The Mycobacterium tuberculosis Phagosome Is a HLA-I Processing Competent Organelle

Jeff E. Grotzke, Melanie Harriff, Anne C. Siler, Dawn Nolt, Jacob Delepine, Deborah Lewinsohn, David Lewinsohn

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Mycobacterium tuberculosis (Mtb) resides in a long-lived phagosomal compartment that resists maturation. The manner by which Mtb antigens are processed and presented on MHC Class I molecules is poorly understood. Using human dendritic lls and IFN-γ release by CD8+ T cell clones, we examined the processing and presentation pathway for two Mtb-derived antigens, each presented by a distinct HLA-I allele (HLA-Ia versus HLA-Ib). Presentation of both antigens is blocked by the retrotranslocation inhibitor exotoxin A. Inhibitor studies demonstrate that, after reaching the cytosol, both antigens require proteasomal degradation and TAP transport, but differ in the requirement for ER-golgi egress and new protein synthesis. Specifically, presentation by HLA-B8 but not HLA-E requires newly synthesized HLA-I and transport through the ER-golgi. Phenotypic analysis of the Mtb phagosome by flow organellometry revealed the presence of Class I and loading accessory molecules, including TAP and PDI. Furthermore, loaded HLA-I:peptide complexes are present within the Mtb phagosome, with a pronounced bias towards HLA-E:peptide complexes. In addition, protein analysis also reveals that HLA-E is enriched within the Mtb phagosome compared to HLA-A2. Together, these data suggest that the phagosome, through acquisition of ER-localized machinery and as a site of HLA-I loading, plays a vital role in the presentation of Mtb-derived antigens, similar to that described for presentation of latex bead-associated antigens. This is, to our knowledge, the first description of this presentation pathway for an intracellular pathogen. Moreover, these data suggest that HLA-E may play a unique role in the presentation of phagosomal antigens.

Original languageEnglish (US)
JournalPLoS Pathogens
Volume5
Issue number4
DOIs
StatePublished - Apr 2009

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Phagosomes
Mycobacterium tuberculosis
Organelles
Antigen Presentation
HLA-B8 Antigen
HLA-A2 Antigen
Antigens
Exotoxins
Microspheres
Cytosol
Proteins
Clone Cells
Alleles
T-Lymphocytes
Peptides
Mycobacterium tuberculosis antigens

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology

Cite this

The Mycobacterium tuberculosis Phagosome Is a HLA-I Processing Competent Organelle. / Grotzke, Jeff E.; Harriff, Melanie; Siler, Anne C.; Nolt, Dawn; Delepine, Jacob; Lewinsohn, Deborah; Lewinsohn, David.

In: PLoS Pathogens, Vol. 5, No. 4, 04.2009.

Research output: Contribution to journalArticle

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