Immune No. 2 Party Intervention In Aids Patients Clinical And Mechanisms Of Immune Reconstitution

AIDS (acquired immune deficiency syndrome) is a lethal disease infected by the HIV (human immunodeficiency virus) and can caused serious harm to human health. Highly active antiretroviral therapy (HAART) is now recognized as a clinically effective treatment, which can inhibit the HIV replication, effectively reduce the plasma viral load, and to some extent, reconstitute the patients' immune function. However, this immune reconstitution is often incomplete, and there is big difference between different individuals. How to solve incomplete immune reconstitution after HAART, and find effective treatment and intervention is essential.Chinese medicine has a unique role in the regulation of human immune system. Promoting immune reconstitution by Chinese medicine has become the focus in field of AIDS research. In this study, we started by reviewing the literature and theories of AIDS and immune reconstituiton to identify research objectives and interventions, followed by clinical epidemiological study to know more about the application of HAART in China, and finally using the randomized, blinded, placebo-controlled RCT to confirm the efficacy of Mianyi No.2. At the same time, we examined the T-cell receptor gene rearrangement and TLR signaling pathways to find the molecular mechanism. The research is devided into2parts:PartⅠ Study on effect of immune reconstitution and mortality by HAART in ChinaObjectiveRelatively little is known regarding HIV-positive antiretoviral-naive patients among Chinese people, and how highly active antiretroviral therapy (HAART) influenced immunological outcomes and mortality of HIV positive patients in China We aimed to examine this by analysis of retrospective dataMethodsWe merged data on demographics, risk behavious, CD4cell counts and dates of death based on clinical records of2644HIV-positive antiretroviral-naive patients in Liuzhou district, Guangxi Province, China from2005to2009. We analyzed the trend of HIV prevalence, and calculated standardized mortality ratios (SMRs) standardized by age and sex. CD4counts at start and the end were also compared. Kaplan-Meier methods were used to assess survival over time.ResultsFrom2005to2009, most follow-up was fiom men (1740,65.81%), and the men age30-39years was the most, which was similar as in women, The incidence of young women (20-29years old) was higher than that of young men.1064patients (53%,1064/2004, not including deaths, lost of follow up) had increased magnitudes of CD4more than10percent of origin count, and262patients (13%,262/2004) had10percent decreased CD4count after treatment. The rest678patients (34%,678/2004) had incomplete information. Incomplete immune reconstitution (CD4count increased less than100/ul after one year treatment) were seen in188patients, accounting for17.42%. SMR was highest in patients age20-29at62.42,(95%Q41.44-83.40), and lowest in patients age>=60at1.55(95%CI0.90-2.20).169(6.39%,169/2644) patients died during follow-up, giving a mortality incidence rate of3.99per100person years.156/2241(7.0%) patients with CD4counts of200cells/μl or less died compared with11/314(3.5%) patients with CD4counts above200cells/μl and below350cell/μl, and2/77(2.6%) patients with CD4counts above350cells/μl (p=0.073).2/475(10.9%) patients infected by drug injection died compared with105/2015(5.2%) patients infected by sexual contact and12/154(7.8%) patients by other ways (p0.000).ConclusionHAART has reduced mortality among patients in China with AIDS, but incomplete immune reconstitution was still seen in188patients, accounting for17.42%. We need more effective means to promote the immune reconstitution in patients after HAART.PartⅡ Effect and related mechanism of Mianyi No.2on the Immune Reconstitution in Patients with AIDS after HAARTObjectiveTo observe the Mianyi No.2on the immune reconstitution in patients with AIDS after HAART, and to investigate the possible targets and mechanisms of Mianyi No.2.MethodsA randomized, double-blind, placebo-controlled clinical trial was designed.264patients failure to immune reconstitution after HAART were randomly divided into treatment group (131cases) and control group (133cases), respectively, using Mianyi No.2plus HAART and placebo combined with HAART for6months. CD4+, CD4CD45RA+, CD4CD45RO+cell numbers, as well as the symptoms, signs and integral improvement rates were observed in order to evaluate the immune reconstitution efficiency. At the same time changes in genetic diversity