A method to detect the early presence of lymphoma cells in the AKR mouse.
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Biomedical subjects
Publications and source records attributed to L J Flinton.
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We previously reported a successful model for treatment of BW 5147 leukemia in AKR mice by adoptive immunotherapy using allogeneic spleen cells from C57BL/6 mice. The leukemia cells were given 3 days before initiation of therapy. Graft-versus-host reaction was prevented by treatment with spleen cells from a second allogeneic strain (CBA), followed by cyclophosphamide and syngeneic spleen cells. We now show that it is not necessary to use syngeneic spleen cells in the final transplant since H-2-compatible, allogeneic CBA cells are as effective. In addition, it is possible to initiate successful therapy 5 days after leukemia implantation providing that the initial cyclophosphamide, given in two doses of 100 mg/kg each and spaced 7 days apart, is administered prior to establishment of graft-versus-host reaction. Higher single doses of drugs were followed by fatal graft-versus-host disease.
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Units of blood were intentionally contaiminated with suspensions of either Aerobacter aerogenes, Escherichia coli, Bacillus subtilis, Enterobacter cloacae, Pseudomonas aeruginosa, Sarcina lutea, Seratia marcesens, Staphylococcus epidermidis, Streptococcus faecalis, Paracolabacterum aerogenoides (Enterobacter hafriae), Mima polymorpha or Acinetobacter- calcoaceticus. When inoculation was made prior to glycerolization, the subsequent glycerolization, freezing, thawing, and deglycerolization resulted in roughly a two log reduction in the number of bacteria. When inoculation was made with a final concentration of between 10(1) and 10(5) organisms per milliliter, immediately following deglycerolization or following washing without glycerolization and freezing, no increase in the number of bacteria was seen after 72 hours storage at 4 C. Three of the 12 organisms studied decreased in number during 72 hours of storage. These data suggest that the current 24-hour limit on the post-thaw storage of frozen red blood cells may be unnecessarily restrictive.
A simple and inexpensive method for the freezing of human blood lymphocytes at -80 C is described. Lymphocytes were frozen in either polystyrene tubes or polyolefin bags. Optimal recovery of cells was obtained when 5% DMSO and 62% plasma were used as cryoprotectants. Viable cell recovery, as assessed by response to phyto hemagglutinin (PHA) and pokeweed mitogen (PWM), was greater than 80% after 60 days of storage. In addition, percentages of T and B lymphocytes were essentially unchanged. The excellent viable cell recovery in polyolefin bags suggests that it is possible to cryopreserve large numbers of lymphocytes in a single container.