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Study on Mamu-A~*01 of Chinese-Origin Rhesus Macaques and Analyses on Its Possible Role in Disease Progression with SIV/SHIV Infection

Author: LiuXiuYing
Tutor: WeiQiang；QinChuan；LiWanBo
School: Peking Union Medical College , China
Course: Zoology
Keywords: Simian immunodeficiency virus (SIV) Chinese rhesus Mamu-A * 01 p11C NASBA
CLC: R-332
Type: Master's thesis
Year: 2008
Downloads: 78
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Abstract

The development of AIDS in patients exist in varying degrees of differences, influencing factors, individual differences, that some of the genes in the genetic background may be involved in the impact of the disease process. Mamu-A ~ * 01 rhesus monkeys in a MHC-I molecules, foreign Mamu-A ~ * 01 gene and infected with SIV Indian rhesus disease process significantly associated with the role of this article to the Chinese rhesus Mamu-A ~ * 01 gene with SIV / SHIV infection susceptibility studies, study aimed to investigate the distribution of Mamu-A ~ * 01 gene in Chinese rhesus its SIV / SHIV infection role can further deepen the understanding of the role in vaccine research in the MHC, with higher targeted to establish more suitable for the evaluation of AIDS vaccine SIV / SHIV / SAIDS animal model experimental basis. This study selected 128 Chinese rhesus Mamu-A ~ * 01 gene screening, amplification of Mamu-A by sequence specific primers PCR (PCR-SSP) method ~~ * 01 gene, cloned and sequenced with India Ganges monkey Mamu-A ~~ * 01 gene homologous comparison; ELISPOT Mamu-A ~ * 01 gene-positive and-negative rhesus monkeys against SIV, SHIV antigen peptide p11C-specific CTL responses were detected. Screening of Mamu-A ~~ * 01 gene-positive rhesus observed in viral load, CD4 ~~ / CD8 ~~ ratio of CD4 ~ T cells absolute difference with Mamu-A ~ * 01 gene negative rhesus to further explore the Chinese Rhesus Mamu-A ~~ * 01 gene in SIV / SHIV rhesus monkey model of the course of. At the same time, we have also some other part of Chinese rhesus Mamu-I molecules allele (Mamu-A ~ * 02, Mamu-A * 04, Mamu-A * 08, Mamu-A ~~ * 13 , Mamu-NA4, Mamu-NA7, Mamu-B ~ * 01, Mamu-B ~ * 07, Mamu-B ~ * 17) screening to further speculated that the synergy between the various alleles or antagonism. The experiments were screened out of the five Mamu-A ~~ * 01 gene-positive rhesus monkeys, sequence analysis shows up to 99.1% of India rhesus gene homology, five were SIV / SHIV infected rhesus , display the ELISPOT result of four SIV-infected monkeys for the p11C the high frequency CTL responses, the number of spots in between 500-1400/10 PBMCs, an SHIV-infected rhesus macaques and five negative monkey no spots appear. Weed out the four SIV-infected Mamu-A ~~ * 01 gene-positive rhesus virus load, CD4-/ CD8 ~~ ratio of CD4 ~ T cells in the absolute value of the retrospective study found, Mamu-A ~ * 01 gene positive rhesus CD4 to / CD8 ~~ ratio is slightly higher than the Mamu-A ~ * 01 gene negative rhesus monkeys, but the ratio is greater than 1.0, does not appear significant proportion of inverted, viral load and absolute CD4 ~ T cells value is no significant difference between this part of the experiment is a retrospective study, the lack of acute infection, and may be affected by factors other experiments, the part of the experimental results are for reference only. Real-time PCR specificity, high degree of automation, can be an effective solution to the problem of PCR contamination and widely used. Build standard real-time quantitative PCR experiments very important and critical step, which determines the accuracy and reliability of the experimental results. Nucleic acid sequence-dependent amplification (Nucleicacid sequence-based amplification, NASBA) is mainly used for the RNA amplification, detection, and sequencing. The reaction has a fast, sensitive and specific characteristics. In this study, NASBA method for the establishment of SIV / SHIV viral RNA load standard, in order to find a convenient, simple and practical standard to establish a method. Experimental results show that we have successfully established a standard measured by the SIV / SHW viral RNA load of the RNA standards has good reproducibility and stability, and therefore can use the standard goods on SIV / SHIV viral RNA quantitative.

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