KDEL修饰的HSV-2CD8~+T细胞表位促进CTL效应的研究

目的:研究赖氨酸天冬氨酸谷氨酸亮氨酸(Lys-Asp-GluLeu,KDEL)修饰的2型单纯疱疹病毒(herpessimplexvirustype2,HSV2)CD8+T细胞表位促进细胞毒T淋巴细胞(cytotoxicityTlymphocyte,CTL)效应。方法:将雄性C57BL/6小鼠随机分为5组,每组5只,用各抗原肽免疫C57BL/6小鼠,采用3HTdR掺入法检测淋巴细胞增殖反应、标准4h51Cr释放试验检测CD8+T细胞特异性CTL效应,观察HSV2CD8+T细胞表位(SSIEFARL,S1)、KDEL修饰的HSV-2CD8+T细胞表位(SSIEFARL-KDEL,S1-KDEL)、4拷贝串联的CTL表位(SSIEFARL)4,S4]及KDEL修饰的串联CTL表位(SSIEFARL)4KDEL,S4-KDEL]的特异性细胞免疫应答。结果:3HTdR掺入试验中,S4组和S4KDEL组cpm值显著高于对照组和S1组、S1KDEL组(P<0.05);S4KDEL组的cpm值显著高于S4组(P<0.05);S1组和S1KDEL组与对照组比较,差异无显著性(P>0.05);S1组与S1KDEL组比较,cmp值亦无显著性差异(P>0.05)。以S1致敏的EL4细胞为靶细胞的杀伤实验中,S4和S4KDEL组诱导的CTL活性明显高于对照组、S1组及S1KDEL组(P<0.05);S4KDEL组诱导的CTL活性明显高于S4组(P<0.05),S1组和S1KDEL组与对照组比较,差异无显著性(P>0.05);S1组与S1KDEL组比较,差异亦无显著性(P>0.05);以EL4细胞为靶细胞的实验中,实验组的杀伤率均在10%以下,与对照组?

CTL Epitopes Modified by KDEL and Recognized by CD8 + T Lymphocytes to Herpes Simplex Virus Type 2 Improve CTL Effect

OBJECTIVE: To study the improvement of CTL effect by CTL epitopes which are modified by KDEL and recognized by CD8+ T lymphocytes to herpes simplex virus type 2. METHODS: Observations were made on the specific immune response induced by the CD8+ CTL epitopes(SSIEFARL, S1), the CD8+ CTL epitopes modified by KDEL(SSIEFARL-KDEL, S1-KDEL), the tandem four copies CD8+ CTL epitopes(SSIEFARL)4, S4] and the tandem four copies CD8+ CTL epitopes modified by KDEL(SSIEFARL)4-KDEL, S4-KDEL], 25 male C57BL/6 mouse were randomly divided into 5 group, 5 mice per group, respectively immunized with istonic Na chloride, S1, S1-KDEL, S4 and S4-KDEL. Lymphocyte proliferation was detected by 3H-TdR and the CTL effect induced by CTL epitopes in vivo was detected by 51Cr. RESULTS: In the 3H-TdR test, compared with the control, the group S1 and group S1-KDEL, the cpm values of Group S4 and Group S4-KDEL were markedly higher (P < 0.05) and the cpm value of Group S4-KDEL was significantly higher than Group S4 (P < 0.05), but the cpm values of Group S1 and Group S1-KDEL were not significantly different from the control (P > 0.05), nor was that of Group S1 from Group S1-KDEL (P > 0.05). In the experiment in which the EL4 cells sensitized by S1 were attacked as target cells, the CTL activities induced in Group S4 and Group S4-KDEL were markedly higher (P < 0.05) compared with the control, Group S1 and Group S1-KDEL, and that induced in Group S4-KDEL was significantly higher than Group S4 (P < 0.05), but the CTL activity induced in Group S1 and Group S1-KDEL were not significantly different from the control (P > 0.05), nor was that induced in Group S1 from Group S1-KDEL (P > 0.05). In the experiment in which the EL4 cells were attacked as target cells, the kill rate was below 10% in every group, not significantly different from the control. CONCLUSION: The tandem four copies CD8+ CTL epitopes, modified by KDEL and recognized by CD8+ T lymphocytes to herpes simplex virus type 2, can improve the CTL effect.