Surface Antigens of Smooth Brucellae

Díaz, Ramón; Jones, Lois M.; Leong, Daniel; Wilson, J. B.

doi:10.1128/jb.96.4.893-901.1968

Cited by 88 publications

(62 citation statements)

References 11 publications

(12 reference statements)

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“…The R-type LPS found in the water phase of the phenol-water extract was not examined further since it did not react in immunodiffusion with bovine antiserum prepared against whole B. abortus cells. The crude LPS recovered in a 1.2% yield from the phenol phase showed strong serological reactivity with the bovine B. abortus antiserum, as recorded by other workers (8,9,23,25), and this material was subjected to further study.…”

supporting

confidence: 59%

Antigenic S-type lipopolysaccharide of Brucella abortus 1119-3

Caroff¹,

Bundle²,

Perry³

et al. 1984

Infect Immun

187

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Antigenic phenol-phase soluble lipopolysaccharide isolated from Brucella abortus 1119-3 by hot phenol-water extraction was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, controlled hydrolysis, periodate oxidation, methylation, and 'H and 1 C nuclear magnetic resonance studies to be an S-type lipopolysaccharide which could be cleaved to yield a lipid A and an 0-chain polysaccharide identified as an unbranched linear homopolymer of 1,2-linked 4,6-dideoxy-4-formamido-ft-D-mannopyranosyl residues. The serological reactivity of bovine antiserum to B. abortus 1119-3 with the lipopolysaccharides of Yersinia enterocolitica serotype 0:9 and Vibrio cholerae species has now been related to the occurrence of 1,2-linked Nacylated 4-amino-4,6-dideoxy-a-D-mannopyranosyl units in the 0-chain polysaccharides of their lipopolysaccharides.The diagnosis of brucellosis in humans and animals is frequently difficult to establish. Often the infection either is subclinical or yields varied responses in the host. Diagnosis, therefore, has frequently been based on the detection and quantification of Brucella antibodies in serum samples by agglutination, precipitin, complement fixation, and other methods (11). More recently, enzyme immunoassay tests with crude or purified antigens have been introduced (4, 20-22). False-negative and nonspecific reactions may be problems in these test systems. This report describes our work on the isolation, purification, and determination of the structure of a unique Brucella surface antigen which could be used both for diagnostic tests and for studies involving the production and use of monoclonal antibodies for the identification of Brucella abortus.The present investigation led to the identification of the antigenic 0-chain polysaccharide of B. abortus S-type lipopolysaccharide (LPS) as a linear homopolymer of 1,2-linked 4,6-dideoxy-4-formamido-a-D-mannopyranosyl units.(This paper was presented previously

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supporting

confidence: 59%

Antigenic S-type lipopolysaccharide of Brucella abortus 1119-3

Caroff¹,

Bundle²,

Perry³

et al. 1984

Infect Immun

187

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“…Smooth-lipopolysaccharides (sLPS) are long chain molecules consisting of a region of several polysaccharide repeating units (the O-chain), a small chain of a few carbohydrates (the core) and lipids (Lipid A) which bind the complex to the cell wall (Luderitz et al, 1971). For BruceUa, it is the sLPS-complex which is involved in almost all serological tests (Diaz et al, 1968). Some controversy exists as to whether the sLPS of BruceUa is complexed to proteins (Perera et al, 1984) or can be isolated (Caroff et al, 1984a).…”

Section: Brucella Antigensmentioning

confidence: 99%

“…Another component of Brucella which has a potential use in ELISA is poly B (also called Component 1, second component or PB ) a low molecular weight carbohydrate released from the rough strain B. melitensis Bl15 as well as other BruceUa strains with trichloroacetic acid (Diaz et al, 1968(Diaz et al, , 1981.…”

Section: Brucella Antigensmentioning

confidence: 99%

“…Its composition is unclear; it is chemically different from native hapten (NH) or acid hapten (AH) (Moreno et al, 1981;Perera et al, 1984); it is purified from rough rather than smooth strains , it is made within the cell of rough strains and it is immunologically distinct as determined by its inability to absorb out all antibodies to NH (Moreno et al, 1981 ). However, its similar cathodal mobility to sLPS (Diaz et al, 1968); partial immunological identity with NH (Fernandez-Lago et al, 1982) and partial identity with AH (Moreno et al, 1981 ) contrast the former differences. Although it does not bind antibodies when adsorbed to polystyrene tubes for an ELISA (Moreno et al, 1981 ) another interpretation might be that this carbohydrate does not adsorb to polystyrene (Diaz et al, 1984 ).…”

Section: Brucella Antigensmentioning

confidence: 99%

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A review of enzyme immunoassay for detection of antibody to Brucella abortus in cattle

Nielsen

Wright

Kelly

et al. 1988

Veterinary Immunology and Immunopathology

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Enzyme immunoassay has gained wide acceptance for serological diagnosis of bovine brucellosis because of its ability to detect antibody of all isotypes unlike the conventional tests. The indirect enzyme immunoassay, however, presents several parameters that require careful analysis. These parameters include the choice of antigen and antiglobulin-enzyme conjugate reagents for use in the assay, dealing with the large amount of data the semi-automatic or automatic assay can generate and the inter- and intralaboratory standardization and quality control. This review considers the various methods described in the literature and, briefly, how some of the problems have been overcome or how they might be dealt with.

