[Isolation and function analysis of rat CD4+ CD25+ regulatory T cells].
OBJECTIVE
To investigate the isolation method and to analyze the function of rat CD4(+) CD25(+) regulatory T cells.


METHODS
Lymphocytes were isolated from the rat spleens and then CD4(+) CD25(+) T cells were sorted by magnetic bead cell sorting (MACS) system. The purity and Foxp3 expression of CD4(+) CD25(+) T cells were analyzed by flow cytometry(FCM) and RT-PCR, respectively. The suppressive effect of CD4(+) CD25(+) T cells on the proliferation of CD4(+) CD25(-) T cells was analyzed by mixed lymphocyte reaction. IL-2, IFN-gamma and IL-10 levels in culture supernatant were detected by ELISA.


RESULTS
The purity of CD4(+) CD25(+) T cells sorted by MACS was 86%-93%. The CD4(+) CD25(+) T cells could specifically express the Foxp3 gene as compared with CD4(+) CD25(-) T cells. In vitro CD4(+) CD25(+) T cells could suppress the proliferation of CD4(+) CD25(-) T cells and IFN-gamma, IL-2 production, but they themselves could secrete IL-10.


CONCLUSIONS
We established an effective procedure for enrichment of CD4(+) CD25(+) regulatory T cells by MACS with satisfactory cell purity, viability and function. CD4(+) CD25(+) T cells can suppress CD4(+) CD25(-) T cells and specifically express Foxp3 gene.
Lu
Ling
L
Center of Liver Transplantation, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. lvling_vip@yahoo.com.cn
Zhang
Feng
F
Wang
Xue-hao
XH
Pu
Li-yong
LY
Yao
Ai-hua
AH
Yu
Yue
Y
D000818:Animals; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D016207:Cytokines / Q000378:metabolism; D005434:Flow Cytometry; D051858:Forkhead Transcription Factors / Q000378:metabolism; D005786:Gene Expression Regulation; D053645:Interleukin-2 Receptor alpha Subunit / Q000276:immunology / Q000378:metabolism*; D008280:Magnetics; D008297:Male; D008863:Microspheres; D051381:Rats; D011917:Rats, Inbred Lew; D050378:T-Lymphocytes, Regulatory / Q000166:cytology* / Q000276:immunology / Q000378:metabolism*
D016207:Cytokines; D051858:Forkhead Transcription Factors; C500522:Foxp3 protein, rat; D053645:Interleukin-2 Receptor alpha Subunit