Decreased capacity of recombinant 45/47-kDa molecules (Apa) of Mycobacterium tuberculosis to stimulate T lymphocyte responses related to changes in their mannosylation pattern.
The Apa molecules secreted by Mycobacterium tuberculosis, Mycobacterium bovis, or BCG have been identified as major immunodominant antigens. Mass spectrometry analysis indicated similar mannosylation, a complete pattern from 1 up to 9 hexose residues/mole of protein, of the native species from the 3 reference strains. The recombinant antigen expressed in M. smegmatis revealed a different mannosylation pattern: species containing 7 to 9 sugar residues/mole of protein were in the highest proportion, whereas species bearing a low number of sugar residues were almost absent. The 45/47-kDa recombinant antigen expressed in E. coli was devoid of sugar residues. The proteins purified from M. tuberculosis, M. bovis, or BCG have a high capacity to elicit in vivo potent delayed-type hypersensitivity (DTH) reactions and to stimulate in vitro sensitized T lymphocytes of guinea pigs immunized with living BCG. The recombinant Apa expressed in Mycobacterium smegmatis was 4-fold less potent in vivo in the DTH assay and 10-fold less active in vitro to stimulate sensitized T lymphocytes than the native proteins. The recombinant protein expressed in Escherichia coli was nearly unable to elicit DTH reactions in vivo or to stimulate T lymphocytes in vitro. Thus the observed biological effects were related to the extent of glycosylation of the antigen.
Horn
C
C
Unité de Physiopathologie de l'Infection, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France.
Namane
A
A
Pescher
P
P
Rivière
M
M
Romain
F
F
Puzo
G
G
Bârzu
O
O
Marchal
G
G
D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000494:pharmacology*; D006023:Glycoproteins / Q000235:genetics / Q000494:pharmacology*; D006031:Glycosylation; D006168:Guinea Pigs; D006968:Hypersensitivity, Delayed; D008213:Lymphocyte Activation / Q000187:drug effects; D008358:Mannose / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009169:Mycobacterium tuberculosis; D011994:Recombinant Proteins / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*
C095715:45-47 kDa antigen complex, Mycobacterium; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D006023:Glycoproteins; D011994:Recombinant Proteins; D008358:Mannose