The Study Of SIV-infected Rhesus Macaque Long-term Non-progressors,and The Effects Of Artesunate On AIDS Rhesus Model

Objective:1.To compare the levels of T lymphocyte immune activation and mitochondrial antiviral protein levels in Chinese rhesus monkeys with different progress types of SIV,and to explore the intrinsic pathologic mechanism of long-term non-progressive SIV-infected monkeys.2.To investigate the effects of recurrent infection on normal and long-term non-proculating SIV-infected monkeys by repeatedly attacking SIVmac239 of Chinese rhesus monkeys with different progress types of SIV.3.To observe the therapeutic effect of artesunate on the immune activation of Chinese rhesus monkeys infected with SIV,and to find the treatment strategy of Chinese medicine intervention in AIDS from the perspective of regulating immune abnormalities.Method:1.A Comparative Study on the Immune Characteristics of Chinese RhesusMonkey Model with Different Progressive Type SIV Infection:17 Chinese rhesus monkeys were divided into healthy control(HC)group according to whether they were infected,the viral load after infection,and the survival time after infection;Typical progressor(TP)group;long-term development(viremic controllor,VC)group;elite control(elite controllor,EC)group.Detection of immune activation regulation related indicators,two weeks after repeated testing to verify the stability of the relevant indicators.2.Chinese Rhesus monkey SIV repeated attack research:Among 8 rhesus infected with SIV,4 are in the normal progress,with the other 4 in elite control group.Four normal progressive rhesus monkeys were randomly divided into two groups:inoculation group and control group.Four elite control rhesus monkeys were randomly divided into two groups:control group and control group.In the challenge group,1 mL of SIVmac239 virus suspension was injected intravenously at 5 weeks,4 weeks and 8 weeks with 5 MID100(5 times 100%infection dose).3.Artesunate on SIV infection in China rhesus monkey model intervention:healthy Chinese rhesus monkey 8,copy AIDS model,to be progressed to the platform stage,divided into two groups,artesunate treatment group 4,saline control Group 4 only.Artesunate was administered for 8 weeks.The immunoregulatory parameters were measured at 2 weeks,4 weeks,6 weeks,8 weeks,and at the respective point of 4 weeks and 8 weeks after drug administration.result:1.Comparison of immunological characteristics of Chinese rhesus monkeys with different progress types of SIV infectionTP group viral load is typical of the incubation period,the average is lg 5.25 copies/ml.VC group was lower than TP group(P<0.05),the average value was lg 4.02 copies/ml.The viral load of the EC group is at an undetectable level for less than 50 copies/ul.HC group,TP group and EC group,the white blood cell count is similar,VC group higher than the other three groups.The proportion of CD4 cells in HC group was the highest,and there was no significant difference between the three groups.CD4 count was the lowest in the TP group,slightly lower than the other three groups.CD8 cells in four groups of rhesus monkeys no significant difference.The ratio of CD4/CD8 in HC group was higher than that in the other three groups.There was no significant difference in CD4/CD8 ratio between TP,VC and EC groups.In the four groups,the proportion of TP mononuclear cells was the highest,followed by VC group and EC group,the lowest in HC group.Monocyte count was the highest in VC group,followed by TP group and EC group,the lowest in HC group.The expression level of CD69 on the CD4 cell surface of the rhesus monkey group was maintained at a low level,and the other three groups had different levels of infection.(P<0.01),while VC group and EC group were lower than TP group,but higher than HC group(P<0.01).The relative levels of CD25 expression in CD4 group were similar to those in CD69 group(P<0.01),and VC group(P<0.05)compared with HC group.HC monkey CD8 cells had lower levels of CD69 expression,while the other three groups of rhesus monkeys were elevated.The expression level of CD25 in CD8 cells was similar to that in CD69,the level of HC group was lower and the other three groups were slightly increased,and the TP group was the most obvious.The expression levels of CD38 in CD4 type rhesus monkey and VC rhesus monkey CD4 cell were higher than that of HC monkey.The expression level of CD38 in EC type rhesus was lower than that in TP group(P<0.01)and VC