Production of a recombinant form of early pregnancy factor that can prolong allogeneic skin graft survival time in rats

H Morton, D A McKay, R M Murphy, M J Somodevilla-Torres, C E Swanson, A I Cassady, Kim Summers, A C Cavanagh

Research output: Contribution to journalArticlepeer-review

Abstract

Early pregnancy factor (EPF), an extracellular chaperonin 10 homologue, has immunosuppressive and growth factor properties. In order to carry out more extensive studies on the in vivo characteristics of EPF, a recombinant form of the molecule has been prepared. Recombinant human EPF (rEPF) was expressed in Escherichia coli using the plasmid pGEX-2T expression system. Potency of rEPF in vitro in the rosette inhibition test, the bioassay for EPF, was equivalent to that of native EPF (nEPF), purified from human platelets, and synthetic EPF (sEPF). However, the half-life of activity (50% decrease in the log value) in serum, following i.p. injection, was significantly decreased (3.2 h, compared with nEPF 6.2 days, sEPF 5.8 days). This was thought to be due to modification of the N-terminus of the recombinant molecule inhibiting binding to serum carrier proteins. Because EPF can modify Th1 responses, the ability of the recombinant molecule to suppress allogeneic graft rejection was investigated. Following skin grafts from Lewis rats to DA rats and vice versa, rEPF was delivered locally at the graft site and the effect on survival time of the allografts noted. Results demonstrated that rEPF treatment significantly prolonged skin graft survival time by as much as 55% in stringent models of transplantation across major histocompatibility barriers.
Original languageEnglish
Pages (from-to)603-7
Number of pages5
JournalImmunology and Cell Biology
Volume78
Issue number6
DOIs
Publication statusPublished - 2000

Keywords

  • Animals
  • Chaperonin 10
  • Escherichia coli
  • Graft Survival
  • Immunosuppressive Agents
  • Injections, Intraperitoneal
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Peptides
  • Pregnancy Proteins
  • Rats
  • Rats, Inbred Lew
  • Recombinant Proteins
  • Rosette Formation
  • Skin Transplantation
  • Suppressor Factors, Immunologic
  • Time Factors
  • Transplantation Tolerance
  • Transplantation, Homologous

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