| Objectuve Tuberculosis(TB)is the most common opportunistic disease in HIV-infected patients and the most common cause of death in AIDS patients.Similarly,HIV infection is one of the most important risk factors increasing the progression of latent TB infection to active TB infection.At present,the mechanism of HIV/Mtb co-infection has not been fully elucidated,while after the occurrence of HIV/Mtb co-infection,it involves two aspects of anti-tuberculosis and anti-HIV treatment,and complex factors such as autoimmune status,drug selection,adverse drug reactions,medication compliance,and drug-drug interactions may affect its therapeutic effect.Therefore,it is very necessary to carry out continuous research on HIV/Mtb co-infection.The aim of this study was to understand the disease process of HIV/Mtb co-infection during anti-TB treatment and long-term antiretroviral therapy,elucidate the influencing factors of immune reconstitution and survival in patients,and explore the possible immune mechanism of innate immune SOCS1/3 pathway in HIV promoting Mtb infection.Methods 1.Study on the disease progression of HIV/Mtb-coinfected patients: A follow-up study was conducted in this study.AIDS patients with pulmonary tuberculosis were enrolled in the Guangxi longtan hospital on October 1,2012 and December 31,2014.All study controls did not receive antiretroviral therapy(ART)or anti-tuberculosis treatment before enrollment.Information on demographic characteristics,laboratory data,ART status,anti-TB status,complications,and treatment outcomes of the study subjects were collected at different time periods.Kaplan-Meier method was used to analyze the cumulative immunological failure rate of the study subjects,life table method was used to analyze their cumulative survival rate,and COX regression model was performed to examine the factors influencing immunological failure and death.2.Study of the role of SOCS-TLRs signaling pathway in HIV-promoted Mtb infection: Blood samples were collected from HIV-infected patients,Mtb-infected patients,and HIV/Mtb-co-infected patients,so as to detect SOCS1/3 protein and Mtb infection-related cytokine expression downstream of the TLR2/4/9 pathway,and to explore the role and mechanism of SOCS1/3 protein in HIV promoting Mtb infection based on the macrophage-HIV-1-Mtb infection model.Results 1.Basic information and clinical examination data of study subjects(1)Basic information of study subjects: in the follow-up study,a total of 90 AIDS patients with pulmonary tuberculosis were included for follow-up and analysis.The baseline age of study subjects was between 20 and 64 years,with an average age of 41.32 ± 10.60 years;most of them were male(74.4%),Han Nationality(57.8%),and infected with HIV through sexual transmission route(94.4%).Most patients had baseline antiretroviral therapy of TDF + 3TC + EFV(62.2%),and 88.9% used first-line drugs for anti-tuberculosis treatment.(2)Auxiliary examination data of study subjects: chest radiography or chest CT showed that 83.9% patients had pulmonary lesions and 75.9% patients had lesions in both lungs.Etiological examination showed that 25.6% of patients were positive for sputum bacteria,16.7% were positive for acid-fast bacilli examination in sputum smears,15.6% were positive for mycobacterial culture in sputum samples,and 3.3% were positive for biopsy in samples.The study subjects had high anemia(69.5% at baseline),and the anemia rate slowly decreased as the duration of ART treatment advanced.The abnormal rates of ALT(19.5%)and AST(23.2%)at baseline were high,the abnormal rates of ALT and AST increased during anti-tuberculosis treatment,and the indicators decreased after the end of anti-tuberculosis treatment.(3)CD4 + T cell count levels of study subjects: most patients had baseline CD4 + T cell counts less than 200 cells/μL(63.3%),and CD4 + cell counts increased with the duration of ART treatment [β(95% CI): 1.82(0.64 – 3.01),P = 0.002].2.Analysis of immunological failure and its influencing factors: immunological failure occurred in 31.1% of patients,with an overall immunological failure rate of 6.42/100 man-year(95% CI: 4.11 – 8.75).The cumulative immunological failure rates at 3,6,12,18,24,30,36,42,48,60,and 84 months after antiviral therapy were 14.6%,22.7%,26.1%,28.5%,29.6%,29.6%,30.9%,30.9%,32.2%,32.2%,and 32.2%,respectively.Most immunological failures occurred within 1 year of receiving ART(82.1%).Treatment time of less than 2 months(HR=2.91,95%CI:1.34-6.30),and baseline CD4 + cell level > 350 cells/μL(HR=4.51,95%CI:1.53-13.27)during the intensive phase of anti-TB were risk factors for immunological failure.3.Survival analysis: A total of 435.5 person-years were followed up in this study,with a mean follow-up