| BackgroundTuberculosis(TB) is an infectious disease caused by mycobacterium tuberculosis. Early diagnosis have extremely vital significance for rapid spread TB treatment and control. The body immune mechanism can be divided into humoral immunity and cellular immunity after infected with TB, so the diagnosis of tuberculosis(TB) is mainly from two aspects of humoral immunity and cellular immunity. At present main laboratory diagnosis of TB is sputum smear, bcterial culture, mycobacterium tuberculosis PCR amplification and rigra test(IGRAs), etc.The sputum smear with low positive rate; bcterial culture takes too long;PCR amplification method with high technical requirements;IGRAs may partly compensate for the insufficiency, with high sensitivity and specificity, but still cannot meet the clinical needs, especially the diagnosis of tuberculosis(TB) in different course. While detecting multiple TB immune humoral immunity and cellular immunity markers, which can effectively improve TB diagnosis positive rate, and may through the different TB progression in the expression of immune molecules spectrum research found the new diagnostic molecular markers of tuberculosis.ObjectiveTo analyze the immune molecule spectrum in different course of TB, determine the characteristic of the biological markers, we defined the regulatory T cells and TH17 cells through the flow cytometry,we also defined the tuberculosis immunity molecular with the liquid chip technology in different stages of TB. Determine different TB infection phase characteristic of the biological markers, for clinical diagnosis, immune function in patients with tuberculosis and prognosis assessment to provide scientific basis.Methods1. One hundred and eighty-one subjects with active tuberculosis(N=66), latent tuberculosis(N=57), or no evidence of tuberculosis infection(N=58) were evaluated.We used the flow cytometry to detect Treg cells and Th17 cells expression levels in peripheral blood in patients with different stages, at the same time IL-10 and IL-17 were detect by ELISA.2. One hundred and forty-nine subjects with active tuberculosis(N=50), latent tuberculosis(N=49), or no evidence of tuberculosis infection(N =50) were evaluated. We used the Luminex Multiplexed Bead Array platform to simultaneously evaluate 38 biomarkers in the supernatant of whole blood samples. We defined the response to stimulation as the difference(within an individual patient) between the response to the pooled tuberculosis antigens and the negative control.Results1. Treg cells and Th17 cells in active TB group were significantly higher than the proportion of TB incubation period group and normal control group(Treg cells: 9.61±2.26% VS 6.31±1.44% and 9.61±2.26% VS 4.16±1.01% P<0.05; Th17 cells: 3.08±0.80% VS 1.31±0.50% and 3.08±0.80% VS 1.15±0.46% P<0.05), the proportion of Treg cells incubation period group were obviously higher than that of normal control group(6.31±1.44% VS4.16±1.01% P<0.05), but the incubation period and the proportion of the group of Th17 cells compared to normal control group no significant difference(1.31±0.50% VS 1.15±0.46% P>0.05).Activity of IL-10 and the expression of IL-17 incubation period is significantly higher in the group and normal control group(P < 0.05), and the expression of IL-10 incubation group was obviously higher than that of normal control group(P < 0.05), but the incubation period of IL-17 expression and normal control group no significant difference(P > 0.05).Treg cells and TH17 cells to differentiate in active TB incubation period and the area under the ROC curve was 0.890 and 0.710 respectively.2. Eotaxin、IL-1α、IP-10、MCP-1、IL-10 and TNF-α in tuberculosis patients are significantly higher expression than the healthy controls(P<0.05), MCP-1 and IL-1α in active TB were significantly higher expression than that in latent tuberculosis(P <0.05). The activity of area under the ROC curve with MCP-1 and IL-1α was used to distinguish between active TB and latent tuberculosis was 0.905 and 0.825 respectively.Conclusions1. Treg cells, Th17 cells, Eotaxin, IL-1α, IP-10, IL-10, MCP-1 and TNF-α cytokines in TB infection are significantly higher expression than the healthy controls(P <0.05), and Treg cells, IL-1α and MCP-1 in active TB are significantly higher expression than that in latent tuberculosis(P <0.05).2. For a variety of immune markers detection at the same time not only effectively enhance the TB diagnosis positive rate, at the same time will help determine different immune status in the period of the course of the disease, tuberculosis for the clinical diagnosis of TB, the immune function of patients with treatment of adjustment and provide a scientific basis for evaluation. |
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