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Fırat Tıp Dergisi
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Comparison of the Effect of Phytohemagglutinin and Interleukin-2 on Human-T-Cells
Pınar ÇETİNALP DEMİRCAN
İstanbul Bilim Üniversitesi, Tıbbi Biyokimya Anabilim Dalı, İstanbul, Türkiye

Objective: In this study, it is aimed to determine the effect of stimulation with different stimuli on CD3+T-cells proliferation and activation mole-cules by stimulating phytohemagglutinin (PHA), Interleukin-2 (IL-2) and PHA+IL-2 together.

Material and Method: Six mL peripheral blood samples were obtained from each 15 healthy women and their 3x106 CD3+T-cells were isolated by the RosetteSep method. CD3+T-cells were counted on a phase-contrast microscope and their isolation purity were determined by immunofluorescen-ce and flow cytometric methods. 2x105 cells were apportioned in a 3-well dish of a 12-well culture dish, applied 10μg/mL PHA, 13μL/mL IL-2 and 10μg/mL PHA+13 μL/mL IL-2 and incubated in incubator for 24 hours. Cells groups of stimulated CD3+T-cells compared with CD3+T-cells(control group) not stimulated with any stimulants; their proliferations measured by spectrophotometric method using WST-1 test and flow cytometry by CFSE test, their expressions of activation molecules measured by flow cytometry with CD25, CD38, CD69, CD71 and HLA-DR antibodies.

Results: When WST-1 and CFSE test values were compared statistically between groups; It was determined that PHA-CD3+T-cells (p <0.001) and PHA + IL-2-CD3+T-cells (p <0.001) were significantly more proliferated, in other words activated than IL-2-CD3+T-cells. When levels of activation molecules evaluated by flow cytometry were compared statistically between groups; in comparison to IL-2-CD3+T-cells, significantly increased % expression values of CD25 (p <0.001), CD38 (p <0.001), CD71 (p <0.001) and HLA-DR (p <0.001) were detected in PHA-CD3+T-cells and PHA+IL-2-CD3+T-cells.

Conclusion: Since the proliferation and activation molecule levels in CD3+T-cells stimulated by PHA and PHA + IL-2 were similar and more than than stimulated by IL-2, it was concluded that the use of PHA as a CD3+T-cell stimulant in invitro studies alone was effective with no need for other stimulants and provided more proliferation and stimulation compared to IL-2 in CD3+T-cells.


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