ofTCRVβ and TLR signaling pathways were detected. ResultsDuring the trial,264cases were enrolled and randomized, among which3cases drew out for adverse reactions,2cases for not taking medicine at all, and25cases quit by themselves. There were233cases that accomplished the clinical trial, of which116cases in the experiment group and117in the control group.Results are as follows:1Efficacy of immune function:after the intervention for1months, the effective rate of experiment group (18.97%) was significantly higher than that in control group (9.40%),(P=0.02);3months after treatment, the effective rate of experiment group (27.59%) is still higher than that of the control group (22.22%)(P=0.31);6months after treatment, the effective rate of experiment group (34.48%) was significantly superior to the control group (21.37%)(P=0.02). CD4+, CD4CD45RA+, CD4CD45RO+count of experiment group increased significantly higher than that of control group (P<0.05).2Efficacy of symptoms and signs:The accumulated points of symptoms and signs in treatment group had a better improvement effect compared with the control group (P=0.02), and the improvement of fatigue, muscle and joint pain, pruritus and shortness of breath in treatment group was better than control group (P<0.05).3Efficacy of viral load:There were no significant difference in viral load between or within groups before or after treatment (p>0.05).4Safety analysis:The incidence of adverse events occurred in some patients in both groups, the adverse event rate of experiment group was3.08%, control group6.25%(P=0.2263). There is no significant difference in changes of blood test, urine test, ECG, B-ultrasound, or chest X-ray.5Results ofTCRVP genetic diversity change indicate that:①Gaussian distribution ofTCR V(3families in patients with incomplete immune reconstitution after one year of HAART, had been broken with the occurrence of the offset TCR lineage. After six months of treatment of Traditional Chinese medicine combined HAART, the TCR lineage has been partially restored.②Evaluated by the D (distance) value calculated by a quantitative analysis software which the kit provides, there were no significant difference in D value change (p>0.05) between the two groups, but with traditional Chinese medicine can reduce the data variability.③CD4+T cell counts had a significant correlation (r=-0.772, P=0.000)with TCRV(3genetic diversity.④Compared with the normal group, there appeared some single or oligoclonal amplification of Vβ CDR3region in the patients, which were improved or recovered after treatment Among them, D value of four families (9,11,21,22) decreased after treatment in both groups. The decrease in family21and22was significant (P<0.05) in treatment group compared with the control group. And family18was decreased in treatment group and increased significantly in control group (P<0.05).6Changes on Toll like Receptor Signalling Pathway:There were5genes involved in the complete immune reconstruction significantly raised more than2times (such as interferon-γ, IL-6, IL-8, etc.), and4genes decreased (such as TLR9, NF-κB etc). Genetic changes in upstream were mainly down regulated (such as TLR9, TOLLIP, TRIF, IRAKI, TAB1, p105), while in downstream were mainly up regulated (such as AP-1, IL-6, IL-8). After treatment of immune No2, there were7related genes significantly raised more than2times (such as IL-2, IL-10, TLR1etc), and no genes significant decreased. Genetic changes in upstream were mainly up regulated (such as TLR1, CD14, TRAF6).Conclusion1Mianyi No.2can effectively improve die numbers of CD4+T cell and its subgroups, as well as the main clinical symptoms and signs of patients after HAART, such as fatigue, muscle and joint pain, skin itching, shortness ofbreath, therefore promoting immune reconstitution.2Mianyi No.2is safe to treat AIDS disease, with no significant adverse reactions.3Study of the mechanism showed oligoclonal of TCRVp family can get recovery in some degrees after treated by Mianyi No.2plus HAART, suggesting that the medicine may promote T-cell receptor gene rearrangement, helping immune cells to effectively identify the virus to reduce T-cell apoptosis; TLR9were expressed lowly in AIDS patients with incomplete immune reconstitution that in healthy person, and also IRAK1,NF-κB, suggesting that abnormal signal pathway mediated by TLR9maybe one of the mechanisms of incomplete immune reconstitution of AIDS. The effect of Immune No.2may be realized by up regulating TLR1and CD14expression, resulting changes of signal molecules in downstream.