Fractionation of feline bone marrow with the soybean agglutinin lectin yields populations enriched for erythroid and myeloid elements: transplantation of soybean agglutinin-negative cells into lethally irradiated recipients.

Feline bone marrow cells treated with the soybean agglutinin (SBA) lectin are separated into two populations, the agglutinated SBA(+) fraction containing predominantly cells of myeloid origin and the nonagglutinated SBA(-) fraction consisting of cells primarily of the erythroid lineage. FACScan analyses revealed a clear distinction of the cells based on their light scattering properties, i.e., large cells and cells with high granularity were found in the SBA(+) fraction, whereas cells having a low forward light scatter and side light scatter were found in the SBA(-) fraction. Colony-forming assays showed colony-forming unit-granulocyte/monocyte (CFU-GM) cells to have a strong affinity for SBA because these were found almost entirely in the SBA(+) fraction; in contrast, burst-forming unit-erythroid (BFU-E)-forming cells were concentrated in the SBA(-) fraction. When the marrow was fractionated by counterflow centrifugal elutriation (CCE), a differential binding to SBA among the CFU-GM forming cells was found. The SBA(-) fractions of cells collected at 21 and 25 ml/min contained primarily BFU-E forming cells, similar to that observed with whole marrow; the later CCE fractions, those collected at 32 ml/min and the rotor off fraction, when treated with SBA showed a small but significant number of CFU-GM cells in the SBA(-) fraction. T lymphocytes were found predominantly in the SBA(+) fractions of whole bone marrow and the CCE fractions. Successful autologous marrow transplants were performed with the early CCE SBA(-) fractions. The latter cells were used for our initial transplant attempts because ongoing studies in our laboratory had shown these cells to be free of any viral-containing cells when the marrow had been obtained from animals infected with the feline immunodeficiency virus. In summary, although SBA treatment of feline marrow yields a marked separation of CFU-GM and BFU-E progenitors, select CCE SBA(-) fractions contain stem cells capable of providing hematopoietic reconstitution of lethally irradiated animals.

[1]  A. Legendre,et al.  SEPARATION OF FELINE BONE MARROW CELLS BY COUNTERFLOW CENTRIFUGAL ELUTRIATION: IDENTIFICATION AND ISOLATION OF PRESUMPTIVE EARLY AND LATE MYELOID/ERYTHROID PROGENITORS , 1995, Transplantation.

[2]  N. Day,et al.  A method of purifying feline T lymphocytes from peripheral blood using the plant lectin from Pisum sativum. , 1994, Journal of immunological methods.

[3]  N. Gengozian,et al.  Functional evaluation of T helper, T suppressor, and B lymphocytes in lethally irradiated rhesus monkeys injected with autologous bone marrow. , 1992, Transplantation.

[4]  N. Day,et al.  Sugar competition assays reveal high affinity receptors for Erythrina cristigalli lectin on feline monocytes. , 1990, Journal of immunological methods.

[5]  N. Day,et al.  Guinea pig and gerbil erythrocytes rosette with different cells in the blood, bone marrow, and thymus of the cat. , 1988, Cellular immunology.

[6]  N. Pedersen,et al.  Isolation of a T-lymphotropic virus from domestic cats with an immunodeficiency-like syndrome. , 1987, Science.

[7]  R. Good,et al.  Enrichment for CFU-C from murine and human bone marrow using soybean agglutinin. , 1982, Blood.

[8]  A. Ravid,et al.  Use of soybean agglutinin for the separation of mouse B and T lymphocytes. , 1976, Biochemical and biophysical research communications.

[9]  A. Morley,et al.  Cyclic leucocytosis as evidence for retention of normal homoeostatic control in chronic granulocytic leukaemia. , 1967, Lancet.

[10]  N. Sharon,et al.  Lectin receptors as lymphocyte surface markers. , 1983, Advances in immunology.