Chikungunya viruses that escape monoclonal antibody therapy are clinically attenuated, stable, and not purified in mosquitoes

• Published in: Journal of virology 2014-08, Vol.88 (15), p.8213-8226
• Main Authors: Pal, Pankaj, Fox, Julie M, Hawman, David W, Huang, Yan-Jang S, Messaoudi, Ilhem, Kreklywich, Craig, Denton, Michael, Legasse, Alfred W, Smith, Patricia P, Johnson, Syd, Axthelm, Michael K, Vanlandingham, Dana L, Streblow, Daniel N, Higgs, Stephen, Morrison, Thomas E, Diamond, Michael S
• Format: Article
• Language: English
• Subjects: Aedes - virology; Aedes albopictus; Alphavirus; Alphavirus Infections - therapy; Alphavirus Infections - virology; Animals; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - therapeutic use; Antibodies, Viral - immunology; Antibodies, Viral - therapeutic use; Chikungunya Fever; Chikungunya virus; Chikungunya virus - immunology; Chikungunya virus - isolation & purification; Chikungunya virus - pathogenicity; Chikungunya virus - physiology; Disease Models, Animal; Female; Homeodomain Proteins; Immunotherapy - methods; Macaca mulatta; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Pathogenesis and Immunity; Survival Analysis; Virulence; Virus Replication
• ISSN: 0022-538X; 1098-5514
• DOI: 10.1128/JVI.01032-14

Chikungunya virus (CHIKV) is a reemerging mosquito-transmitted alphavirus that causes epidemics of debilitating polyarthritis in humans. A prior study identified two anti-CHIKV monoclonal antibodies ([MAbs] CHK-152 and CHK-166) against the E2 and E1 structural proteins, which had therapeutic efficacy in immunocompetent and immunocompromised mice. Combination MAb therapy was required as administration of a single MAb resulted in the rapid selection of neutralization escape variants and treatment failure in mice. Here, we initially evaluated the efficacy of combination MAb therapy in a nonhuman primate model of CHIKV infection. Treatment of rhesus macaques with CHK-152 and CHK-166 reduced viral spread and infection in distant tissue sites and also neutralized reservoirs of infectious virus. Escape viruses were not detected in the residual viral RNA present in tissues and organs of rhesus macaques. To evaluate the possible significance of MAb resistance, we engineered neutralization escape variant viruses (E1-K61T, E2-D59N, and the double mutant E1-K61T E2-D59N) that conferred resistance to CHK-152 and CHK-166 and tested them for fitness in mosquito cells, mammalian cells, mice, and Aedes albopictus mosquitoes. In both cell culture and mosquitoes, the mutant viruses grew equivalently and did not revert to wild-type (WT) sequence. All escape variants showed evidence of mild clinical attenuation, with decreased musculoskeletal disease at early times after infection in WT mice and a prolonged survival time in immunocompromised Ifnar1(-/-) mice. Unexpectedly, this was not associated with decreased infectivity, and consensus sequencing from tissues revealed no evidence of reversion or compensatory mutations. Competition studies with CHIKV WT also revealed no fitness compromise of the double mutant (E1-K61T E2-D59N) neutralization escape variant in WT mice. Collectively, our study suggests that neutralization escape viruses selected during combination MAb therapy with CHK-152 plus CHK-166 retain fitness, cause less severe clinical disease, and likely would not be purified during the enzootic cycle. Chikungunya virus (CHIKV) causes explosive epidemics of acute and chronic arthritis in humans in Africa, the Indian subcontinent, and Southeast Asia and recently has spread to the New World. As there are no approved vaccines or therapies for human use, the possibility of CHIKV-induced debilitating disease is high in many parts of the world. To this end, our laboratory re