A flow cytometric crossmatch test for simultaneous detection of antibodies against donor lymphocytes and endothelial precursor cells
| Autor: | Mats Alheim, P. Grufman, Jan Holgersson, Dan Hauzenberger, S. M. Johansson |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Serum
T-Lymphocytes CD3 Lymphocyte Immunology Population Human leukocyte antigen Biochemistry Antibodies CD19 Flow cytometry HLA Antigens Genetics medicine Humans Immunologic Factors Immunology and Allergy Lymphocytes education B-Lymphocytes education.field_of_study biology medicine.diagnostic_test Chemistry Histocompatibility Antigens Class I Histocompatibility Antigens Class II Endothelial Cells General Medicine Flow Cytometry Donor Lymphocytes Molecular biology Tissue Donors medicine.anatomical_structure biology.protein CD8 |
| Zdroj: | Tissue Antigens. 75:269-277 |
| ISSN: | 1399-0039 0001-2815 |
| Popis: | Complement-dependent cytotoxicity or flow cytometric lymphocyte crossmatch (LXM) tests may fail to detect clinically significant antibodies (Abs) against non-human leukocyte antigen (HLA). A flow cytometric endothelial precursor cell crossmatch (EPCXM) test (XM-ONE) is available for detection of Abs against donor endothelial precursor cells (EPCs). We showed that lymphocytes co-purified with EPCs can be used in LXM tests allowing simultaneous detection of Abs reactive with donor EPCs and lymphocytes. The lymphocyte population co-purified with EPCs on anti-Tie-2 Ab-coupled magnetic beads contained CD 8(+) and CD 4(+) T-cells, B-cells, and natural killer (NK)- and natural killer T (NKT)-cells. HLA class I antigen expression was slightly higher on CD 3(+) lymphocytes co-purified on Tie-2 Ab beads than on unseparated lymphocytes, whereas HLA class I and II antigen levels on CD 19(+) lymphocytes were not significantly different. Sera from 10 patients with panel-reactive Abs were tested on cells from nine donors using flow cytometric LXM and EPCXM tests. There was a very good correlation (R(2) = 0.94) between the channel shift values obtained on unseparated and Tie-2 Ab bead-isolated T-lymphocytes, whereas the correlation between the channel shift values obtained on the two B-lymphocyte populations was lower (R(2) = 0.71). T- and B-lymphocytes co-purified with EPCs can be used in LXM tests enabling simultaneous detection of donor lymphocyte- and EPC-reactive Abs in a single-tube XM-ONE assay. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|