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...

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Published in:	Microbiology and immunology Vol. 63; no. 7; pp. 261 - 268
Main Authors:	Tanabe, Taishi, Fukuzawa, Hiroki, Amatatsu, Yuki, Matsui, Keigo, Ohtsuka, Hiromichi, Maeda, Yousuke, Sato, Hisaaki
Format:	Journal Article
Language:	English
Published:	Australia Wiley Subscription Services, Inc 01-07-2019 John Wiley and Sons Inc
Subjects:	Affinity chromatography Amino Acid Sequence Animal diseases Animals Anion exchange Anion exchanging Bacterial Outer Membrane Proteins - genetics Bacterial Outer Membrane Proteins - metabolism Bacterial Outer Membrane Proteins - pharmacology bovine respiratory disease Cation exchanging Cattle Cell Line - drug effects Cell proliferation Cell Proliferation - drug effects Chromatography Escherichia coli - genetics Gel electrophoresis Genetic transformation Humans Hydroxyapatite Interleukin-2 - metabolism Lymphocytes Lymphocytes T Mice Molecular Weight Original Pasteurella Pasteurella multocida Pasteurella multocida - genetics Pasteurella multocida - metabolism pathogenicity factor Phosphorylase Phosphorylases Recombinant Proteins Sodium lauryl sulfate Spleen T-Lymphocytes - drug effects Uridine Uridine phosphorylase Uridine Phosphorylase - genetics Uridine Phosphorylase - isolation & purification Uridine Phosphorylase - metabolism Uridine Phosphorylase - pharmacology Virulence factors Pasteurella bovine respiratory disease uridine phosphorylase pathogenicity factor
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0385-5600 1348-0421
DOI:	10.1111/1348-0421.12720