The Role Of CCR5 In The Development Of Related Diseases & Analysis Of The Polymorphism In Human CDNA Sequence Of Prothymosin-α

CC chemokine receptor 5 is a kind of cell membrane protein, which is a kind of subtype of chemokine receptor. CCR5 is expressed mostly on white blood cells, such as monocytes, T cells, et al. It is recently demonstrated to be the surface marker of activated Th1 cells. We investigated the expression of CCR5 in experimental autoimmune myocarditis (EAM) mice and human follicular thyroid carcinoma to find the role of CCR5 in the development of diseases. RT-PCR, immunohistochemical analysis and flow-cytometry analysis were applied in these experiments. Furthermore, we established the monoclonal antibody blocked mice model and transferred mice model to investigate the critical role of CCR5 in EAM mice and human follicular thyroid carcinoma from the different levels of molecular, cellular and animal models. We found that CCR5-positive cells predominate in infiltrated inflammatory cells in cardiac tissue of EAM mice and CCR5-positive T cells in peripheral blood increased markedly in EAM mice compared with controls. Moreover, we demonstrated that the severity of myocarditis was significantly reduced when CCR5-negative T cells from EAM mice were adoptively transferred. When administrated with CCR5-positive T cells, the myocarditis was significantly aggravated. We also demonstrated that blockade of CCR5 with monoclonal antibodies significantly reduced severity of myocarditis in EAM mice. We also found that the expression of CCR5 was detected positively in follicular thyroid carcinoma (the positive rate was 73.33%) and was not detected in follicular thyroid adenoma and the normal controls. The concentration of CCL3 and CCL5 in the serum of follicular thyroid carcinoma patients increased significantly (P> 0.01) compared to those in follicular thyroid adenoma patients and the normal controls. So we think that CCR5 plays critical role in the development of EAM mice, and CCR5 might play an important role in the pathogenesis of follicular thyroid carcinoma.By sequencing the cDNA of prothymosin-α, we analyze the polymorphism in the cDNA sequence of prothymosin-α. The encoding cDNA sequence of human ProTα was amplified from cells of peripheral blood and cord blood by RT-PCR.The product of RT-PCR was purified and digested with EcoRⅠ/ SalⅠ. After cloned and sequenced, the sequence of cDNA from prothymosin-αwas compared with standard sequence to analyze the polymorphism in the cDNA sequence of prothymosin-α. The cloned cDNA sequence of prothymosin-αwas different from that of the standard. There are two kinds of variation. (1) The nucleotide in 107 was varied, with the deletion of nucleotides in 110-121 and 191-205. (2) The nucleotide in 306 was deleted, which is significantly more in the 60-80 years old group than the other groups. The two kinds of variation do not refer to the 28 peptides in the N-terminal of prothymosin-α. So the variation do not influence the function of prothymosin-α. It provides evidence for the structure, function and evolution in prothymosin-α.