A murine monoclonal antibody defines a unique epitope shared by Klebsiella lipopolysaccharides

Abstract

A hybridoma secreting a monoclonal antibody (MAb) directed against Klebsiella lipopolysaccharide (LPS) was derived from spleen cells of mice immunized with a smooth, nonencapsulated Klebsiella strain (Friedlander 201; serogroup 01). The MAb, called V/9-5 (immunoglobulin G2a), cross-reacted with LPS preparations produced from reference strains for the Klebsiella 0 serogroups 01, 02ab, 02ac, 03, 04, 05, and 012. Furthermore, the MAb reacted with LPSs from serogroup reference strains 06/08, 09, and 011, which are regarded as being identical to 01, 02, and 04, respectively. When testing the supernatant of clinically isolated Kkbsiella strains by means of an inhibition enzyme-linked immunosorbent assay, we found that 86 (92.4%) of 93 Klebsiella pneumoniae subsp. pneumoniae isolates and 24 (96.0%) of 25 K. oxytoca isolates harbored the cross-reactive epitope. By contrast, two laboratory strains of K. pneumoniae subsp. rhinoscleromatis did not react with MAb V/9-5. The MAb proved to be specific for the genus Klebsiella, since it did not react with any of a total of 73 strains belonging to other gram-negative bacterial genera. In conjunction with other LPS-specific MAbs, MAb V/9-5 might become a useful reagent for rapid identification of klebsiellae in clinical specimens. Furthermore, the epitope recognized by MAb V/9-5 might serve as a target epitope for the production of human MAbs for immunotherapeutic purposes.

O-antigen seroepidemiology of Klebsiella clinical isolates and implications for immunoprophylaxis of Klebsiella infections

“…These findings confirm earlier observations by Ørskov, who showed that strains in which only parts of the O2a antigen are expressed may exist (36). Three hundred thirty-eight of 378 isolates (89.4%) reacted with MAb V/9-5, which confirms our previous observation that ϳ10% of clinical Klebsiella strains lack the genus-specific core epitope (45). Nineteen of 40 nonreactive strains were found in the O1 serogroup (Table 4).…”

“…MAbs. MAb Ru-O1, specific for Klebsiella O1 LPS, and MAb V/9-5, specific for a Klebsiella genus-specific antigen located in the outer core region of LPS, have been described previously (39,45). MAb IV/4-5 was obtained by the same methods except that strain 7380 (O2ab:KϪ) was used for immunization of mice.…”

“…One of the gels was stained with silver according to the method of Tsai and Frasch (47), while the LPS bands from the second were transblotted to an Immobilon-P membrane with a transblotting cell (Bio-Rad, Richmond, Calif.). The membrane was reacted with rabbit antisera or murine MAbs as described previously, and the reactions were developed by adding alkaline phosphatase-conjugated secondary antibodies and the appropriate substrates (45).…”

The structure of the core region of the lipopolysaccharide from Klebsiella pneumoniae O3 3-Deoxy-alpha-D-manno-octulosonic acid (alpha-Kdo) residue in the outer part of the core, a common structural element of Klebsiella pneumoniae O1, O2, O3, O4, O5, O8, and O12 lipopolysaccharides

“…Preliminary NMR data show that O-specific polysaccharides, obtained after deamination of LPSs from K. pneumoniae serotypes O1±O5, O8, and O12, contained an aHep-(134)-a-Kdo-(236)-anh-Man fragment at the reducing end, where a-Hep (residue P) is present in the same proportion as in the nonsubstituted core and the O-chain is linked to O-4 of the Kdo residue L (data will be published elsewhere). The presence of the common fragment between the O-chain and core parts of the LPS may be responsible for the interaction with a monoclonal antibody, specifically recognizing Klebsiella LPSs [25,26], since other structural fragments of the LPS are present only in some serotypes or not unique for Klebsiella [26].…”

Aquisição passiva de anticorpos IgG maternos reativos com os lipopolissacarídeos de enterobactérias incidentes em infecções neonatais por recém-nascidos pré-termos e a termo.

“…Já as linhagens de Klebsiella e Pseudomonas são isoladas com freqüência do trato intestinal de neonatos nos primeiros dias de vida, e vão sendo substituídas por outras espécies, durante a colonização pela microbiota, tornando-se incomuns no trato intestinal de adultos (Bezirtzoglou, 1997 (Luderitz, et al, 1973;Freudenberg, et al,2001). O core também compartilha epítopos entre as diversas bactérias Gram-negativas, o que poderia explicar a reatividade cruzada entre os anticorpos IgM dirigidos às espécies de E. coli, como também à outras espécies de bactérias Gram-negativas analisadas no presente trabalho (Currie, et al, 2001;Zapata-Quintanilla, et al, 2006) As (Rukavina, et al, 2005), que compartilham um epítopo comum localizado no core da molécula do LPS (Trautmann, et al, 1994). (sorotipos 1, 3, 4, 5, 6, 10, 11 e 16), que mostrou eficácia em estudos retrospectivos realizados em pacientes com fibrose cística (Cryz, et al, 1994(Cryz, et al, , 1997Lang, et al, 2004;Langford and Hiller, 1984;Schaadet, al., 1990).…”