of 4.84 person-years,with a maximum follow-up of 7 years and a minimum follow-up of 1 month in this study.A total of 10 study subjects died of AIDS-related diseases during the study period,with a mortality rate of 11.1% and a death density of 2.30/100 man-year(95% CI: 0.91 – 3.68).In AIDS patients with pulmonary tuberculosis,the cumulative survival rate decreased from 93.22% to 88.27% within 0 ~ 84 months after starting treatment,and the probability of death within 1 year after starting treatment was the highest(6.78%).The risk of death was higher in patients with less than 2 months of intensive anti-TB treatment than in those with 2 to 9 months of intensive anti-TB treatment(HR=4.70,95% CI:1.31-16.89).Patients who self-discontinued during medication had a higher risk of death than those who adhered to medication(HR = 3.58,95% CI: 1.02 – 12.57).4.Study of the role of SOCS-TLRs signaling pathway in HIV-promoted Mtb infection(1)Expression levels of SOCS1/3 protein and Mtb infection-related cytokines downstream of TLR2/4/9 pathway: The results of flow cytometry showed that the relative protein expression levels of SOCS3,TLR9,IL-6 and IL-10 in peripheral blood PBMCs of HIV-1-infected group were higher than those of healthy control group(P < 0.05),and IFN-γ,IL-1β,and IL-12 in peripheral blood PBMCs of Mtb single-infected group were higher than those of healthy control group(P < 0.05)(2)Verification of the effect of HIV-1/Mtb infection on macrophage SOCS-TLR signaling pathway in vitro: In the macrophage-HIV-Mtb infection model,both HIV-1 infection and Mtb infection can up-regulate macrophage SOCS1 protein expression,which in turn inhibits macrophage TLR2 expression,up-regulates IL-6 and IL-10 expression,and down-regulates IFN-γ expression in the late stage of infection;HIV-1/Mtb co-infection will inhibit macrophage TLR2 expression and up-regulate IL-6 expression,and the colony count of the HIV/Mtb co-infection group is higher than that of the Mtb mono-infection group.(3)Effect of overexpression of SOCS1 protein on the expression of TLR pathway cytokines associated with Mtb infection: In the case of overexpression of SOCS1 protein,TLR2 agonist could not elevate the relative expression level of TLR2 m RNA in macrophages;on the contrary,TLR2 agonist could significantly inhibit the relative m RNA expression level of SOCS1 in macrophages overexpressing SOCS1 protein,which further verifying the direct negative feedback regulation of SOCS1 and TLR2.IL-6 m RNA expression was higher in the HIV-1 infected group and HIV-1/Mtb co-infected group with overexpression of SOCS1 protein than in the control group,this upregulation was significantly inhibited in the HIV-1/Mtb co-infected group after the addition of TLR2 agonist.It was suggested that both HIV-1infection and HIV-1/Mtb co-infection and activation of TLR2 pathway could upregulate SOCS1 expression in macrophages,and the high expression of SOCS1 negatively regulated TLR2 pathway,leading to the increase of IL-6 expression of downstream TB infection-related factors.(4)Effect of overexpression of SOCS1 protein on the infectivity of H37Rv: Lysed cells were harvested and CFU experiments were carried out on days 1,2 and 3 after the establishment of co-infection model.The results showed that the Mtb colony counts in the HIV-1/Mtb co-infected group were higher than those in the Mtb infection group at all different infection times.The Mtb colony counts in the overexpression SOCS1 protein group were higher than those in the non-overexpression SOCS1 protein group.The results suggested that the high expression of SOCS1 ultimately promoted the infective ability of Mycobacterium tuberculosis.Conclusion 1.AIDS patients with pulmonary tuberculosis have a high immunological failure rate and mortality rate.Too short treatment time in anti-tuberculosis intensive phase and medication compliance are the main risk factors of immunological failure and death.It is necessary to strengthen the management of HIV/Mtb co-infection and standardize medication;at the same time,attention should be paid to nutritional supplementation and liver function protection of patients during treatment.2.HIV-1 infection may inhibit the cellular innate immune TLR2 signaling pathway by up-regulating the expression of SOCS1 in macrophages,which results in up-regulation of downstream IL-6 and IL-10 expression,and in turn promotes Mtb infection.The results will help to better understand the natural process of HIV-1 promoting Mtb infection when cellular immunity is not yet significantly defective at the early stage of infection as well as provide new ideas for innate immune-based prevention and treatment of HIV-1/Mtb dual infection. |
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