Rapid GMP-compliant expansion of SARS-CoV-2-specific T cells from convalescent donors for use as an allogeneic cell therapy for COVID-19

Abstract

COVID-19 disease caused by the SARS-CoV-2 virus is characterized by dysregulation of effector T cells and accumulation of exhausted T cells. T cell responses to viruses can be corrected by adoptive cellular therapy using donor-derived virus-specific T cells. Here we show that SARS-CoV-2-exposed blood donations contain CD4 and CD8 memory T cells specific for SARS-CoV-2 spike, nucleocapsid and membrane antigens. These peptides can be used to isolate virus-specific T cells in a GMP-compliant process. These T cells can be rapidly expanded using GMP-compliant reagents for use as a therapeutic product. Memory and effector phenotypes are present in the selected virus-specific T cells, but our method rapidly expands the desirable central memory phenotype. A manufacturing yield ranging from 1010 to 1011 T cells can be obtained within 21 days culture. Thus, multiple therapeutic doses of virus-specific T cells can be rapidly generated from convalescent donors for treatment of COVID-19 patientsOne Sentence SummaryCD4+ and CD8+ T cells specific for SARS-CoV-2 can be isolated from convalescent donors and rapidly expanded to therapeutic doses at GMP standard, maintaining the desired central memory phenotype required for protective immune responses against severe COVID-19 infections.

Rapid GMP-Compliant Expansion of SARS-CoV-2–Specific T Cells From Convalescent Donors for Use as an Allogeneic Cell Therapy for COVID-19

“…However, the donor HLA range seen in Table 1 indicates HLA subtypes with strong peptide-presenting capacity such as A*02:01 and DRB*15:01, so supplementation of immune response with donor T cells from a matched donor could be a therapeutic option where few others exist. Therefore, we demonstrate the feasibility of generating large quantities of virus-specific T cell products for clinical trials in support of severe SARS-CoV-2 infections where the endogenous T cell response is compromised, representing a potentially significant advance in therapy for COVID-19 ( 46 ).…”

“…We thank Dr Anne Richter of Miltenyi Biotec GmbH for scientific advice around the use of the peptide pools. This manuscript has been released as a pre-print at bioRxiv ( 46 ).…”

Cytometric analysis of T cell phenotype using cytokine profiling for improved manufacturing of an EBV-specific T cell therapy.

“…A key feature of this work was to identify a robust panel of surface and intracellular markers which could effectively classify the T cell differentiation status and development from initial material through to final product. Our approach supplies clear data for this, and demonstrates the utility of this approach for T cell therapies [37]. In addition, the use of t-SNE dimensionality reduction was very effective at condensing multiple parameters into a single image which could be used to identify the status of the material at any stage of manufacture.…”

“…Robust, validated flow cytometric assays are a cornerstone of effective reproducible cell therapy manufacture [21]. The use of flow cytometric analysis and functional profiling of EBV-specific T cells through cytokine expression in this study resulted in improved characterization of both start material and final product, and effective assessment of inprocess culture optima, which has been used for analysis of other T cell therapeutics including a SARS-CoV-2 T cell product for COVID-19 treatment [37].…”

DataSheet_1.docx

“…We thank Dr Anne Richter of Miltenyi Biotec GmbH for scientific advice around the use of the peptide pools. This manuscript has been released as a preprint at bioRxiv (46).…”