CD40-CD40 Ligand Interactions in the Production of IL-12 and IFN-gamma by Tuberculous Pleural Mononuclear Cells

BACKGROUND: Our previous study showed that purified protein derivative (PPD)- stimulated pleural mononuclear cells (PMC) from tuberculous pleurisy (Tbp) produced significantly more IFN-gamma (10- to 70-fold) after in vitro PPD stimulation than freshly isolated pleural cells from malignant pleurisy. The present study was designed to determine whether blocking the CD40-CD40 ligand (CD40L) interaction decreases IFN-gamma production by altering IL-12 levels. METHODS: IL-12 and IFN-gamma production after neutralizing anti-CD40L antibody treatment was compared to the efficacy of anti-CD80, anti-CD86, and a combination of anti-CD80 and CD86 (CD80+86) monoclonal antibodies (mAb). These activities were measured by enzyme-linked immunosorbent assays (ELISAs) and reverse transcription-polymerase chain reaction (RT-PCR), after in vitro stimulation with PPD antigen (Ag). RESULTS: Neutralization of CD80, CD86 and CD80+86 did not decrease IFN-gamma and IL-12 production in Tbp-PMC, whereas neutralization of CD40L significantly depressed IL-12 p40 and IFN-gamma. In addition, neutralization of CD40L completely inhibited IL-12 p40 and IFN-gamma mRNA expression. CONCLUSION: The CD40-CD40L interaction might play a major role in IL-12 and IFN-gamma production in Tbp-PMC, thus contributing to protective immunity in human tuberculosis.

CD40-CD40 Ligand Interactions in the Production of IL-12 and IFN-gamma by Tuberculous Pleural Mononuclear Cells

BACKGROUND: Our previous study showed that purified protein derivative (PPD)- stimulated pleural mononuclear cells (PMC) from tuberculous pleurisy (Tbp) produced significantly more IFN-gamma (10- to 70-fold) after in vitro PPD stimulation than freshly isolated pleural cells from malignant pleurisy. The present study was designed to determine whether blocking the CD40-CD40 ligand (CD40L) interaction decreases IFN-gamma production by altering IL-12 levels. METHODS: IL-12 and IFN-gamma production after neutralizing anti-CD40L antibody treatment was compared to the efficacy of anti-CD80, anti-CD86, and a combination of anti-CD80 and CD86 (CD80+86) monoclonal antibodies (mAb). These activities were measured by enzyme-linked immunosorbent assays (ELISAs) and reverse transcription-polymerase chain reaction (RT-PCR), after in vitro stimulation with PPD antigen (Ag). RESULTS: Neutralization of CD80, CD86 and CD80+86 did not decrease IFN-gamma and IL-12 production in Tbp-PMC, whereas neutralization of CD40L significantly depressed IL-12 p40 and IFN-gamma. In addition, neutralization of CD40L completely inhibited IL-12 p40 and IFN-gamma mRNA expression. CONCLUSION: The CD40-CD40L interaction might play a major role in IL-12 and IFN-gamma production in Tbp-PMC, thus contributing to protective immunity in human tuberculosis.

Increased IL-12 , but Depressed IL-18 Production after In Vitro Stimulation with a 30-kDa Mycobacterial Antigen in Tuberculous Pleural Mononuclear Cells

In this study, we investigated interleukin (IL)-18 and IL-12 following in vitro stimulation with either the 30-kDa or purified protein derivative (PPD) antigens (Ag) of pleural mononuclear cells from 12 cases of tubercular pleurisy (TB-PMC) and 8 cases of malignant pleurisy (MG-PMC). Ag-stimulated TB-PMC produced significantly more IL-12 than did MG-PMC and the levels correlated with those of IFN - gamma. Although elevated IL-18 levels were found in freshly isolated pleural fluids, in vitro IL-18 production in response to either Ag was dramatically decreased in TB-PMC. Pro-IL-18 mRNA was detected before and after Ag stimulation in TB patients. Supernatants from the Ag-stimulated TB-PMC significantly suppressed IL-18 production in normal peripheral blood mononuclear cells (PBMC) and primary malignant cells over an 18 h incubation period. In addition, this suppressive activity was not inactivated by either heat or trypsin. Our findings imply that modulation of IL-12 and IL-18 levels may contribute to the Th1 elevation induced in human TB-P VIC by the 30-kDa and PPD antigens.

Cell-mediated Immune Responses in Syphilitic Patients after In vitro Stimulation with the 47 kDa Antigen of Treponema pallidum / 대한면역학회지

Present study was aimed to investigate the immunological activities of the 47 kDa glycoprotein antigen from Treponema pallidum and conducted on 24 patients with syphilis, (early, late, spontaneously healed, congenital and treated patients) and on 17 normal healthy controls. Two opposite lymphoproliferative manifestations to the 47 kDa antigen were observed in syphilis patients by H-thymidine incorporation assay. Ten responders (stimulation index [Sl] >4) showed a 3-fold-higher proliferation than the nonresponders, and four of those responders were spontaneously healed patients. Furthermore, analysis by flow cytometry indicated a preferential expansion of CD4' T lymphocytes by the 47 kDa antigen in the spontaneously healed syphilis patients. Stimulation of PBMCs of spontaneously healed syphilis patients with the 47 kDa antigen for greater than 72 hrs resulted in piogressive augmentation of IFN-r, IL-2Ra and IL-2 mRNA measured by RT-PCR, but considerably reduced IL-4 and IL-10 mRNA expression. However, patients with late latent syphilis exhibited more increased IL-4 and IL-10 mRNA expressions in response to the 47 kDa antigen than spontaneously healed syphilis group. In contrast to other groups, when cultured with the 47 kDa antigen very low IFN-#y, IL-2Ra and IL-2 mRNA expressions were shown in early syphilis group. These data suggest that the Th1-predominant cellular responses induced by the 47 kDa antigen may be involved in the clinical outcome of syphilis and provide the immunologic basis for further functional studies regarding the role of the 47 kDa in the immunopathogenesis of syphilis.