A Case of Biliary Cast Syndrome in a Non-liver Transplatation Patients / 대한소화기내시경학회지

The development of total biliary casts is very unusual, and especially in patients who have not undergone liver transplantation. There are only a few reports of total biliary casts in non-liver transplantation patients who have antiphospholipid antibody syndrome, B-cell non-Hodgkin's lymphoma, cholecystectomy or allogenic hematopoietic stem cell transplantation. Here we present the case of a previously well 77-year-old man who developed a total biliary casts without any risk factors and there was no obvious liver insult. The casts were managed endoscopically.

The Effect of Intravenous Immunoglobulin on Hyperacute and Acclerated Rejection in Heart Transplantation of the Rat / 대한이식학회지

Hyperacute or acute accelerated rejection caused by preformed antibody in sensitized patients resulted in increased waiting period and complicated posttransplant hospital course. Intravenous immunoglobulin (IVIG) has known to have anti cytotoxic effect by blocking the anti HLA antibody. PURPOSE: We investigated the effect of IVIG on hyperacute and acclerated rejection of the heart graft in the presensitized rat. METHODS: Recipients (Wistar) were sensitized from repeated allo (Lewis) skin graft and followed by heterotopic allo cardiac transplantation. A guinea pig was used for the xenotransplantation model. IVIG (Green Cross kappa, 400 mg/kg in allotransplantation, 800 mg/kg in xenotransplantation) was given just before heart transplantation. Graft survival and donor specific IgG, IgM and complement were measured. RESULTS: Graft survival was 7.2 days in non sensitized allogenic heart transplantation (n=9), 1.3 days in sensitized allogenic recipients (n=7). Graft survival was prolonged from 1.3 days to 4.4 days with IVIG treatment (n=5). As for xenogenic transplantation, graft survival was prolonged from 30 min to 7.4 hr with IVIG treatment (n=5). Donor specific IgG and IgM and complement increment were blocked by IVIG during the IVIG treatment. Donor specific IgG and Ig M and complement were increased after the cessation of IVIG treatment. CONCLUSION: IVIG was able to prolong the graft survival of the sensitized allograft and xenograft. Suppression of the donor specific IgG, IgM and complement might be one of the underlying mechanisms. A further studies have to follow to clarify the more detailed mechanism.

Study for the Improvement of Early Implantation and Long Term Graft Survival in Pancreatic Islet Cell Transplantation by Induction of Angiogenesis with Gene Transfection of Vascuar Endothelial Growth Factor / 대한이식학회지

PURPOSE: Transplantation of pancreas islet has been worldwidely studied as a one of therapeutic modalities to achieve the insulin independence. We studied whether the expression of vascular endothelial growth factor (VEGF) on pancreas islets with liposomal VEGF gene transfer could improve the efficacy of early implantation and long term graft survival in pancreatic islet cell transplantation. METHODS: Syngenic pancreas islets were transplanted beneath the renal capsule. Islets were transfected with plasmid VEGF c-DNA using cationic liposome DMRIE-C. Glucose metabolism and histologic findings were compared between the groups transplanted with VEGF DNA containing islets (n=5) and the control group with (n=5) or without (n=4) local recombinant VEGF adminstration during islet transplant. RESULTS: Glucose was controlled at 5.5 days after transplantation in control group without r-VEGF adminstration, at 4 days in group with recombinant VEGF adminstration, and at 6.6 days in group with VEGF DNA transfected islets. Euglycemia was maintained over 150 days in control group. However, graft failure was developed in 22 days after transplantation in group with VEGF DNA transfected islet. Histologically there were severe infiltrations of neutrophil and lymphocyte in VEGF DNA transfected grafts from 5 days after transplantation. CONCLUSION: Although VEGF could be a favorable angiogenic factor in pancreas islet transplantation, VEGF expression following VEGF DNA transfection into islets could not increase the graft survival due to inflammatory process. More investigations are needed to clarify the mechanism on destructive process of islets after gene transfection into islets, and another approaches to get the effect of gene transfection should be followed.

