The Expression Of IFN-γ、IP-10and MCP-1in Peripheral Blood And The Diagnosis Value Of Active Tuberculosis

Purpose: The study aimed to explore the expression of Interferon-γ （IFN-γ）,γ-interferon-inducible protein10（IP-10）and monocyte chemoattractant protein-1（MCP-1）in peripheral blood and the diagnosis value of active tuberculosis.Methods:292individuals were prospectively enrolled,40healthy controls,95non-tuberculous respiratory disease (17Lung cancers，44Pneumonia,25Chronic obstructivepulmonary disease and9others) and157active-TB (108Pulmonary TB,5Tuberculousmeningitis,26Tuberculous pleurisy,3Tuberculous peritonitis,5Tuberculous polyserositis,4Cervical tuberculous lymphadenitis,1Epididymis TB,4Bone TB and1Left renal TB).292human whole blood samples stimulated by PHA and purified fusion proteinCFP10/ESAT6of mycobacterium tuberculosis respectively to produce IFN-γ, IP-10andMCP-1. The level of IFN-γ, IP-10and MCP-1were evaluated by chemiluminescenceenzyme immunoassay. This study compared different antigen concentration and the culturesupernatant dilution of the impact on the level of cytokine, ultimately determine theexperimental methods. Stimulating the concentration of PHA and CFP-10/ESAT-6fusionprotein were both20ug/mL. In this study, the original times the culture supernatant level ofdetection of A,10-fold dilution of culture supernatant level of B and C was detected. Thenthe receiver operating characteristic curve was drawn to determine the threshold of IFN-γ,IP-10and MCP-1for diagnosis of active tuberculosis and to evaluate its diagnosticperformance.Results:1. The content of IFN-γ and IP-10in the supematant unstimulated (negative controlhole) had no significant difference in three group (P>0.05). But the content of MCP-1in thesupematant unstimulated in active tuberculosis group was significantly higher than that ofnon-tuberculous respiratory disease group (P<0.05), the content of MCP-1innon-tuberculous respiratory disease group was significantly higher than that of the healthycontrol group (P<0.05).2. The production of IFN-γ, IP-10and MCP-1stimulated by PHA in active tuberculosisgroup and non-tuberculous respiratory disease group were all significantly lower than that of the healthy control group (P<0.05)，but active tuberculosis group and non-tuberculousrespiratory disease group were not different (P>0.05).3. The production of IFN-γ, IP-10and MCP-1stimulated by CFP-10/ESAT-6in activetuberculosis group were significantly higher than that of non-tuberculous respiratory diseasegroup (P<0.01) and that of the healthy control group (P<0.01)，but non-tuberculousrespiratory disease group and healthy control group were not different (P>0.05).4. Observing the influence of factors(1) The production of IFN-γ, IP-10and MCP-1stimulated by CFP-10/ESAT-6with TB patiens in in lymphocyte count group of<1.0×109were significantly lower than that of in lymphocyte count group of≥1.0×109(P<0.01);(2) The production of IFN-γ, IP-10and MCP-1stimulated by CFP-10/ESAT-6with TB patiens in different age groups, acid-fast bacilli smear positive and negative groups,pulmonary tuberculosis and extrapulmonary tuberculosis among groups, there were nosignificant differences between them (P>0.05);(3) The production of specific IFN-γstimulated by CFP-10/ESAT-6with TB patiens in PPD skin test positive group was higherthan that of the PPD-negative group (P<0.05), but IP-10and MCP-1were not different(P>0.05).5. The study applied SPSS17.0and drew the ROC curve. The production of IFN-γ,IP-10and MCP-1stimulated by CFP-10/ESAT-6, cut off value were256pg/mL,190pg/mL, and389pg/mL as active tuberculosis diagnosis standard. The sensitive of IFN-γ, IP-10and MCP-1was57.3%,68.2%and92.8%, respectively; specificity was80%,80%and80.7%, respectively; the positive predictive value was76.9%,78.7%and84.9%, respectively;and negative predictive value was61.7%,67.9%and78.7%, respectively; accuracy rateswas76.9%,78.7%and84.9%, respectively. Three kinds of cytokines of IFN-γsensitivity waslowest, MCP-1maximum; the specific of three cytokines were similar; positive predictivevalue and the negative predictive value of MCP-1maximum, IP-10was slightly higher thanIFN-γ.Conclusions: The level of IFN-γ, IP-10and MCP-1stimulated by CFP-10/ESAT-6were evaluated by chemiluminescence enzyme immunoassay, which can be used for activetuberculosis diagnosis and differential diagnosis. Detection of content of whole blood IP-10 and MCP-1on the active tuberculosis compared IFN-γ, it can obtain higher sensitivity anddiagnostic accuracy. Therefore, detection the levels of IP-10and MCP-1in whole blood canaplly on clinical practice as amethod of diagnosis of active tuberculosis.