sq-23377 and Tuberculosis--Pulmonary

The roles of multifunctional CD4 T cells in human tuberculosis are not well defined. In this study, we found that patients with tuberculosis had decreased PMA/ionomycin stimulated multifunctional CD4 T cells, and increased Mycobacterium tuberculosis antigen-specific multifunctional CD4 T cells, when compared to individuals with latent tuberculosis infection and healthy controls. PMA/ionomycin stimulated IFN-γ+IL-2+TNF-α+ CD4 T cell responses were decreased in patients with smear-positive tuberculosis compared to those with smear-negative tuberculosis. The percentage of IFN-γ+IL-2+TNF-α+ CD4 T cells in smear positive tuberculosis patients negatively correlated with the grade of sputum smear Acid-Fast Bacilli and high-resolution computed tomography score. Therefore, our findings argue against the notion that Mycobacterium tuberculosis antigen-specific multifunctional Th1 responses in peripheral blood can serve as correlates of protective immunity against tuberculosis; they suggest that the decrease in PMA/ionomycin stimulated IFN-γ+IL-2+TNF-α+ CD4 T cells may be applied for clinical diagnosis of active tuberculosis.

Topics: Adult; Case-Control Studies; CD4-Positive T-Lymphocytes; Female; Flow Cytometry; Humans; Interferon-gamma; Interleukin-2; Ionomycin; Male; Tetradecanoylphorbol Acetate; Tomography, X-Ray Computed; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

The possible immunological relationship between the pattern of Th1/Th2 cytokine production and tuberculin reactivity was assessed in patients with active Mycobacterium tuberculosis infection. The production of the intracellular cytokines interferon (IFN)-gamma and interleukin-4 (IL-4) was measured in CD4(+) and CD8(+) T cells obtained from peripheral blood and bronchoalveolar lavage fluid (BALF) of 20 tuberculin skin-positive patients and compared with the findings recorded in nine tuberculin skin-negative patients with active pulmonary tuberculosis. Upon stimulation with phorbol 12-myristate acetate/ionomycin for 6 h, tuberculin-negative patients had a significantly higher proportion of IFN-gamma-producing CD4(+) T lymphocytes in BALF than in peripheral blood, while both CD4(+) and CD8(+) T-lymphocyte subsets in BALF of tuberculin-positive patients secreted more IFN-gamma than their peripheral blood counterparts. Tuberculin-negative patients had a significantly higher proportion of IFN-gamma-producing CD4(+) T lymphocytes in peripheral blood than tuberculin-positive patients. There was no significant difference in the production of IFN-gamma by BALF CD4(+) T lymphocytes, or by either peripheral blood or BALF CD8(+) T lymphocytes. In two tuberculin-negative patients, peripheral blood CD4(+) T lymphocytes produced IL-4. Study results suggested a higher immune activity in the blood of tuberculin-negative patients, with an increased lymphocyte activity in BALF versus peripheral blood in both patient groups.

Topics: Adult; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Female; Flow Cytometry; Humans; Interferon-gamma; Interleukin-4; Ionomycin; Male; Middle Aged; T-Lymphocytes; Tetradecanoylphorbol Acetate; Tuberculosis, Pulmonary

An understanding of T cell responses that are crucial for control of Mycobacterium tuberculosis (MTB) has major implications for the development of immune-based interventions. We studied the frequency of purified protein derivative (PPD)-specific CD3) cells expressing interleukin-2 (IL)-2, gamma interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha and IL-10 in HIV-negative pulmonary tuberculosis patients (TB, n=30) as well as in healthy individuals (controls, n=21) from Central Africa. Increased frequencies of PPD-stimulated CD3+ cells expressing IL-2, IFN-gamma, and TNF-alpha in TB were seen when compared with frequencies of controls. The presence of type 1 cytokine biased responses in TB patients was supported by a shift in the distribution pattern of cytokine expression from exclusively IL-2 or TNF-alpha expression seen in controls towards an increased frequency of IFN-gamma/IL-2 or IFN-gamma/TNF-alpha co-expression in TB. Higher levels of PPD-induced IFN-gamma in the supernatants from TB patients than from controls were found, which correlated with its intracellular expression. PPD was a weak inducer of IL-10 in T cells and insufficient in promoting cytokine production in TCRgammadelta+CD3+ cells. Non-specific stimulation with PMA and ionomycin revealed increased frequencies of CD4+ cells expressing IFN-gamma in controls, while expression of IL-2, IL-4, IL-10, IL-13, and TNF-alpha was not different. Non-specific cytokine responses of TCRgammadelta+CD3+ cells were similar in all groups. Pulmonary TB in Central Africa is associated with enhanced expression and secretion of specifically induced cytokines that are frequently implicated in host defense against MTB.

