XII International Workshop on Alloantigenic Systems in the RatImmunization with Escherichia coli-produced soluble MHC class I proteins induces accelerated rejection or prolonged survival of rat cardiac allografts
Section snippets
Production of soluble class I MHC proteins
Production of RT1.An(donor) and RT1.Ac(recipient) class I MHC proteins was accomplished by using the pET vector-based IPTG-inducible E coli protein production system. The class I MHC proteins were purified in a one-step process using a Ni2+ chromatography affinity column utilizing a 6-Histidine tag at the C-terminus region of the recombinant protein.
Animals
Adult male inbred Brown-Norway (BN, RT1.An), PVG (RT1.Ac), and Lewis (LEW, RT1.Al) rats were purchased from Harlan Sprague Dawley (Indianapolis,
Results
Untreated PVG (RT1.Ac) recipients reject BN (RT1.An) cardiac allografts at a mean survival time (MST) of 9.1 ± 0.5 days. Subcutaneous (SC) immunization (day −7) with 50 and 100 μg of RT1.An protein induced accelerated rejection of BN cardiac allografts at MSTs of 7.8 ± 0.8 days (P = .004) and 6.1 ± 1.0 days (P = .0004), respectively. SC immunization of RT1.Ac protein did not cause accelerated rejection of BN cardiac allografts in PVG recipients, MST = 9.0 ± 0.0 days, P = NS. SC immunization
Discussion
Class I MHC sequences are well conserved from rodents up through primates and humans. The ability to determine the tolerogenic amino acid determinants in all species can result in their clinical use as potent immune modulators, including the potential for diminution or complete cessation of immunosuppressive therapy. Bacteria-produced rat class I MHC proteins possess both immunogenic and potent tolerogenic properties. The ability to produce large amounts of eukaryotic protein in a prokaryotic
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