group(P<0.01).The expression of HLA-DR in CD4 group was significantly different from that in TP group(P<0.05)and VC group(P<0.05).The expression of CD38 in EC group was lower than that in HC monkey,the average value was below 10%,but the difference was not statistically significant.There was significant difference between CD group(P<0.01)and VC group(P<0.05).The expression of HLA-DR in CD8 cells was significantly different from that in TP group(P<0.01)and VC group(P<0.01).The expression levels of PD-1 in CD4 group were similar in the three groups,and the HC group was slightly higher than that of the three infection groups.The level of PD-1 expression in CD8 cells was similar to that in the three infection groups,but decreased compared with the HC group and decreased more than that of the CD4 cells.There was no statistically significant difference between groups.The ratio of SIV-specific CD8 cells in the three infected groups decreased,and the EC group decreased the least,followed by the VC group.TP monkey SIV specific CD8 ratio decreased the most serious,and the difference was significant(P<0.01).EC group was significantly higher than TP group(P<0.01).The absolute value of SIV-specific CD8 cells was lower in HC group than that in HC group,while VC group and EC group were higher than HC group,among which EC group was the highest,but statistically significant.The expression levels of PD-1 in SIV-specific CD8 cells,VC and EC were not the same as those of HC at the same level,and the mean of TP group was higher than that of the other three groups.The expression level of CD38 and the expression of HLA-DR in R8 monkey CD8 cells were inversely proportional to the ratio of SIV-specific CD8 cells.The Treg ratios of VC and EC were not consistent with those of HC group,but TP was significantly higher than that of the other three groups.The difference was statistically significant.There was no significant difference in the absolute number of Tregs in the four groups of rhesus monkeys,and the mean value of TP group was slightly higher than that of the other three groups.The expression rate of Ki67 in Treg was significantly higher than that in non-Treg CD4 cells in all 17 rhesus monkeys,the median of which was about 11 times larger than that of the latter.The expression rate of CTLA-4 on Treg was significantly higher than that in non-Treg cells in CD4 cells of all four groups of rhesus monkeys.Compared with HC group and TP group,CTLA-4 expression level was down-regulated in both CE and EC groups,and there was significant difference between TP and VC and EC group(P<0.05).The expression level of CTLA-4 on non-Treg was higher than that of the other three groups.2.Experimental Study on Repeated Infection of Chinese Rhesus Monkey SIVTP-A rhesus monkeys of normal development showed the emergence of fatigue,chills,weight loss,in the first attack and died on the 22nd day.No obvious abnormalities were found after the systematical postmortem.The other rhesus showed no obvious symptoms.EC-B did not change the EC-A virus load,EC-B increased to measurable level after the third challenge,but the level remained below 3 for at least 8 weeks,and 16 weeks after the third challenge,the result can not be measured.TP-A monkey after the first attack showed an obvious decrease in the white blood cells,and the value reached the lowest point in the second week.In the third week,the valued increased.TP-B monkeys in the second attack had an increase in the white blood cells,after 3 weeks the result returned to pre-experimental level.The result decreased after 16 weeks.EC-A and EC-B after the repeated attacks of white blood cells repeated fluctuations,the highest level of 12 × 103/μ1,the lowest as low as 12 × 103/μ1.4 weeks after the third attack,the result stabilized.TP-B monkeys after the second attack showed an increase in lymphocyte ratio fluctuations,while after the third attack,the value rose slowly.The ratio of EC-A and EC-B in the lymphocyte ratio after the first and third challenge was fluctuated,and the value after ECL was higher than that before the experiment.TP-B monkey pure CD4 cell ratio fluctuated after the first attack,the second and third attack after the fluctuations were significantly reduced,but the values are slightly lower than before the attack.TP-A monkey increased the level of CD4 cell proliferation after the first challenge,and decreased after 2 weeks.TP-B monkey pure CD4 cells were lower than the pre-challenge levels after