Membrane-bound or soluble truncated RT1.Aa rat class I major histocompatibility antigens induce specific alloimmunity.
Academic Article
Overview
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 alpha 1, alpha 2, and alpha 3, transmembrane and intracellular domains. The TR-RT1.Aa cDNA insert included only the extracellular alpha 1, alpha 2, and alpha 3 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; RT1u) rats induced accelerated rejection of ACI (RT1a) but not third-party Brown Norway (RT1n) 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 alloantigens induce potent sensitization against alloantigens.
