本文選題：系統(tǒng)性紅斑狼瘡 + Sm抗原　； 參考：《中南大學》2007年碩士論文

【摘要】： 目的 本文利用Smith抗原模擬表位肽對自行制備的系統(tǒng)性紅斑狼瘡(systemic lupus erythematosus，，SLE)樣模型小鼠誘導免疫耐受進行初步探討，旨在為發(fā)展基于Sm抗原模擬表位的SLE肽耐受疫苗提供理論依據(jù)。 方法 應用ELISA法對我們前期研究中獲得的20個Sm模擬抗原表位噬菌體單克隆(命名為Sm-1~#～Sm-20~#)進行特異性鑒定，并對4個與Sm單抗呈明顯結(jié)合的噬菌體克隆進行DNA序列分析；然后通過尾靜脈途徑分別于第0-14-28-42-49-56天共6次注射上述4條單個模擬表位肽于SLE樣模型小鼠(注射劑量為0.8×10~(18)肽滴度／鼠)，并注射原始肽庫作為陰性對照組；70天后處死小鼠，分別檢測處理組和對照組小鼠血中自身抗體(雙鏈DNA抗體和抗核抗體)、測定各組小鼠尿蛋白含量以及觀察各組小鼠腎臟的病理改變。 結(jié)果 1．4個陽性噬菌體單克隆Sm-3~#、Sm-6~#、Sm-9~#、Sm-11~#插入的抗原表位的氨基酸序列分別為：Sm-3~#：SCFLCEIVIRSQ；Sm-6~#：NCYPLTIPHNIR；Sm-9~#：IRSQFHTTYEPE；Sm-11~#：KYPPNSYASQSE。同源性分析顯示上述4個表位肽段氨基酸序列與Genbank中已知的氨基酸序列相比無同源性。 2．Sm-11~#處理組小鼠血清中ANA和dsDNA抗體均有不同程度的明顯下降，與原始肽庫對照組小鼠比較，差異有顯著性(P＜0.05)；Sm-11~#處理組小鼠蛋白尿含量顯著降低，與原始肽庫對照組小鼠比較，差異有顯著性(P＜0.05)；與原始肽庫對照組小鼠比較，Sm-11~#處理組小鼠腎臟病理變化顯著改善，免疫復合物沉積明顯減少。而其他三個處理組小鼠的自身抗體水平、蛋白尿含量以及腎臟病理改善程度與原始肽庫對照組比較差異沒有顯著性。 結(jié)論 尾靜脈注射Sm抗原模擬表位單克隆噬菌體可以推遲或緩解狼瘡樣綜合癥小鼠病變的發(fā)生與發(fā)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.
【學位授予單位】：中南大學
【學位級別】：碩士
【學位授予年份】：2007
【分類號】：R392

【參考文獻】

相關期刊論文 前8條

1 高蓉,謝艷華,王四旺,樊春梅,王劍波;SLE小鼠血清中自身抗體檢測[J];第四軍醫(yī)大學學報;2003年10期

2 謝紅付,樹葉,張慧,施為,馮浩;運用噬菌體隨機肽庫分析系統(tǒng)性紅斑狼瘡的Sm抗原模擬表位[J];臨床皮膚科雜志;2004年10期

3 孫濤,陸蘋,王欣;一種實用的篩選病毒抗原表位方法的建立(英文)[J];生物化學與生物物理進展;2004年03期

4 ;A mimotope of Pre-S_2 region of surface antigen of viral hepatitis B screened by phage display[J];Cell Research;2001年03期

5 郝進,郝飛,鐘白玉,唐書謙,潘峰,茍俊,徐云升;核小體誘導BALB/c小鼠產(chǎn)生狼瘡腎的研究[J];中華皮膚科雜志;2004年01期

6 歐陽理,易新元Department of Pathogen Biology,曾憲芳Department of Pathogen Biology,周金春Department of Pathogen Biology,王慶林Department of Pathogen Biology,Larry Mc Reynolds;Partial protection induced by phage library-selected peptides mimicking epitopes of Schistosoma japonicum[J];Chinese Medical Journal;2003年01期

7 陽曉,葉任高,陳偉英,魏毅,許韓師,李幼姬,關偉民,余學清;慢性移植物抗宿主病狼瘡樣腎炎小鼠模型的研制[J];中華腎臟病雜志;2000年05期

8 謝紅付;馮浩;曾海燕;李吉;施為;易梅;伍斌;;利用Sm抗原模擬表位肽構(gòu)建狼瘡樣鼠模型[J];中國醫(yī)學科學院學報;2007年02期

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