Manufacture of CD22 CAR T cells following positive versus negative selection results in distinct cytokine secretion profiles and γδ T cell output

Hannah W Song; Mehdi Benzaoui; Alka Dwivedi; Sarah Underwood; Lipei Shao; Sooraj Achar; Vesna Posarac; Victoria A Remley; Michaela Prochazkova; Yihua Cai; Ping Jin; Robert P Somerville; David F Stroncek; Grégoire Altan-Bonnet; Nirali N Shah; Christopher D Chien; Naomi Taylor; Steven L Highfill

doi:10.1016/j.omtm.2023.101171

Manufacture of CD22 CAR T cells following positive versus negative selection results in distinct cytokine secretion profiles and γδ T cell output

Mol Ther Methods Clin Dev. 2023 Dec 11;32(1):101171. doi: 10.1016/j.omtm.2023.101171. eCollection 2024 Mar 14.

Authors

Hannah W Song¹, Mehdi Benzaoui^{2

3}, Alka Dwivedi², Sarah Underwood¹, Lipei Shao¹, Sooraj Achar⁴, Vesna Posarac⁵, Victoria A Remley¹, Michaela Prochazkova¹, Yihua Cai¹, Ping Jin¹, Robert P Somerville¹, David F Stroncek¹, Grégoire Altan-Bonnet⁴, Nirali N Shah², Christopher D Chien², Naomi Taylor^{2

3}, Steven L Highfill¹

Affiliations

¹ Center for Cellular Engineering, Department of Transfusion Medicine, National Institutes of Health, Bethesda, MD, USA.
² Pediatric Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
³ Institut de Génétique Moléculaire de Montpellier, University of Montpellier, CNRS, Montpellier, France.
⁴ Laboratory of Integrative Cancer Immunology, NCI, Bethesda, MD, USA.
⁵ STEMCELL Technologies, Vancouver, Canada.

Abstract

Chimeric antigen receptor T cells (CART) have demonstrated curative potential for hematological malignancies, but the optimal manufacturing has not yet been determined and may differ across products. The first step, T cell selection, removes contaminating cell types that can potentially suppress T cell expansion and transduction. While positive selection of CD4/CD8 T cells after leukapheresis is often used in clinical trials, it may modulate signaling cascades downstream of these co-receptors; indeed, the addition of a CD4/CD8-positive selection step altered CD22 CART potency and toxicity in patients. While negative selection may avoid this drawback, it is virtually absent from good manufacturing practices. Here, we performed both CD4/CD8-positive and -negative clinical scale selections of mononuclear cell apheresis products and generated CD22 CARTs per our ongoing clinical trial (NCT02315612 NCT02315612). While the selection process did not yield differences in CART expansion or transduction, positively selected CART exhibited a significantly higher in vitro interferon-γ and IL-2 secretion but a lower in vitro tumor killing rate. Notably, though, CD22 CART generated from both selection protocols efficiently eradicated leukemia in NSG mice, with negatively selected cells exhibiting a significant enrichment in γδ CD22 CART. Thus, our study demonstrates the importance of the initial T cell selection process in clinical CART manufacturing.

Keywords: B-ALL; CAR T cell manufacturing; CD22 CAR; T cell selection; chimeric antigen receptor; cytokines; negative selection; positive selection; γδ T cells.

Abstract

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