ABSTRACT
Until now, the exactly effect of Kupffer cells (KCs) on inducing immune tolerance or aggravating acute rejection is still unknown. Activated by various way after liver transplantation, have the ability of phagucytosis the apoptositic T cells, up-regulated expression of FasL and many Th2/Th3 cytokines, such as interleukin-10 and transforming growth factor-lB. These up-regulated cytokines could induce the apoptosis of Th1 cells and enhance the proliferation and differentiation of the Th2 cells, finally induce the immune tolerance, However, the activated KCs also have the ability of expression many cytokine-dependent molecules,such as class Ⅱ major histocompatibility antigens, adhesion molecule and costimulatory molecules which could enhance the function of the antigen presentation, increase the expression of Thl cytokine and aggravate the acute rejection after liver transplantation. It maybe relate to the ratio of theTh1/Th2 cells determined by the complicated net of the cytokine produced by the activated Kupffer cells: the predominance of Th2 cells could induce the immune tolerance, on the contrary, the acute rejection proceed.
ABSTRACT
<p><b>OBJECTIVE</b>To establish a rapid specific method to identify the single-nucleotide polymorphisms (SNPs) of HBV polymerase gene region which are the methionine residue of the conserved YMDD motif.</p><p><b>METHODS</b>Two specific primers were designed to amplify interested gene region involved in SNPs which were also used as HBV DNA identification. Specific primers of SNaPshot were designed to detect 741A-G (YVDD), 743G-T (YIDD). The different fluorescent dye labeled ddNTP was used to further extend the strand of PCR product and was detected by ABI PRISM 310 Genetic Analyzer. Sera from 13 patients with chronic hepatitis B after lamivudine treatment were analyzed.</p><p><b>RESULTS</b>Aside from mutation of YMDD, there were mutations of 514C-A, 523C-A, 562T-A, 667C-A. The 13 samples were simultaneously tested with SNaPshot and DNA sequencing, the same results were obtained. The method of SNaPshot showed high specificity.</p><p><b>CONCLUSION</b>Mutation of YMDD results in the changes of ATG codon, and there are new ATG codon in the upper strand of YMDD. SNaPshot technique is rapid, specific and accurate for the SNPs monitoring of HBV DNA mutation during lamivudine therapy. Two samples were determined by SnaPshot technique, identifying the co-existence of the mixed wild type and mutant type HBV infection.</p>