Shigella flexneri transformants expressing type 1 (mannose-specific) fimbriae bind to, activate, and are killed by phagocytic cells.
Shigella flexneri M90T (invasive) and BS176 (noninvasive) are typical nonfimbriated organisms that do not bind to or activate phagocytic cells. We demonstrate that S. flexneri M90Tp and BS176p, obtained by transformation of the strains named above with the cluster of genes encoding type 1 (mannose-specific) fimbriae of Escherichia coli, express the functional fimbriae, as shown by electron microscopy, by binding of antifimbria antibodies and by yeast cell aggregation. The transformants, but not the parental strains, bound to human granulocytes and mouse peritoneal macrophages. This binding was inhibited by methyl alpha-D-mannoside but not by methyl alpha-D-galactoside. The bound bacteria induced oxidative burst activation and degranulation of the granulocytes in vitro. With mouse peritoneal macrophages, the binding of the fimbriated bacteria induced degranulation in vitro. Injection of the bacteria into mouse peritoneum also induced degranulation of the macrophages in vivo; no such effect was observed with the nonfimbriated strains. The bound fimbriated transformants were effectively killed by the human granulocytes in vitro in the absence of opsonins or after opsonization with human anti-S. flexneri antiserum. The nonfimbriated strains were killed only after opsonization. These results provide further evidence for the role of type 1 fimbriae in lectin-mediated nonopsonic phagocytosis.
Gbarah
A
A
Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Mirelman
D
D
Sansonetti
P J
PJ
Verdon
R
R
Bernhard
W
W
Sharon
N
N
D000818:Animals; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001770:Blood Bactericidal Activity; D015550:Cell Degranulation; D004926:Escherichia coli / Q000235:genetics; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000502:physiology*; D005798:Genes, Bacterial; D006098:Granulocytes / Q000276:immunology*; D006801:Humans; D007111:Immunity, Cellular; D066298:In Vitro Techniques; D008163:Luminescent Measurements; D008297:Male; D008358:Mannose / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D010587:Phagocytosis; D011994:Recombinant Proteins / Q000378:metabolism; D016897:Respiratory Burst; D012763:Shigella flexneri / Q000378:metabolism / Q000472:pathogenicity* / Q000648:ultrastructure; D014170:Transformation, Genetic
D001425:Bacterial Outer Membrane Proteins; D011994:Recombinant Proteins; D036781:Fimbriae Proteins; D008358:Mannose