Refinement of molecular approaches to improve the chance of identification of hematopoietic-restricted minor histocompatibility antigens
| Autor: | Theo de Witte, Elly van de Wiel-van Kemenade, Harry Dolstra, Björn de Rijke, Agnes van Horssen-Zoetbrood, Hanny Fredrix, Sharon Veenbergen |
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| Rok vydání: | 2008 |
| Předmět: |
Male
Genetic Linkage Immunology Single-nucleotide polymorphism Lymphocyte Activation Transfection Polymorphism Single Nucleotide HLA-B44 Antigen Minor Histocompatibility Antigens HLA-B7 Antigen Interferon-gamma Gene Frequency Antigen Immune Regulation [NCMLS 2] Translational research [ONCOL 3] Chlorocebus aethiops MHC class I Minor histocompatibility antigen Animals Humans Iron metabolism [IGMD 7] Immunology and Allergy Cytotoxic T cell Cloning Molecular Allele Gene Library Genetics biology cDNA library Reproducibility of Results Immunotherapy gene therapy and transplantation [UMCN 1.4] Intercellular Adhesion Molecule-1 Pedigree Transplantation HLA-B Antigens COS Cells B7-1 Antigen biology.protein Female K562 Cells T-Lymphocytes Cytotoxic |
| Zdroj: | Journal of Immunological Methods, 329, 125-37 Journal of Immunological Methods, 329, 1-2, pp. 125-37 |
| ISSN: | 0022-1759 |
| DOI: | 10.1016/j.jim.2007.10.001 |
| Popis: | Contains fulltext : 71202.pdf (Publisher’s version ) (Closed access) Minor histocompatibility antigens (mHAgs) constitute the target antigens of the T cell-mediated graft-versus-leukemia response after HLA-identical allogeneic stem cell transplantation (SCT). Several human mHAgs have been identified, but only a few are selectively expressed by hematopoietic cells representing potential targets for specific immunotherapy. Molecular approaches including cDNA library screening and genetic linkage analysis have been successfully applied to identify T cell-defined mHAgs, but each approach has its drawbacks which may lead to mis-identification of the mHAg of interest. We improved both molecular strategies to facilitate more robust identification of hematopoietic-restricted mHAgs. First, we adapted cDNA library cloning by using 293T cells with stable expression of the relevant MHC class I allele, CD80 and CD54. We demonstrated that cDNA library screening using this 293T expression system results in strong activation of cytotoxic T lymphocytes, which significantly contributes to improvement of the assay sensitivity. Second, we refined genetic linkage analysis using single nucleotide polymorphism (SNP) genotyping to narrow down the defined genetic region that holds the mHAg-encoding gene. We showed that SNP marker analysis provides additional information about the genetic position of the antigen-encoding gene. Application of these optimized molecular approaches will lead to more rapid and reliable molecular identification of hematopoietic-restricted mHAgs. |
| Databáze: | OpenAIRE |
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