Characterization of an immunogenic cellulase secreted by Cryptococcus pathogens

Med Mycol. 2020 Nov 10;58(8):1138-1148. doi: 10.1093/mmy/myaa012.

Abstract

Members of the C. neoformans/C. gattiii species complex are an important cause of serious humans infections, including meningoencephalitis. We describe here a 45 kDa extracellular cellulase purified from culture supernatants of C. neoformans var. neoformans. The N-terminal sequence obtained from the purified protein was used to isolate a clone containing the full-length coding sequence from a C. neoformans var. neoformans (strain B-3501A) cDNA library. Bioinformatics analysis indicated that this gene is present, with variable homology, in all sequenced genomes of the C. neoformans/C. gattii species complex. The cDNA clone was used to produce a recombinant 45 kDa protein in E. coli that displayed the ability to convert carboxymethyl cellulose and was therefore designated as NG-Case (standing for Neoformans Gattii Cellulase). To explore its potential use as a vaccine candidate, the recombinant protein was used to immunize mice and was found capable of inducing T helper type 1 responses and delayed-type hypersensitivity reactions, but not immune protection against a highly virulent C. neoformans var grubii strain. These data may be useful to better understand the mechanisms underlying the ability C. neoformans/C. gattii to colonize plant habitats and to interact with the human host during infection.

Keywords: cellulase; cryptococcal infection; immune responses.

MeSH terms

  • Animals
  • Carboxymethylcellulose Sodium / metabolism
  • Cellulase / chemistry
  • Cellulase / genetics
  • Cellulase / immunology*
  • Cellulase / metabolism
  • Cryptococcosis / immunology
  • Cryptococcosis / microbiology
  • Cryptococcus / enzymology*
  • Cryptococcus / genetics
  • Cryptococcus / immunology
  • Cryptococcus / metabolism
  • Cryptococcus neoformans / enzymology
  • Cryptococcus neoformans / genetics
  • Cryptococcus neoformans / immunology
  • Cryptococcus neoformans / metabolism
  • Culture Media, Conditioned
  • Cytokines / immunology
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics
  • Fungal Proteins / immunology*
  • Fungal Proteins / metabolism
  • Immunization
  • Mice
  • Molecular Weight
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Th1 Cells / immunology

Substances

  • Culture Media, Conditioned
  • Cytokines
  • Fungal Proteins
  • Recombinant Proteins
  • Cellulase
  • Carboxymethylcellulose Sodium