Jannick Prentoe1, Rodrigo Velázquez-Moctezuma1, Steven K H Foung2, Mansun Law3, Jens Bukh1. 1. Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Hvidovre Hospital and Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. 2. Department of Pathology, Stanford University School of Medicine, Stanford, CA. 3. Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, CA.
Abstract
There are 3-4 million new hepatitis C virus (HCV) infections yearly. The extensive intergenotypic sequence diversity of envelope proteins E1 and E2 of HCV and shielding of important epitopes by hypervariable region 1 (HVR1) of E2 are believed to be major hindrances to developing universally protective HCV vaccines. Using cultured viruses expressing the E1/E2 complex of isolates H77 (genotype 1a), J6 (2a), or S52 (3a), with and without HVR1, we tested HVR1-mediated neutralization occlusion in vitro against a panel of 12 well-characterized human monoclonal antibodies (HMAbs) targeting diverse E1, E2, and E1/E2 epitopes. Surprisingly, HVR1-mediated protection was greatest for S52, followed by J6 and then H77. HCV pulldown experiments showed that this phenomenon was caused by epitope shielding. Moreover, by regression analysis of HMAb binding and neutralization titer of HCV we found a strong correlation for HVR1-deleted viruses but not for parental viruses retaining HVR1. The intergenotype neutralization sensitivity of the parental viruses to HMAb antigenic region (AR) 2A, AR3A, AR4A, AR5A, HC84.26, and HC33.4 varied greatly (>24-fold to >130-fold differences in 50% inhibitory concentration values). However, except for AR5A, these differences decreased to less than 6.0-fold when comparing the corresponding HVR1-deleted viruses. Importantly, this simplified pattern of neutralization sensitivity in the absence of HVR1 was also demonstrated in a panel of HVR1-deleted viruses of genotypes 1a, 2a, 2b, 3a, 5a, and 6a, although for all HMAbs, except AR4A, an outlier was observed. Finally, unique amino acid residues in HCV E2 could explain these outliers in the tested cases of AR5A and HC84.26. CONCLUSION: HVR1 adds complexity to HCV neutralization by shielding a diverse array of unexpectedly cross-genotype-conserved E1/E2 epitopes. Thus, an HVR1-deleted antigen could be a better HCV vaccine immunogen. (Hepatology 2016;64:1881-1892).
There are 3-4 million new hepatitis C virus (HCV) infections yearly. The extensive intergenotypic sequence diversity of envelope proteins E1 and E2 of HCV and shielding of important epitopes by hypervariable region 1 (HVR1) of E2 are believed to be major hindrances to developing universally protective HCV vaccines. Using cultured viruses expressing the E1/E2 complex of isolates H77 (genotype 1a), J6 (2a), or S52 (3a), with and without HVR1, we tested HVR1-mediated neutralization occlusion in vitro against a panel of 12 well-characterized human monoclonal antibodies (HMAbs) targeting diverse E1, E2, and E1/E2 epitopes. Surprisingly, HVR1-mediated protection was greatest for S52, followed by J6 and then H77. HCV pulldown experiments showed that this phenomenon was caused by epitope shielding. Moreover, by regression analysis of HMAb binding and neutralization titer of HCV we found a strong correlation for HVR1-deleted viruses but not for parental viruses retaining HVR1. The intergenotype neutralization sensitivity of the parental viruses to HMAb antigenic region (AR) 2A, AR3A, AR4A, AR5A, HC84.26, and HC33.4 varied greatly (>24-fold to >130-fold differences in 50% inhibitory concentration values). However, except for AR5A, these differences decreased to less than 6.0-fold when comparing the corresponding HVR1-deleted viruses. Importantly, this simplified pattern of neutralization sensitivity in the absence of HVR1 was also demonstrated in a panel of HVR1-deleted viruses of genotypes 1a, 2a, 2b, 3a, 5a, and 6a, although for all HMAbs, except AR4A, an outlier was observed. Finally, unique amino acid residues in HCV E2 could explain these outliers in the tested cases of AR5A and HC84.26. CONCLUSION: HVR1 adds complexity to HCV neutralization by shielding a diverse array of unexpectedly cross-genotype-conserved E1/E2 epitopes. Thus, an HVR1-deleted antigen could be a better HCV vaccine immunogen. (Hepatology 2016;64:1881-1892).
