ABSTRACT
OBJECTIVE: 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. METHODS: 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. RESULTS: (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). CONCLUSION: 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.
Subject(s)
CD4-Positive T-Lymphocytes/immunology , Th1 Cells/immunology , Th2 Cells/immunology , Tuberculosis, Pulmonary/immunology , Adolescent , CD4 Lymphocyte Count , CD4-Positive T-Lymphocytes/metabolism , Cells, Cultured , Female , Flow Cytometry , Humans , Interferon-gamma/metabolism , Interleukin-4/metabolism , Ionomycin/pharmacology , Male , Middle Aged , Monensin/pharmacology , Tuberculosis, Pulmonary/pathologyABSTRACT
OBJECTIVE: To analyze the association between HLA-DR genes and pulmonary tuberculosis and to explore susceptible genes associated with pulmonary tuberculosis in a population of Han nationality from southern China. METHODS: Using case-control study, we detected by polymerase chain reaction-sequence specific primers (PCR-SSP) technique the 23 alleles of HLA-DR gene sites in 110 tuberculosis cases and 101 controls from Guangdong, Guangxi, Hunan, Hubei, Jiangxi and Fujian provinces. Gene frequency (GF) and odds ratio (OR) were calculated and compared. RESULTS: The frequency of DR16 allele in pulmonary tuberculosis cases was strikingly higher than that in the healthy controls (chi2=5.915, Pc< 0.05). Their GFs were 12.62% and 5.60% respectively, and the OR was 2.53. Significantly decreased frequency of DR1 and DR13.3 alleles in cases were found (chi2 values were 17.847 and 14.258 respectively, Pc < 0.01). Their ratios of GFs were 8.08% vs. 23.57% and 29.29% vs. 50.24%, and their ORs were 0.26 and 0.33 respectively. CONCLUSIONS: The results suggest that DR16 allele is closely correlated to incidence of pulmonary tuberculosis in this population of Han nationality from southern China, or linked to susceptible genes which are functional. It is also suggested that expression of DR1 and DR13.3 alleles may be associated with an antagonist effect in the pathogenesis of pulmonary tuberculosis in this population.