H84T-Banana Lectin (BanLec) CAR-NK cells bind high mannose glycosites that decorate the SARS-CoV-2 envelope, thereby decreasing cellular infection in a model of SARS-CoV-2. H84T-BanLec CAR-NK cells are innate effector cells, activated by virus. This novel cellular agent is a promising therapeutic, capable of clearing circulating SARS-CoV-2 virus and infected cells. Banana Lectin (BanLec) binds high mannose glycans on viral envelopes, exerting an anti-viral effect. A point mutation (H84T) divorces BanLec mitogenicity from antiviral activity. SARS-CoV-2 contains high mannose glycosites in proximity to the receptor binding domain of the envelope Spike (S) protein. We designed a chimeric antigen receptor (CAR) that incorporates H84T-BanLec as the extracellular moiety. Our H84T-BanLec CAR was devised to specifically direct NK cell binding of SARS-CoV-2 envelope glycosites to promote viral clearance. The H84T-BanLec CAR was stably expressed at high density on primary human NK cells during two weeks of ex vivo expansion. H84T-BanLec CAR-NK cells reduced S-protein pseudotyped lentiviral infection of 293T cells expressing ACE2, the receptor for SARS-CoV-2. NK cells were activated to secrete inflammatory cytokines when in culture with virally infected cells. H84T-BanLec CAR-NK cells are a promising cell therapy for further testing against wild-type SARS-CoV-2 virus in models of SARS-CoV-2 infection. They may represent a viable off-the-shelf immunotherapy for patients suffering from COVID-19.
【저자키워드】 COVID-19, SARS-CoV-2, Immunotherapy, glycobiology, CAR-NK cells, Banana Lectin, 【초록키워드】 ACE2, therapy, spike, SARS-COV-2 infection, Infection, NK cell, SARS-CoV-2 virus, antiviral activity, virus, viral clearance, Receptor binding domain, Anti-viral, Protein, Culture, therapeutic, Patient, receptor, chimeric antigen receptor, Inflammatory cytokine, glycan, binding, cellular, S-protein, Point mutation, lectin, Ex vivo, infected cells, wild-type SARS-CoV-2, circulating, effector cells, banana, pseudotyped, SARS-CoV-2 envelope, Cell, CAR, Extracellular, bind, reduced, activated, expressed, promote, expressing, 293T cell, lentiviral, BanLec, divorce, secrete, the SARS-CoV-2,