The Implication of Monocyte/Macrophage into the Graft Following Heart Transplantation in Rat

BACKGROUND: In organ transplantation, the cellular immune reaction, namely T-cell immunity, plays a major role in rejecting the graft. While T & B cell activities in organ transplantation have been studied extensively, monocytes/macrophages have not because of their a minor role in innate immunity. Monocytes act as immunologically active cells in several aspects in organ transplantation, such as antigen-presenting cells, cells releasing many substance, such as IL-1, IL-2, TNF-alpha, and many growth factors, and cells phagocytosing foreign antigens and tissues in the effector phase of immune reaction. METHODS: We attempted to study the role of monocytes/ macrophages in graft rejection following allogenic organ transplantation in rodents. RESULTS: While graft survivals following a cardiac allograft were more then 100 days in all the singenic Wistar to Wistar transplants, the graft survival for Lewis to Wistar allografts were 7 to 12 days with a mean of 9.2 days. In the histology of the transplanted hearts, cellular infiltration developed from posttransplantation day 1, and all the histologic findings, such as myocardial ischemia, interstitial bleeding, and endocardial changes, were more progressive around the days of graft rejection. Macrophage infiltration analyzed by immunohistochemstry using the spectific antibody ED1, was noticed from postoperative day 1, and the macrophages were distributed all through the layer of the heart. In the study on the intragraft monokine gene by using RT-PCR, mRNA of IL-1 expressed on day 1 and reappearedon day 7. mRNA of TNF-alphaexpressed on day 3 and MCP-1 on day 1. All the monokine gene expressions progressed up to the days of rejection. CONCLUSION: From these results showing the concurrent pattern of cell infiltration and intragraft cytokine gene expression of monocytes/macrophages with the lymphocyte, we suggest that intervention of monocytes in organ transplantation may prolong graft survival with or without the anti T cell strategy.

In Vitro Analysis of Co-cultured Pancreatic Islet with Lymphocyte / 대한이식학회지

BACKGROUND: Transplantation of Pancreatic islet represents one of the most exciting treatment modalities for Type I diabetes mellitus. To achieve better graft survival, it is important to protect the graft from alloantigen-specific immune response. It was emphasized that islets, as with other forms of cellular transplants, have the potential advantage of being immunologically altered before transplantation, resulting in tolerance to the host without using long-term, nonspecific immunosuppression. The mixed islet-lymphocyte culture is an excellent tool to evaluate the immunogenicity of a pancreatic islets. Therefore, we co-cultured pancreatic islet and lymphocyte to investigate cytokine gene expression from the lymphocyte, and to investigate pancreatic islet viability and functional changes after co-culture. MATERIALS AND METHODS: Lewis rat islets were purified from pancreas by collagenase type XI and dextran gradient method. Afterwards, Lewis rat islets were co-cultured as a stimulator and Wistar rat lymphocyte as a reponder. As a control group lymphocyte alone and islet alone were cultured in a same condition. For estimating islets viability, we counted islet viability as an IEQ (Islet equivalent). For evaluation of islets function, insulin release assay was perfomed by RIA under the glucose challenge. used by RIA. We studied cytokines gene expression of cultured cells by reverse transcriptase polymerase chain reaction (RT-PCR). RESULT AND CONCLUSION: 1) Islet yield was 1229.8 420.1 per rat, and counted as 2615.4 548.2 IEQ per rat. 2) Islet viability was decreased gradually with the lapse of time, more rapid in allogenic co-culture group than islet alone or isogenic co-culture group. 3) Release of insulin increased until day 3, and then decreased gradually. Insulin release was positively correlated with glucose gradient. The amount of released insulin was greater in co-cultured group than islet alone group. 4) Interleukine-2 and interferron-gamma gene expression increased in allo co-culture group, however transforming growth factor-beta gene expression was not affected by co-culture.

In Vitro Analysis of Co-cultured Pancreatic Islet with Lymphocyte / 대한이식학회지

BACKGROUND: Transplantation of Pancreatic islet represents one of the most exciting treatment modalities for Type I diabetes mellitus. To achieve better graft survival, it is important to protect the graft from alloantigen-specific immune response. It was emphasized that islets, as with other forms of cellular transplants, have the potential advantage of being immunologically altered before transplantation, resulting in tolerance to the host without using long-term, nonspecific immunosuppression. The mixed islet-lymphocyte culture is an excellent tool to evaluate the immunogenicity of a pancreatic islets. Therefore, we co-cultured pancreatic islet and lymphocyte to investigate cytokine gene expression from the lymphocyte, and to investigate pancreatic islet viability and functional changes after co-culture. MATERIALS AND METHODS: Lewis rat islets were purified from pancreas by collagenase type XI and dextran gradient method. Afterwards, Lewis rat islets were co-cultured as a stimulator and Wistar rat lymphocyte as a reponder. As a control group lymphocyte alone and islet alone were cultured in a same condition. For estimating islets viability, we counted islet viability as an IEQ (Islet equivalent). For evaluation of islets function, insulin release assay was perfomed by RIA under the glucose challenge. used by RIA. We studied cytokines gene expression of cultured cells by reverse transcriptase polymerase chain reaction (RT-PCR). RESULT AND CONCLUSION: 1) Islet yield was 1229.8 420.1 per rat, and counted as 2615.4 548.2 IEQ per rat. 2) Islet viability was decreased gradually with the lapse of time, more rapid in allogenic co-culture group than islet alone or isogenic co-culture group. 3) Release of insulin increased until day 3, and then decreased gradually. Insulin release was positively correlated with glucose gradient. The amount of released insulin was greater in co-cultured group than islet alone group. 4) Interleukine-2 and interferron-gamma gene expression increased in allo co-culture group, however transforming growth factor-beta gene expression was not affected by co-culture.

