The aim of this study was to evaluate microchimerism after human liver transplantation (LT). This study included 13 female recipients who received hepatic allograft from male donors at Asan Medical Center. A nested PCR specific for Y-chromosome gene (DYZ3) was used to analyze the small number of male cells in the peripheral blood mononuclear cells of the female recipients. Microchimerism was observed in 6 of 13 recipients and 16 out of 35 samples. Only 3 patients showed microchimerism 3 months after LT. There was no statistical difference between the presence of microchimerism and clinical findings such as type of donor, type of immunosuppression, episode of rejection and age of recipient. This study did not show any clinical relevance of microchimerism and further larger study are needed to confirm the results.
Adolescence ; Adult ; Animal ; Child ; Child, Preschool ; Female ; Human ; Infant ; Liver Transplantation* ; Lymphocytes/immunology* ; Male ; Transplantation Chimera/immunology* ; Transplantation Chimera/genetics ; Y Chromosome

The aim of this study was to evaluate microchimerism after human liver transplantation (LT). This study included 13 female recipients who received hepatic allograft from male donors at Asan Medical Center. A nested PCR specific for Y-chromosome gene (DYZ3) was used to analyze the small number of male cells in the peripheral blood mononuclear cells of the female recipients. Microchimerism was observed in 6 of 13 recipients and 16 out of 35 samples. Only 3 patients showed microchimerism 3 months after LT. There was no statistical difference between the presence of microchimerism and clinical findings such as type of donor, type of immunosuppression, episode of rejection and age of recipient. This study did not show any clinical relevance of microchimerism and further larger study are needed to confirm the results.
Adolescence ; Adult ; Animal ; Child ; Child, Preschool ; Female ; Human ; Infant ; Liver Transplantation* ; Lymphocytes/immunology* ; Male ; Transplantation Chimera/immunology* ; Transplantation Chimera/genetics ; Y Chromosome

OBJECTIVETo explore the feasibility of transplanting the skin from chimeric rats to rabbits.

METHODSChimeric rats were produced by transplanting the haematopoietic stem cells (HSCs) from rabbit marrows into fetal rats in uterus and followed by injecting the HSCs into the livers of the rats at newborn stage. After six weeks, the skin from chimeric rats was transplanted to the rabbits. In group A, the skin grafts from chimeric rat donors were transplanted to the HSCs donating rabbits, with the skin from non-chimeric rat to normal rabbits were used as control. In group B, the skin grafts from chimeric and non-chimeric rats were transplanted to the HSCs donating rabbits at the same time. Gross observation and the surviving time of heterogenic-skin graft were observed. The wound healing time was also recorded.

RESULTSIn group A, the surviving time and the wound healing time of non-chimeric grafts were (9.3 +/- 1.8) days and (20.9 +/- 2.1) days, respectively, while those in chimeric grafts were (15.1 +/- 2.6) and (18.5 +/-1.3) days, respectively. In group B, the surviving time and the wound healing time of non-chimeric grafts were similar to those of group A. Compared with those in non-chimeric grafts, the surviving time of chimeric grafts in both groups were prolonged (P < 0.01), and the wound healing time shortened (P < 0.05 or 0.01). Most of the wounds healed quickly after rejection of chimeric grafts, while the wounds with non-chimeric grafts were re-opened with exudation and some necrotic tissue.

CONCLUSIONImmunologic tolerance for skin graft can be induced by the skin from chimeric donors, which can prolong the surviving time of skin grafts and shorten the wound healing time.

Animals ; Immune Tolerance ; Male ; Myeloid Progenitor Cells ; transplantation ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; immunology ; Transplantation Chimera ; immunology ; Transplantation, Homologous

Child ; Graft Survival ; HLA Antigens ; genetics ; immunology ; Hematopoietic Stem Cell Transplantation ; Humans ; Polymerase Chain Reaction ; Tissue Donors ; Transplantation Chimera ; genetics ; Transplantation Conditioning ; methods ; Transplantation, Homologous ; immunology

