Immunogenic properties of the M. leprae recombinant 18-kDa antigen purified from Saccharomyces cerevisiae; enhancement of delayed-type hypersensitivity after gamma-irradiation

Int J Lepr Other Mycobact Dis. 1995 Sep;63(3):381-90.

Abstract

In this paper we report the purification and study of the immunogenic properties of the Mycobacterium leprae 18-kDa protein antigen produced and secreted by the yeast Saccharomyces cerevisiae, using an expression system we recently described [Biotech. Lett. 16 (1994) 1241-1246]. The 18-kDa protein was purified from the yeast culture media by precipitation, ion exchange chromatography (MonoQ) and exclusion size chromatography (Sephacryl S-100). The biological properties of the recombinant protein, previously irradiated with gamma rays, were assayed by immunization of mice. Humoral and cellular responses, monitored by antibody production and delayed-type hypersensitivity, respectively, were obtained. Furthermore, gamma-irradiation of the recombinant protein prior to the administration was shown to significantly potentiate the T-cell response. The data suggest that this irradiated recombinant antigen could be used in a more sensitive standardized skin test to monitor M. leprae infection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood
  • Bacterial Proteins / immunology*
  • Bacterial Proteins / isolation & purification
  • Bacterial Proteins / radiation effects
  • Gamma Rays*
  • Hypersensitivity, Delayed / immunology*
  • Mice
  • Mice, Inbred CBA
  • Mycobacterium leprae / immunology*
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / radiation effects
  • Saccharomyces cerevisiae*

Substances

  • 18 kDa protein, Mycobacterium leprae
  • Antibodies, Bacterial
  • Bacterial Proteins
  • Recombinant Proteins