Dissertation > Excellent graduate degree dissertation topics show

Construction of a New Kind of PR1-HLA-A~*0201-SCT Tetramer and Preliminary Identification

Author: ZhaoYanXia
Tutor: JiMingChun
School: Yangzhou University
Course: Immunology
Keywords: Of MHC Ⅰ class of molecules Tetramer Single chain trimer Cytotoxic T lymphocyte Chronic myeloid leukemia
CLC: R392
Type: Master's thesis
Year: 2010
Downloads: 13
Quote: 0
Read: Download Dissertation

Abstract


MHC Ⅰ tetramer (major histocompatibility complex Ⅰ tetramer) antigen-specific cytotoxic T lymphocytes (cytotoxic T lymphocytes, CTLs) detection and screening, the understanding of the cellular immune function, an important tool to explore the mechanism and evaluation of disease-specific immunotherapy one. Due to the sensitive, specific, and efficient, cell damage etc., MHC Ⅰ tetramer technology was born to become the gold standard for detection of CTLs quantitative. But the the traditional MHC I tetramers were prepared process cumbersome, protein renaturation of MHC Ⅰ and β2-microglobulin (β2m) two peptide chains and antigenic peptide while the correct folding, refolding inefficient antigen peptide affinity requirements affinity low antigen peptide is difficult to form a stable tetramer. Because of these shortcomings, limiting of the of MHC Ⅰ tetramer technology, large-scale applications, many scholars are trying to improve the technology. Single-chain trimer (single chain trimer, SCT) antigenic peptide, β2m and MHC Ⅰ molecules in order to the appropriate connector (linker) series is more conducive to a significant improvement of the traditional tetramer technology, antigen-specific the detection of CTLs. Chronic myeloid leukemia (Chronic myeloid leukemia, CML) is a malignant proliferative disease that originated in hematopoietic stem cells. PR1 from the CML associated antigens protease 3 HLA-A * 0201-limiting 9 peptide (VLQELNVTV), the induction of CTLs can kill leukemia cells, but on the normal bone marrow cells without toxicity. The purpose of this study is the preparation of an HLA-A * 0201-restricted PR1-HLA-A * 0201-SCT (hereinafter referred PR1-SCT) tetramer PR1-specific CTLs, to the joint flow cytometry studies, for CML The adoptive immunotherapy lay the foundation. In this study, we first use of genetic engineering technology will PR1 peptide, β2m and HLA-A * 0201 n flexible linker (G4S) followed by series build the PR1-SCT fusion gene, and then cloned to pQE31 vector carried prokaryotic expression by washing the inclusion bodies get a large number of PR1-SCT inclusion body proteins. Then we use the the pQE31 vector 6 × His tag the PR1-SCT inclusion body proteins purified by refolding it to restore the native conformation. By the introduction of 2 mol / L urea in the refolding system, greatly improving the efficiency of renaturation. We further the PR1-SCT Protein Renaturation biotinylated, and in accordance with the efficiency of the biotinylated, and PE-labeled streptavidin and prime appropriate rate mixing, to prepare a new PR1-SCT tetramer and its Preliminary identification, expected to provide a powerful tool for the CML clinical and basic research.

Related Dissertations

  1. The Research of the Expression Level of Hexokinasel1 in the Chronic Phase and Blastic Phase of Patients with Chronic Myeloid Leukemia,R733.7
  2. ABCG2 Expression and the Study of Methylaion in Gleevec-resistant Chronic Myeloid Leukemia Cell Line,R733.7
  3. Modeling Cml by Human Cells,R733.7
  4. Research of Susceptibility Genes of Pediatric Diabetes Mellitus,R725.8
  5. Study on the Usage of T-Cell Receptor α and β Chains of the HBV-Specific CD8~+ T Lymphocytes in the Patients with Acute Hepatitis B,R392
  6. The Effects of 6-gingerol on Proliferation, Apoptosis of Human Leukemia Molt4 Cells and K562 Cells,R733.7
  7. In Vivo Effect of Recombinant Protein PTD-OD-HA in the Mouse Model of Chronic Myeloid Leukemia,R733.7
  8. Measurement of Chromosome-Specific Relative Telomere Lengths in Leukocytes of Normal Individuals and CML Patients,R733.7
  9. Association of the CTLA-4 Gene Polymorphism with the Recurrent Spontaneous Abortion,R714.21
  10. The Association of CTLA-4、 VDR and MIC-A Gene Polymorphism with Latent Autoimmune Diabetes in Adults,R587.1
  11. Chronic myeloid leukemia cell genetics,R733.7
  12. Expression and Clinical Significance of CDX2 in Acute Leukemia,R733.7
  13. A Qualitative Study of Simian Immunodeficiency Virus Model,O175.13
  14. Virological and Immunological Response in AIDS Patients Treated with Highly Active Antiretroviral Therapy,R512.91
  15. The Research and Development of Catalyst of 1-Octene by Ethylene Tetramerization and the Performance Analysis of Catalyst System,TQ221.21
  16. Preliminary Study of the Development of Bcr Promotor-driven EGFP Expressing Transgenic Mice,Q78
  17. The Detection and Mechanism of the Factor Ⅷ Inhibitor with Hemophilia A,R554.1
  18. Pumpkin protein in the anti- human chronic myelogenous leukemia cell line K562 role in research,R733.72
  19. The effect of lamivudine treatment on cellular immune responses in patients with hepatitis B,R512.62
  20. The Levels of Serum Granzyme B and sFasL Are Involved in the Development of Virus Myocarditis in Children,R725.4

CLC: > Medicine, health > Basic Medical > Medical Immunology
© 2012 www.DissertationTopic.Net  Mobile