Depressed CCL5 Expression in Human Pulmonary Tuberculosis

CCL5/regulated on activation, normal T expressed and secreted production (RANTES) is a principal CC chemokine, and can activate macrophages and Th1 lymphocytes, however, little is known about the CCL5 profiles associated with active tuberculosis (TB). In this study, we investigated the production of CCL5 by the peripheral blood mononuclear cells (PBMCs) of patients with active pulmonary TB after stimulation with Triton X-100 soluble proteins (TSP) or the 30-kDa antigen. The profiles of cytokines/chemokines [CXCL8/interleukin (IL)-8, IL-12 p40, and interferon (IFN)-gamma] were also examined by PBMCs from TB patients, and compared with those obtained from healthy tuberculin reactors (HTR). Concordant with earlier studies, IFN-gamma production was significantly depressed in the PBMCs from TB patients compared with those from HTR. In addition, the CCL5, but not CXCL8, levels in the PBMCs from TB patients were significantly depressed after stimulation for 18 hr compared to those in the PBMCs from HTRs. The CCL5 release was not significantly correlated with the release of IFN-gamma in the cells from TB patients and HTRs. Further, inhibitor studies show that the 30-kDa- or TSP-induced CCL5 mRNA expression is sensitive to inhibitors of mitogen-activated protein kinase kinase (MEK) 1/2 and Janus kinase (JAK) 2, but not p38, pathway activation, suggesting a MEK1/2- or JAK2-based mechanism is responsible for modulating of the CCL5 expression in human PBMCs. Collectively, these data suggest that TB patients show depressed production of CCL5 secretion, which can be modulated by MEK- and JAK2-based transcriptional regulatory mechanisms, in response to the mycobacterial antigens.

Identification and Functional analysis of Gene Expression in Mycobacterium tuberculosis-infected Human Monocytic Cells Under Hypoxic Conditions

Mycobacterium tuberculosis-induced granulomatous lesions, particularly those undergoing central caseation, are known as hypoxic. To analyze the host genes associated with hypoxic conditions from cells infected with M. tuberculosis, we performed GeneChip analyses on mRNA from M. tuberculosis H37Rv-treated human monocytic THP-1 cells cultured in oxygen-depleted status for 18 h. The expression of 99 genes was altered, including those involved in intracellular signaling, energy production, and protein metabolism, as revealed by stringent microarray data analysis. Most notably, mRNA expression of chemokine macrophage inflammatory protein 3alpha/CC chemokine ligand 20 (CCL20) was significantly up-regulated in M. tuberculosis-infected cells under hypoxic conditions. We further analyzed the CCL20 expression in peripheral blood mononuclear cells (PBMCs) and monocyte derived macrophages (MDMs) from healthy controls and TB patients. A comparative analysis has revealed that the mRNA and protein expression of CCL20 were prominently up-regulated in PBMCs, and MDMs from TB patients, compared with healthy controls. Collectively, these data show that the gene expression of CCL20 was up-regulated in M. tuberculosis H37Rv-infected human monocytic THP-1 cells cultured in hypoxic conditions. In addition, the production of CCL20 is substantially increased in cells from TB patients than in healthy controls, suggesting an important role of CCL20 in the immunopathogenesis during TB infection.

Differential Roles of Toll-like Receptor (TLR) 2 and 4 between PPD- and 38-kDa-induced Proinflammatory Cytokine Productions in Human Monocytes

In this study, we investigated the role of toll-like receptor (TLR) and mitogen-activated protein kinase (MAPK) pathways involved in the tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 expression after stimulation with purified protein derivatives (PPD) or native 38-kDa protein antigen (Ag) of Mycobacterium tuberculosis H37Rv in human primary monocytes. Both PPD and 38-kDa Ag significantly induced TNF-alpha and IL-6 in human primary monocytes. MAPK [extracellular signal-regulated kinase (ERK) 1/2 and p38] are rapidly phosphorylated in human monocytes stimulated with the PPD or 38-kDa Ag. Both p38 and ERK 1/2 activation are essential for PPD- or 38-kDa-induced TNF-alpha and IL-6 production. The inhibition of TLR2 and TLR4 by specific antibodies significantly abrogated the 38-kDa-induced secretion of TNF-alpha and IL-6, whereas blockade of TLR2, but not TLR4, was responsible for the PPD-induced TNF-alpha and IL-6 production in human monocytes. Collectively, these data suggest that the PPD and 38-kDa Ag differentially interact with TLR2 and TLR4, which in turn mediate an essential role for the early inflammatory immune responses during human tuberculosis.

