The direct killing of infected cells or invasive pathogens is the Patients infected with human immunodeficiency virus (HIV) exhibitedДЄntiviral potential through their cytolytic functions. Wallace et al ( 13) reported that ОіОґ T cells from Such as influenza viruses, which include human seasonal H1N1 andДЄvian H5N1 and H9N2 viruses ( 10вЂ“ 12). Of cytolytic activity in response to various types of pathogens, Killer cells, macrophages and dendritic cells) ( 9). Tumor necrosis factor-О±) and chemokines, rapid cytotoxicity orД¬rosstalk with other cell types that are involved (such as natural Potent, soluble proinflammatory molecules (such as interferon-Оі and Infection through multiple pathways, including the production of О“Оґ T cells participate in early immunity against This report, the suppression of ОіОґ T cells by chronic tuberculosis Which indicates the importance of ОіОґ T cells in the control of Host response to mycobacterial infections was described as early asДЄctivation of ОіОґ T cells was reported during M.tb invasion, Make up only a small proportion (3вЂ“10%) of the circulatingД¬D3 + T cells in peripheral blood, ОіОґ T cells haveД®ssential roles in host defenses against invasive pathogens Population with a specific tissue distribution and antigen (Ag) Human ОіОґ T cells are a subset of T cells thatД®xpress the ОіОґ T cell receptor (TCR) and are a unique lymphocyte Understanding of the details underlying the host protective immune There are ~1.5 million deaths worldwide from this disease annuallyДЄnd an increasing number of drug-resistant cases are reported eachД¬oncerning global health issue, which is partially due to a limited With Mycobacterium tuberculosis ( M.tb), the agent Organization (WHO), one-third of the global population is infected Therefore, the present study may provide valuable information concerning the methods employed to investigate the function of ОіОґ T cells and potentially other lymphocyte subsets.ДЄccording to a 2014 report by the World Health In conclusion, the flow cytometricвЂ‘based assay described in the current study is a simple, sensitive and reliable tool to determine the cytolytic activity of ОіОґ T lymphocytes against mycobacteria. The present study describes a simple and convenient assay that is based on the direct quantitative and qualitative analysis of cell damage at a single cell level utilizing a twoвЂ‘color flow cytometric assay. The percentage of cytotoxicity in the CFSEвЂ‘gated cell population was calculated by subtracting the value obtained for nonвЂ‘specific PIвЂ‘positive target cells, which was measured in a control group that did not contain effector cells. CFSE and PI double positive cells were recognized as the dead target cells. During sample acquisition, target cells were gated on the CFSE positivity and examined for cell death based on the uptake of PI. After 4В h of inВ vitro coвЂ‘culture between ОіОґ T cells and CFSEвЂ‘labeled infected or uninfected THPвЂ‘1 cells, staining with propidium iodide (PI) was performed to distinguish between vital and dead cells. Staining of target THPвЂ‘1 cells with 8В ВµM CFSE revealed high and stable loading of fluorescence and no effect of the viability of cells. In the present study, target cells were stained with 5вЂ‘(andвЂ‘6)вЂ‘carboxyfluorescein diacetate succinimidyl ester (CFSE), which is a stable integrated fluorescent probe that allows target and effector cells to be distinguished from one another. In addition to this indirect method, fluorescence assays have been described for the assessment of inВ vitro cellвЂ‘mediated cytotoxicity. The effector function of natural killer, lymphokine-Вactivated killer cells and T lymphocytes is commonly evaluated by radioactive chromiumвЂ‘release cytotoxicity assays.

T cell functional assay