Ten-color flow cytometry for measuring T cell subpopulation and its activation state in the peripheral blood of HIV/AIDS patients

doi:10.3877/cma.j.issn.1674-1358.2018.01.006

Abstract

Abstract:

Objective

To establish the ten-color flow cytometry for measuring T cell subpopulations and their activated state in the peripheral blood of HIV/AIDS patients.

Methods

The monoclonal antibodies targeting cell surface antigen CD3, CD4, CD8, CD45RA, CD25, CCR7, CD28, CD38 and HLA-DR to establish the ten-color panel were used. One peripheral blood PBMC sample from a HIV/AIDS patient was applied for adjustment of voltage, compensation and FMO control. After that peripheral blood samples, of 5 patients with HIV/AIDS were analyzed by ten-color flow cytometry.

Result

The ten-color flow cytometric staining method was established for the measurement of T cell subpopulations and activation state in human peripheral blood. This method was applied to detect peripheral blood samples from 5 patients with HIV/AIDS and was able to evaluate the immune status of these patients. And then the percentage of T lymphocyte subgroup and the activation level were detected, respectively. The expression patterns of HLA-DR, CD38 and CD28 in different subpopulations were with significant difference.

Conclusions

The ten-color flow cytometry was a simple and reliable method for the analysis of T cell subpopulations and their activation in HIV/AIDS patients.

Key words: Muti-color flow cytometry, Human immunodeficiency virus, Acquired immuno deficiency syndrome, T cell, Activated status

Yangzi Song, Yaxian Kong, Yu Hao, Yu Jiang, Di Wang, Ling Xu, Xuejing Chong, Guorui Dai, Hongxin Zhao, Hui Zeng. Ten-color flow cytometry for measuring T cell subpopulation and its activation state in the peripheral blood of HIV/AIDS patients[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2018, 12(01): 28-34.

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Figures/Tables 5

References 25

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细胞亚群	细胞亚群比例	HLA-DR	CD38	CD28
CD4⁺	33.06 ± 11.68	18.81 ± 10.06	6.00 ± 1.61	93.32 ± 7.75
CD4⁺T_N	42.20 ± 9.65	2.29 ± 0.85	4.65 ± 2.19	99.90 ± 0.17
CD4⁺T_CM	40.90 ± 7.85	19.23 ± 7.58	9.29 ± 2.20	98.50 ± 1.98
CD4⁺T_EM	16.14 ± 7.13	58.90 ± 22.44	4.05 ± 2.44	71.58 ± 24.11
CD4⁺T_EMRA	0.79 ± 0.47	52.34 ± 25.77	19.98 ± 11.39	74.82 ± 30.10
CD4⁺rTreg（Fr Ⅰ）	2.55 ± 0.41	3.99 ± 3.00	2.23 ± 1.14	99.62 ± 0.50
CD4⁺aTreg（Fr Ⅱ）	0.71 ± 0.21	71.50 ± 14.55	12.03 ± 5.58	99.84 ± 0.36
CD4⁺ Treg Fr Ⅲ	7.29 ± 2.85	29.80 ± 12.36	6.63 ± 2.50	99.86 ± 0.17

细胞亚群	细胞亚群比例	HLA-DR	CD38	CD28
CD4⁺	33.06 ± 11.68	18.81 ± 10.06	6.00 ± 1.61	93.32 ± 7.75
CD4⁺T_N	42.20 ± 9.65	2.29 ± 0.85	4.65 ± 2.19	99.90 ± 0.17
CD4⁺T_CM	40.90 ± 7.85	19.23 ± 7.58	9.29 ± 2.20	98.50 ± 1.98
CD4⁺T_EM	16.14 ± 7.13	58.90 ± 22.44	4.05 ± 2.44	71.58 ± 24.11
CD4⁺T_EMRA	0.79 ± 0.47	52.34 ± 25.77	19.98 ± 11.39	74.82 ± 30.10
CD4⁺rTreg（Fr Ⅰ）	2.55 ± 0.41	3.99 ± 3.00	2.23 ± 1.14	99.62 ± 0.50
CD4⁺aTreg（Fr Ⅱ）	0.71 ± 0.21	71.50 ± 14.55	12.03 ± 5.58	99.84 ± 0.36
CD4⁺ Treg Fr Ⅲ	7.29 ± 2.85	29.80 ± 12.36	6.63 ± 2.50	99.86 ± 0.17

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