O-chain expression in the rough Brucella melitensis strain B115 : induction of O-polysaccharide-specific monoclonal antibodies and intracellular localization demonstrated by immunoelectron microscopy
| Autor: | Jean-Claude Nicolle, Axel Cloeckaert, J. N. Limet, Gérard Dubray, Michel S. Zygmunt |
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| Přispěvatelé: | Unité de Pathologie Infectieuse et Immunologie [Nouzilly] (PII), Institut National de la Recherche Agronomique (INRA), Unité de recherche Physiologie de la reproduction des mammifères domestiques, Nouzilly, ProdInra, Migration |
| Jazyk: | angličtina |
| Rok vydání: | 1992 |
| Předmět: |
medicine.drug_class
Immunoelectron microscopy BACTERIOLOGIE [SDV]Life Sciences [q-bio] Brucella abortus [INFO] Computer Science [cs] Monoclonal antibody Microbiology Epitope 03 medical and health sciences Antibody Specificity Immunoblot Analysis medicine [INFO]Computer Science [cs] Microscopy Immunoelectron ComputingMilieux_MISCELLANEOUS 030304 developmental biology 0303 health sciences Hybridomas biology Strain (chemistry) 030306 microbiology Polysaccharides Bacterial Antibodies Monoclonal O Antigens Immunogold labelling biology.organism_classification Proteinase K Brucella Immunohistochemistry [SDV] Life Sciences [q-bio] POLYSACCHARIDE biology.protein lipids (amino acids peptides and proteins) Brucella melitensis |
| Zdroj: | Journal of general microbiology Journal of general microbiology, Society for General Microbiology, (Journal Sales), 1992, 138, pp.1211-1219 |
| ISSN: | 0022-1287 |
| Popis: | Spleen cells from mice infected with the rough Brucella melitensis strain B115 were fused with NSO myeloma cells. Hybridoma supernatants were screened in ELISA with cell walls (CW), sonicated cell extracts (CE) and rough lipopolysaccharide (R-LPS) of B. melitensis strain B115 and whole B. melitensis B115 cells. Surprisingly, 22 monoclonal antibodies (mAbs) reacting in ELISA with both CW and CE but not with R-LPS and bacterial cells were shown by immunoblot analysis and ELISA to react with smooth lipopolysaccharide (S-LPS). These mAbs also reacted in ELISA with O polysaccharides (OPS) from the smooth Brucella abortus strain 99 and the smooth B. melitensis strain 16M and thus recognize epitopes present on the O-chain. Proteinase K LPS preparations from B. melitensis B115 analysed by immunoblotting with one mAb (12G12) recognizing S-LPS of both A and M specificity displayed the typical S-LPS high-molecular-mass ladder pattern but no S-LPS was detected in the phenol/water/chloroform/light petroleum LPS preparation of the same strain. mAb 12G12, specific for S-LPS, and a mAb (A68/03F03/D05) specific for R-LPS were used to localize the O-chain and R-LPS expressed in B. melitensis strain B115 by immunoelectron microscopy. Immunogold labelling was observed at the surface of B. melitensis B115 cells with the anti-R-LPS mAb but not with the anti-S-LPS mAb. In ultrathin sections, immunogold labelling with the S-LPS specific mAb was observed in the cytoplasm and in the periphery of the cytoplasm, probably at the cytoplasmic membrane.(ABSTRACT TRUNCATED AT 250 WORDS) |
| Databáze: | OpenAIRE |
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