Production of TNF-alpha and IL-6 in Macrophages by Mycobacterial Protein Antigens

BACKGROUND: Mycobacterial antigens released as PIM, LM, LAM, lipoproteins and other cellular factors may contribute to macrophage and dendritic cell activation through pattern recognition receptors such as TLRs. In this study, we assessed cytokine production and ERK activation with stimulation of several major mycobacterial antigens. METHODS: Purified mycobacterial antigens (10, 22, 30, 38kappaDa) and recombinant antigens (6, 16, 19, 38kappaDa, Ag85A antigen) were studied. The production of cytokines (TNF-alpha, IL-12, IL-6) was measured by ELISA. The ERK activation was detected by western blotting. The expression of TLR2 or TLR4 was measured by flow cytometry. RESULTS: Among purified antigens only 30kappaDa antigen induced production of IL-6 or TNF-alpha in THP-1 macrophage cells. When THP-1 macrophage cells were treated with 30kappaDa antigen, phosphorylation of ERK was detected. ERK activation also occurred in TLR2 transfectant HEK293 cells with 30kappaDa antigen stimulation. CONCLUSION: 30kappaDa antigen is one of the major mycobacterial antigens inducing cytokine production and MAP kinases phosphorylation in macrophages.

Elevated Levels of Interferon-inducible Protein-10 (IP)-10/CXCL10, but not of Interferon-gamma, in Patients with Pulmonary Tuberculosis

Mycobacterial strains are potent inducers of cytokines/chemokines by mononuclear phagocytes, which constitute an important cellular component of the first line of defense in the innate immune system. Interferon (IFN)-gamma-inducible protein (IP-10 or CXCL10) is a potent chemoattractant; however, little is known about the IP-10 profiles attributable to the Th1 regulation associated with active tuberculosis (TB). In this study, we investigated the production of IP-10, interleukin (IL)-12 p40, and IFN-gamma by the peripheral blood mononuclear cells (PBMCs) of patients with active pulmonary TB in response to in vitro stimulation with Triton X-100 soluble proteins (TSPs) or the 30-kDa antigen. The TSP antigens used in the present study were isolated and purified from Mycobacterium tuberculosis H37Rv (virulent strain), M. tuberculosis H37Ra (avirulent strain), and Mycobacterium bovis BCG. The results were compared with those obtained for healthy tuberculin reactors (HTRs). Concordant with earlier studies, IFN-gamma production was significantly depressed in the PBMCs from TB patients compared with those in the HTR group. However, the IP-10 levels in the PBMCs from TB patients were significantly elevated 18 h after stimulation compared to those in the PBMCs from HTRs. IP-10 release was correlated in a significant manner with the release of IFN-gamma in the HTRs, but this was not the case for the TB patients. Collectively, these data suggest that TB patients show altered regulation of Th1-driving cytokine and chemokine production in response to a variety of mycobacterial antigens.

Purification of Native Ag85 Complex, 38-kDa and MTB12 Protein Antigens from the Culture Filtrate of Mycobacterium tuberculosis