Topics: Adolescent; Adult; Aged; Brefeldin A; CD3 Complex; CD4-Positive T-Lymphocytes; Cells, Cultured; Cytokines; Female; Gabon; Humans; Interferon-gamma; Interleukin-10; Interleukin-2; Ionomycin; Lymphocyte Activation; Male; Middle Aged; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocyte Subsets; Tuberculin; Tuberculosis, Pulmonary; Tumor Necrosis Factor-alpha

To study the change of peripheral blood CD4+ helper T cells and T helper subtype I and II (Th1/Th2) in patients with pulmonary tuberculosis and to explore their changes during anti-tuberculosis chemotherapy.. CD4+ T cells from 105 patients with pulmonary tuberculosis and 25 normal controls were counted by Flow Cytometry. Peripheral blood cells were stimulated in vitro by phorbol myristate acetate (PMA) (25 ng/ml) and ionomycin (250 ng/ml). Cytokines were confined to the cells by using the protein transporting inhibitor containing monensin (2 micro mol/ml). These cells were incubated in 5% CO(2) for 4 h-4.5 h. The membrane and plasma of CD4+ helper T cells were marked by CD3 -PC5 + CD8 -FITC + INF-gamma -PE/CD3 -PC5 + CD8 -FITC + IgG1 -PE, CD3 -PC5 + CD8 -FITC + IL-4 -PE/CD3 -PC5 + CD8 -FITC + IgG1 -PE monoclonal antibodies respectively. Th1 and Th2 cells were counted and the ratio of Th1/Th2 cells was calculated. The levels of Th1 and Th2 cells in the 67 patients with pulmonary tuberculosis were detected at the end of intensive chemotherapy and at the sixth month of chemotherapy.. (1) The levels of CD4+ T and Th1 cells in patients with tuberculosis were significantly lower than those of controls. Their values were (663 +/- 160)/ microl vs (735 +/- 156)/ microl and (9.56 +/- 3.60)% vs (18.7 +/- 5.03)% respectively (P < 0.05). (2) The levels of CD4+ T and Th1 cells in patients with severe pulmonary tuberculosis were lower than those in patients with moderate or mild pulmonary tuberculosis. Their values were (579 +/- 120)/ microl vs (726 +/- 166)/ microl, (684 +/- 192)/ microl and (5.43 +/- 2.33)% vs (12.2 +/- 1.81)% and (10.9 +/- 2.30)% respectively (P < 0.05). The level of Th2 cells was in contrast to Th1 cells and their values were (5.63 +/- 1.26)% vs (2.93 +/- 0.87)% and (3.22 +/- 1.01)% (P < 0.01). (3) The level of Th1 cells increased while that of Th2 decreased in the 61 patients who gradually recovered. (4) The level of Th2 cells in patients with smear positive tuberculosis was strikingly higher than that in patients with smear negative tuberculosis and their values were (5.20 +/- 0.97)% vs (2.77 +/- 1.96)% (P < 0.05).. The detection of CD4+ helper T cells and Th1/Th2 cells in the peripheral blood cells from patients with pulmonary tuberculosis is useful in evaluating the state of disease and the effect of chemotherapy.

Topics: Adolescent; CD4 Lymphocyte Count; CD4-Positive T-Lymphocytes; Cells, Cultured; Female; Flow Cytometry; Humans; Interferon-gamma; Interleukin-4; Ionomycin; Male; Middle Aged; Monensin; Th1 Cells; Th2 Cells; Tuberculosis, Pulmonary