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“…The lipid-protein moiety seems to differ from that of other gram-negative bacteria ( MARX et al, 1983;MORENO et al, 1979). The core region (H-antigen) is similar in both smooth and rough Brucella strains, while the polysaccharide side chain (0-antigen), which carries the main antigenic determinants recognized by the antisera of infected or vaccinated animals (DIAZ et al, 1968;MORENO et al, 1984;RAYBOULD, 1982), is missing in rough mutants ( MARX et al, 1983). Moreover, due to the similar chemical composition of the polysaccharide moieties of LPS, extensive serological cross-reactions between smooth Brucella species and other gram-negative bacteria have been reported (CIOGLIA, 1950;FEELEY, 1969;HURVELL, 1972).…”

Section: Introductionmentioning

confidence: 99%

Characterization of monoclonal antibodies against Brucella melitensis

Greiser‐Wilke

Moennig

Thon

et al. 1985

Zentralblatt für Veterinärmedizin Reihe B

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Summary Eight specific monoclonal antibodies were isolated from fusions of myeloma cells with spleen‐cells from mice immunized with Brucella melitensis 16 M. An Enzyme‐linked immunosorbent assay was used to screen the hybridoma supernatants and to determine the cross‐reactivity of the monoclonal antibodies with other bacteria. Five monoclonal antibodies reacted with smooth and rough Brucella strains and with all gram‐negative bacteria tested. One monoclonal antibody reacted with smooth Brucella strains and with Yersinia enterocolitica 0:9, and two monoclonal antibodies were specific for smooth Brucella strains. The latter two antibodies belonged to the antibody subclasses IgG 2 a (BM/38) and IgG 1 (BM/40). Both cultures were grown in serum‐free medium and were subsequently concentrated by AMICON ultrafiltration. The protein concentrations ranged between 1 and 3.5 mg/ml, the main contamination being residues of bovine serum albumin (40—60%). Samples of the concentrated antibodies were used for peroxidase‐labelling and were found to have high titres as determined by ELISA. A double antibody sandwich ELISA with both monoclonal antibodies was devised which was highly specific for Brucella melitensis 16 M. The applicability of such a test for diagnostic purposes is discussed. Zusammenfassung Brucella melitensis 16 M wurde zur Immunisierung von Mäusen verwendet. Durch die Fusion von Lymphozyten immuner Tiere mit Myelomzellen wurden acht Zellinien etabliert, die spezifische Antikörper produzieren. Ein „Enzyme‐linked Immunsorbent Assay” (ELISA) wurde zum Nachweis der Antikörper in den Hybridoma Überständen und zum Nachweis von Kreuzreaktionen mit anderen Bakterien verwendet. Fünf monoklonale Antikörper reagierten mit glatten und rauhen Brucella sp. und mit allen untersuchten gramnegativen Bakterien. Ein monoklonaler Antikörper reagierte mit glatten Brucella sp. und mit Yersinia enterocolitica 0 : 9, während zwei monoklonale Antikörper spezifisch für glatte Brucella‐Stämme waren. Die beiden letzteren gehörten zur IgG Subklasse 2 a (BM/38) bzw. zur Subklasse 1 (BM/40). Beide Antikörper wurden in serumfreiem Kulturmedium produziert und durch Amicon Ultrafiltration konzentriert. Die Proteinkonzentrationen lagen zwischen 1 und 3.5 mg/ml. Die Hauptkontamination bestand aus Resten von fetalem Kälberserum (40—60%). Ein Teil der konzentrierten Antikörper wurde mit Peroxidase markiert. Diese Antikörper zeigten einen hohen Titer im ELISA. Mit markiertem BM/40 und nicht‐markiertem BM/38 wurde ein ELISA zum Antigennachweis etabliert. Der Test war spezifisch für Brucella melitensis 16 M. Die mögliche Anwendung einer solchen Nachweismethode für diagnostische Zwecke wird erörtert. Résumé Caractérisation d'anticorps monoclonaux contre Brucella melitensis Brucella melitensis 16 M a été utilisée pour immuniser des souris. Par la fusion de lymphocytes des animaux immunisés avec des cellules de myélome, on a établi huit lignées cellulaires qui produisaient des anticorps. Un test ELISA (Enzyme‐linked‐Immunosorbent Assay) fut utilisé pour la m...

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Surface Antigens of Smooth Brucellae

Cited by 88 publications

References 11 publications

Antigenic S-type lipopolysaccharide of Brucella abortus 1119-3

Antigenic S-type lipopolysaccharide of Brucella abortus 1119-3

A review of enzyme immunoassay for detection of antibody to Brucella abortus in cattle

Characterization of monoclonal antibodies against Brucella melitensis

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