Immune Netw.  2003 Dec;3(4):287-294. 10.4110/in.2003.3.4.287.

Inhibition of Graft Versus Host Disease Using CD4+ CD25+ T Cells Induced with Interleukin-2 in Mismatched Allogeneic Murine Hematopoietic Stem Cell Transplantation

• Hyun JH
• Jeong DC
• Chung NG
• Park SJ
• Min WS
• Kim TG
• Choi BO
• Kim WI
• Han CW
• Kim HK

• KMID: 2054233
• DOI: http://doi.org/10.4110/in.2003.3.4.287

Abstract

BACKGROUND
In kidney transplantation, donor specific transfusion may induce tolerance as a result of some immune regulatory cells against the graft. In organ transplantation, the immune state arises from a relationship between the immunocompromised graft and the immunocompetent host. However, a reverse immunological situation exists between the graft and the host in hematopoietic stem cell transplantation (HSCT). In addition, early IL-2 injections after an allogeneic murine HSCT have been shown to prevent lethal graft versus host disease (GVHD) due to CD4+ cells. We investigated the induction of the regulatory CD4+CD25+ cells after a transfusion of irradiated recipient cells with IL-2 into a donor. METHODS: The splenocytes (SP) were obtained from 6 week-old BALB/c mice (H-2(d)) and irradiated as a single cell suspension. The donor mice (C3H/ He, H-2(k)) received 5x10(6) irradiated SP, and 5,000 IU IL-2 injected intraperitoneally on the day prior to HSCT. The CD4+CD25+ cell populations in SP treated C3H/He were analyzed. In order to determine the in vivo effect of CD4+CD25+ cells, the lethally irradiated BALB/c were transplanted with 1x10(7) donor BM and5x10(6) CD4+CD25+ cells. The other recipient mice received either 1x10(7) donor BM with 5x10(6) CD4+ CD25- cells or the untreated SP. The survival and GVHD was assessed daily by a clinical scoring system. RESULTS: In the MLR assay, BALB/c SP was used as a stimulator with C3H/He SP, as a responder, with or without treatment. The inhibition of proliferation was 30.0 13% compared to the control. In addition, the MLR with either the CD4+CD25+ or CD4+CD25- cells, which were isolated by MidiMacs, from the C3H/He SP treated with the recipient SP and IL-2 was evaluated. The donor SP treated with the recipient cells and IL-2 contained more CD4+CD25+ cells (5.4+/-1.5%) than the untreated mice SP (1.4+/-0.3%)(P<0.01). There was a profound inhibition in the CD4+CD25+ cells (61.1+/-6.1%), but a marked proliferation in the CD4+CD25- cells (129.8+/-65.2%). Mice in the CD4+CD25+ group showed low GVHD scores and a slow progression from the post-HSCT day 4 to day 9, but those in the control and CD4+CD25- groups had a high score and rapid progression (P<0.001). The probability of survival was 83.3% in the CD4+CD25+ group until post-HSC day 35 and all mice in the control and CD4+CD25- groups died on post-HSCT day 8 or 9 (P=0.0105). CONCLUSION: Donor graft engineering with irradiated recipient SP and IL-2 (recipient specific transfusion) can induce abundant regulatory CD4+CD25+ cells to prevent GVHD.

Keyword

CD4+ CD25+ T cells; interleukin-2; recipient specific transfusion; GVHD

MeSH Terms

Animals
Graft vs Host Disease*
Hematopoietic Stem Cell Transplantation*
Hematopoietic Stem Cells*
Humans
Interleukin-2*
Kidney Transplantation
Mice
Organ Transplantation
T-Lymphocytes*
Tissue Donors
Transplants*
Interleukin-2