Potentiating Vγ9Vδ2â
T cell proliferation and assessing their cytotoxicity towards adherent cancer cells at the single cell level.
Biol Open
; 11(1)2022 02 15.
Article
em En
| MEDLINE
| ID: mdl-34994391
ABSTRACT
Vγ9Vδ2â
T cells is the dominant γδ T cell subset in human blood. They are cytotoxic and activated by phosphoantigens whose concentrations are increased in cancer cells, making the cancer cells targets for Vγ9Vδ2â
T cell immunotherapy. For successful immunotherapy, it is important both to characterise Vγ9Vδ2â
T cell proliferation and optimise the assessment of their cytotoxic potential, which is the aim of this study. We found that supplementation with freshly thawed human serum potentiated Vγ9Vδ2â
T cell proliferation from peripheral mononuclear cells (PBMCs) stimulated with (E)-4-Hydroxy-3-methyl-but-2-enyl diphosphate (HMBPP) and consistently enabled Vγ9Vδ2â
T cell proliferation from cryopreserved PBMCs. In cryopreserved PBMCs the proliferation was higher than in freshly prepared PBMCs. In a panel of short-chain prenyl alcohols, monophosphates and diphosphates, most diphosphates and also dimethylallyl monophosphate stimulated Vγ9Vδ2â
T cell proliferation. We developed a method where the cytotoxicity of Vγ9Vδ2â
T cells towards adherent cells is assessed at the single cell level using flow cytometry, which gives more clear-cut results than the traditional bulk release assays. Moreover, we found that HMBPP enhances the Vγ9Vδ2â
T cell cytotoxicity towards colon cancer cells. In summary, we have developed an easily interpretable method to assess the cytotoxicity of Vγ9Vδ2â
T cells towards adherent cells, found that Vγ9Vδ2â
T cell proliferation can be potentiated by media-supplementation and how misclassification of non-responders may be avoided. Our findings will be useful in the further development of Vγ9Vδ2â
T cell immunotherapy.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Receptores de Antígenos de Linfócitos T gama-delta
/
Neoplasias
Limite:
Humans
Idioma:
En
Revista:
Biol Open
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
Suécia