人工设计的HIV-1重组鸡痘病毒诱导恒河猴产生免疫应答的研究  

Immune reponses induced by a designed HIV-1 recombinant fowlpox virus in Macaca mulatta

作　　者：张立树[1] 金宁一[1] 宋英[2] 洪宝庆[3] 马鹤雯 尚玉璞[2] 王宏[5] 孙岩松[2] 金洪涛[1] 战大伟[2] 李昌[1] 

机构地区：[1]军事医学科学院11所全军基因工程重点实验室,长春130062 [2]天津大学农业与生物工程学院,天津300072 [3]军事医学科学院实验动物中心,北京100071 [4]Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, USA 74078 [5]暨南大学生命科学院生物系,广州510632

出　　处：《高技术通讯》2007年第11期1198-1203,共6页Chinese High Technology Letters

基　　金：863计划（2003AA219051）资助项目.

摘　　要：将人工设计的HIV-1多表位-p24嵌合基因重组鸡痘病毒rFPV-MEGp24大量制备并纯化后,分别在0周、8周、18周肌肉注射至实验恒河猴中,在第2次免疫后2周和第3次免疫后2周采集血液,分离血清和外周血淋巴细胞（PBMC）,ELISA法检测血清HIV-1抗体和PBMC培养上清Th1类细胞因子的含量,MTT法测定PBMC的增殖能力,乳酸脱氢酶（LDH）释放法检测PBMC特异性CTL杀伤活性。结果表明：HIV-1多表位-p24嵌合基因重组鸡痘病毒可刺激恒河猴产生HIV-1特异性抗体;免疫猴的PBMC在HIV-1抗原肽刺激下细胞增殖能力加强,针对HIV-1靶细胞的特异性CTL杀伤活性产生,分泌Th1类细胞因子IFN-γ、IL-2的水平升高。研究提示,HIV-1多表位-p24嵌合基因重组鸡痘病毒能诱导恒河猴产生特异性细胞和体液免疫应答。A recombinant fowlpox virus （rFPV） containing a designed HIV-1 multi-epitope p24 chimeric gene was prepared on a large scale, and purified. Monkeys were then inoculated muscularly with the rFPV for three times in 0w, 8w and 18w. Whole blood was collected two weeks after the second and third inoculation, followed by serum and peripheral blood mononuclear cell （PBMC） separation. Anti-HIV-1 antibody in serum and Thl cytokins in supernatant of cultured PBMC stimulated by Macaca muiatta-specific HIV-1 peptide were assayed by enzyme linked immunosorbent assay （ELISA）. The proliferation and CTL activity of PBMC stimulated by Macaca mulatta-specific HIV-1 peptide were analyzed by MrIT and LDH release assay, respectively. The results showed that HIV-1 specific antibody in serum of immunized monkeys was higher than that in controls; Promoted proliferation, CTL target-killing activity, and secretion of Thl cytokines （IFN- 7 and IL-2） of monkey PBMC stimulated by HIV-1 CTL peptide were detected in the immunization group inoculated with rFPV for two times, which were further strengthened by the third inoculation. It is concluded that the designed rFPV may induce HIV-1 specific cellular and humoral immune responses in monkeys.

关 键 词：人免疫缺陷病毒I型 人工设计 重组鸡痘病毒 恒河猴 免疫应答 

分 类 号：S852.65[农业科学—基础兽医学]