER, FDA Second WHO Integrated Meeting on Development and Clinical Trials of Influenza Vaccines that Induce broadly Protective and Long-Lasting Immune Responses May Respiratory Syncytial Virus (RSV) Seronegative children administered FI-RSV vaccine followed by exposure to wild type RSV demonstrated enhanced respiratory disease. INFLUENZA During 29 pandemic, several reports from Canada suggested increased risk of medically attended ph1n1 illness in recipients of 28 9 TIV vaccination Animals (Pigs) vaccinated with inactivated influenza vaccines showed enhanced respiratory disease following mismatched Influenza virus challenge. Gauger, et al.(211) Vaccine. Mar 24;29(15): VACCINATION & ENHANCED RESPIRATORY DISEASE ROLE OF ANTIBODY SPECIFICITY IN DISEASE ENHANCEMENT WIV-H1N2 IMMUNIZED PIGS SHOW ENHANCED DISEASE FOLLOWING MISMATCHED CHALLENGE WITH ph1n1 VIRUS Non-vaccinated ph1n1 challenged WIV-H1N2 vaccinated ph1n1 challenged Group1 = Whole Inactivated (WIV)-UV H1N2 Vaccine (A/Swine/Minnesota/211/28) Group2 = Whole Inactivated (WIV)-UV ph1n1 Vaccine (A/California/4/9) Group3 = Mock PBS (Non-vaccinated) IM in Emulsigen-D adjuvant (4:1 ratio) n=1 CHALLENGED INTRANASALLY Day Day 21 Day 42 1st Dose IM 2 nd Dose IM 2 x 1 5 TCID 5 (ph1n1) LRT Lesions WIV-H1N2-IM Sham-IM Amy Vincent, USDA 3 % Macroscopic Lesion Microscopic Lung Score (-21)
2 Absorbance 7/14/214 REACTIVITY OF ANTIBODIES ELICITED BY WIV-H1N2 & WIV-H1N1 VACCINE AGAINST THE ph1n1 CHALLENGE VIRIONS IN ELISA WIV-H1N2 DOES NOT GENERATE ph1n1-neutralizing ANTIBODIES IgG ELISA with ph1n1 virion WIV-H1N1 Pigs WIV-H1N2 Pigs Post-Vaccination Pig Sera (1:8 dilution) 5 6 PRODUCTION OF ph1n1- HA1 AND HA2 DOMAINS IN E. coli WIV-H1N2 IMMUNIZED PIG SERA ONLY REACT WITH ph1-ha2 (BUT NOT WITH ph1-ha1 DOMAIN) ph1n1 vs H1N2 Amino acid Identity: HA: aa % HA1: aa % HA2: aa % rha1 rha2 EXPRESSION IN E. coli (Inclusion Bodies) DENATURATION NATIVE CONDITIONS RENATURATION & DIALYSIS (6 D) Ni-NTA PURIFICATION (ph7.2) GEL FILTRATION (ph7.2) 7 WIV-H1N1-immunized (HA1) #1-Pre 14 #1 12 #2 1 #3 8 #4 6 #5 4 #6 2 #7 # WIV-H1N2-immunized (HA1) #11-Pre #11 #12 #13 #14 #15 #16 #17 #18 #19 WIV-H1N1-immunized (HA2) WIV-H1N2-immunized (HA2) #1-Pre #11-Pre 2 #1 2 #11 16 #2 #12 16 #13 12 #3 12 #14 #4 8 #5 8 #15 #16 4 #6 4 #17 #7 #18 #8 # Khurana S, et al.(213) Science Trans. Medicine. Vol 5 (2) 2ra
3 7/14/214 MDCK-BASED INFLUENZA VIRUS INFECTION ASSAY CROSS-REACTIVE ANTI-HA2 ANTIBODIES CAUSE ENHANCEMENT OF ph1n1 INFECTION OF MDCK CELLS Antibodies NP-Based ELISA- To Quantify Virus Infection 9 1 ELUCIDATION OF POST-VACCINATION ANTIBODY EPITOPE REPERTOIRE USING ph1n1 GENOME FRAGMENT PHAGE DISPLAY SPECIFICITY AND MECHANISM OF ENHANCEMENT OF ph1n1 INFLUENZA VIRUS INFECTION
