Effect Of PPD On HIV Replication And Relationship To Cytokines

PrefaceAIDS (Acquired Immunodeficiency Syndrome) , which is caused by HIV( Human Immunodeficiency Virus) , is one of the extremely life - threatening infectious diseases. HIV invades immune system, so AIDS patients almost died of opportunistic infection. Tuberculosis ( TB) is the most common opportunistic infection in HIV infected persons and AIDS patients. Worldwide, nearly 30% of AIDS patients died of TB.Purified protein derivative (PPD) of tuberculin could induce secretion of TNF -α and IL - 10, which was same but less than My co-bacterium tuberculosis ( MTB). Delia Golett et al and C. J. C. Dehaas et al reported PPD and MTB increased replication of HIV in peripheral blood mononuclear cells ( PBMC ) from PPD (+) healthy donors. Tumor necrosis factor ( TNF) -α increased replication of HIV in monocyte - macrophage and T cell line in vitro. IL - 10 inhibited replication of HIV in monocyte - macrophage and PBMC in vitro. Because of difference in immune situation, it was necessary to study the effect of tuberculosis on HIV replication in Chinese persons. Now, the effect of PPD on secretion of cytokines in HIV - infected PBMC and the correlation between MTB - induced HIV replication and secretion of cytokines were not clear. The related foreign reports were rare, nodomestic reports.Our research incubated PPD with normal PBMC or HTV - 1 infected PBMC from Chinese persons, then measured the concentration of p24, IL - 10 and TNF - a in supernatant fluid. We evaluated the effect of PPD on HIV replication and the production of cytokines in normal PBMC and HIV -1 infected PBMC in order to provide a new thought for the prevention and treatment of AIDS and TB.MaterialsPeripheral blood mononuclear cells ( PBMC) were extracted from fresh anticoagulated blood collected from four PPD ( -f ) healthy persons and three PPD ( - ) healthy persons. National AIDS Reference Laboratory, Ministry of Health, China kindly provided HTV -1 SF -33 virus strain. PPD was obtained from national institute for the control of pharmaceutical and biological products. HIV - 1 p24 antigen testing kit produced by Holland Organon Teknika Company. IL - 10 and TNF - a ELISA kit distributed by Shenzhen Jingmei Company, produced by American Genzyme Company. RPMI 1640 produced by GIBCO BRL Company.Method1. Effect of PPD on replication of HIVPBMC from four PPD( + ) persons and three PPD( - ) persons were isolated by Ficoll -hypaque density gradient centrifugation, then PBMC were infected with HIV - 1 SF - 33 (125 TCID50/lml) and cultured in 48 - well plates(1 x 106 cells/ml) in the presence or ab-sence of 10 and 1 ug/ml PPD. Culture supernatant fluid was collected after 72 hours, and assessed HIV -1 p24 by a quantitative E1ISA kit.2. Effect of PPD on the secretion of cytokines in normal PBMC PBMC from PPD ( + ) healthy persons were cultured with orwithout PPD for 72 hours. Supernatant fluid was collected and assessed TNF - a and IL -10 by a quantitative ELISA kit.3. Effect of PPD on cytokines secretion in HIV -1 infected PBMCHIV -1 SF -33 infected PBMC from PPD ( + ) healthy persons were cultured with PPD for 72 hours. Supernatant fluid was collected and assessed TNF-α and IL - 10 and negative control was introduced simultaneously.4. Statistical analysis t -testResults1. Effect of PPD on HIV replication.PBMC from four PPD ( + ) healthy persons were infected with HIV - 1 SF - 33 (125 TCID50ml) and cultured for 72 hours in the presence of PPD (10ug/ml). The production of HIV -1 p24 was significantly increased compared with negative control (P <0. 05) ; PPD couldn't significantly increase HIV -1 p24 in PBMC from three PPD ( -) persons (P>0.05) (Figl).2. Effect of PPD on the secretion of cytokines in normal PBMC. PBMC from four PPD ( + ) persons were stimulated with PPD(10ug/ml) for 72 hours. PPD could promote PBMC of PPD ( + ) healthy persons to secrete TNF - a and IL -10(P < 0.05) ( Fig 2).3. Effect of PPD on cytokines secretion in HIV - 1 infected PB-MC.HIV...