Molecularcloning and mRNA expression of porcine interleukin-12

Dennis L. Foss, Michael P Murtaugh

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

Interleukin (IL)-12 is a heterodimeric cytokine consisting of 35 and 40 kDa subunits, produced primarily by phagocytic cells in response to bacteria or bacterial products. IL-12 is important in the regulation of both innate and antigen-specific immunity through its stimulatory effects on NK cells and cytotoxic lymphocytes. Reverse transcriptase-polymerase chain reaction with primers derived from human sequence was used to clone the p35 and p40 subunits of porcine IL-12. Predicted amino acid sequences for both subunits are approximately 85% homologous to their human cognates but contain a 3 aa addition and a 4 aa deletion in p35 and p40 subunits, respectively. The high degree of similarity indicates the proteins may be cross reactive, an important consideration in pig-human xenotransplantation. Both subunits of pIL-12 are constitutively expressed in a variety of porcine tissues. Highest levels of the p40 subunit were found in lymphoid tissues including inguinal and mesenteric lymph nodes, Peyer's patches, spleen and thymus. The p35 subunit was also detected in these tissues. Levels of mRNA encoding the p40 subunit, but not the p35 subunit, were rapidly increased in alveolar macrophages stimulated with lipopolysaccharide or killed Staphylococcus aureus. Thus, the heterodimeric subunits appear to be differentially regulated at the transcriptional level. Since p40 also self-associates to form inactive homodimers, differential expression may be a mechanism for regulating IL-12 activity.

Original languageEnglish (US)
Pages (from-to)121-134
Number of pages14
JournalVeterinary immunology and immunopathology
Volume57
Issue number1-2
DOIs
StatePublished - Jun 1997

Bibliographical note

Funding Information:
We appreciatet he helpful comments and discussion provided by Mary Pampusch. D.L.F. is supportedb y a USDA Animal Biotechnology fellowship. Research funding was provided by the Minnesota Agricultural Experiment Station.

Copyright:
Copyright 2004 Elsevier Science B.V., Amsterdam. All rights reserved.

Keywords

  • Alveolar macrophage
  • Cytokine
  • Lipopolysaccharide
  • Peyer's patch
  • Staphylococcus aureus

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