Dissertation > Medicine, health > Basic Medical > Medical Immunology

Immunity Associated Gene Expression Profile Analysis of Human Lymphocytes in Renal Transplantation by cDNA Microarray

Author ChenHai
Tutor ZuoZhiLian
School Second Military Medical University
Course Surgery
Keywords Kidney Transplantation Gene Expression Lymphocytes Reactive antibody Transplantation immunology Microarray Immunosuppressant Methylprednisolone
CLC R392
Type PhD thesis
Year 2004
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Currently, kidney transplants have become all ESRD (ESRD) patients the best choice for renal replacement therapy. While allograft rejection in organ transplantation is still the biggest problem facing the field. Facing kidney transplant rejection, including hyperacute rejection (HAR), accelerated rejection, acute rejection (AR) and chronic rejection (CR). Among them, acute rejection after renal transplantation is a major complication. In addition, it is also the result of chronic rejection or chronic allograft nephropathy (CAN), chronic graft dysfunction is an important risk factor. Acute and chronic rejection in organ transplantation an impact on short-term and long-term effects of the most important reason. In addition to acute and chronic rejection, kidney transplantation also face hyperacute rejection and the risk of accelerated rejection, these two are associated with immune rejection in highly sensitized about. Immune highly sensitized patients in which patients have higher levels of stored body reactive antibody (PRA). Currently under the PRA values ??sensitized patients were divided into mild (<10%), moderate sensitization (10% -50%) and highly sensitized (> 50%). High sensitivity in patients with a higher risk of transplant, intraoperative or postoperative hyperacute, accelerated, acute and chronic rejection episodes were high, often become transplant taboo. High sensitivity of the immune molecules mechanism is not clear, from the genetic level it is important to study the causes of both. In addition to rejection, long term use of immunosuppressive infection lead to graft recipients, cardiovascular complications, cancer, liver and kidney toxicity and other problems significantly increased chance. It is important to get the immunosuppressive drug efficacy and strike a balance between the risk to avoid immunosuppressed recipients insufficient or excessive immunosuppression. If you can reveal the genetic level effects of immunosuppressants target genes will be clinically more effective drugs to provide new theoretical and experimental basis. Transplant rejection is a complex process that is not yet fully understood. In order to better study of renal transplant rejection, allogeneic immune response in a comprehensive understanding of the basic processes and no clinical transplantation rejection performance of the immune response when it is necessary, is also important. After transplantation immune response genes involved in more than one, a very complex process, T cells and antigen presenting cells (APC) in which activation plays an important role. With the rapid molecular immunology research development, people gradually realized that the body's immune response to the transplanted kidney and its regulation strictly controlled by different genes. Studies suggest that in T cell activation process, the activation and expression of genes involved in about 70 species, divided into three categories according to the function of these genes: Cellular oncogenes, cytokine / cytokine receptor genes and other surface molecules genes, according to the the time required for activation or expression sequence is divided into three categories: Instant gene (15-30 minutes after receiving stimulation cells express), early gene (0.5-24 hours expression) and late genes (expressed within a few days). T cells in the row lymphocyte immune-related gene expression profile of kidney transplant study denounced Second Military Medical University doctoral dissertation in the mechanism of the reaction process is affected by many genes and signaling pathways, which involves changes in lymphocyte expression of many genes and regulation. Application of traditional methods in molecular biology, transplantation immunology unable to clarify the complex process of multiple genes and their interaction mechanisms and regulatory relations. Recent development of high-throughput gene expression analysis platforms eleven groups prisoners, the chip meets the need for surgery in support of this claim, can simultaneously sentence {Bu ten million from the study because of the expression, and to understand gene expression profiles for the whole changes provided technical assurance. IiI ago, microarray technology has become a biomedical research _ on a powerful tool. With traditional {research methods, based prisoners, chip has; tube flux, highly parallel, high sensitivity, high speed and so on. Therefore, the present study using gene chip technology that current advanced gene expression analysis methods, design of a kidney transplant lymphocyte immune-related gene expression studies, including three parts. Mount 24 after transplantation {payment early to L gene expression, and to the after 7 days, f majority of cell activation gene (including late gene) has been activated, so the experiment both first and second parts Motion detection after renal transplantation were 24 hours and 7 days subject to changes in gene expression in peripheral blood lymphocytes, will help to understand the basic process of transplantation immunity and facilitate studies of allograft rejection and immune inhibitors mechanism. Also studied immunosuppressants such as methylprednisolone (MP) pulse therapy and triple immunosuppressive therapy recipients lymphocyte immune related genes, understand their target genes exert pharmacological effects, in order for the development and immunosuppressants provide some new clinical drug reference. The third part of the experiment highly sensitized immune gene expression profiles in peripheral blood lymphocytes, screening and analysis of differentially expressed genes in order to understand the immune hypersensitivity-related genes and their regulatory networks from gene levels in the immune lymphocytes revealed the occurrence of high-sensitivity The molecular mechanisms. In short, wood noninvasive method of research applications through high-throughput, high-sensitivity gene chip technology, in vivo studies lymphocyte immune-related gene expression profile of kidney transplant. At home and abroad yet to see the report. I hope to contribute to the further understanding of wood study the basic process of transplantation immunity, the occurrence of highly sensitized immune mechanisms, and thereby contribute to further study the mechanisms of rejection and immunosuppressive regimen. The first part of the renal allograft recipients peripheral blood lymphocytes early response gene expression microarray Objective: To study after renal transplantation recipients early response gene expression changes in peripheral blood lymphocytes, to understand the basic process of transplantation immunology, gene level Learn lymphocyte immune response in renal transplantation in rats. Also studied methylprednisolone (MP) pulse therapy for post-transplant lymphocyte gene expression profiles, to understand the molecular mechanism of action related kidney transplant lymphocyte immune gene expression language Second Military Medical University doctoral dissertation research system. Methods: 16 cases of renal transplant recipients before transplantation as a control group, 24 hours after transplantation as the experimental group. Each patient in the preoperative and postoperative 24 hours of fresh blood was extracted IOml isolated peripheral blood lymphocytes, the same group of specimens mixed. Application of gene chip technology, according to one-step extraction experimental and control groups and total cellular RNA was purified mRNA, were synthesized by reverse transcription with Cy3 dUTP and a Cys an dUT, labeled cDNA probes?

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