Abstract
Monoclonal antibodies (MAbs) were generated against lysates of clinical Mycoplasma hominis isolates. Three of these, designated BG2, BA10, and FE6, recognized an integral membrane protein of M. hominis with an apparent molecular weight of 50,000 (p50). Electron microscopy studies demonstrated that this protein is distributed evenly over the cell surface. These anti-p50 MAbs were species specific for M. hominis; they reacted with 42% of 126 tested clinical M. hominis isolates and showed no reactivity to heterologous mycoplasma species. Immunoblot analysis after limited proteolysis of purified p50 demonstrated that the three MAbs reacted with different epitopes of the protein. Unlike BA10 and FE6, MAb BG2 induced a decrease in arginine metabolism and a reduction of CFU in metabolic inhibition tests. F(ab)2 fragments of MAb BG2 showed the same inhibitory effect as the intact MAb molecule, while Fab and Fc fragments had no influence on vital functions. Preincubation of the mycoplasmas with MAb BG2 followed by trypsin treatment yielded the same amount of CFU as the control without antibodies. In conclusion, the cell aggregates were resolved by the trypsin treatment. These experiments and tests with the antibody fragments led to the conclusion that only the intact MAb structure or the F(ab)2 structure had metabolic inhibition potential and that the observed metabolism inhibition as well as the apparent decrease in viability were a result of agglutination by MAb BG2.
MeSH Terms
Agglutination/immunology
Antibodies, Monoclonal/immunology
Antibody Specificity
Arginine/metabolism
Bacterial Outer Membrane Proteins/immunology,isolation & purification,metabolism
Dose-Response Relationship, Immunologic
Electrophoresis, Polyacrylamide Gel
Epitopes/analysis
Immunoblotting
Immunodiffusion
Immunoglobulin Fab Fragments/immunology,isolation & purification
Microscopy, Immunoelectron
Mycoplasma/immunology,metabolism
Chemicals
Antibodies, Monoclonal
Bacterial Outer Membrane Proteins
Epitopes
Immunoglobulin Fab Fragments
Arginine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Feldmann R C
Institute for Medical Microbiology and Virology, Heinrich-Heine-Universität, Düsseldorf, Germany.
Henrich B
Kolb-Bachofen V
Hadding U
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