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2.8. Mixed lymphocyte reaction assay (MLR)

JR Jofer Andree Zamame Ramirez
GR Graziela Gorete Romagnoli
BF Bianca Francisco Falasco
CG Carolina Mendonça Gorgulho
CF Carla Sanzochi Fogolin
DS Daniela Carvalho dos Santos
JJ João Pessoa Araújo Junior
ML Michael Thomas Lotze
RU Rodrigo Portes Ureshino
RK Ramon Kaneno
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The functional activity of DCs was first evaluated through their ability to stimulate the proliferation of normal allogeneic T lymphocytes. DCs from six different donors were cocultured with allogeneic T lymphocytes (previously marked with carboxyfluorescein succinyl ester (CFSE)) in flat-bottomed 96-well plates in a 1:10 (104:105) DCs: lymphocyte ratio. Cells were harvested five days later, and the lymphocyte proliferation was analyzed by flow cytometry based on the dilution of CFSE in the replicant cells. We also analyzed the expression of PD-1 and CD69 on CD3+ cells using anti-PD-1-PE and CD69-APC-H7 (BD Pharmingen).

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