Attempts to enrich and characterize human marrow T-cell precursors have been performed using discontinuous Percoll gradient centrifugation, phenotypic analysis of cells with monoclonal antibodies (Mabs), and T-cell colony-forming capacity. Marrow cells were extensively depleted of T cells and separated into seven fractions. The depletion was performed with the following Mabs: CD6 (MBG6 or RFT12) + CD8 (RFT8), CD2 (D66) + CD8 (RFT8), and CD6 (RFT12) + CD8 (RFT8) + CD7 (RFT2). A peak of cells with the capacity to differentiate into mature CD2+CD4+ T-cell agar colonies (TL-CFU) was obtained in a fraction with a density 1.063 less than d less than 1.069 g/ml. This peak was associated with the presence of cells expressing RFB1 and OKT10, two markers shared by hemopoietic precursors. Cells in this fraction were negative for CD3, CD4, CD1, CD8, and CD2 antigens. Their treatment by complement-dependent cytotoxicity with the CD7 Mab resulted in a loss of T-cell colony-forming capacity together with a reduction of T10-, RFB1-, and My10-positive cells.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Elimination of T cells from human peripheral blood and bone marrow using a cocktail of three anti-T cell immunotoxins.
Br J Haematol
December 1987
Department of Haematology, Hospital for Sick Children, London.
Four anti-T cell monoclonal antibodies were coupled to ricin-A and tested for their ability to kill T cells in peripheral blood and bone marrow using a clonogenic assay to quantify T cell survival. The immunotoxins (IT) prepared from RFT11 (CD2) and WT1 (CD7) antibodies were the most toxic to peripheral blood T cells. The immunotoxin prepared from RFT1 (CD5) was the next most efficient toxin and the immunotoxin prepared from RFT8 (CD8) was the least toxic.
View Article and Find Full Text PDFAttempts to enrich and characterize human marrow T-cell precursors have been performed using discontinuous Percoll gradient centrifugation, phenotypic analysis of cells with monoclonal antibodies (Mabs), and T-cell colony-forming capacity. Marrow cells were extensively depleted of T cells and separated into seven fractions. The depletion was performed with the following Mabs: CD6 (MBG6 or RFT12) + CD8 (RFT8), CD2 (D66) + CD8 (RFT8), and CD6 (RFT12) + CD8 (RFT8) + CD7 (RFT2).
View Article and Find Full Text PDFStandardization of T-cell depletion in HLA matched bone marrow transplantation.
Br J Haematol
September 1986
The IBM 2991 Blood Cell Processor has been used to isolate a mononuclear cell (MNC) fraction from the marrow of 31 allogeneic donors. The MNC fraction was then incubated with a combination of two murine monoclonal antibodies MBG6 (CD6) and RFT8 (CD8) followed by two rounds of treatment with rabbit complement resulting in a marrow inoculum significantly reduced in the number of T-lymphocytes. We report here new specifications for the use of Ficoll-Metrizoate, the method used to calculate T-lymphocyte depletion and the details of our attempts to improve T-depletion.
View Article and Find Full Text PDFThe regeneration of T cell subsets was studied with double immunofluorescence marker methods in 37 patients who received HLA matched T lymphocyte depleted bone marrow transplants (BMT) as part of the treatment for their haematological disease. A cocktail of anti-pan-T (CD6: MBG6) and anti-suppressor/cytotoxic-T cell (CD8: RFT8) monoclonal antibodies was used with rabbit serum as a source of cytolytic complement to achieve selective T cell lysis. The T8+ cells reached low normal values around 60 days post-transplant and remained within the normal range throughout the study (greater than 150 days).
View Article and Find Full Text PDFAllogeneic bone marrow transplantation has been undertaken within this centre on 62 patients with acute or chronic leukaemia. Employing the standard separation protocol described, all bone marrows were processed on the 2991 Blood Cell Processor to isolate the 'buffy coat' cells and subsequently the mononuclear cell component using a density separation medium of Ficoll-metrizoate. Following the mononuclear cell separation, the cells were identified for their T lymphocyte component using a combination of two murine monoclonal antibodies, MBG6 reacting with a pan T cell antigen (CD6) and RFT8 detecting the 'cytotoxic/supressor' cell antigen (CD8).
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!