体外鉴定的禽流感病毒T细胞表位的免疫效果研究

畜牧兽医学报 ›› 2008, Vol. 39 ›› Issue (4): 460-465.doi:

体外鉴定的禽流感病毒T细胞表位的免疫效果研究

李新生，陈红英，高凤山，夏春，崔保安

1．河南农业大学牧医工程学院，郑州 450002；2．中国农业大学动物医学院农业部预防兽医学重点实验室，北京 100094；3大连大学生物工程学院，大连 116622

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-04-24 发布日期:2008-04-24

The Immunologic Effect of Identified Avian Influenza Viruses T Cell Epitopes in vitro

LI Xin-sheng,CHEN Hong-ying, GAO Feng-shan, XIA Chun,CUI Bao-an

1. College of Husbandry and Veterinary, Henan Agricultural University, Zhengzhou 450002,China；2. The Key Laboratory for Preventive Veterinary Medicine of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100094,China; 3. Bioengineering College, Dalian University,Dalian 116622, China

Received:1900-01-01 Revised:1900-01-01 Online:2008-04-24 Published:2008-04-24

摘要/Abstract

摘要： 为验证来源于禽流感病毒的、可与鸡MHC Ⅰ类分子结合的九肽KILTIYSTV和LLLAIVSLV的免疫原性，使用TIGECPKYV、LLLAIVSLV和KILTIYSTV 3条多肽免疫BALB/c小鼠，加强免疫时和加强免疫1、2、3、4、5周后分别采血，流式细胞术测定免疫前后小鼠外周抗凝血中CD8+淋巴细胞的变化情况；加强免疫后14 d进行MTT试验；首次免疫3周后进行DTH试验；ELISA检测免疫前后小鼠分泌IFNγ的变化情况。结果表明，KILTIYSTV、LLLAIVSLV、TIGECPKYV免疫后分别引起CD8⁺ T淋巴细胞4.2%、4.0%和0.2%的额外增殖。KILTIYSTV和LLLAIVSLV免疫组的IFNγ的增长和迟发型变态反应均显著高于对照组。简言之，与重建的MHC Ⅰ类分子结合的多肽KILTIYSTV和LLLAIVSLV可以刺激小鼠产生特异性CTL反应，而不结合的多肽TIGECPKYV刺激小鼠未产生特异性CTL反应。

Abstract: To identify whether avian influenza viruses (AIV) derived peptides KILTIYSTV and LLLAIVSLV are authentic T cell epitopes, 6 weeks old BALB/c mice were immunized with KILTIYSTV, LLLAIVSLV and KILTIYSTV respectively. The blood samples were collected at the 1st, 2nd, 3rd, 4th and 5th week after reimmunization. CD8⁺ T cell was detected by fluorescence activated cell sorting (FACS). MTT test was carried out at 14 days after boost immunization; Peptide-specific delayed-type of hypersensitivity assay was done on third weeks after the first immunization. Proliferation of secreted IFN-γ in serum of mice was detected by ELISA. The results showed that the peptides KILTIYSTV, LLLAIVSLV and TIGECPKYV stimulated 4.2%, 4.0% and 0.2% growth of peptide-specific CD8⁺ T lymphocyte; Secreted IFN-γ level, specific delayed-type of hypersensitivity (DTH) response and MTT test in KILTIYSTV and LLLAIVSLV group were all significantly higher than that of control groups. In a word, animal immunity experiment revealed that two T cell epitopes KILTIYSTV and LLLAIVSLV could lead to specified CTL reaction, but the peptide TIGECPKYV that don′t bind with reconstructed MHC Ⅰ couldn′t.

李新生;陈红英;高凤山;夏春;崔保安. 体外鉴定的禽流感病毒T细胞表位的免疫效果研究[J]. 畜牧兽医学报, 2008, 39(4): 460-465.

LI Xin-sheng;CHEN Hong-ying;GAO Feng-shan;XIA Chun;CUI Bao-an. The Immunologic Effect of Identified Avian Influenza Viruses T Cell Epitopes in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2008, 39(4): 460-465.

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