CHARACTERIZATION OF URATE MONOSODIUM CRYSTALS EFFECT ON GAMMA/DELTA T CELLS

TOWSTYKA N; SABBIONE F; KEITELMAN I; GEFFNER JR; TREVANI AS; JANCIC C

Congreso; Reunión anual Sociedad Argentina de Inmunoogía; 2016

Uric acid is released from dying cells and functions as an adjuvant that promotes the generation of adaptive immune responses. The inflammatory effect of uric acid depends on its precipitation into monosodium urate crystals (MSUC). MSUC activate different intracellular pathways, i.e. Syk kinase and PI3K. Gamma/delta T cells act as sensors of cellular stress and infection. They recognize danger and pathogen-associated molecular patters, such as phosphoantigens. We have previously reported that MSUC activate gamma/delta T cells. Now, we aim to investigate whether MSUC modulate gamma/delta T cell activation triggered by phosphoantigens, and the mechanism involved in gamma/delta T cell activation by MSUC. Gamma/delta T cells were purified from human peripheral blood by using an anti-TCR gamma/delta MicroBead isolation kit. After purification, gamma/delta T cells were incubated or not with MSUC (200 ug/ml, 48 h). Then, we analyzed CD69 expression by flow cytometry, and IFN-gamma and TNF-alpha production by ELISA. To investigate the co-stimulatory effect of MSUC, we treated gamma/delta T cells with the phosphoantigen HMBPP (10 uM) and then we incubated cells with MSUC. Finally, to study the mechanism involved in gamma/delta T cell activation by MSUC, we treated cells with or without a protein kinase Syk inhibitor, GS-9973 (0.1 uM Gamma/delta T cells co-stimulated with MSUC and HMBPP produced higher levels of IFN-gamma and TNF-alpha compared to gamma/delta T cells activated by MSUC or HMBPP alone (p