Limits...
Antibodies against Escherichia coli O24 and O56 O-Specific Polysaccharides Recognize Epitopes in Human Glandular Epithelium and Nervous Tissue.

Korzeniowska-Kowal A, Kochman A, Gamian E, Lis-Nawara A, Lipiński T, Seweryn E, Ziółkowski P, Gamian A - PLoS ONE (2015)

Bottom Line: The results indicate that various human tissues are recognized by anti-O24 and anti-O56 antibodies.Interestingly, only a single specific reactivity could be found in the anti-O56 antibody preparation.Therefore, the most relevant observation is that the epitope recognized by anti-O56 antibodies is a new marker specific for glandular epithelium and nervous tissue.

View Article: PubMed Central - PubMed

Affiliation: Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114, Wrocław, Poland.

ABSTRACT
Lipopolysaccharide (LPS), the major component of the outer membrane of Gram-negative bacteria, contains the O-polysaccharide, which is important to classify bacteria into different O-serological types within species. The O-polysaccharides of serotypes O24 and O56 of E. coli contain sialic acid in their structures, already established in our previous studies. Here, we report the isolation of specific antibodies with affinity chromatography using immobilized lipopolysaccharides. Next, we evaluated the reactivity of anti-O24 and anti-O56 antibody on human tissues histologically. The study was conducted under the assumption that the sialic acid based molecular identity of bacterial and tissue structures provides not only an understanding of the mimicry-based bacterial pathogenicity. Cross-reacting antibodies could be used to recognize specific human tissues depending on their histogenesis and differentiation, which might be useful for diagnostic purposes. The results indicate that various human tissues are recognized by anti-O24 and anti-O56 antibodies. Interestingly, only a single specific reactivity could be found in the anti-O56 antibody preparation. Several tissues studied were not reactive with either antibody, thus proving that the presence of cross-reactive antigens was tissue specific. In general, O56 antibody performed better than O24 in staining epithelial and nervous tissues. Positive staining was observed for both normal (ganglia) and tumor tissue (ganglioneuroma). Epithelial tissue showed positive staining, but an epitope recognized by O56 antibody should be considered as a marker of glandular epithelium. The reason is that malignant glandular tumor and its metastasis are stained, and also epithelium of renal tubules and glandular structures of the thyroid gland are stained. Stratified epithelium such as that of skin is definitely not stained. Therefore, the most relevant observation is that the epitope recognized by anti-O56 antibodies is a new marker specific for glandular epithelium and nervous tissue. Further studies should be performed to determine the structure of the tissue epitope recognized.

No MeSH data available.


Related in: MedlinePlus

Structures of the O-specific polysaccharides from E. coli O24 and O56.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4472344&req=5

pone.0129492.g001: Structures of the O-specific polysaccharides from E. coli O24 and O56.

Mentions: Bacterial cells of E. coli serotypes O24 and O56 were grown in a liquid medium for the preparation of lipopolysaccharides. Isolation of LPS was performed using a standard hot phenol-water protocol. The method with proteinase K was used for SDS-PAGE analysis. The structures of the sialic acid-containing polysaccharides have been established previously [3] and are shown in Fig 1. Purified LPS was immobilized on a solid phase for the affinity purification of LPS specific antibody. Rabbit polyclonal serum was obtained after immunization with whole bacterial cells. Before affinity purification, the immunoglobulin fraction was obtained by ammonium sulfate precipitation from whole serum. Antibodies bound to immobilized LPS were eluted with KNCS. Fig 2 shows results of SDS-PAGE and immunoblotting analysis of the purified antibody. Clear reaction of antibody with LPS transferred to a membrane is visible. It is noteworthy that anti-E. coli O24 antibodies showed cross-reactivity with LPS O56, while anti-E. coli O56 antibodies recognized only homologous LPS. Anti-E. coli O56 antibodies recognize long chain LPS molecules present in low quantities which were difficult to visualize with silver staining. Interestingly, despite the close structural similarity between both O-antigens and the polyclonal character of serum, anti-O56 antibodies seem to be very specific to an epitope present exclusively on O56 O polysaccharide. The results confirm our previous studies on sialic acid containing lipopolysaccharides isolated from: Salmonella toucra O48, Citrobacter freundii O37, Hafnia alvei PCM 2386, Escherichia coli O24 and Escherichia coli O104, where we observed cross-reactivity only for anti-O24 serum and O56 LPS [9].


