Background. CD4+CD28null T cells are expanded in peripheral blood of patients with chronic kidney disease and associated with subclinical atherosclerosis. However, triggers for the oligoclonal expansion and activation of these cells are not clear. Methods. We investigated twenty-five stage V-IV chronic kidney disease (CKD) patients and eight healthy subjects (HC). Peripheral mononuclear cells were isolated and incubated with heat shock protein- (HSP) 60 and 70. CD4+CD28null and CD4+CD28+ cells were sorted by flowcytometry and antigen specific response was assessed by the mRNA and protein expression of interferon (IFN)- , perforin, and granzyme B using qRT-PCR and Elispot. Results. The basal mRNA expression of IFN- , perforin, and granzyme B in CD4+CD28null cells was higher in subjects with CKD compared to that in HC ( ). Subjects with CKD also showed expression of IFN- , perforin, and granzyme B in the CD4+CD28+ subset, but this was much weaker than that seen in the CD4+CD28null population ( ). We did not note the expression of these molecules at mRNA or protein level in either subset of CD4 cells in HC. After incubation with HSP60 and HSP70, CD4+CD28null cells showed increased expression at mRNA ( ) and protein level ( ). CD4+CD28+ cells also showed a weak increase in expression. No antigen-specific response was noted in HC. Conclusion. These data show that CD4+CD28null cells in subjects with CKD react with HSP60 and HSP70 by upregulating the expression of IFN- , perforin and granzyme B. Increased circulating level of HSP60 and HSP70 might play a role in initiation and/or progression of atherosclerosis in CKD subjects through perturbation of CD4+CD28null cells. 1. Introduction Patients with chronic kidney disease (CKD) exhibit an expansion in the circulating CD4+CD28null cell population [1, 2]. In contrast to the classical CD4+ T helper cells, cells lacking CD28 molecule are cytotoxic and proinflammatory [3–5]. The ability of CD4+CD28null cells to interact with the endothelial cells through fractalkine-CX3CR interaction and the demonstration of these cells in atherosclerosis plaque, in combination with their ability to produce high level of IFN- and cytotoxic molecules leading to plaque destabilization, suggests their role in atherosclerotic disease [6]. The exact mechanism of their activation and expansion, however, is unclear. We have shown that this population is expanded in CKD even before development of clinically overt atherosclerotic coronary artery disease (CAD), and the degree of expansion correlates with common carotid artery intima media
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