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Objective To study the dendritic cells (dendritic cell, DC) changes in content and maturity of different TB patients to explore DC in tuberculosis immunity mechanism of action is based on DC's new tuberculosis vaccine development and anti-TB immunotherapy program to provide experimental basis. Method 1. Extracted peripheral blood of patients with tuberculosis, three color marker flow cytometry (FCM) determination of the content of peripheral blood DC. 2 isolated peripheral blood mononuclear cells induced by DC in vitro cultured, rhGM-CSF and rhIL-4, rhTNF-alpha induced by the combined culture 11 days the cells were harvested. Using an inverted microscope, scanning electron microscopy and transmission electron microscopy to observe the outside of the cell morphology and internal ultrastructure. 4 fluorescence quantitative polymerase chain reaction (RT-PCR) to detect cell IL-12p40, IL-12 p35 and CCR7 mRNA expression. 5 enzyme-linked immunosorbent assay (ELISA) detection of cell culture supernatant of IL-12p70 content. 1 the total number of TB patients in peripheral blood DC and DC1 subsets were (0.42 ± 0.19)% and (0.28 ± 0.13)%, significantly lower than the healthy control group, the difference was statistically significant (P lt; 0.01 ), DC2 subsets was no significant difference; sputum smear-positive patients, the total number of peripheral blood DC and DC1 subsets were (0.24 ± 0.06)%, (0.16 ± 0.04)%, were lower than in patients with smear-negative, the difference has statistically significant (P lt; 0.05), the DC2 subpopulations was no significant difference; untreated patients with retreatment patients the total number of DC, DC1 and DC2 there was no significant difference (P GT; 0.05). 2 in rhGM-CSF, rhIL-4, and rhTNF-alpha induced by the combined culture, cultured for 11 days, by an inverted microscope, scanning electron microscope, transmission electron microscopy can be seen with typical dendritic morphology of dendritic cells. RT-PCR results show TB patient group, IL-12p40, IL-12 p35, CCR7 mRNA expression level was significantly lower than that in normal healthy population (P lt; 0.05) ELISA results showed that the group of patients with tuberculosis cell culture supernatant IL-12p70 was lower than normal healthy control group, and the secretion peak delay. Conclusion 1. Peripheral blood of normal healthy individuals and patients with tuberculosis after separation, adding rhGM-CSF, rhIL-4, rhTNF-alpha induced cultured to 11 days, can be obtained with typical morphology and function of DC. 2 TB patients peripheral blood mononuclear cells derived DC level of secretion of IL-12p70 and IL-12p40, IL-12 p35, CCR7 mRNA expression level was significantly lower than that in normal healthy people, suggesting that DC mature obstacles, thereby initiating immune responses weakened. 3 TB patients, the total number of peripheral blood DC and DC1 subsets decrease in the body infected with Mycobacterium tuberculosis, prompting the the Th0 cells differentiation into Th1 cells ability weakened mediated cellular immune response reduced ability.
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