| Tuberculosis(TB)remains one of the most severe health problems in China as well as worldwide.To define diagnostic methods and evaluation criteria for treatment responses with high accuracy and sensitivity will facilitate the prevention and control of TB,which in turn reduces the economic and social burden for public.At present anti-TB chemotherapy includes two months' intensive period and four months' consolidate period.In the process of anti-TB treatment,along with the decrease of bacteria load TB-specific cellular immune responses might be altered.On the other hand,the alleviation of clinical symptoms might also contribute to the dynamic changes of metabolic due to the consumptive character of TB.Therefore,systematical investigation on immune and metabolic signatures during anti-TB treatment will provide new clues to identify biomarkers for diagnosis and treatment efficacy.This might improve the concept of personalized therapy for TB as well.In the present study,24 onset TB patients have been tracked to monitor TB specific T cell responses as well as metabolomic profiles during the treatment.Peripheral blood samples were collected before the treatment and at 2,4,6 months of the treatment.Antigen specific IFN-? secreting cells were determined by enzyme-linked immune assay spots(ELISPOT).Meanwhile,antigen specific cytokine-producing T cells were detected by intracellular cytokine staining and monoand multifunctional T cell proportions were analyzed.Our results showed that along with the treatment,the numbers of IFN-? secreting cells significantly decreased upon ESAT-6 and CFP-10 stimulation.This was consistent with the results from flow cytometry that the percentage of IFN-?+ T cells showed a decreased after 6-monthtreatment.Antigen specific IFN-? release might become the immunological biomarkers in the process of anti-TB treatment.In parallel,we have detected the metabolomic profiles via Nuclear Magnetic Resonance Spectroscopy(NMR)assay.Multivariate statistical analyses were further performed to identify the biomarkers for efficacy evaluation.Forty-two metabolites have identified in the plasma of TB patients,including those mainly involved in glycometabolism,lipid metabolism and amino acid metabolism.The results from orthogonal partial least squares discriminant analysis(OPLS-DA)revealed that total metabolites could clearly distinguish those before treatment and those at 2,4,6 month treatment,respectively.Among the total metabolites,15 amino acids were significant in defining the treatment course whereas metabolites involved in glycometabolism and lipid metabolism were not.More precisely,5 amino acids including isoleucine,arginine,threonine,4-aminobenzoate and phenylalanine decreased dramatically along with the treatment course.The levels of 5 amino acids after the treatment for 6 months were comparable to those in healthy controls.Results from receiver operating characteristic curve(ROC curve)analysis showed that the area under ROC curve(AUC)value of combined 5 amino acids was 0.670 with certain diagnostic potential in evaluating the efficacy of anti-TB treatment.At the same time,four among five periphery amino acid levels were higher in TB patients than in healthy controls.The AUC value of the ROC curve analysis based on the combination of these four amino acids was 0.867,which meant high diagnostic potential for active TB.More importantly,the decrease of 5 amino acids contents was consistent with the tendency of IFN-? releasing cells in the periphery,which suggest that the metabolites may directly or indirectly influence anti-TB immune function.In summary,the immunological and metabolomic characters during anti-TB treatment were analyzed simultaneously for the first time in this study.A panel of amino acids are defined for distinguish TB.With the consistency between TB-specific immune signature and metabolomic profiles,the elucidation of immune-metabolic interaction module may become a novel strategy for regulating TB-specific immune responses. |
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