The Distribution Changes Of Immune Memory Subtypes Of Different T-cell Subpopulations In Peripheral Blood Of Pulmonary Tuberculosis Patients

Background: Tuberculosis(TB), caused by Mycobacterium tuberculosis(Mtb), is one of the most prevalent and commonest serious infectious diseases worldwide.Previous studies have proved that CD4+ T cells, CD8+ T cells and γδ T cells play an important role in protection against Mtb infection. T cells can be divided into naive T cells(Tnaive), central memory T cells(Tcm), effector memory T cells(Tem) and effector T cells(Teff). Tcm cells are able to home to secondary lymphoid tissues,produce high amounts of IL-2 but low levels of other effector cytokines, while their CD27 counter parts, Tem cells, are able to produce high levels of cytokines, exert rapid effector functions and home to peripheral tissues. Teff have the effector function,such as the production of cytokines or cytotoxic activity. Vγ9Vδ2 T cells represent a major peripheral blood γδT-cell subpopulations in humans. Recent data have shown that Vγ9Vδ2 T cells also comprise naive, memory and effector cells that can be distinguished on the basis of surface marker expression and effector functions.However, the profile of memory type cells in TB patients has seldomly reported.Objective:To investigate of the distribution changes of immune memory subtpyes of different T-cell subpopulations of peripheral blood in healthy adult and TB patients.Methods:1. A certain amount of 5ml venous blood in heparin anticoagulant tube from healthy Adults and pulmonary TB(PTB) patients, were stained with different fluorochrome-conjugated m Abs for staining of surface molecule(anti-CD45RA-FITC,anti-TCRγδ-APC-Vio770, ani-CD4-APC-Vio77, anti-CD27-APC, anti-Vδ1-PE,anti-Vδ2-Per CP, anti-CD8-PE-Cy7, anti-CD3-BV421, anti-CD27-BV510). T cells can be divided into Tnaive(CD45RA+CD27+), Tcm(CD45RA—CD27+), Tem(CD45RA—CD27—) and Teff(CD45RA+CD27—). The proportions of immune memory subtypes of T-cell subpopulations(CD4, CD8, Vδ1, Vδ2 and Vδ3-8) were detected by flow.2. Peripheral blood samples of PTB patients and health donors(HD) were obtained and stimulated with PMA and ionomycin for 2 hours and followed by addition of monensin and incubation for more 4 hours. Cells were collected and stained with different fluorochrome-conjugated m Abs for staining of surface molecule and intracellular cytokines(anti-CD45RA-FITC, anti-TCRγδ-APC-Vio770,ani-CD4-APC-Vio77, anti-CD27-APC, anti-Vδ1-PE, anti-Vδ2-Per CP,anti-CD8-PE-Cy7, anti-CD3-BV421, anti-CD27-BV510, anti-IL-17-APC,anti-IFN-γ-APC). The IL-17-producting or IFN-γ-producting T cells can be divided into Tnaive(CD45RA+CD27+), Tcm(CD45RA—CD27+), Tem(CD45RA—CD27—) and Teff(CD45RA+CD27—). The proportions of immune memory subtpyes of IFN-γproducing cells and IL-17 producing cells were detected by flow cytometry.Results:1.The percentages of immune memory subtpyes(Tnaive, Tcm, Tem, Teff)of CD4+ and CD8+ T cells of health donors were 57.53±13.59, 33.18±11.40, 6.03±3.26, 3.26 ± 4.09 and 63.33 ± 16.63, 15.46 ± 6.86, 6.56 ± 4.25, 14.60 ± 11.75,respectively. The percentages of memory subtpyes of CD4+ and CD8+ T cells of PTB patients were 36.49±17.65,38.32±10.09,24.44±10.76,0.75±0.58 and 24.53±17.58, 11.26±6.84, 39.38±18.31, 24.83±12.90, respectively.2. The percentage of memory subtpyes(Tnaive,Tcm,Tem,Teff) of Vδ1, Vδ2 and Vδ3-8 T cells of health donors were 32.74±21.66, 17.48±15.51, 4.21±3.66, 45.56±29.20; 5.46±4.05, 77.90±10.60, 14.65±10.90, 2.00±2.21 and 38.64±26.64,13.90±10.50, 4.18±3.76, 43.02±27.55, respectively. The percentages of memory subtpyes of Vδ1, Vδ2 and Vδ3-8 T cells of PTB patients were 18.90±17.79, 10.23±4.43, 19.02±20.75, 51.85±21.55; 5.43±3.03, 61.05±22.98, 28.27±23.13, 5.24±2.30 and 29.69±26.22, 17.33±10.15, 8.86±7.27, 44.14±29.76, respectively.3. The percentages of memory subtpyes(Tnaive,Tcm,Tem,Teff) of IFN-γ –producting CD4+ and CD8+ T cells of health donors were 6.96±4.37, 74.03±5.68, 16.22±3.02,2.81±2.61 and 17.75±6.61, 48.99±13.72, 13.70±6.61, 19.57±11.67, respectively.The percentages of memory subtpyes of IFN-γ –producting or CD4+ and CD8+ T cells of PTB patients were 4.17±2.23, 64.20±15.13, 27.33±14.34, 4.32±3.89 and 12.19±3.76, 33.66±14.49, 17.00±9.17, 42±13.52, respectively.4. The percentages of memory subtpyes(Tnaive,Tcm,Tem,Teff) of IFN-γ-producting Vδ1, Vδ2 and Vδ3-8 T cells of health donors were 32.03±19.58, 12.22±14.84, 2.85±2.29, 42.92±28.4; 15.07±7.99, 63.30±1.22, 9.85±5.71, 11.24±15.03 and 13.22±10.48, 57.51±19.87, 14.14±7.19, 15.16±18.70, respectively. The percentages of memory subtpyes of IFN-γ producting Vδ1, Vδ2 and Vδ3-8 T cells of PTB patients were 14.86±9.98, 10.11±10.96, 2.48±2.65, 72.58±18.76; 11.09±6.25, 67.71±10.45, 16.09±7.11, 5.09±3.97 and 6.15±3.51, 36.13±21.43, 23.25±15.93, 34.49±27.78, respectively.5. The percentages of memory subtpyes(Tnaive,Tcm,Tem,Teff) of IL-17-producting CD4+ and CD8+ T cells of health donors were 4.82±3.33, 59.86±9.14, 34.58±9.58,0.76±1.21 and 38.24±20.84, 33.88±15.22, 24.52±13.22, 4.35±4.30, respectively.The percentages of memory subtpyes of IL-17-producting CD4+ and CD8+ T cells of PTB patients were 3.27±2.49, 66.58±10.08, 29.94±8.75, 0.22±0.55 and 27.06±13.10, 38.24±18.74, 29.30±14.75, 5.42±10.40, respectively.Conclusions:1. The CD4+T、CD8+T and Vδ2 cells of Patients with PTB had significantly changes of immune subtype distribution proportion. Except Vδ2 T cell, the IFN-γ-producting CD4+T, CD8+T and γδT cells of PTB patients had changes of immune subtype distribution proportion. These changes which may have a protective effect, may be associated with the immune response against Mtb infection,.2. The proportions of immune subtpyes of IL-17-producting αβ T cells had no difference between PTB patients and health donors. Mtb infection may not change the immune subtype distribution proportion of IL-17-producting αβ T cells.