Molecular basis for the lack of mimicry of Brucella polysaccharide antigens by Ab2gamma antibodies.
The crystal structure of the complex of an anti-Id Fab with an Fab specific for a Brucella polysaccharide antigen has previously been reported (Evans et al., 1994, J. Mol. Biol. 241, 691-705). To complement this study, the binding characteristics and immunological properties of this Ab2 and two others raised with a second anti-Brucella antibody were investigated, including quantitative kinetic measurements by surface plasmon resonance. The affinities of the Fabs from the Ab2s for the Ab1s were three orders of magnitude greater than those estimated for the antigen, but the Ab2s failed to induce antigen-binding Ab3s, that is, they were of the Ab2gamma type. The avidities of the Ab1s for antigen were however within one order of magnitude of their avidities for Ab2. Tests of 16 other anti-Brucella polysaccharide antibodies showed that the two idiotopes were not present in them, and in confirmation of the lack of a dominant idiotope, N-terminal sequencing of their H and L chains showed a wide variety of V genes were employed in the immune response to the Brucella polysaccharides. The failure of the Ab2 to induce antigen-reactive Ab3 thus appears to be due to neither intrinsic affinity nor idiotope frequency, but arises instead from structural reasons, for example, the incomplete penetration of the Ab2 into the binding-site cleft of the Ab1. The surface topography of polysaccharide antigens and their binding-sites thus appears to be especially difficult for Ab2s to mimic and will restrict their routine use as surrogates for T-cell independent polysaccharide antigens.
Young
N M
NM
Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario.
Gidney
M A
MA
Gudmundsson
B M
BM
MacKenzie
C R
CR
To
R
R
Watson
D C
DC
Bundle
D R
DR
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000737:chemistry / Q000276:immunology*; D002002:Brucella / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology; D007130:Immunoglobulin Idiotypes / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000276:immunology*; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D020349:Surface Plasmon Resonance
D000907:Antibodies, Bacterial; D007140:Immunoglobulin Fab Fragments; D007130:Immunoglobulin Idiotypes; D011135:Polysaccharides, Bacterial