The Study Of Characteristics And Function Of Tissue Resident Memory T Cells In Tuberculosis Patients

T cells have critical roles in mediating the host defense against Mycobacterium tuberculosis(Mtb)infection but are insufficient to clear the infection.No other suitable immune correlates for protective immunity against TB,however,have been identified,perhaps because immune responses measured ex vivo in the peripheral blood might not faithfully reflect the immune response that occurs at the site of infection.In recent years,a new subset of cells which are called for resident memory T cells(TRM),are found in the tissues.TRM cells are considered as the first line of defense against pathogens.Recent studies performed in mice have indicated that TRMs in the lung are critical for TB protection;their human counterparts,however,remain relatively uncharacterized in TB.In this study,we collected different types of tissues of TB patients and detected the protein and gene profile of TRM cells by whole genome sequencing,mass spectrometry flow cytometry and cell flow cytometry.Furthermore,the impact of TRM cells on macrophages and its mechanism were studied in vitro.The results of this study revealed the expression,phenotype and function of TRM cells in the tissues of patients with tuberculosis,and provided us with a new understanding of the local immune environment of tuberculosis infection.METHOD1.The distribution of TRM cells in different tissues from the patients of tuberculosis was detected by Flow cytometry.2.The expression of 28 molecules in different types of tissues from patients of tuberculosis was detected by CyTOF.3.The whole gene of TRM cells was used detected by Gene sequencing.4.Eighteen cytokines and chemokines of TRM cells were detected by CyTOF.5.TRM cells from TB patients were isolated and co-cultured with tubercul-infected macrophages,and bactericidal ability,cytokines expression and polarization of macrophages were detected.RESULT1.A high frequency of TRMs in the CD4+and CD8+T-cell population is present at infection sites in patients with TB.These TRM cells exhibit a TEM-like phenotype.2.TRM cells express high levels of activation markers,chemotaxis and exhausting markers.3.TRM cells of tuberculosis patients have unique gene expression profile.4.TRM cells found in the pleural fluid in patients with TB exhibit an Mtb antigen-specific Thl-cell response.5.TRM cells in patients with TB exhibit a multifunctional phenotype based on multiple cytokine production6.TRMs limit intracellular Mtb replication in human macrophages and induce macrophages to differentiate into M1 by expressing high levels of IFN ? and TNF?.CONCLUSION1.TRM cells are highly expanded in non-lymphoid tissues but not in the peripheral blood in the patents of tuberculosis.2.TRMs found at TB infection sites exhibit memory and an activated phenotype;3.CD4+TRMs exhibit a cytotoxic phenotype similar to CD8+T cells;4.Mtb-specific TRMs exhibit a multifunctional phenotype based on multiple cytokine expression;5.TRMs limit intracellular Mtb replication in macrophages by inducing high level of cytokines expression and macrophage differentiation to M1.