Authors: Jason Alexander Ji-Xhin Wong; Rakesh Bhat; Darren Hockman; Michael Logan; Chao Chen; Aviad Levin; Sharon E Frey; Robert B Belshe; D Lorne Tyrrell; John Lok Man Law; Michael Houghton Journal: J Virol Date: 2014-10-01 Impact factor: 5.103
Authors: K G Hadlock; R E Lanford; S Perkins; J Rowe; Q Yang; S Levy; P Pileri; S Abrignani; S K Foung Journal: J Virol Date: 2000-11 Impact factor: 5.103
Authors: Michelle C Sabo; Vincent C Luca; Jannick Prentoe; Sharon E Hopcraft; Keril J Blight; Minkyung Yi; Stanley M Lemon; Jonathan K Ball; Jens Bukh; Matthew J Evans; Daved H Fremont; Michael S Diamond Journal: J Virol Date: 2011-05-04 Impact factor: 5.103
Authors: Zhen-yong Keck; Christine Girard-Blanc; Wenyan Wang; Patrick Lau; Adam Zuiani; Felix A Rey; Thomas Krey; Michael S Diamond; Steven K H Foung Journal: J Virol Date: 2016-01-06 Impact factor: 5.103
Authors: Alexander W Tarr; Ania M Owsianka; Dhanya Jayaraj; Richard J P Brown; Timothy P Hickling; William L Irving; Arvind H Patel; Jonathan K Ball Journal: J Gen Virol Date: 2007-11 Impact factor: 3.891
Authors: Troels K H Scheel; Judith M Gottwein; Tanja B Jensen; Jannick C Prentoe; Anne M Hoegh; Harvey J Alter; Jesper Eugen-Olsen; Jens Bukh Journal: Proc Natl Acad Sci U S A Date: 2008-01-14 Impact factor: 11.205
Authors: Zhen-yong Keck; Jinming Xia; Yong Wang; Wenyan Wang; Thomas Krey; Jannick Prentoe; Thomas Carlsen; Angela Ying-Jian Li; Arvind H Patel; Stanley M Lemon; Jens Bukh; Felix A Rey; Steven K H Foung Journal: PLoS Pathog Date: 2012-04-12 Impact factor: 6.823
Authors: John Lok Man Law; Chao Chen; Jason Wong; Darren Hockman; Deanna M Santer; Sharon E Frey; Robert B Belshe; Takaji Wakita; Jens Bukh; Christopher T Jones; Charles M Rice; Sergio Abrignani; D Lorne Tyrrell; Michael Houghton Journal: PLoS One Date: 2013-03-19 Impact factor: 3.240
Authors: Jan M Pestka; Mirjam B Zeisel; Edith Bläser; Peter Schürmann; Birke Bartosch; Francois-Loïc Cosset; Arvind H Patel; Helga Meisel; Jens Baumert; Sergei Viazov; Kay Rispeter; Hubert E Blum; Michael Roggendorf; Thomas F Baumert Journal: Proc Natl Acad Sci U S A Date: 2007-03-28 Impact factor: 11.205
Authors: Holly Freedman; Michael R Logan; Darren Hockman; Julia Koehler Leman; John Lok Man Law; Michael Houghton Journal: J Virol Date: 2017-03-29 Impact factor: 5.103
Authors: Chaturaka Rodrigo; Melanie R Walker; Preston Leung; Auda A Eltahla; Jason Grebely; Gregory J Dore; Tanya Applegate; Kimberly Page; Sunita Dwivedi; Julie Bruneau; Meghan D Morris; Andrea L Cox; William Osburn; Arthur Y Kim; Janke Schinkel; Naglaa H Shoukry; Georg M Lauer; Lisa Maher; Margaret Hellard; Maria Prins; Fabio Luciani; Andrew R Lloyd; Rowena A Bull Journal: Infect Genet Evol Date: 2017-01-05 Impact factor: 3.342
Authors: Madeleine C Mankowski; Valerie J Kinchen; Lisa N Wasilewski; Andrew I Flyak; Stuart C Ray; James E Crowe; Justin R Bailey Journal: Proc Natl Acad Sci U S A Date: 2017-12-18 Impact factor: 11.205
Authors: Jannick Prentoe; Rodrigo Velázquez-Moctezuma; Elias H Augestad; Andrea Galli; Richard Wang; Mansun Law; Harvey Alter; Jens Bukh Journal: Proc Natl Acad Sci U S A Date: 2019-04-30 Impact factor: 11.205
Authors: Michelle D Colbert; Andrew I Flyak; Clinton O Ogega; Valerie J Kinchen; Guido Massaccesi; Mayda Hernandez; Edgar Davidson; Benjamin J Doranz; Andrea L Cox; James E Crowe; Justin R Bailey Journal: J Virol Date: 2019-06-28 Impact factor: 5.103