بررسی بیان ژنهای CXCL8 ، CCL4 ، CCL3 در نوتروفیل های مواجهه یافته با انگل لیشمانیا اینفانتوم در محیط کشت سلولی

Background: Studies have shown that neutrophils in effect by parasitic triggers can produce many different chemokines.These chemokines are called leukocytes,T lymphocytes, dendritic cells, monocytes, lethal cells and neutrophils to the site of infection. The aim of this study is to investigate the expression of neutrophils exposure with leishmania infantum. Methods: During this experimental-laboratorial study the blood was collected from 30 healthy individuals. Peripheral blood neutrophils were isolated by HYSTOPAQUE 1077, dextran and centrifuge. Then cells were exposed to the promastigote forms of the Leishmania infantum parasite, which was prepared from the immunology department of the Pasteur Institute in stationary phase and was incubated for one hour in co2 incubator, not stimulated neutrophils were used as control. After extracting RNA, RNA was converted to cDNA by enzyme complex. cDNA was used for Real Time PCR. Selected gene been gene of CCL3,CCL4 and CXCL8 chemokines and β-Actin was used as references gene and then gene expression change was evaluated by ANOVA statistical methods. Results: The results indicated that CXCL8 gene expression were significantly increased in the face of leishmania compared to control neutrophils (p<0.001). On the other hand CCL3 and CCL4 genes expression weren’t significant change in the face of leishmania compared to the control neutrophils. Conclusion: The results show that exposure with leishmania parasite caused no change in expression of CCL3 and CCL4 mRNA genes among of simulation neutrophilsbut lead to increases high expression of CXCL8. On this basis measuring of CXCL8 can consider as a criterion for detecting leishmania infection level.