Immunosuppressive Effects of Tautomycetin on T Cells / 대한면역학회지

T cell activation is a critical event for initiation and regulation of immune responses and inhibitors of such signaling pathways are clinically useful for the treatment of patients received allogratt and autoimmune disease. In the course of screening soil microorganisms from the forest of Cheju island in Korea for new immunosuppressive agent, one of Streptomyces species (CK-95441) was found to produce a new immunosuppressant, tautomycetin which also had antifungal activity. Tautomycetin showed the inhibition of T cell proliferation in murine mixed lymphocyte reaction (MLR) and T cell activation induced by concanavalin A. Tautomycetin also blocked the induction of IL-2 gene expression which was examined in Jurkat TAg cell line in which multiple NFAT-binding sites and minimal IL-2 promoter drive the production of B-galactosidase. Also, the level of inhibition in activation-induced IL-2 receptor expression by tautomycetin was greater than those by cyclosporin A measured by flow cytometry. But, Fas ligand-induced apoptosis in Jurkat cells was unaffected by tautomycetin which was measured by DNA fragmentation assay. These results suggested that tautomycetin will be able to be used as a potent immunosuppressive drug following organ transplantation.

Okadaic Acid, RK682 and Calyculin Modulate TcR - Mediated Signaling Events / 대한면역학회지

The T cell antigen receptor (TcR) in combination with costimulatory signals triggered by accessory molecules present on the surface of the antigen-presenting cells (APC) regulates the activation and growth of T lymphocytes. Calyculin A and Okadaic acid is known to be an inhibitor of serine/threonine phosphatase and RK-682 specifically blocks functions of tyrosine phosphatase. To investigate roles of these inhibitors in TcR-mediated signaling cascade, chimeric molecule CD8-5 which contains the extracellular and transmembrane domains of the human CD8a molecule and the cytoplasmic tail of TcR 5 chain were stably expressed in Jurkat cell line. CD8-5 chimeric protein induced tyrosine phosphorylation of various cytoplasmic substrates and IL-2 gene expression in a NFAT dependent manner by stimulation with anti-CD8 mAb OKT8 as seen in TcR stimulation. When CD8-5 transfectants were preincubated with Okadaic acid, Calyculin or RK682, they differentially affected tyrosine phosphorylation of signaling mediators including CD8-5 molecule. When Jurkat Tag cell line was used where SV40 T antigen is stably expressed and the expression of p-galactosidase is driven by the multiple NFAT binding sites plus minimal IL-2 promoter, these phosphatase inhibitors -RK682, Calyculin A, Okadaic acid- effectively inhibited IL-2 gene expression at the concentration of 1.2832 x 10 ' M, 3.9924 x 10 M, 7.2707 x 10 M respectively. These results suggested that Okadaic acid, Calyculin or RK682 modulate TcR-proximal as well as TcR-distal signaling events during T cell activation.

A Novel Cell Line for Screening of Immunosuppressor Specific to T Lymphocytes / 대한면역학회지

The systematic study of products from bacteria and fungi has led to the development of two immunosuppressive drugs, cyclosporin A and FK 506 (tacrolimus) which are useful to suppress adaptive immune responses to the grafted tissue. However, they affect all immune responses indiscriminately and are both toxic to kidneys and other organs. To facilitate the development of immunosuppressor to block the T cell receptor (TcR)-mediated signaling cascade specifically, a novel Jurkat T cell transfectants, JK NFAT-SEAP were generated in which the expression of the secreted alkaline phosphatase (SEAP) is driven by the multiple NFAT binding sites plus minimal IL-2 promoter. Upon stimulation with ionomycin or anti-TcR mAb OKT3 in the presence of PMA, these transfectants secreted high level of SEAP into the medium, which was conveniently analyzed by SEAP analysis. The secretion of SEAP was effectively inhibited by cyclosporin A or FK 506 at the concentration of [10 ' ug/ml], [10 ug/ml] respectively. JK NFAT-SEAP transfectants will provide two major advantages for the development of a novel immunosuppressor. First, analysis of SEAP secreted into the culture medium by SEAP analysis enables us to test a large number of samples within a short period of time. Second, Usage of IL-2 promoter for the expression of SEAP makes us identify bioproducts to target specifically on TcR-mediated signaling pathway.