The purification of immunodominant native protein antigens from the culture filtrates of Mycobacterium tuberculosis is needed for the development of new vaccines and immunodiagnostic reagents against tuberculosis. In the present study, we conducted large scale purification of well-known secreted antigens, Ag85 complex, 38-kDa, and MTB12, from the culture filtrate proteins (CFPs) prepared from M. tuberculosis H37Rv grown as a surface pellicle on synthetic Sauton medium. The protein and antigen concentrations of culture filtrates were sufficiently increased after 6 week of culture. The MTB12 antigen was detected as early as 1 week of culture, and Ag85 complex and 38-kDa antigen were detected after 2 and 3 week of culture, respectively, by immunodiffusion with specific antiserum against 100-fold concentrated culture filtrates. For large-scale purification, the six-week-culture filtrates of M. tuberculosis H37Rv diluted 2.5-fold with 20 mM Tris-HCl, (P)H 8.3 were subjected to anion-exchange chromatography. The CFPs were eluted with 100 mM NaCl-20 mM Tris-HCl, pH 8.3 and concentrated by ultrafiltration. The concentrated CFPs were fractionated with ammonium sulfate, and followed by hydrophobic interaction chromatography and anion-exchange chromatography (FPLC). Eventually, 10 mg of Ag85 complex, 0.56 mg of 38-kDa, and 1.81 mg of MTB12 antigens were purified from 1 liter of the six-week-culture filtrates of M. tuberculosis H37Rv which contained 307.81 mg of protein of culture filtrate.

The Phospholipase-Protein Kinase C-MEK-ERK Pathway is Essential in Mycobacteria-induced CCL3 and CCL4 Expression in Human Monocytes

BACKGROUND: Little information is available on the identification and characterization of the upstream regulators of the signal transduction cascades for Mycobacterium tuberculosis (M. tbc)-induced ERK 1/2 activation and chemokine expression. We investigated the signaling mechanisms involved in expression of CCL3/MIP-1 and CCL4/MIP-1 in human primary monocytes infected with M. tbc. METHODS: MAP kinase phosphorylation was determined using western blot analysis with specific primary antibodies (ERK 1/2, and phospho-ERK1/2), and the upstream signaling pathways were further investigated using specific inhibitors. RESULTS: An avirulent strain, M. tbc H37Ra, induced greater and more sustained ERK 1/2 phosphorylation, and higher CCL3 and CCL4 production, than did M. tbc H37Rv. Specific inhibitors for mitogen-activated protein kinase (MAPK) kinase (MEK; U0126 and PD98059) significantly inhibited the expression of CCL3 and CCL4 in human monocytes. Mycobacteria-mediated expression of CCL3 and CCL4 was not inhibited by the Ras inhibitor manumycin A or the Raf-1 inhibitor GW 5074. On the other hand, phospholipase C (PLC) inhibitor (U73122) and protein kinase C (PKC)- specific inhibitors (GO6976 and Ro31-8220) significantly reduced M. tbc-induced activation of ERK 1/2 and chemokine synthesis. CONCLUSION: These results are the first to demonstrate that the PLC-PKC-MEK-ERK, not the Ras-Raf-MEK-ERK, pathway is the major signaling pathway inducing M. tbc-mediated CCL3 and CCL4 expression in human primary monocytes.

Induction of TNF-alpha, IL-12, IL-10, MCP-1 and IL-8 in Human Peripheral Blood Monocytes in Response to the Cell Wall-Associated Triton X-100 Soluble Protein (TSP) Antigens of Mycobacterium tuberculosis

