Prevention From Murine Acute Graft-versus-host Disease By Recipient-derived Paired Immunoglobin-like Receptor B Lentivirus Transfected Dendritic Cells

Object:To establish a murine model of acute graft-versus-host disease (aGVHD) induced by allogeneic bone marrow transplantation (allo-BMT) for providing experimental studies of aGVHD and transplantation tolerance.Methods:All recipients female BALB/C (H-2d) mice were received total body irradiation 8.5Gy at 4 hours prior to transplantation. Bone marrow cells mixed with spleen cells of donors male C57BL/6J (H-2b) mice based upon different proportions (1:1 and 1:2) were injected into tail vein. The chimeras were evaluated as well as the recipients'survival time, clinical and pathologic manifestations of aGVHD.Result:All mice received allo-BMT with different ratios of bone marrow cells and spleen cells showed aGVHD evidence. However, there were significant differences in the incidence and severity of aGVHD. The group with co-transplantation of bone marrow cells and spleen cells in ratio of 1:2 showed typical clinical and pathological manifestations of aGVHD at a relatively concentrated time, and died within 14-24d. Mice in group infused with bone marrow cells and spleen cells in ratio of 1:1 showed insignificant aGVHD, and were all alived (4 weeks after allo-BMT).Conclusion:The group with co-transplantation of bone marrow cells and spleen cells in ratio of 1:2 showed more significant manifestations and more concentrated time of incident and death than the group that of 1:1. Thus, infusion of bone marrow cells and spleen cells in ration of 1:2 is perfect as to establish aGVHD murine model induced by allogeneic bone marrow transplantation for successoral experimental studies. Object:To explore the influence of recipient-derived dendritic cells induced by different methods infused after allogeneic bone marrow transplantation (allo-BMT) on acute graft-versus-host disease (aGVHD) and hematopoietic reconstitution of mice.Methods:Recipients were female BALB/c (H-2kd) mice, donors were male C57BL/6 (H-2kb) mice. Recipients'bone marrow cells were isolated sterilely. ImDCs were prepared by culturing BM cells with GM-CSF. IL-10-DCs were generated from BM cells cultured with GM-CSF and recombinant murine interleukin 10. PIR-B-DCs were obtained by Paired immunoglobin-like receptor B lentivirus transfected imDCs. The recipient mice were randomly divided into four groups, and received intravenous injections of various additional DCs from BALB/C mice (imDCs,IL-10-DCs,PIR-B-DCs) or PRMI 1640 at the time of BMS transplantation respectively. Incidence of GVHD, pathological lesion of liver,small intestine,skin, survival time and hematopoietic reconstitution in the recipients were observed after allo-BMT.Result:The mean survival time of the four groups (PIR-B-DCs group, IL-10-DCs group, imDCs group and BMT group) was (46.0±13.6) days,(36.4±13.0) days,(21.6±2.8) days and (17.4±3.6) days, p<0.01. The GVHD clinical score at 15 days after transplantation in the four groups was (5.28±0.27),(5.26±0.31),(2.46± 0.18) and (0.86±0.21),P<0.05. Histopathologic analyses of BMT group showed extensive lymphocytic infiltration of the liver, small intestine and skin, bile duct damaged, intestinal villus atrophy or denaturation. Samples from the liver and skin of a mouse did not show clinical or histological signs of GVHD following the injection of PIR-B-DCs.Conclusion:Recipient-derived paired immunoglobin-like receptor B lentivirus transfected dendritic cells infusion can mitigate the aGVHD so as to prolong survival time after allo-BMT in murine.