1、Immunological Features And Metabolic Classificationof Tuberculosis Complicated By Type 2 Diabetes Mellitus Patients 2、Part Ⅱ Protection Of Memory Th17 Cells Against Secondary Pneumococcal Pneumonia Following Influenza Infection

Part I:Immunological Features and Metabolic Classification ofTuberculosis Complicated by Type 2 Diabetes Mellitus PatientsAlthough type 2 diabetes mellitus(DM)has long been recognized as a risk factor for tuberculosis(TB),the biological mechanisms underlying this interaction remain unclear Persistent hyperglycemic environment favors defects in immune function of DM patients in which Mycobacterium tuberculosis(Mtb)-specific T cells are mostly affected.We thus hypothesize that chronic hyperglycemia leads to impaired Mtb-specific T cells in TB with DM(TB~+DM)patientsIn this study,IFN-γ,TNF-α and IL-2 producing Mtb-specific CD4~+ and CD8~+ T cell of TB~+DM and TB patients were detected by in vitro stimulation of PBMCs from TB or TB~+DM patients with Mtb-specific antigens 6 kDa early secretory antigenic target(ESAT-6)or 10 kDa culture filtrate protein(CFP-10)with subsequent intracellular cytokine staining(ICS).Frequencies of Mtb-specific T cells were compared between TB~+DM and TB patients.Clinical manifestation indicated that TB~+DM patients were older with slightly higher bacillus load,similar cavity rate and higher fasting blood sugar level compared with TB patients.Compared with TB patients,frequencies of ESAT-6 and CFP-10 specific IFN-y and TNF-α producing CD4~+ and CD8~+ T cells decreased in TB~+DM patients.More significantly,AFB positive TB~+DM patients showed decreased frequencies of ESAT-6 and CFP-10 specific IFN-y producing CD4~+ cells compared with age matched AFB positive TB patients as well.ESAT-6 and CFP-10 specific IFN-y and TNF-α producing CD4~+ cells decreased with the increase in the duration and comorbidity of DM rather than HbAlcSince activation and differentiation of immune cells are tightly regulated by energy metabolism,whether the defect of T cell responses in TB~+DM patients is related to abnormal metabolism is worthy of investigation,In addition,little is known about the peripheral metabolomics of TB~+DM patients.1H nuclear magnetic resonance(NMR)spectroscopy-based metabolomics was applied to study metabolite profiles in the plasma of TB and TB~+DM patients.Multivariate statistical analysis was utilized to construct a model to subgruoup TB~+DM patients with different metabolic and clinical features.26 metabolites were detected with significant difference between TB~+DM and TB.Among them,glucose level increased in TB~+DM group,which was consistent with clinical biochemical analysis.However,other glycolysis-related metabolites such as lactate,pyruvate and succinate decreased in TB~+DM patients.In addition,amino acids such as glutamine,BCAAs and fatty acid also decreased in the plasma of TB~+DM patients.The change of plasma metabolomics in TB~+DM patients indicated that main energy metabolism including glycolysis,amino acids and lipids metabolism was impaired in TB~+DM when compared to TB only patients.Interestingly,TB~+DM patients were able to be divided into two groups with TB-like and DM-like metabolic patterns TB~+DM patients with DM-like glucose metabolic pattern displayed lower level of T cell response than those with TB-like metabolic pattern.Similar properties were observed when different lipid metabolic patterns in TB~+DM patientsTo be concluded,Mtb-specific T cells are impaired in TB~+DM patients compared with TB patients,which might be associated with worse prognosis of TB~+DM patients.Mtb-specific T cell responses decreased in TB~+DM patients with long duration of DM and more diabetes-related comorbidity,which indicates that persistent hyperglycemia impairs Mtb-specific T cell responses In addition,plasma metabolomic alteration might also contribute to dysfunction of Mtb-specific T cells observed in TB~+DM patients.Therefore,metabolic intervention in these patients might become an alternative strategy for the amelioration of TB~+DM pathogenesisPart II:Protection of Memory Th17 Cells against Secondary Pneumococcal Pneumonia Following Influenza InfectionSecondary Streptococcus pneumonia(Sp)infection after influenza A virus(IAV)is a significant clinical complication resulting in morbidity and mortality.Prior IAV infection has been demonstrated to impair the IL-17 immunity against bacterial infection.In this study we showed that preceding IAV causes persistent Sp infection and suppression of Sp specific Th17 cell response in lung of mice.Sp immunization provided protection against IAV/Sp co-infection by clear bacterium in the lung and blood of mice.Sp immunization induced Sp specific Th17 responses in lung of co-infected mice and blockage of IL-17 increased bacterial lord the mice,which indicated that protection of Sp immunization was depend on IL-17.Sp immunization induced cross-active Th17 cells and provided cross protection against co-infection induced by heterologous strain of Sp.These results indicate that Sp immunization induced memory Th17 cells had a key role in providing protection against secondary Sp infection after IAV infection and suggest the feasibility of developing a broadly protective vaccine against IAV/Sp co-infection by targeting Th17 T cells.