摘要
目的筛查中国恒河猴Mamu-A*01基因,比较中国恒河猴和印度恒河猴的Mamu-A*01基因序列和功能是否相同。方法PCR方法检测128只中国恒河猴,用特异性引物扩增Mamu-A*01基因,将PCR扩增后的产物克隆测序后与印度恒河猴的Mamu-A*01基因进行同源比对;酶联免疫斑点检测(ELISPOT)方法分别检测5只Mamu-A*01基因阳性和5只阴性恒河猴针对SIV、SHIV抗原肽p11C的特异性CTL反应。结果共筛查出5只Mamu-A*01基因阳性恒河猴(3.91%),经测序分析后与印度恒河猴的同源性可达99.1%。这5只均为SIV/SHIV感染恒河猴,其中四只SIV感染的猴的ELISPOT结果显示针对p11C的高频CTL反应,斑点数在500-1400/106PBMCs之间,而另1只SHIV感染的恒河猴及5只阴性猴没有斑点出现。结论中国恒河猴含有Mamu-A*01基因,基因频率有区域性差异,中国恒河猴的Mamu-A*01可提呈特异性抗原肽p11C。
Objective To screen Mamu-A^＊01 positive Chinese rhesus macaques and compare the possible differences in Mamu-A^＊01 between the rhesus macaques from China and India. Methods 128 Chinese rhesus macaques were included in this study. PCR was applied with specific primers amplified Mamu-A^＊01 gene to detect Mamu-A^＊01 positive monkeys. PCR products were cloned and sequenced, and used for the homology comparison with Indian rhesus macaques. ELISPOT method was used to detect the specific CTL response targeting the peptide p11C. Results A total of 5 rhesus monkeys with Mamu-A^＊01 positive genes were found in the 128 Chinese rhesus macaques, and the sequence homology comparison with Indian rhesus macaques showed a 99.1% identity. Except one monkey inoculated with SHIV ,the other four SIV-infected monkeys showed that the high-frequency CTL response in spots between 500- 1400/106 PBMCs,and the 5 Mamu-A^＊01 negative monkeys did not show any CTL response spot. Conclusion 3.91 % of monkeys with Mamu-A^＊01 gene has been found from 128 Chinese rhesus macaques,and that gene can present the specific antigen p11C in Chinese rhesus macaques.
出处
《中国实验动物学报》
CAS
CSCD
2008年第2期100-104,共5页
Acta Laboratorium Animalis Scientia Sinica
基金
中国CIPRA项目
美国NIH资助(编号:U19AI51915-04)
中央级公益性科研院所基本科研业务费专项资金(编号:DWS200712)