The aim of study was to explore the feasibility of quantitative chimerism analysis of regulatory T (Treg) cells using immune sorting coupling short tandem repeat (STR) method. 14 sets of artificial chimera samples were prepared by mixed lymphocytes according to different proportion. The CD4(+)CD25(+) Treg cells were harvested by negative and positive selection of immunomagnetic beads, then the STR polymorphisms of 16 loci in sorted Treg cells was analyzed. The results showed that the DNA amount extracted from sorted Treg cells was fit for STR detection. All STR alleles specific for recipient or donor could be detected and the quantitative results were consistent with theoretic values in over 10% recipient chimeras. But only partial recipient alleles could be detected and the quantitative results were different from theoretic values in less then 1% recipient chimeras. It is concluded that a quantitative chimerism analysis of Treg cell based on immune sorting is established. The sensitivity and accuracy for chimera detection are 1% to 10%, and this method can be used to monitoring hematopoietic chimerism following allogeneic hematopoietic stem cell transplantation.
Chimerism ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Immunomagnetic Separation ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Transplantation Chimera ; genetics ; immunology

Transplantation would be the only way to cure the end-stage organ failure involving heart, lung, liver, kidney and pancreas. The replacement of the parts of the body damaged to lose its function or lost to trauma must be a dream of human-being. Human history is replete with chimeras, from sphinxes to mermaids, making one wonder if the ancients might actually have dreamed of what now is called 'xenotransplantation'. In the 20th century, the transplantation of organs and tissues to cure disease has become a clinical reality. The development in the fields of surgical techniques, physiology and immunology attributed to the successful transplantation in human. In the center of the successful transplantation lies the progress in understanding the cellular and molecular biology of immune system which led to the development of immunosuppressive drugs and the invention of the concept of immunological tolerance. The mandatory side effects of immunosuppressive drugs including infection and cancer forced us to search alternative approaches along with the development of new immunosuppressive agents. Among the alternative approaches, the induction of a state of immunologic tolerance would be the most promising and the most generic applicability as a future therapy. Recent reports documenting long-term graft survival without immunosuppression suggest that tolerance-based therapies may become a clinical reality. Last year, we saw the epoch making success of overcoming hyperacute rejection in porcine to primate xenotransplantation which will lead porcine to human xenotransplantation to clinical reality. In this review, I dare to summarize the development of transplantation immunology from the perspective of history.
Allergy and Immunology ; Chimera ; Graft Survival ; Heart ; Humans ; Immune System ; Immunosuppression ; Immunosuppressive Agents ; Inventions ; Kidney ; Liver ; Lung ; Molecular Biology ; Pancreas ; Physiology ; Primates ; Transplantation Immunology* ; Transplantation, Heterologous

OBJECTIVETo investigate whether the fetal immune tolerance induction could replace the HLA typing for hematopoietic stem cell transplantation.

METHODSImmune tolerance of SD rats was induced by injecting host Wistar rats peripheral blood mononuclear cells into yolk sac of the embryo, afterward the mature male offsprings were used as donor. The host female recipients received lethal dose irradiation and bone marrow transplantation(BMT). The Wistar rats transplanted with bone marrow from donor and unrelated SD rats as well as the rats which received radiation alone were used as control. The survival, histopathologically GVHD, the mental status, food and water intake, coat characteristics, activities were observed. Forty days after BMT, autologous and allogenous skin transplantation between donor and recipient rats was performed to observe the engraftment of solid organ.

RESULTSThe survival of the rats received bone marrow grafts from the immune tolerant donor was significantly longer than that of control groups (30 day survival rates were 86.7%, 6.7%, 0%, and 0% respectively), and there was no histopathologically GVHD observed, while in the sham group, the manifestations of GVHD was clearly visible. The skin engraftment rate between the host and the immune tolerant donor was significantly higher than that among non-related rats (84.6% and 0% respectively).

CONCLUSIONThe induction of immune tolerance in embryo can overcome the HLA barrier and provide a good donor for hematopoietic stem cell and solid organ transplantation.

Animals ; Embryo, Mammalian ; immunology ; Female ; Graft vs Host Disease ; immunology ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; Immunosuppression ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Transplantation Chimera

This study was aimed to explore the feasibility of transplanting human cord blood stem cells (HSC) into pre-immune fetal and neonatal pigs, and to investigate the self-renewal of HSC in the recipient pigs. The fetus and neonate were manipulated in sterile separated room and human donor cells were injected into fetus via fetus muscle or umbilical vein (dissectted womb) or into neonate via umbilical vein before cutting it. Human CD45(+) cells s were detected by labeling with human anti-CD45 antibody and analyzed by fluorescence activated cell sorting (FACS). The results showed that tested pigs developed as well as control and a definite proportion of human cells existed in peripheral blood of chimeric pig on day 60 after transplantation. In conclusion, the fetus and neonate pigs can tolerate a definite proportion of human antigens, and to establish the human/pig model of hematopoietic chimerism is possible.
Animals ; Animals, Newborn ; Cord Blood Stem Cell Transplantation ; methods ; Fetus ; Flow Cytometry ; Humans ; Leukocyte Common Antigens ; blood ; Models, Animal ; Pilot Projects ; Swine ; Transplantation Chimera ; blood ; immunology ; Transplantation, Heterologous

OBJECTIVETo investigate the effect of allogene hepatic nonparenchymal cell (NPC) on the survival of grafted skin in mice and its underlying mechanism.

