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Suppression of alloimmunity in mice by regulatory T cells converted with conditioned media.
J Surg Res 2011; 171(2):797-806JS

Abstract

BACKGROUND

Compared with rare CD4+CD25+ regulatory T cells (Tregs), CD4+CD25- T cells are abundant in peripheral blood. Studies have demonstrated that tumors can convert naive CD4+CD25- T cells into CD4+CD25+Foxp3+ Tregs; however, the potential application of the converted Tregs in transplant medicine has not well demonstrated.

MATERIALS AND METHODS

CD4+CD25- T cells isolated cultured for 5 d in various combinations of cell culture medium and conditioned medium from RenCa cells. Foxp3 levels were determined by flow cytometry and real-time polymerase chain reaction. BALB/c mouse mononuclear cells (responder) combined with different ratios of the converted CD4+CD25+ Tregs were co-cultured with inactivated C57BL/6 mononuclear cells (stimulator) in one-way mixed lymphocyte reaction (MLR). In addition, the converted Tregs were transferred into a mouse skin transplant model, and graft histology, survival time, and delayed-type hypersensitivity were assessed.

RESULTS

CD4+CD25- T cells cultured in media supplemented with increasing concentrations of RenCa conditioned medium were partially converted into CD4+CD25+ Tregs; conversion increased as the percentage of conditioned medium increased. Moreover, the converted CD4+CD25+ Tregs possessed characteristics similar to those of naturally occurring Tregs, including Foxp3 expression and the ability to suppress T cell proliferation in one-way MLR in a concentration-dependent manner. Immunosuppressive effects of the converted CD4+CD25+ Tregs were also similar to those of naturally occurring Tregs in vivo, suppressing allograft rejection and prolonging survival time of allogeneic skin grafts following adoptive transfer in mice (P<0.05).

CONCLUSION

Our findings indicate that CD4+CD25- T cells cultured with conditioned media from tumor cells may be useful for obtaining sufficient numbers of Tregs, which may help suppress acute rejection and prolong graft survival.

Authors+Show Affiliations

Department of Organ Transplantation, First Affiliated Hospital, Sun Yat-sen University, Guangzhou City, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20538297

Citation

Teng, Lichen, et al. "Suppression of Alloimmunity in Mice By Regulatory T Cells Converted With Conditioned Media." The Journal of Surgical Research, vol. 171, no. 2, 2011, pp. 797-806.
Teng L, Liu L, Su Y, et al. Suppression of alloimmunity in mice by regulatory T cells converted with conditioned media. J Surg Res. 2011;171(2):797-806.
Teng, L., Liu, L., Su, Y., Yuan, X., Li, J., Fu, Q., ... Wang, C. (2011). Suppression of alloimmunity in mice by regulatory T cells converted with conditioned media. The Journal of Surgical Research, 171(2), pp. 797-806. doi:10.1016/j.jss.2010.03.044.
Teng L, et al. Suppression of Alloimmunity in Mice By Regulatory T Cells Converted With Conditioned Media. J Surg Res. 2011;171(2):797-806. PubMed PMID: 20538297.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Suppression of alloimmunity in mice by regulatory T cells converted with conditioned media. AU - Teng,Lichen, AU - Liu,Longshan, AU - Su,Yajuan, AU - Yuan,Xiaopeng, AU - Li,Jun, AU - Fu,Qian, AU - Chen,Siyang, AU - Wang,Changxi, Y1 - 2010/04/15/ PY - 2009/10/23/received PY - 2010/03/02/revised PY - 2010/03/19/accepted PY - 2010/6/12/entrez PY - 2010/6/12/pubmed PY - 2012/1/10/medline SP - 797 EP - 806 JF - The Journal of surgical research JO - J. Surg. Res. VL - 171 IS - 2 N2 - BACKGROUND: Compared with rare CD4+CD25+ regulatory T cells (Tregs), CD4+CD25- T cells are abundant in peripheral blood. Studies have demonstrated that tumors can convert naive CD4+CD25- T cells into CD4+CD25+Foxp3+ Tregs; however, the potential application of the converted Tregs in transplant medicine has not well demonstrated. MATERIALS AND METHODS: CD4+CD25- T cells isolated cultured for 5 d in various combinations of cell culture medium and conditioned medium from RenCa cells. Foxp3 levels were determined by flow cytometry and real-time polymerase chain reaction. BALB/c mouse mononuclear cells (responder) combined with different ratios of the converted CD4+CD25+ Tregs were co-cultured with inactivated C57BL/6 mononuclear cells (stimulator) in one-way mixed lymphocyte reaction (MLR). In addition, the converted Tregs were transferred into a mouse skin transplant model, and graft histology, survival time, and delayed-type hypersensitivity were assessed. RESULTS: CD4+CD25- T cells cultured in media supplemented with increasing concentrations of RenCa conditioned medium were partially converted into CD4+CD25+ Tregs; conversion increased as the percentage of conditioned medium increased. Moreover, the converted CD4+CD25+ Tregs possessed characteristics similar to those of naturally occurring Tregs, including Foxp3 expression and the ability to suppress T cell proliferation in one-way MLR in a concentration-dependent manner. Immunosuppressive effects of the converted CD4+CD25+ Tregs were also similar to those of naturally occurring Tregs in vivo, suppressing allograft rejection and prolonging survival time of allogeneic skin grafts following adoptive transfer in mice (P<0.05). CONCLUSION: Our findings indicate that CD4+CD25- T cells cultured with conditioned media from tumor cells may be useful for obtaining sufficient numbers of Tregs, which may help suppress acute rejection and prolong graft survival. SN - 1095-8673 UR - https://www.unboundmedicine.com/medline/citation/20538297/Suppression_of_alloimmunity_in_mice_by_regulatory_T_cells_converted_with_conditioned_media_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-4804(10)00243-X DB - PRIME DP - Unbound Medicine ER -