Abstract
Morphological, phenotypic and functional analyses were made of cells obtained at each step after successive treatments of 23 separate human bone marrow suspensions with soybean lectin and sheep erythrocytes (SRBC). The average total number of nucleated cells harvested was 1.9 X 10(10) and the final cell suspensions contained a mean of 1.9 X 10(9) nucleated cells or 9.2 +/- 4.8% of the initial counts. Monoclonal antibody analyses revealed that both T and B lymphocytes were present in every cell fraction in percentages similar to those found initially until after the first SRBC rosette-depletion. Moreover, both soy lectin agglutinated and non-agglutinated cells exhibited vigorous proliferative responses to phytohaemagglutinin and allogeneic cells. Following the SRBC depletions, no cells having T lymphocyte phenotypes or functions could be detected, whereas 5% of the cells reacted with a monoclonal antibody to B lymphocytes. The final fraction was composed predominantly of immature myeloid cells and blasts and was depleted of erythroid elements, lymphocytes and essentially all mature cells. It contained cells reactive with monoclonal antibodies recognizing undifferentiated T cell precursors (3A1), the transferrin receptor (5E9), and a human progenitor cell antigen (My-10). The final fraction was also enriched 10-100-fold for CFU-C and 5-10-fold for CFU-GEMN colonies. These studies fail to demonstrate selective removal of T lymphocytes from human bone marrow cells by soybean lectin agglutination.
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Selected References
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- Blazar B. R., Quinones R. R., Heinitz K. J., Sevenich E. A., Filipovich A. H. Comparison of three techniques for the ex vivo elimination of T cells from human bone marrow. Exp Hematol. 1985 Feb;13(2):123–128. [PubMed] [Google Scholar]
- Buckley R. H., Schiff S. E., Sampson H. A., Schiff R. I., Markert M. L., Knutsen A. P., Hershfield M. S., Huang A. T., Mickey G. H., Ward F. E. Development of immunity in human severe primary T cell deficiency following haploidentical bone marrow stem cell transplantation. J Immunol. 1986 Apr 1;136(7):2398–2407. [PubMed] [Google Scholar]
- Burgess A. W., Wilson E. M., Metcalf D. Stimulation by human placental conditioned medium of hemopoietic colony formation by human marrow cells. Blood. 1977 Apr;49(4):573–583. [PubMed] [Google Scholar]
- Civin C. I., Strauss L. C., Brovall C., Fackler M. J., Schwartz J. F., Shaper J. H. Antigenic analysis of hematopoiesis. III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG-1a cells. J Immunol. 1984 Jul;133(1):157–165. [PubMed] [Google Scholar]
- Fauser A. A., Messner H. A. Granuloerythropoietic colonies in human bone marrow, peripheral blood, and cord blood. Blood. 1978 Dec;52(6):1243–1248. [PubMed] [Google Scholar]
- Fauser A. A., Messner H. A. Identification of megakaryocytes, macrophages, and eosinophils in colonies of human bone marrow containing neurtophilic granulocytes and erythroblasts. Blood. 1979 May;53(5):1023–1027. [PubMed] [Google Scholar]
- Harden E. A., Haynes B. F. Phenotypic and functional characterization of human malignant T cells. Semin Hematol. 1985 Jan;22(1):13–26. [PubMed] [Google Scholar]
- Haynes B. F. Human T lymphocyte antigens as defined by monoclonal antibodies. Immunol Rev. 1981;57:127–161. doi: 10.1111/j.1600-065x.1981.tb00445.x. [DOI] [PubMed] [Google Scholar]
- Kurtzberg J., Bigner S. H., Hershfield M. S. Establishment of the DU.528 human lymphohemopoietic stem cell line. J Exp Med. 1985 Nov 1;162(5):1561–1578. doi: 10.1084/jem.162.5.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reinherz E. L., Geha R., Rappeport J. M., Wilson M., Penta A. C., Hussey R. E., Fitzgerald K. A., Daley J. F., Levine H., Rosen F. S. Reconstitution after transplantation with T-lymphocyte-depleted HLA haplotype-mismatched bone marrow for severe combined immunodeficiency. Proc Natl Acad Sci U S A. 1982 Oct;79(19):6047–6051. doi: 10.1073/pnas.79.19.6047. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reisner Y., Kapoor N., Hodes M. Z., O'Reilly R. J., Good R. A. Enrichment for CFU-C from murine and human bone marrow using soybean agglutinin. Blood. 1982 Feb;59(2):360–363. [PubMed] [Google Scholar]
- Reisner Y., Kapoor N., Kirkpatrick D., Pollack M. S., Cunningham-Rundles S., Dupont B., Hodes M. Z., Good R. A., O'Reilly R. J. Transplantation for severe combined immunodeficiency with HLA-A,B,D,DR incompatible parental marrow cells fractionated by soybean agglutinin and sheep red blood cells. Blood. 1983 Feb;61(2):341–348. [PubMed] [Google Scholar]
- Reisner Y., Kapoor N., O'Reilly R. J., Good R. A. Allogeneic bone marrow transplantation using stem cells fractionated by lectins: VI, in vitro analysis of human and monkey bone marrow cells fractionated by sheep red blood cells and soybean agglutinin. Lancet. 1980 Dec 20;2(8208-8209):1320–1324. doi: 10.1016/s0140-6736(80)92394-6. [DOI] [PubMed] [Google Scholar]
- Reisner Y., Pahwa S., Chiao J. W., Sharon N., Evans R. L., Good R. A. Separation of antibody helper and antibody suppressor human T cells by using soybean agglutinin. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6778–6782. doi: 10.1073/pnas.77.11.6778. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ritz J., Pesando J. M., Notis-McConarty J., Schlossman S. F. Modulation of human acute lymphoblastic leukemia antigen induced by monoclonal antibody in vitro. J Immunol. 1980 Oct;125(4):1506–1514. [PubMed] [Google Scholar]
- Saxon A., Feldhaus J., Robins R. A. Single step separation of human T and B cells using AET treated srbc rosettes. J Immunol Methods. 1976;12(3-4):285–288. doi: 10.1016/0022-1759(76)90050-8. [DOI] [PubMed] [Google Scholar]
- Schiff R. I., Buckley R. H., Gilbertsen R. B., Metzgar R. S. Membrane receptors and in vitro responsiveness of lymphocytes in human immunodeficiency. J Immunol. 1974 Jan;112(1):376–386. [PubMed] [Google Scholar]
- Woolfrey A. E., Neudorf S., Filipovich A. H. Comparison of two immuno-mechanical methods of T-cell depletion of human bone-marrow for prevention of graft-versus-host disease: soybean lectin agglutination and sheep erythrocyte rosette depletion versus triple rosette depletion. Thymus. 1985;7(6):327–334. [PubMed] [Google Scholar]
- van Dongen J. J., Hooijkaas H., Comans-Bitter M., Hählen K., de Klein A., van Zanen G. E., van't Veer M. B., Abels J., Benner R. Human bone marrow cells positive for terminal deoxynucleotidyl transferase (TdT), HLA-DR, and a T cell marker may represent prothymocytes. J Immunol. 1985 Nov;135(5):3144–3150. [PubMed] [Google Scholar]