METHODSSixty-five C(3)H and fifty-eight C(57)BL/6 mice were employed in the study. Twenty C(3)H mice were used as skin donor and forty as the source of hepatic NPC. The rest five served as the stimulators of mixed lymphocyte culture (MLC) before and on the 7th, 18th, 30th, and 60th day after NPC infusion with 1 at each time point. MLC was determined and expressed as count per minute (CPM). Fifty-eight C(57)BL/6 mice were further divided into experimental (E, n = 50) and control groups (C, n = 8). The mice in C group only underwent skin grafting without NPC infusion. The mice in E group received with 2 x 10(7) NPC via caudal vein, followed by peritoneal injection of cytoxan (200 mg/kg) 48 hours later; They were grafted with skin donated from C(3)H mice 18 days after injections. The survival time of the mice in the two groups was observed. The serum levels of interleukin-4, chimera and MLC in the two groups were determined before and on 7th, 18th, 30th, 60th days after NPC infusion, and micro-chimera were aslo assessed on the 1st and 3rd day after NPC infusion. Five mice were sacrificed at each time point.

RESULTSThe survival time of skin graft in E group (70.0 +/- 17.2 day) was obviously longer than that in C group. The serum levels of IL-4, chimera in E group were increased gradually, while MLC response decreased gradually. The serum IL-4 level reached 251.5 +/- 11.0 ng/L and splenic chimera level to 26.30 +/- 1.04% on the 60th day after NPC infusion.

CONCLUSIONThe high levels of IL-4 and chimera might play important roles in inducing and maintaining immune tolerance.

Animals ; Female ; Graft Survival ; immunology ; Hepatocytes ; immunology ; Immune Tolerance ; Interleukin-4 ; metabolism ; Male ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Skin Transplantation ; immunology ; Surgical Flaps ; Transplantation Chimera ; Transplantation, Homologous ; immunology

BACKGROUNDNonmyeloablative allogeneic bone marrow transplantation has been used since the 1990s as a new hematological stem cell transplantation strategy for treating hematological diseases. The purpose of this study was to explore the graft-versus-leukemia (GVL) effects of donor lymphocyte infusions (DLIs) after nonmyeloablative allogeneic bone marrow transplantations, while assessing the declines in treatment-associated morbidity, mortality, and graft-versus-host disease (GVHD).

METHODSA total of 615 (H-2k) mice were injected with L615 tumor cells and received 500 cGy (60Co gamma-ray) irradiation three days later, followed by an allogeneic bone marrow transplantation (allo-BMT). The allo-grafts consisted of 3 x 10(7) bone marrow cells and 1 x 10(7) spleen cells from BALB/C (H-2d) donor mice. Two days after the allo-BMT, the recipient mice were given 200 mg/kg of cyclophosphamide. Subsequently, recipient mice were infused with either donor spleen cells (2 x 10(7)) on day 14 or 21, or donor spleen cells (5 x 10(7)) pretreated with hydrocortisone and cyclosporin A (CsA) in vitro on day 14 post-BMT.

RESULTSThe median survival time of mice that received DLI on day 21 and pretreated DLI on day 14 post-BMT was longer than that of controls and the day 14 DLI group (P < 0.01). No evidence of severe GVHD was observed in the day 21 DLI group nor in the day 14 treated DLI group. Mixed chimerism was confirmed in the day 14 DLI group, the day 14 treated DLI group, and the day 21 DLI group on the thirteenth day post-transplantation; full donor chimerism was observed two weeks after DLI.

CONCLUSIONDonor lymphocyte infusion after nonmyeloablative bone marrow transplantation may reduce transplantation-associated morbidity and mortality while strengthening graft-versus-leukemia effects.

Animals ; Bone Marrow Transplantation ; immunology ; Cyclosporine ; pharmacology ; Female ; Graft vs Host Disease ; etiology ; Graft vs Leukemia Effect ; immunology ; Hydrocortisone ; pharmacology ; Lymphocyte Activation ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred BALB C ; Transplantation Chimera ; Transplantation, Homologous