Mycobacterium tuberculosis is a potent inducer of cytokine production by mononuclear phagocytes, which are an important cellular component in the first line immune defence. In this study, the cell wall-associated Triton X-100 soluble protein (TSP) antigens, TSP-H37Rv, TSP-H37Ra, TSP-K, and TSP-BCG, were isolated from M. tuberculosis H37Rv, M. tuberculosis H37Ra, M. tuberculosis K-strain, and M. bovis BCG, respectively. The monocytes were isolated from the peripheral blood mononuclear cells of healthy individuals and were co-cultured with each TSP antigens and the secretory proteins of M. tuberculosis (PPD and 30-kDa antigen) to measure the production of cytokines; tumor necrosis factor (TNF)-a, interleukin (IL)-12, IL-8 and monocyte chemotactic protein-1 (MCP-1). The TSP-H37Rv antigen- stimulated monocytes showed higher level of TNF-a and IL-12 production compared to those of other TSP antigens and PPD. Especially, IL-12 production in response to the TSP-H37Rv antigen was significantly elevated in comparison with that of PPD-stimulated monocytes (TSP-H37Rv, 255.5+/-256.9 pg/ml; PPD, 55.7+/-55.4 pg/ml). However, the 30-kDa antigen did not induce TNF-alpha expression and also showed the lowest level of cytokine and chemokine production by monocytes. MCP-1 and IL-8 production were similarly increased in response to all TSP antigens and the PPD antigen. The production of IL-12 by the TSP-H37Rv antigen stimulation was significantly increased in PPD reactors than that in the non-reactor group, while the levels of other cytokines stimulated with each TSP antigens, 30-kDa and PPD antigen were not significantly different between the tuberculin reactor and the non-reactor groups. These results suggest that the cell wall-associated TSP antigen isolated from M. tuberculosis H37Rv acts as a more potent IL-12 inducer than the PPD antigen in innate immune response and thus it could further activate the Th1-mediated immune responses effectively against M. tuberculosis infection.

Mycobacteria-induced Interleukin-12 Expression by Human Monocyte-derived Macrophages Is Negatively Regulated by Phosphatidylinositol 3-kinase and ERK 1/2 Pathways

Both interleukin (IL)-12, an important cytokine skewing the immune response towards a Th1 cytokine profiles, and tumor necrosis factor (TNF)-alpha, are thought to be critical factors in defenses against mycobacteria. In this study, we evaluated the roles of phosphatidylinositol 3-kinase (PI 3-K), and extracellular signal-regulated kinase (ERK) 1/2 pathways in the expression of IL-12 in human monocyte-derived macrophages (MDMs) after stimulation with Mycobacterium tuberculosis H37Rv (M. tbc) or the Triton X-114 solublized proteins (TSP) of M. tbc. Both M. tbc and TSP rapidly phosphorylated ERK 1/2, and Akt in human MDMs. Inhibition of PI 3-K-Akt pathway by specific inhibitors (LY294002 and wortmannin) dramatically increased M. tbc- or TSP-induced IL-12 p40 and p35 mRNA and IL-12 production. In addition, blockade of ERK 1/2 pathway by specific inhibitors (PD98059 and U0126) significantly increased the mRNA levels and cytokine production in M. tbc- or TSP-treated MDMs. On the contrary, M. tbc- or TSP-induced TNF-a production was significantly depressed in human MDMs by pretreatment with inhibitors of PI 3-K or ERK pathways. The M. tbc or TSP stimulation decreased ERK 1/2 phosphorylation by 70% in the presence of wortmannin or LY294002, suggesting that some cross-talk between the PI 3-K-Akt and mitogen-activated protein kinase kinase (MEK)-ERK pathways may be operating in human monocytes during mycobacterial infection. PI 3-K activity is partially required for the M. tbc- or TSP-induced ERK 1/2 phosphorylation. Collectively, these data suggest that the PI 3-K and ERK 1/2 pathways play a central role in the negative regulation of IL-12, but not TNF-a, production by M. tbc.

Isolation of a Specific Antigen from the Triton X-100 Soluble Protein (TSP) Of Mycobacterium tuberculosis

Tremendous efforts have been made to develop better vaccines and diagnostic markers for the effective control of tuberculosis. Recently, we reported that the Triton X-100 soluble protein (TSP) of Mycobacterium tuberculosis induced strong T-cell proliferation and IFN-gamma production in humans, and also conferred a significant level of protection against tuberculosis in a mouse model. In this study, the TSP was prepared by Triton X-100 extraction of Mycobacterium tuberculosis bacilli, which was followed by Triton X-114 phase partitioning. Western blot analysis using sera of 177 active pulmonary tuberculosis patients and 323 healthy individuals revealed that the TSP contained a immunodominant 40-kDa antigen specifically reacting with some sera from pulmonary tuberculosis patients. The 40-kDa antigen was purified by ion-exchange chromatography, and partially characterized by two-dimensional gel electrophoresis and N-terminal sequencing. Results of this study suggest that 40-kDa molecule of the TSP antigen from the cell suface of Mycobacterium tuberculosis can be used as a serodiagnostic marker as well as a potential vaccine candidate against tuberculosis.

