摘要
目的:针对同种异体细胞移植的免疫反应主要由HLA-Ⅱ类抗原介导,而CⅡTA基因对HLA-Ⅱ分子的表达起严格且专一性的调控作用,拟通过CⅡTA的反义cDNA抑制细胞表面HLAⅡ分子的表达。方法:以RT-PCR从HLAⅡ抗原组成型表达的Raji细胞株克隆CⅡTA反义片段(arⅡ),插入腺相关病毒载体psNAV中(pAarⅡ)。将该质粒通过脂质体稳定转染Heda细胞(pAar Ⅱ H),检测其表面HLAⅡ分子表达,并以RT-PCR检测其CⅡTA、HLA-Ⅱ(DR、DP及DQ)及Ii分子的mRNA水平。结果:pAarⅡH在重组人IFN-γ诱导下,DR、DP及DQ抗原表达分别降低了79±12%、90±15%及40±8%;同时HLA-Ⅱ及Ii分子的mRNA被明显抑制。结论:提示CⅡTA反义基因片段抑制了自身组成型及诱导型表达量,并相应阻止了CⅡTA所调控的基因(HLA-Ⅱ及Ii)的表达,从而为进一步探讨免疫耐受形成及其在组织工程中的应用奠定了基础。
Objective: For the alloantigen-specific immune response is mainly mediated by MHC class Ⅱ (MHCⅡ) molecules, this paper investigate the feasibility of using an antisense cDNA sequence of CⅡTA (ar Ⅱ),which is a major regulator of HLAⅡ expression, to suppress the alloantigen-specific immune response. Methods: The ar Ⅱ was obtained by RT-PCR, and then inserted into the psNAV plasmid (pAar Ⅱ). Stable transfection of Hela cell line with pAar Ⅱ (pAar Ⅱ H) were tested for class Ⅱ HLA induction by recombinant human interferon- gamma(IFN-γ). mRNA abundance of HLA-Ⅱ, Ii and C Ⅱ TA was measured by RT-PCR. Results: When induced with IFN-γ, the expression of HLA-DR, -DP and -DQ on the cell surface of pAar Ⅱ H was almost totally inhibited, while the mRNA contents of CⅡTA, HLA-DR, -DP, -DQ and Ii were reduced significantly. Conclusion: The ar Ⅱ downregulates CⅡTA and thus inhibits the family of genes (HLA-Ⅱ and Ii) regulated by CⅡTA. These results provide insight into the future application of antisense CⅡTA for the antigen-specific tolerance induction in allo-transplantation of the tissue engineering.
出处
《中国临床医学》
2004年第2期174-176,共3页
Chinese Journal of Clinical Medicine
基金
上海市科技发展基金资助(批准号 OODJ14001-8)