the first and second challenge.TP-B pure CD8 cells were significantly lower than before the second challenge.The Treg ratio of TP-A monkey after the first attack increased significantly.The expression level of CTLA-4 in TP-A monkey Treg cells increased rapidly after the first challenge,and began to decline after reaching the highest point after 2 weeks.The expression level of CD38 in CD4-A monkey increased rapidly after the first challenge,and it was about 3 times the valve before the experiment in 3 weeks,while TP-B monkey decreased after each challenge.The level of HLA-DR expression in TP-A monkey CD4 cells continued to rise after the first challenge until the third week.The expression level of CD38 in TP-B monkey CD8 cells increased significantly after 1 week of the second challenge,and then maintained at a relatively high level to maintain relative fluctuation.The level of HLA-DR expression in CD8 cells decreased significantly 3 weeks after the attack on TPE-1 monkeys.3.Experimental study on the treatment of SIV-infected Chinese rhesus monkeys with artesunateIn the 8 weeks after administration,the white blood cell count began to decrease from the 4th week and began to rise after the 6th week fell to the lowest point.White blood cells continued to rise after withdrawal,withdrawal 4 weeks before the rise to the level before the fall.The proportion of lymphocytes increased to the highest point at the 6th week,which was significantly higher than that of the control group(P<0.05).In the 8 weeks,the proportion of neutrophils in the treatment group decreased from the 4th week to the 6th week,which was significantly lower than that of the control group(P<0.05)。The percentage and count of CD8 cells in the administration group were lower than those in the control group at the time of withdrawal,and the difference was significant(P<0.05).The expression level of CD69 in CD8 cells was significantly lower than that in the control group(P<0.05).After administration,the levels of HLA-DR and CD38 in the CD8 cells of the treatment group began to decrease,and both were significantly lower than those in the control group at the 4th week(P<0.05).The two indicators rebounded from the 6th week,the two groups disappeared.The percentage of HLA-DR + CD8 cell count decreased significantly after 2 weeks of administration(P<0.01),and the percentage of 4 weeks and 8 weeks was significantly higher than that of the control group(P<0.05).The percentage of CD38 + CD8 cell count was significantly higher than that of the control group at the 2nd week and the fourth week after drug withdrawal(P<0.05).4 weeks after dosing,the expression of CD107a in CD8 cells decreased.Conclusion1:Compared with monkeys of the normal progress,long-term non-progress ones have a better CD8 cell function.CD4/CD8 ratio can not evaluate the type of progress of SIV infection in Chinese rhesus monkeys.Long-term non-progressive SIV infection in Chinese rhesus monkey T lymphocyte activation was significantly lower than normal progression monkeys,elite control rhesus monkeys exist in addition to anti-viral immunity other than inhibition of lymphocyte abnormal activation mechanism.2:Reducing the immune system Self-regulated cell apoptosis is one of the causes of long-term non-progressive SIV infection in Chinese rhesus monkeys that maintain high levels of CD4 cells for a long time,but the main reason for SIV infection in Chinese rhesus CD4 cell reduction is not spontaneous cell apoptosis.The higher the abnormal activation level of CD8 cells become,the lower the proportion of cells with cytotoxicity to SIV will be.3:The expression level of CXCR4 in monkey CD4 cells was maintained at a low level.Reducing the immune regulation of Treg is one of the important mechanisms of long-term non-progression of SIV-infected monkeys,and Treg’ s transmission of viral DNA is not a key point for long-term progression of monkeys.4:Repeated challenge can inhibit the renewal of TP monkey CD4 cells and CD8 cells,accelerate the progression of TP monkeys,can increase the amount of EC type rhesus virus.Repeated challenge increased the immune activation of CD4 monkey and CD8 cells in monkey monkeys,and had no effect on immune activation of EC monkeys.MBL and MAVS are one of the mechanisms by which EC monkeys control the virus.5:Artesunate improves SIV infection in Chinese rhesus monkey cytotoxic T cell immune abnormalities.The presence of artesunate in the treatment of neutropenia is reversible.