4 % Hemolysis of Control Resonance (RU) 7/14/214 WIV-H1N2 INDUCED CROSS-REACTIVE ph1n1-ha2 ANTIBODIES BIND EPITOPE (aa 32-77) ADJACENT TO THE FUSION PEPTIDE ph1n1 HA2 sequence (1-223): GLFGAIAGFIEGGWTGMVDGWYGYHHQNEQGSGYAADLKSTQNAIDEITNKVNSVIEKMNTQFTAVGKEFNHLEKRIE NLNKKVDDGFLDIWTYNAELLVLLENERTLDYHDSNVKNLYEKVRSQLKNNAKEIGNGCFEFYHKCDNTCMESVKNGT YDYPKYSEEAKLNREEIDGVKLESTRIYQILAIYSTVASSLVLVVSLGAISFWMCSNGSLQCRICI HA2-epitope Antibodies ANTIBODY MEDIATED ENHANCEMENT OF INFLUENZA VIRUS INFECTION Fetuin- Receptor binding Assay Time (sec) HA2 fusion peptide Khurana S, et al.(213) Science Trans. Medicine. Vol 5 (2) 2ra No change in ph1n1 virus receptor binding in presence of WIV-H1N2 immune sera 14 HEMOLYSIS ASSAY FOR VIRUS FUSION: ACID ACTIVATION OF INFLUENZA VIRUS VIRUS FUSION-PROMOTING ACTIVITY OF HA2-TARGETING ANTIBODIES CORRELATES WITH ENHANCED LUNG PATHOLOGY IN VAERD ANIMALS 35% Correlation between % Macroscopic lesion and Hemolysis by HA2-Eluate antibodies 3% 25% 2% ph 5.2 ANTI-HA2 ANTIBODIES IN WIV-H1N2 IMMUNIZED PIG SERA ENHANCED ph1n1-mediated VIRUS FUSION 15 15% r=.68 p=.4 1% % Macroscopic Lesion Khurana S, et al.(213) Science Trans. Medicine. Vol 5 (2) 2ra
5 % of Control 7/14/214 POST-H1N2 VACCINATION SERA COMPETE WITH NEUTRALIZING STEM MAbs FOR BINDING TO H1N1pdm9 VIRUS 1 8 SUMMARY AND CONCLUSIONS: VACCINE SAFETY WIV-H1N2 VACCINE IMMUNIZED PIGS SHOWED ENHANCED LUNG DISEASE FOLLOWING MISMATCHED CHALLENGE WITH ph1n1 VIRUS. Possibly Mediated By Cross-reactive HA2-specific Serum Antibodies in absence of HA1 specific neutralizing antibodies Bound to site adjacent to the fusion peptide in HA2. Enhanced ph1n1 virus fusion # PBS #11-Pre #11-Post #18-Post Crowe (213) Sci Trans. Med. Vol 5 (2) 2fs34. C179 CR6261 F1 Increased virus fusion in presence of vaccine induced anti-ha2 antibodies correlated with the magnitude of observed pathology in VAERD-affected lungs. SUMMARY AND CONCLUSIONS: RELEVANCE FOR UNIVERSAL VACCINES ACKNOWLEDGEMENTS / COLLABORATORS THE ANTI-HA2 ANTIBODIES FROM VAERD PIGS BOUND STRONGLY TO PRE- FUSION HA FORM AND TO ph1n1 VIRUS PARTICLES THE POLYCLONAL HA-2 TARGETING ANTIBODIES FROM VAERD PIGS COMPETED WITH BINDING OF SEVERAL STEM-SPECIFIC BROADLY NEUTRALIZING MONOCLONAL ANTIBODIES THESE FINDINGS SHOULD BE CONSIDERED DURING DEVELOPMENT OF UNIVERSAL INFLUENZA VACCINES BASED ON THE HA2 STEM REGION: NEED TO MONITOR FOR FUSION-ENHANCING ANTIBODIES IN ADDITION TO NEUTRALIZING ANTIBODIES IN APPROPRIATE ASSAYS NEED TO COMPARE PERSISTANCE/LONGEVITY OF NEUTRALIZING VS. ENHANCING ANTIBODIES DVP, CBER, FDA SURENDER KHURANA LISA KING JODY MANISCHEWITZ ELIZABETH COYLE USDA AMY VINCENT CRYSTAL LOVING PHILLIP GAUGER 5
6 7/14/214 PIG AS AN INTERMEDIATE HOST FOR INFLUENZA TRANSMISSION TO HUMANS 21 6
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