Immunological characterization of canine hematopoietic progenitor cells.

Canine hematopoietic progenitor cells were characterized by separation with monoclonal antibodies. Depleted and enriched fractions were studied for growth of CFU-GM in semisolid agar and for repopulating capacity of lethally irradiated dogs. CFU growth was not reduced by depletion of marrow using monoclonal antibodies F 3-20-7 (anti-dog Thy-1), MT606 (anti-human CD 6), and IOT2a (anti-human DR). CFU growth was variable following treatment with the anti-canine T-cell antibody MdT-P 1 and immunomagnetic bead separation. It was regularly enriched when MdT-P 1 treatment was followed by immunorosetting with staphylococcal protein A-loaded sheep red blood cells and density gradient separation. Lethally irradiated dogs were reconstituted by autologous marrow depleted of MdT-P 1-positive cells using immunorosetting and density gradient centrifugation, whereas immunomagnetic bead-depleted marrow was ineffective. Fluorescence-activated cell sorting showed enrichment of hematopoietic progenitor cells in the weakly MdT-P 1-positive fraction.

Dose rate and fractionation of total body irradiation in dogs: short and long term effects.

Variations of regimens of total body irradiation (TBI) were investigated in the dog as a preclinical model for bone marrow transplantation. Inactivation of hemopoietic precursor cells (CFU-GM) was studied following irradiation of marrow in vitro, following TBI at sublethal doses in vivo and following autologous transplantation of marrow obtained after sublethal TBI. Inactivation and recovery of CFU-GM as well as restoration of hemopoiesis following autologous transplantation was independent of the dose rate, but nadirs of blood counts were lower following sublethal TBI with the higher dose rate. Acute non-hemopoietic toxicity of TBI depended on the dose, the dose rate and the total treatment time and not on the fractionation regimen. At a total dose of 25 Gy acute mortality was prevented by prophylactic administration of oral, non-absorbable antibiotics. Late mortality was due to degenerative and autoimmune-like disorders with or without infections and to malignant tumors. Evaluation of long-term survival is still preliminary, since surviving dogs of two groups (10 Gy as single dose, 25 Gy as hyperfractionated TBI) have not yet reached the median survival time of their group. So far, long-term survival depended on the total dose (p = 0.05) and, possibly, the fractionation regimen (p = 0.12). The latency period until development of malignant tumors was influenced by the total doses given in the same treatment time (p = 0.05) and by the total treatment time for equal doses (p = 0.04). It was concluded that TBI at a low dose rate may give the best therapeutic ratio of inactivation of hemopoietic precursor cells to acute toxicity.(ABSTRACT TRUNCATED AT 250 WORDS)

Revised classification of the DLA loci by serological studies.

Peripheral blood mononuclear cells from DLA typed dogs were treated with rabbit-anti-dog-beta 2-microglobulin and subsequently with goat-anti-rabbit-immunoglobulin in order to aggregate the DLA class I molecules on the cell membrane (lysostrip). Utilizing a panel of 70 defined DLA-A and DLA-B antisera, lymphocytes treated in this way showed resistance to complement dependent lysis with monospecific DLA-A sera only, whereas reactivity of DLA-B antisera was not blocked; on the contrary, complete lympholysis with each DLA-B antiserum was recognized. Thus, the DLA-B antigens, evidently not associated with beta 2-microglobulin, are designated as candidates for class II gene products. The different reactions of DLA-C antisera after lysostrip did not allow a precise assignment of this antigen series as yet.