Sm抗原表位肽诱导狼疮样鼠免疫耐受的初步实验研究

发布时间：2018-05-08 17:37

本文选题：系统性红斑狼疮 + Sm抗原　；参考：《中南大学》2007年硕士论文

【摘要】： 目的本文利用Smith抗原模拟表位肽对自行制备的系统性红斑狼疮(systemic lupus erythematosus，，SLE)样模型小鼠诱导免疫耐受进行初步探讨，旨在为发展基于Sm抗原模拟表位的SLE肽耐受疫苗提供理论依据。方法应用ELISA法对我们前期研究中获得的20个Sm模拟抗原表位噬菌体单克隆(命名为Sm-1~#～Sm-20~#)进行特异性鉴定，并对4个与Sm单抗呈明显结合的噬菌体克隆进行DNA序列分析；然后通过尾静脉途径分别于第0-14-28-42-49-56天共6次注射上述4条单个模拟表位肽于SLE样模型小鼠(注射剂量为0.8×10~(18)肽滴度／鼠)，并注射原始肽库作为阴性对照组；70天后处死小鼠，分别检测处理组和对照组小鼠血中自身抗体(双链DNA抗体和抗核抗体)、测定各组小鼠尿蛋白含量以及观察各组小鼠肾脏的病理改变。结果 1．4个阳性噬菌体单克隆Sm-3~#、Sm-6~#、Sm-9~#、Sm-11~#插入的抗原表位的氨基酸序列分别为：Sm-3~#：SCFLCEIVIRSQ；Sm-6~#：NCYPLTIPHNIR；Sm-9~#：IRSQFHTTYEPE；Sm-11~#：KYPPNSYASQSE。同源性分析显示上述4个表位肽段氨基酸序列与Genbank中已知的氨基酸序列相比无同源性。 2．Sm-11~#处理组小鼠血清中ANA和dsDNA抗体均有不同程度的明显下降，与原始肽库对照组小鼠比较，差异有显著性(P＜0.05)；Sm-11~#处理组小鼠蛋白尿含量显著降低，与原始肽库对照组小鼠比较，差异有显著性(P＜0.05)；与原始肽库对照组小鼠比较，Sm-11~#处理组小鼠肾脏病理变化显著改善，免疫复合物沉积明显减少。而其他三个处理组小鼠的自身抗体水平、蛋白尿含量以及肾脏病理改善程度与原始肽库对照组比较差异没有显著性。结论尾静脉注射Sm抗原模拟表位单克隆噬菌体可以推迟或缓解狼疮样综合症小鼠病变的发生与发展。
[Abstract]:Purpose In this paper, Smith antigen mimic epitope peptide was used to study the induction of immune tolerance in systemic lupus erythematosus model mice. The aim of this study was to provide a theoretical basis for the development of SLE peptide tolerance vaccine based on Sm antigen mimic epitope. Method ELISA method was used to identify 20 Sm mimic antigen epitopes (named Sm-1 #Sm-20 #A), and four phage clones with obvious binding to Sm McAbs were sequenced by DNA. Then the mice were injected with the four single mimic epitope peptides in SLE like model mice (0. 8 脳 10 ~ 0 ~ (18) peptide titer by tail vein route on the 0-14-28-42-49-56 days, respectively. The mice were killed after 70 days of injection of the primordial peptide library as a negative control group. The autoantibodies (double-stranded DNA antibody and anti-nuclear antibody) were detected in the blood of the treated group and the control group respectively. The urine protein content and the pathological changes of kidney in each group were measured. Result 1.The amino acid sequence of Sm-9 Sm-11 ~ # inserted from Sm-6 #Sm-6, #Sm-6, a phage positive phage, is: 1: Sm-3 #Sm-3 #CEIVIRSQSm-6 #1: NCYPLTIPIPHNIRN Sm-9 #IRSQFHTTYEPEEPE Sm-11 #KYNSYASQSE. The amino acid sequences of the epitopes are: 1: Sm-3 #Sm-3 #Sm-3 #Sm-3 #Sm-6 #Sm-6 #Sm-9 #Sm-9 #Sm-11: #KYPNPPNSYASQSE. Homology analysis showed that the amino acid sequences of the four epitope peptides had no homology compared with the known amino acid sequences in Genbank. The levels of ANA and dsDNA antibody in serum of 2.Sm-11~# treated mice were significantly decreased compared with the control group of the original peptide library. Compared with the control group of the original peptide library, the proteinuria content in the treated group was significantly lower than that in the original peptide library control group (P < 0.05 Sm-11 ~ #), and compared with the control group of the original peptide library, the content of proteinuria in the treated group was significantly lower than that in the control group. The difference was significant (P < 0.05), compared with the control group, the pathological changes of kidney and the deposition of immune complex in Sm-11 ~ # treated mice were significantly improved. However, there was no significant difference in autoantibody level, proteinuria content and renal pathological improvement between the other three treatment groups and the control group of the original peptide library. Conclusion Monoclonal phage injection of Sm antigen mimic epitopes in tail vein can delay or alleviate the occurrence and development of pathological changes in mice with lupus like syndrome.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392

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