Antibodies against Escherichia coli O24 and O56 O-Specific Polysaccharides Recognize Epitopes in Human Glandular Epithelium and Nervous Tissue.

Korzeniowska-Kowal A, Kochman A, Gamian E, Lis-Nawara A, Lipiński T, Seweryn E, Ziółkowski P, Gamian A - PLoS ONE (2015)

Structures of the O-specific polysaccharides from E. coli O24 and O56.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4472344&req=5

pone.0129492.g001: Structures of the O-specific polysaccharides from E. coli O24 and O56.
Mentions: Bacterial cells of E. coli serotypes O24 and O56 were grown in a liquid medium for the preparation of lipopolysaccharides. Isolation of LPS was performed using a standard hot phenol-water protocol. The method with proteinase K was used for SDS-PAGE analysis. The structures of the sialic acid-containing polysaccharides have been established previously [3] and are shown in Fig 1. Purified LPS was immobilized on a solid phase for the affinity purification of LPS specific antibody. Rabbit polyclonal serum was obtained after immunization with whole bacterial cells. Before affinity purification, the immunoglobulin fraction was obtained by ammonium sulfate precipitation from whole serum. Antibodies bound to immobilized LPS were eluted with KNCS. Fig 2 shows results of SDS-PAGE and immunoblotting analysis of the purified antibody. Clear reaction of antibody with LPS transferred to a membrane is visible. It is noteworthy that anti-E. coli O24 antibodies showed cross-reactivity with LPS O56, while anti-E. coli O56 antibodies recognized only homologous LPS. Anti-E. coli O56 antibodies recognize long chain LPS molecules present in low quantities which were difficult to visualize with silver staining. Interestingly, despite the close structural similarity between both O-antigens and the polyclonal character of serum, anti-O56 antibodies seem to be very specific to an epitope present exclusively on O56 O polysaccharide. The results confirm our previous studies on sialic acid containing lipopolysaccharides isolated from: Salmonella toucra O48, Citrobacter freundii O37, Hafnia alvei PCM 2386, Escherichia coli O24 and Escherichia coli O104, where we observed cross-reactivity only for anti-O24 serum and O56 LPS [9].

Bottom Line: The results indicate that various human tissues are recognized by anti-O24 and anti-O56 antibodies.Interestingly, only a single specific reactivity could be found in the anti-O56 antibody preparation.Therefore, the most relevant observation is that the epitope recognized by anti-O56 antibodies is a new marker specific for glandular epithelium and nervous tissue.

View Article: PubMed Central - PubMed

Affiliation: Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114, Wrocław, Poland.

ABSTRACT
Lipopolysaccharide (LPS), the major component of the outer membrane of Gram-negative bacteria, contains the O-polysaccharide, which is important to classify bacteria into different O-serological types within species. The O-polysaccharides of serotypes O24 and O56 of E. coli contain sialic acid in their structures, already established in our previous studies. Here, we report the isolation of specific antibodies with affinity chromatography using immobilized lipopolysaccharides. Next, we evaluated the reactivity of anti-O24 and anti-O56 antibody on human tissues histologically. The study was conducted under the assumption that the sialic acid based molecular identity of bacterial and tissue structures provides not only an understanding of the mimicry-based bacterial pathogenicity. Cross-reacting antibodies could be used to recognize specific human tissues depending on their histogenesis and differentiation, which might be useful for diagnostic purposes. The results indicate that various human tissues are recognized by anti-O24 and anti-O56 antibodies. Interestingly, only a single specific reactivity could be found in the anti-O56 antibody preparation. Several tissues studied were not reactive with either antibody, thus proving that the presence of cross-reactive antigens was tissue specific. In general, O56 antibody performed better than O24 in staining epithelial and nervous tissues. Positive staining was observed for both normal (ganglia) and tumor tissue (ganglioneuroma). Epithelial tissue showed positive staining, but an epitope recognized by O56 antibody should be considered as a marker of glandular epithelium. The reason is that malignant glandular tumor and its metastasis are stained, and also epithelium of renal tubules and glandular structures of the thyroid gland are stained. Stratified epithelium such as that of skin is definitely not stained. Therefore, the most relevant observation is that the epitope recognized by anti-O56 antibodies is a new marker specific for glandular epithelium and nervous tissue. Further studies should be performed to determine the structure of the tissue epitope recognized.

No MeSH data available.


Related in: MedlinePlus