Membrane-bound or soluble truncated RT1.Aa rat class I major histocompatibility antigens induce specific alloimmunity

R. Mark Ghobrial, Takashi Hamashima, Malgorzata Kloc, Laurence Etkin, Stanislaw M. Stepkowski, Barry D. Kahan

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Transfectants that express membrane-bound (MB) or secrete soluble truncated (TR) rat class I RT1.Aa major histocompatibility (MHC) antigens induce alloimmunity in vivo. The MB-RT1.Aa was produced by transfecting the full-length RT1.Aa cDNA, including the α1, α2, and α3, transmembrane and intracellular domains. The TR-RT1.Aa cDNA insert included only the extracellular α1, α2, and α3 domains; a stop codon was placed in front of the transmembrane domain. Following full-length sequencing, MB-RT1.Aa and TR-RT1.Aa cDNAs were translated in vitro into glycosylated MB-RT1.Aa (45 kDa) and TR-RT1.Aa (36 kDa) proteins, respectively. Each cDNA construct was individually subcloned into the pSG5 vector before transfection into Buffalo (BUF; RT1b) hepatoma cells. FACscan analysis with anti-RT1.Aa-specific R2/15S monoclonal antibody (MAb) confirmed surface expression of RT1.Aa molecules on the MB-RT1.Aa, but not on the TR-RT1.Aa, transfectants. In contrast, enzyme-linked immunoadsorbent assays documented the presence of soluble RT1.Aa molecules in supernates from cells transfected with the TR- RT1.Aa, but not from cells transfected with the MB-RT1.Aa, cDNA. Subcutaneous injection of MB-RT1.Aa or TR-RT1.Aa transfectants to BUF or Wistar Furth (WF; RT1(u)) rats induced accelerated rejection of ACI (RT1a) but not third-party Brown Norway (RT1(n)) heart allografts. Furthermore, supernates of TR-RT1.Aa, but not of MB-RT1.Aa, transfectants immunized WF hosts toward ACI hearts. Thus, both intact MB-RT1.Aa and soluble TR-RT1.Aa class I alloantigensinduce potent sensitization against alloantigens.

Original languageEnglish (US)
Pages (from-to)602-610
Number of pages9
JournalTransplantation
Volume60
Issue number6
DOIs
StatePublished - Jan 1 1995

ASJC Scopus subject areas

  • Transplantation

Fingerprint Dive into the research topics of 'Membrane-bound or soluble truncated RT1.A<sup>a</sup> rat class I major histocompatibility antigens induce specific alloimmunity'. Together they form a unique fingerprint.

Cite this