Role of Mitogen-Activated Protein Kinase Pathways in the Production of TNF-alpha, IL-10, and MCP-1 by Mycobacterium tuberculosis H37Rv-Infected Human Blood Monocytes

Clinical manifestations of tuberculosis are closely associated with the initial responses of macrophages to mycobacteria. In this study, we investigated the signal transduction pathways for the secretion of cytokines and chemokines [tumor necrosis factor (TNF)-alpha, interleukin (IL)-10, IL-8, and monocyte chemotactic protein-1 (MCP-1)] in human blood monocytes infected with Mycobacterium tuberculosis H37Rv. M. tuberculosis H37Rv infection induced the secretion of significant amounts of TNF-alpha, IL-10, IL-8, and MCP-1 from human blood monocytes. Analysis of mitogen-activated protein kinase (MAPK) activation [extracellular signal-regulated kinase 1/2 (ERK) and p38 kinase] showed rapid phosphorylation of both subfamilies in response to M. tuberculosis H37Rv. Using highly specific inhibitors of p38 (SB203580) and of MEK-1 (U0126 and PD98059), we found that both p38 and ERK were essential for M. tuberculosis H37Rv-induced TNF-alpha production, whereas activation of the p38 pathway, but not that of ERK, was essential for M. tuberculosis H37Rv-induced IL-10 production. Interestingly, the ERK pathway, but not that of p38, was critical for MCP-1 secretion from human blood monocytes infected with M. tuberculosis H37Rv. However, IL-8 secretion was regulated neither by ERK1/2 nor p38 MAPK. Collectively, these results suggest that induction of the MAPK pathway is required for the expression of TNF-alpha. IL-10, and MCP-1 by human blood monocytes during M. tuberculosis H37Rv infection.

Dysregulated Production of IFN-g, IL-12, and IL-10 by Peripheral Blood Mononuclear Cells from Early Active Pulmonary and Multidrug-Resistant Tuberculosis Patients

In this study, we investigated profiles of the cytokines IFN-g, IL-12, and IL-10 in active pulmonary tuberculosis (EAPTB) patients, HIV-negative patients with multidrug-resistant tuberculosis (MDR-TB) and in healthy tuberculin reactors (HTR). We studied the responses of peripheral blood mononuclear cells (PBMC) from 12 EAPTB patients and 15 MDR-TB patients to stimulation with a purified protein derivatives (PPD) antigen (Ag), and compared them with those from 14 HTR. Using ELISA, IFN-g production was found to be significantly depressed, while IL-10 was significantly elevated in both MDR-TB and EAPTB after in vitro stimulation with PPD, compared with those in HTR. Although there was no significant difference in IL-12 production among the three groups, mean IL-12 production was highest in patients with MDR-TB. In these patients, IL-12 production was significantly correlated with IL-10 expression, but not IFN-g production. In addition, neutralization of endogenous IL-10 led to enhanced IFN-g and IL-12Rb2 mRNA expression in TB patients. Our findings suggest that both groups of TB patients may have a similar disregulated pattern of IL-12, IL-10, and IFN-g production during M. tuberculosis infection. Furthermore, the results suggest a potentially pathogenic role for IL-10 in impaired Th1 immune responses in TB patients.

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.

Depression of Both Interferon-gamma and Tumor Necrosis Factor-alpha Production by Peripheral Blood Mononuclear Cells from Chronic Refractory Tuberculosis Patients

