In Vitro Expansion of Vδ1+ T Cells from Cord Blood by Using Artificial Antigen-Presenting Cells and Anti-CD3 Antibody

Abstract

γδ T cells have the potential for adoptive immunotherapy since they respond to bacteria, viruses, and tumors. However, these cells represent a small fraction of the peripheral T-cell pool and require activation and proliferation for clinical benefits. In cord blood, there are some γδ T cells, which exhibit a naïve phenotype, and mostly include Vδ1+ T cells. In this study, we investigated the effect of CD3 signaling on cord blood γδ T-cell proliferation using K562-based artificial antigen presenting cells expressing costimulatory molecules. There were significantly more Vδ1+ T cells in the group stimulated with anti-CD3 antibody than in the group without. In cultured Vδ1+ T cells, DNAM-1 and NKG2D were highly expressed, but NKp30 and NKp44 showed low expression. Among various target cells, Vδ1+ T cells showed the highest cytotoxicity against U937 cells, but Daudi and Raji cells were not susceptible to Vδ1+ T cells. The major cytokines secreted by Vδ1+ T cells responding to U937 cells were Granzyme B, IFN-γ, and sFasL. Cytotoxicity by Vδ1+ T cells correlated with the expression level of PVR and Nectin of DNAM-1 ligands on the surface of target cells. Compared to Vδ2+ T cells in peripheral blood, cord blood Vδ1+ T cells showed varying cytotoxicity patterns depending on the target cells. Here, we determined the ideal conditions for culturing cord blood Vδ1+ T cells by observing that Vδ1+ T cells were more sensitive to CD3 signals than other subtypes of γδ T cells in cord blood. Cultured cord blood Vδ1+ T cells recognized target cells through activating receptors and secreted numerous cytotoxic cytokines. These results are useful for the development of tumor immunotherapy based on γδ T cells.

Advancements in γδT cell engineering: paving the way for enhanced cancer immunotherapy

“…Over time, Vγ9Vδ2 T cells emerge as the predominant subset (over 75%) of the γδT cell population in peripheral blood by adulthood, with less than 10% being Vδ1+ ( 80 ). Therefore, cord blood has been explored for its predominant expansion of Vδ1+ cells, or occasional viable expansion of Vδ2+ cells ( 81 – 83 ). However, there are several challenges associated with the in vitro expansion of γδT cells from cord blood, including a low number of γδT cells (less than 1% of cord blood lymphocytes), phenotypically and functionally immature γδT cells, and a poor response to IL-2 and phosphoantigen stimulation ( 80 ).…”

Expansion and Polarization of Human γδT17 Cells in vitro from Peripheral Blood Mononuclear Cells

“…In humans, γδ T cells can be distinguished by the δ chain, including Vδ1, Vδ2, Vδ3, and Vδ5 ( Ling et al, 2022 ). While previous studies have reported methods for in vitro expansion of human γδ T cells ( Ness-Schwickerath et al, 2010 ; Caccamo et al, 2011 ; Hur et al, 2023 ), the majority of peripheral blood γδ T cells in humans are Vδ2 + subsets that mainly produce IFN-γ. It is difficult to investigate human γδT17 cells as they are scarce, and little has been done to expand human γδT17 cells in vitro.…”