Optimal T cell responses to Cryptococcus neoformans mannoprotein are dependent on recognition of conjugated carbohydrates by mannose receptors

J Immunol. 2002 Mar 15;168(6):2872-9. doi: 10.4049/jimmunol.168.6.2872.

Abstract

Cryptococcosis is a leading cause of death among individuals with compromised T cell function. Soluble Cryptococcus neoformans mannoproteins (MP) have emerged as promising vaccine candidates due to their capacity to elicit delayed-type hypersensitivity and Th type 1-like cytokines, both critical to the clearance of this pathogenic yeast. In this study, the mechanisms responsible for the potent immunostimulatory properties of MP were explored. Using Chinese hamster ovary cells expressing human macrophage mannose receptor (MMR), we determined that MP is a MMR ligand. Functionally, competitive blockade of multilectin mannose receptors (MR) on APCs diminished MP-dependent stimulation of primary T cells from immunized mice and the MP-reactive CD4(+) T cell hybridoma, P1D6, by 72 and 99%, respectively. Removal of O-linked saccharides from MP by beta-elimination inhibited MP-dependent stimulation of P1D6 and primary T cells by 89 and 90%, respectively. In addition, MP-dependent stimulation of P1D6 was abrogated after digestion with proteinase K, suggesting the protein core of MP contributed the antigenic moiety presented by APC. Stimulation of P1D6 by MP also was abolished using APC obtained from invariant chain-deficient mice, demonstrating Ag presentation was MHC class II restricted. Our data suggest that MP is a ligand for the MMR and that T cell stimulation is functionally inhibited either by competitive blockade of MR or by removal of carbohydrate residues critical for recognition. The demonstration that efficient T cell responses to MP require recognition of terminal mannose groups by MMR provides both a molecular basis for the immunogenicity of cryptococcal MP and support for vaccination strategies that target MR.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antigens, Differentiation, B-Lymphocyte / genetics
  • Antigens, Fungal / chemistry
  • Antigens, Fungal / immunology
  • CHO Cells / metabolism
  • Carbohydrate Conformation
  • Cricetinae
  • Cryptococcus neoformans / immunology*
  • Epitopes, T-Lymphocyte / immunology
  • Fungal Proteins / chemistry
  • Fungal Proteins / immunology*
  • Fungal Proteins / metabolism
  • Fungal Proteins / physiology
  • Glycoconjugates / immunology*
  • Histocompatibility Antigens Class II / genetics
  • Humans
  • Hybridomas / immunology
  • Hybridomas / microbiology
  • Immunologic Deficiency Syndromes / genetics
  • Immunologic Deficiency Syndromes / immunology
  • Lectins, C-Type*
  • Lymphocyte Activation / immunology
  • Male
  • Mannose Receptor
  • Mannose-Binding Lectins*
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / immunology*
  • Membrane Glycoproteins / metabolism
  • Membrane Glycoproteins / physiology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Receptors, Cell Surface / antagonists & inhibitors
  • Receptors, Cell Surface / biosynthesis
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / immunology*
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / microbiology*

Substances

  • Antigens, Differentiation, B-Lymphocyte
  • Antigens, Fungal
  • Epitopes, T-Lymphocyte
  • Fungal Proteins
  • Glycoconjugates
  • Histocompatibility Antigens Class II
  • Lectins, C-Type
  • Mannose Receptor
  • Mannose-Binding Lectins
  • Membrane Glycoproteins
  • Receptors, Cell Surface
  • invariant chain
  • mannoproteins