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Rapid production of clinical-grade SARS-CoV-2 specific T cells #MMPMID32838213
Leung W; Soh TG; Linn YC; Low JG; Loh J; Chan M; Chng WJ; Koh LP; Poon ML; Ng KP; Kuick CH; Tan TT; Tan LK; Seng MS
Adv Cell Gene Ther 2020[Oct]; 3 (4): e101 PMID32838213show ga
OBJECTIVES: To determine whether the frequencies of SARS-CoV-2-specific T cells are sufficiently high in the blood of convalescent donors and whether it is technically feasible to manufacture clinical-grade products overnight for T-cell therapy and assessment of COVID-19 immunity. METHODS: One unit of whole blood or leukapheresis was collected from each donor following standard blood bank practices. The leukocytes were stimulated using overlapping peptides of SARS-CoV-2, covering the immunodominant sequence domains of the S protein and the complete sequence of the N and M proteins. Thereafter, functionally reactive cells were enriched overnight using an automated device capturing IFNgamma-secreting cells. RESULTS: From 1 x 10(9) leukocytes, a median of 0.98 x 10(6) (range 0.56-2.95) IFNgamma + T cells were produced from each of the six donors, suggesting a high frequency of SARS-CoV-2 reactive T cells in their blood, even though only one donor had severe COVID-19 requiring mechanical ventilation whereas the other five donors had minor symptoms. A median of 57% of the enriched T cells were IFNgamma+ (range 20%-74%), with preferential enrichment of CD56+ T cells and effector memory T cells. TCRVbeta-spectratyping confirmed distinctively tall oligoclonal peaks in final products. With just six donors, the probability that a recipient would share at least one HLA allele with one of the donors is >88% among Caucasian, >95% among Chinese, >97% among Malay, and >99% among Indian populations. CONCLUSIONS: High frequencies of rapid antigen-reactive T cells were found in convalescent donors, regardless of severity of COVID-19. The feasibility of clinical-grade production of SARS-CoV-2-specific T cells overnight for therapeutics and diagnostics is revealed.