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In vitro amplification of CD4(+) CD25(+) regulatory T cells and identification of amplified T cell immunosuppressive function / 中国实验血液学杂志
Journal of Experimental Hematology ; (6): 1266-1270, 2013.
Article in Chinese | WPRIM | ID: wpr-283940
ABSTRACT
This study was purposed to compare the effect of 3 different cell components for expanding CD4(+) CD25(+) Treg in vitro, and identify their immunosuppressive function. CD4(+) T cells, CD4(+) CD25(-)T cells and CD4(+) CD25(+)T cells were isolated from mouse splenocytes by MACS and then expanded in vitro. Phenotype of the T cell lines and expression of the FOXP3 was determined by flow cytometry. The inhibitory effect of expanded CD4(+) CD25(+) T cells on CD4(+) CD25(-)T cells was tested by MLR method. The results showed that the Treg cells from all the three groups were expanded significantly after culture for 2 weeks. In the CD4(+) T cells group, the proliferation rate was (77.8 ± 5.32) folds with a percentage of Treg cells increasing from (6.61 ± 1.00)% to (15.33 ± 1.31)%. The proliferation rate in the CD4(+) CD25(-) T cells group was (95.20 ± 7.67) folds, with the percentage of CD4(+) CD25(+) T cells raising from (0.37 ± 0.13)% to (9.84 ± 0.98)%. The proliferation rate in the CD4(+) CD25(+) T cells group was (41.20 ± 6.92) folds, the proportion of Treg cells decreased from (86.75 ± 1.25)% to (85.32 ± 1.62)%, and the expression of Foxp3 decreased from (76.92 ± 1.72)% to (75.33 ± 2.11)% during the culture, there were not significant differences in the cell purity and the expression of Foxp3, compared with pre-amplification. The inhibitory test showed that the expanded CD4(+) CD25(+) T cells could inhibit the proliferation of CD4(+) CD25(-) T cells in vitro in a cell dose-dependent manner. It is concluded that the amplification of CD4(+) CD25(+) Treg cells is successful in vitro, especially in the CD4(+) CD25(+) T cells group, the cell purity and Foxp3 gene is not obviously changes after amplification.
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Full text: Available Index: WPRIM (Western Pacific) Main subject: Cells, Cultured / T-Lymphocytes, Regulatory / Lymphocyte Culture Test, Mixed / Cell Biology / Cell Proliferation / Allergy and Immunology / Forkhead Transcription Factors / Interleukin-2 Receptor alpha Subunit / Flow Cytometry / Metabolism Type of study: Diagnostic study / Prognostic study Limits: Animals Language: Chinese Journal: Journal of Experimental Hematology Year: 2013 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Cells, Cultured / T-Lymphocytes, Regulatory / Lymphocyte Culture Test, Mixed / Cell Biology / Cell Proliferation / Allergy and Immunology / Forkhead Transcription Factors / Interleukin-2 Receptor alpha Subunit / Flow Cytometry / Metabolism Type of study: Diagnostic study / Prognostic study Limits: Animals Language: Chinese Journal: Journal of Experimental Hematology Year: 2013 Type: Article