Understanding human immune responses in chronic refractory tuberculosis (CRTB) is important for developing immunotherapy against the disease. The aim of this study was to examine cytokine responses [interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-10] by peripheral blood mononuclear cells (PBMCs) in CRTB patients after in vitro stimulation with the 30-kDa or purified protein derivative (PPD) antigen (Ag). Most of the CRTB cases were multidrug-resistant (MDR) TB. The results were compared with those from early TB (E-TB) patients and healthy tuberculin reactors (HTR). IFN-gamma production was significantly depressed in both CRTB and E-TB groups compared with HTR. In response to the 30-kDa Ag, TNF-alpha levels were significantly depressed only in CRTB patients, while greatly increased in E-TB patients. In addition, IL-10 production was significantly increased in E-TB patients, and PBMC from both E-TB and CRTB patients secreted more IL-6 than HTR. IL-10 neutralization significantly increased TNF-alpha levels, whereas anti-TNF-alpha did not alter IL-10 induction significantly in PBMC from HTR and CRTB patients. Our findings suggest that CRTB patients have depression in both IFN-gamma and TNF-alpha reponses, which might play important roles during chronic M. tuberculosis infection.

Identification and Functional Characterization of Differentially Expressed Genes in Human-derived Monocytic Cell Line U937 Infected with Mycobacterium tuberculosis H37Rv and Mycobacterium marinum: Comparative Evaluation of IL-8

Although Mycobacterium marinum is closely related to M. tuberculosis H37Rv (M. tbc) genomically, clinical outcome of human infection is quite different. The role of the host macrophage in determining differential pathologic responses was analyzed using an in vitro model of macrophage infection. By using subtractive hybridization, thirty-two differentially expressed genes were identified in the monocytic cell line U937 infected with M. tbc or M. marinum. Among them, IL-8 mRNA expression was more prominent in the M. tbc-infected U937 cells by Northern hybridization than in those infected with M. marinum. The IL-8 production was significantly lower in M. marinum-infected U937 or monocytes when compared with those infected by other strains of mycobacteria, such as M. tuberculosis H37Ra, M. bovis BCG or M. smegmatis. To identify possible mechanisms underlying these differences, changes in the expression of molecules such as nuclear factor-kappaB (NF-kappaB) involved in the signaling pathway activated by mycobacteria were assessed. U937 cells infected with M. tbc showed a significant degradation of IkBa proteins compared with M. marinum-infected U937 cells. Collectively, these results implicate distinct differences in IL-8 production in human macrophages infected with M. tbc or M. marinum, and suggest important role of IL-8 in the immunopathogenesis of tuberculosis.

IL-12 and TNF-alpha productions from human peripheral blood mononuclear cells in untreated patients with active pulmonary tuberculosis stimulated with 30-kDa or TSP antigen of Mycobacterium tuberculosis H37Rv

No abstract available.

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.

Increased IL-12 and Interferon-Hamma, But Not IL-18 Production, After In Vitro Stimulation with a 30-kDa Mycobacterial Antigen in Patients with Tuberculous Pleurisy

No Abstract Available.

Follow-up Study on 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 protein antigen from Treponema pallidum and conducted on 12 patients with syphilis (early, late, spontaneously healed, congenital and treated patients) followed by therapy. Peripheral blood mononuclear cells (PBMC) were obtained three times from each patient, on admission before the initiation of therapy, 1 and 6 months later. Eleven (96.7%) of the patients prior to therapy, showed depressed lymphoproliferative responses to the 47 kDa antigen (stimulation index <4) by 3H-thymidine incorporation assay. However, these T cell responses were seemed to be transient because most of the patients (63.6%) exhibited significantly higher lymphoproliferation after therapy. Before therapy, PBMC from spontaneously healed syphilis patients resulted in significantly increased gene expression of IFN- and proinflarnmatory cytokines, such as TNFa, IL-1B and IL-6, in response to the 47 kDa. Patients with late latent and late congenital syphilis exhibited lower IFN-r and proinflammatory cytokine mRNA expression than spontaneously healed syphilis group did. After therapy, IFN-r and proinflammatory cytokine mRNA expressions were gradually reduced in these groups. On the other hand, IFN- and proinflammatory cytokine gene expressions were considerably depressed in early syphilis patients, but these patients went on to express prominent IFN-r and proinflamrnatory cytokine mRNA with treatment. These data suggest that the pattern of cellular immune response in response to the 47 kDa antigen may be involved in the evaluation of the clinical course and outcome of syphilis followed by therapy.