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Infection of Chinese Original Rhesus Macaques by Three Routes with Subtype C SHIV-1157ipd3N4 and Analysis of Viral Variation

Author: JinGuang
Tutor: WeiQiang；QinChuan；LiWanBo
School: Beijing Union Medical College
Course: Zoology
Keywords: SHIV-1157ipd3N4 Primate model AIDS Variation Multiple low dose
CLC: R512.91
Type: Master's thesis
Year: 2011
Downloads: 3
Quote: 0
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Abstract

Currently, the main subtype C HIV-1 is still in the world widely popular, there is an urgent need to develop a safe and effective AIDS vaccine, thus the need for appropriate animal models of AIDS vaccine evaluation. Subtype C SHIV and non-human primate model study of HIV-1 infection and pathogenesis and evaluation of candidate vaccine efficacy of subtype C env region as a target effective tool. Evaluation of vaccine candidates, in order to establish the appropriate subtype C SHIV animal model chosen for this study the SHIV-built RMRuprecht Professor 1157ipd3N4 strains of SHIV encoding HIV-1 C subtype env gene, assisted by a high degree of ability to replicate and single CCR5 body tropism, mucosal routes infected rhesus monkeys and induced AIDS similar symptoms and slow disease progression in infected monkeys is similar to the human HIV-1 infection. This study is first prepared in SHIV-1157ipd3N4 China rhesus monkey cell adapted strain and its sequence and the SHIV-1157ipd3N4 clones of similarity is found to 99%. Subsequently, in order to provide a reference to the subsequent animal model inoculation dose of the SHIV-1157ipd3N4 the Chinese rhesus monkey cells adapted strains and India rhesus cell adapted strains carried the cellular level evaluation, the results of the former virus concentration 500TCID50/ml, while the latter is 1000TCID50/ml. Simulation of blood-borne and sexually transmitted establish SHIV-1157ipd3N4 / rhesus monkey model to reflect the spread of AIDS, this study were used to infect Chinese rhesus the SHIV-1157ipd3N4 Chinese rhesus monkey cells adapted strains intravenous and rectal routes, SHIV- Indian rhesus cells adapted strains vaginal route 1157ipd3N4 infected Chinese rhesus monkeys. Meanwhile, in order to better reflect the natural modes of transmission of HIV in the crowd, rectum and vagina pathway model using a small dose of infection. Results SHIV-1157ipd3N4 the Chinese rhesus monkey cells adapted strains can be successfully infected through intravenous and rectal routes Chinese rhesus SHIV-1157ipd3N4 Indian rhesus monkey cell adapted strains through the vaginal route successfully infect Chinese rhesus establish three Ways to of SHIV-1157ipd3N4 / Chinese rhesus monkey model. Subsequently, the study compared the biological characteristics of the different routes of infection animal models, susceptibility and occurrence regularity. The results found that the three ways to establish the SHIV-1157ipd3N4 / rhesus monkey model biological characteristics differ. Plasma viral RNA load the intravenous group of infected monkeys in a peak when the 21d rapid decline into the platform of rectal group and vaginal infections monkey seroconversion plasma viral RNA load and peak time of the same time, part of the infected monkeys peak is not obvious, after all small dose infection gradually into the plateau; peripheral blood CD4 / CD8 T lymphocyte ratio of the intravenous group of infected monkeys 21d decline, followed by a rapid return to normal, rectal group and vaginal infections monkey presents several small amplitude fluctuations in the process of multiple infection ratio in small doses, part monkey apparent decline in the process, three groups of monkeys infected with the ratio generally decreased with the infection time; vein group and rectum groups throughout the infection process and foreign weeks blood CD4 T lymphocyte absolute number gradually increased vaginal group remained stable; three groups of infected monkey plasma combined with a steady increase in antibody titers, late intravenous group was significantly higher than rectal group and vagina group. The Chinese rhesus three ways different the SHIV-1157ipd3N4 susceptibility, intravenous route seeded 50 TCID50 and 5 TCID50SHIV-1157ipd3N4, of Chinese rhesus cells adapted strains can be successfully infected, while 0.5 TCID50 and 0.05 TCID50 not, infection. Rectal and vaginal routes were small doses of the experimental monkeys were infected 13 times and 11 times after inoculation. Only 49w to infection, all infected monkeys were obtained significantly similar clinical symptoms of AIDS. In addition, in order to reveal the Variation of the AIDS virus, this study further variation trend SHIV-1157ipd3N4 Chinese rhesus monkeys infected by various means to explore. Found that the nucleotide and amino acid sequences of gp120 was amplified from the plasma of infected monkeys in the short term (s) exists only the point of change of a few bits, no changes in the N-glycosylation sites did not appear obvious variation trend. HIV-1 p24 and SIV p27 two ELISA kit, in order to prove the reliability of the results of HIV-1 p24 ELISA kit for detection of SIV p27 antigen in the process of preparation of virus strains, this study compared the virus liquid and plasma SIV p27 antigen test results. The results showed that the HIV-1p24 and SIV p27 two ELISA kit for the detection of SIV p27 antigen the sensitivity 150pg/mL and 62.5 pg / ml; the two kits detect viruses in fluid and plasma SIV p27 antigen qualitative results; quantitative results on statistical analysis of the virus solution linear regression coefficient of determination R2 = 0.857, correlation coefficient r = 0.926, P lt; 0.01; plasma linear regression coefficient of determination R2 = 0.512, correlation coefficient r = 0.716, P lt; 0.05. To sum up, the subject is first prepared the in SHIV-1157ipd3N4 China rhesus monkey cells adapted strains then simulate natural human infected with HIV the way and the three main routes of infection are intravenous, rectal and vaginal route SHIV-1157ipd3N4 / Ganges monkey model, and biological characteristics of the different routes of infection animal model susceptible to infection and morbidity of the law, further virus infections the Chinese rhesus variation trend by various means to explore the other compared the HIV-1 p24 the two SIV p27 ELISA kit detection of SIV P27 antigen results. These studies reveal HIV-1 infection and pathogenesis reference, and provides an effective tool for the evaluation of candidate vaccine targets subtype C env region.

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