| Objective:HIV-related immune activation and inflammation are closely related to the occurrence of incomplete immune reconstitution and serious non-AIDS events,which is a major obstacle for the cure of HIV Currently,ART treatment combined with immunomodulatory drugs has been attempt to improve the abnormal immune response related to HIV/AIDS.However,the ideal therapeutic effect has not been achieved.5-hydroxy-triptolide(LLDT-8),as a modified compound of Triptolide,is an effective immunomodulator and is widely used in rheumatic diseases.This study aims to investigate the effect of LLDT-8 on T cell activation and proliferation in healthy people and its effect on abnormal immune activation and inflammation in Chinese rhesus monkey model infected by SIV for the first time.Methods:The toxicity and inhibitory effect of LLDT-8 and triptolide on NF-?B signal pathway were compared in vitro,and then the effects on T cell activation and proliferation of peripheral blood mononuclear cells(PBMCs)in healthy people after non-specific stimulation were further discussed.In the animal experiment,6 Chinese rhesus monkeys infected with SIV mac239 were used,and antiretroviral therapy(ART)was initiated at 14 weeks after infection.Among them,2 monkeys in the control group received ART treatment,and the other 4 monkeys in the experimental group were treated with ART and LLDT-8.ART treatment was terminated at 33 weeks after infection,and the experimental group continued to receive LLDT-8 until the end of the experiment(week 48).The differences of immune activation and inflammatory factors(CRP,IL-6,TNF-?,IP-10)were compared between the two groups during treatment.The rebound time of plasma viral load and the difference of SIV reservoir between two groups were also evaluated after interrupting ART treatment.Results:1.The cell survival rate was more than 60%when the concentration of LLDT-8 was 1000 ng/mL,which was similar to that of triptolide at the concentration of 100 ng/mL.Both LLDT-8 and Triptolide at 100 ng/mL could inhibit the activity of NF-?B reporter gene[(7.04±3.79)vs(0.29±0.07)],and there was no significant difference between them(P=0.12).2.Compared with the control group,LLDT-8 significantly reduced CD38+CD4+/CD4+%(56.53±5.14%vs 28.6±8.5%,P=0.06),HLA-DR+CD4+/CD4+%(18.55±1.38%vs 5.37±1.62%,P=0.00),PD1+CD4+/CD4+%(58.05±5.05%vs 27.85±2.5%,P=0.03).There was no significant difference between LLDT-8 and triptolide(P=0.58,0.63,0.32,respectively).After PHA stimulation,the proliferation of CD4+ T cells in PBMCs of healthy people reached 17.44±1.93%.After the addition of triptolide and LLDT-8,the proliferation of CD4+T cells decreased to 1.79±0.64%and 5.2±1.1%,respectively,and the effect of LLDT-8 was slightly weaker than that of triptolide(P=0.03).3.The activation level of T cells in Chinese rhesus monkeys after 14 weeks infected with SIV mac239 was significantly higher than that before infection(CD38+CD8+/CD8+%[83.1%(IQR:81.6-86.5)vs 20.8%(IQR:18.7-28.8)],HLA-DR+CD8+/CD8+%[15.6%(IQR:13.3-21.7)vs 0.3%(IQR:0.1-0.6)]).However,there were no significant changes in CRP,IL-6,IP-10 and TNF-? level compared with those before infection(P=0.47,0.99,0.53,0.97,respectively).One month after ART treatment,the expression of CD38+HLA-DR+double positive cells on CD4+T and CD8+T cells and CRP level in both groups were significantly higher than those in 14 weeks after infection.ART combined with LLDT-8 had no significant effect on T cell activation and inflammation.After the termination of ART,the viral load of rhesus monkeys in both groups rebounded at 2 weeks,and LLDT-8 had no effect on SIV reservoir.Conclusions:1.At the same dose,the cytotoxicity of LLDT-8 was lower than that of triptolide,and the inhibition of NF-?B signaling pathway was similar between them.2.LLDT-8 can inhibit the activation of CD4+T cells,there was no significant difference between the ability of LLDT-8 to inhibit the activation of CD4+ T cells and triptolide at the same dose;But the effect of inhibiting lymphocytes proliferation was weaker than that of triptolide.3.The level of T cell activation in Chinese rhesus monkeys infected with SIV mac239 was significantly higher than that before infection,whether or not LLDT-8 intervention had no effect on T cell activation and inflammatory response during treatment.4.After the termination of ART treatment,the plasma viral load rebounded within 2 weeks,and LLDT-8 had no significant effect on the time of virus rebound and SIV reservoir in rhesus monkeys. |
PDF Full Text Request