SUCCESSFUL MANUFACTURING OF CLINICAL GRADE SARS CoV 2 SPECIFIC T CELLS FOR ADOPTIVE CELL THERAPY

Background Adoptive therapy with SARS-CoV-2 specific T cells for COVID-19 has not been reported. The feasibility of rapid clinical-grade manufacturing of virus-specific T cells from convalescent donors has not been demonstrated for this or prior pandemics. Methods One unit of whole blood was collected from each convalescent donor following standard blood bank practices. After the plasma was separated and stored separately, 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. Thereaftesr, functionally reactive cells were enriched overnight using an automated device capturing IFNγ-secreting cells. Findings From 1×109 leukocytes, 0.56 to 1.16×106 IFNγ+ T cells were produced from each of the first two donors. Most of the T cells (64% to 71%) were IFNγ+, with preferential enrichment of CD56+ T cells, effector memory T cells, and effector memory RA+ T cells. TCRVβ spectratyping revealed oligoclonal distribution, with over-representation of subfamilies including Vβ3, Vβ16 and Vβ17. With just two donors, the probability that a recipient in the same ethnic group would share at least one donor HLA allele or one haplotype could be as high as >90% and >30%, respectively. Interpretations This study is limited by small number of donors and absence of recipient data; however, crucial first proof-of-principle data are provided demonstrating the feasibility of clinical-grade production of SARS-CoV-2 specific T cells for urgent clinical use, conceivably with plasma therapy concurrently. Our data showing that virus-specific T cells can be detected easily after brief stimulation with SARS-CoV-2 specific peptides suggest that a parallel diagnostic assay can be developed alongside serology testing. Funding The study was funded by a SingHealth Duke-NUS Academic Medicine COVID-19 Rapid Response Research Grant. ### Competing Interest Statement WL is a part-time advisor to Miltenyi Biomedicine. ### Clinical Trial NCT04351659 ### Funding Statement The study was funded by a SingHealth Duke-NUS Academic Medicine COVID-19 Rapid Response Research Grant. The funder had no role in study design, data collection, data analysis, data interpretation, or writing of the report. ### Author Declarations All relevant ethical guidelines have been followed; any necessary IRB and/or ethics committee approvals have been obtained and details of the IRB/oversight body are included in the manuscript. Yes All necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines and uploaded the relevant EQUATOR Network research reporting checklist(s) and other pertinent material as supplementary files, if applicable. Yes The corresponding authors had full access to all the data in the study and had final responsibility for the decision to submit for publication.