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Department of Medicine, Division of Infectious Diseases,
Biotechnology Laboratory, Departments of
Medical Genetics,
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Microbiology and Immunology, and
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Zoology,
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Department of Advanced Therapeutics of The British Columbia Cancer Agency, Faculties of Medicine and Science, University of British Columbia; and
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Research Institute of Vancouver Hospital and Health Sciences Center, Vancouver, British Columbia, Canada.
MHC class II expression was examined in macrophages infected with
Mycobacterium tuberculosis. IFN-
increased the surface
expression of class II molecules in THP-1 cells and this was markedly
reduced in cells infected with M. tuberculosis. Despite
this effect, steady state levels of HLA-DR
, HLA-DRß, and invariant
(Ii) chains were equivalent in control and infected cells. Metabolic
labeling combined with pulse-chase experiments and biochemical analysis
showed that the majority of class II molecules in infected cells became
resistant to endoglycosidase H, consistent with normal Golgi
processing. However, results of intracellular staining and dual color
confocal microscopy revealed a significant defect in transport of newly
synthesized class II molecules through the endocytic compartment. Thus,
compared with findings in control cells, class II molecules in infected
cells colocalized to a minimal extent with a lysosomal-associated
membrane protein-1+ endosomal compartment. In addition, in
contrast to control cells, class II molecules in infected cells failed
to colocalize with endocytosed BSA under conditions where this marker
is known to label late endosomes, lysosomes, and the MHC class II
compartment. Consistent with defective transport along the endocytic
pathway, the maturation of SDS-stable class II ß dimers—dependent
upon removal of Ii chain and peptide loading of class II dimers in the
MHC class II compartment—was markedly impaired in M.
tuberculosis-infected cells. These findings indicate that
defective transport and processing of class II molecules through the
endosomal/lysosomal system is responsible for diminished cell surface
expression of MHC class II molecules in cells infected with M.
tuberculosis.
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