Use of antilymphocyte globulin after cadaveric renal transplantation

Antilymphocyte globulin (A.L.G.) was prepared by injecting fresh frozen splenic cells subcutaneously into horses. The IgG fraction of the serum was concentrated by a batch technique using diethylaminoethanol-Sephadex. Fourteen patients given this material by intramuscular injection after cadaveric renal transplants, in addition to azathioprine and prednisone, had less evidence of rejection compared with patients previously treated with azathioprine and prednisone only, despite a reduction of the mean daily prednisone dose from 65 to 45 mg. Toxicity, especially local reaction, fever, and hypotension, limited the amount of A.L.G. that was given.     

Identification of an antilymphocyte transformation substance from Pasteurella multocida

Pasteurella multocida is one of the most important bacteria responsible for diseases of animals. Crude extracts from sonicated P. multocida strain Dainai‐1, which is serotype A isolated from bovine pneumonia, were found to inhibit proliferation of mouse spleen cells stimulated with Con A. The crude extract was purified by cation and anion exchange chromatography and hydroxyapatite chromatography. Its molecular weight was 27 kDa by SDS‐PAGE and it was named PM27. PM27 was found to inhibit proliferation of mouse spleen cells stimulated with Con A as effectively as did the crude extract; however, its activity was lost after heating to 100°C for 20 min. PM27 did not directly inhibit proliferation of HT‐2 cells, which are an IL‐2‐dependent T cell line, nor did it modify IL‐2 production by Con A‐stimulated mouse spleen cells. The N‐terminal amino acid sequence of PM27 was determined and BLAST analysis revealed its identity to uridine phosphorylase (UPase) from P. multocida. UPase gene from P. multocida Dainai‐1 was cloned into expression vector pQE‐60 in Escherichia coli XL‐1 Blue. Recombinant UPase (rUPase) tagged with His at the C‐terminal amino acid was purified with Ni affinity chromatography. rUPase was found to inhibit proliferation of mouse spleen cells stimulated with Con A; however, as was true for PM27, its activity was lost after heating to 100°C for 20 min. Thus, PM27/UPase purified from P. multocida has significant antiproliferative activity against Con A‐stimulated mouse spleen cells and may be a virulence factor.