Analysis of CCL5 expression in classical Hodgkin's lymphoma L428 cell line

Abstract

Abstract. CCL5 is one of the chemoattractant cytokines involved in inflammatory observed in both diffuse large B-cell lymphoma (DLBCL) and classical Hodgkin's lymphoma (CHL). However, the pathological effects of CCL5 remain unclear. To gain a better understanding of the role of CCL5 in CHL and DLBCL, we examined the expression of CCL5 in the CHL cell line L428 and the DLBCL cell lines Ly1 and Ly8, as well as its chemotactic effect on CD4 + T cells. CCL5 mRNA expression was detected by real-time quantitative RT-PCR. Intracellular CCL5 protein expression was analyzed using confocal microscopy, and CCL5 protein secretion was detected by ELISA. The chemotactic function of CCL5 was assessed using a Transwell coculture system, and the number of migrated CD4 + T cells was counted. Moreover, the p-iкBα and p65 levels of NF-кB signaling molecules in these lymphoma cell lines were detected by Western blotting. The results showed that CCL5 mRNA and protein expression in the L428 cells was significantly higher than in Ly1 and Ly8 cells (p<0.05). L428 cells secreted more CCL5 than the Ly1 or Ly8 cells, and the secreted CCL5 was capable of inducing CD4 + T cell migration. The expression levels of the NF-кB transcription factors p65 and p-iкBα were examined in these lymphoma cells. L428, Ly1 and Ly8 cells expressed similar levels of p65, while p-iкBα expression was higher in the L428 cells than in the Ly1 or Ly8 cells, indicating that a high CCL5 expression may be related to the increased activity of the NF-кB signaling pathway in L428 cells.

Microenvironment Cell Contribution to Lymphoma Immunity

“…The T cells surrounding Reed-Sternberg cells express CCL5, which acts as a chemo-attractant for monocytes, eosinophils, basophils and mast cells as well as CD4 positive T cells ( 30 , 31 ). C-C chemokine receptor type 3 (CCR3) + Th2 cells and eosinophils are attracted by the CCL1(eotaxin) produced by fibroblasts surrounding RS cells ( 32 , 33 ).…”

Migration Properties Distinguish Tumor Cells of Classical Hodgkin Lymphoma from Anaplastic Large Cell Lymphoma Cells

“…Several studies have been performed to investigate the expression of chemokine receptors and their ligands in the HRS cells of cHL [15,16,17,18,19,20] These studies found that the chemokine receptors CCR5, CXCR3, CXCR4, CXCR5 and CCR7 are expressed in the HRS cells of cHL in a significant number of cases [15,18,19]. Furthermore, the ligands CXCL10, CXCL12, CXCL13, CCL21, CX3CL1, CCL5 and CCL17 are expressed in cHL cell lines and primary HRS cells [17,18,19,21,22]. CCL17 and CCL5 are capable of attracting T helper 2 (Th2) and regulatory T (Treg) cells, which interact with HRS cells and form what has been named rosettes around them [17,23].…”