Characterization of a 78-kilodalton outer membrane protein of Haemophilus somnus

Kania, Stephen A.; Gogolewski, R P; Corbeil, Lynette B.

doi:10.1128/iai.58.1.237-244.1990

Cited by 24 publications

(12 citation statements)

References 28 publications

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“…The position of p76 was confirmed on other blots with an additional lane containing recombinant p76 ( Fig. 1) and could be differentiated from the p78 OMP; the latter was found mainly in the Sarkosyl-insoluble fraction of the bacterial pellet, confirming previous studies suggesting that p78 is an integral membrane protein (31). The major OMP, p41, reacts poorly with convalescent-phase serum (59) but can be seen as a large pale staining band which displaced the immunodominant p40 in the Sarkosyl-insoluble fraction of the bacterial pellet (especially in the 20-l lane of Fig.…”

Section: Resultssupporting

confidence: 87%

“…The E. coli recombinant pHS138 expresses large amounts of p76 as well as truncated antigen. This recombinant was included to define the location of p76 because H. somnus has an immunodominant integral OMP with a molecular mass of ϳ78 kDa (16,20,31) which can be confused with p76. In this case, both p76 and the smaller peptides bind IgG2 anti-DNP.…”

Section: Resultsmentioning

confidence: 99%

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Haemophilus somnus immunoglobulin binding proteins and surface fibrils

1997

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The high-molecular-weight (HMW) immunoglobulin binding proteins (IgBPs) of Haemophilus somnus and a 76-kDa surface protein (p76) are found in serum-resistant virulent strains but not in several serum-sensitive strains from asymptomatic carriers. For the first time, p76 was shown to be an IgBP also. This was done by competitive inhibition studies with affinity-purified antidinitrophenol (anti-DNP) and DNP to ensure that binding was not antigen specific. The HMW IgBPs, but not the p76 IgBP, were partially purified from concentrated culture supernatant in detergent by fluid-phase liquid chromatography with a gel filtration column. Membrane extraction studies showed that p76 predominated in the Sarkosyl-soluble fraction of the bacterial cell pellet. Since integral outer membrane (OM) proteins are Sarkosyl insoluble, this is consistent with our previous finding that implicated p76 as a peripheral OM protein. The HMW IgBPs were found predominantly in the Sarkosyl-soluble fraction of the culture supernatant. This suggests that they were not integral membrane proteins and that their presence in the supernatant was not due to OM blebbing. We then showed that two IgBP-positive serum-resistant virulent strains have a surface fibrillar network but that two IgBP-negative serum-sensitive H. somnus strains from asymptomatic preputial carriers do not. Fibrils on the surfaces of IgBP ؉ strains bound gold-labelled bovine immunoglobulin G2 (IgG2) anti-DNP, indicating that these fibrils have IgG2 binding activity. Therefore, this study shows that H. somnus has two IgBPs, including a peripheral membrane protein and a fibrillar surface network.

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Section: Resultssupporting

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Haemophilus somnus immunoglobulin binding proteins and surface fibrils

1997

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“…Currently, the use of serological assays for diagnostic purposes, based on antibodies to whole H. somni, is of limited value; moreover, such assays are not widely available. Their value is small due to H. somni inhabiting mucosal sites in asymptomatic cattle, because of the possibility of subclinical infections, cross-reactive antibodies with other bacteria, or the confounding effects of vaccination (Kania et al 1990;Corbeil 2007;Pan et al 2014). The recent development of an enzyme-linked immunosorbent assay (ELISA) using antibodies to H. somni exopolysaccharide (EPS) (described in a later chapter), a major component of the H. somni biofilm matrix, holds promise of having an application in diagnostic settings (Pan et al 2014).…”

Section: Laboratory Diagnosis and Antimicrobial Resistance Testingmentioning

confidence: 99%

Histophilosis as a Natural Disease

O’Toole

Sondgeroth

2015

Current Topics in Microbiology and Immunology

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Histophilus somni is responsible for sporadic disease worldwide in cattle and, to a lesser extent, in small ruminants, bighorn sheep (Ovis canadensis), and North American bison (Bison bison). The importance of H. somni diseases can be attributed to improved clinical and laboratory recognition, combined with the growth in intensive management practices for cattle. Although outbreaks of bovine histophilosis can occur year-round, in northern and southern hemispheres, it is most frequent in late fall and early winter. Weather, stress, dietary changes, and comingling of cattle are likely to be major triggers for outbreaks. The most frequent clinical expressions of histophilosis include undifferentiated fever, fibrinosuppurative pneumonia, encephalitis-leptomeningitis, necrotizing myocarditis, and diffuse pleuritis. Neurological disease occurs either as thrombotic meningoencephalitis (TME) or as suppurative meningitis with ventriculitis. Acute myocarditis is characteristically necrotizing and generally involves one or both papillary muscles in the left ventricular myocardium. Biofilm-like aggregates of bacteria occur in capillaries and veins in myocardium, in the central nervous system, and on endocardial surfaces. H. somni is a component of bovine respiratory disease (BRD) complex. In our experience, it is most commonly diagnosed in subacute-to-chronic polymicrobial pulmonary infections in combination with Mannheimia haemolytica, Trueperella pyogenes, Pasteurella multocida, or Mycoplasma bovis. Other, less common forms of H. somni disease present as polyarthritis/tenosynovitis, abortion with placentitis and fetal septicemia, epididymitis-orchitis, and ocular infections. It is likely that H. somni is under-recognized clinically and diagnostically. Most state and provincial laboratories in North America rely on bacterial isolation to confirm infection. The use of more sensitive detection methods on field cases of histophilosis will help resolve the pathogenesis of H. somni in natural outbreaks, and whether the disease is as common elsewhere as it is in Canada.

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“…Isolates of H. somnus and other gram-negative bacteria were frozen at -70°C in 40% phosphate-buffered saline (PBS) (pH 7.2)-60% glycerol. For consistency, we included the same isolates of H. somnus (9,27) and gramnegative bacteria (27) as those used in our previous studies. The H. somnus isolates included five from TME (43826, 0289, 8025, 91-1, and 109B), five from pneumonia (1542, 2336, 3581, 3415-2, and 1297), four from reproductive failure (1030, 2069, 570, and B83-145), four from vaginal carriers (41VC, 202V, 208V, and 221V), and four from preputial carriers (iP, 24P, 127P, and 129Pt).…”

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Characterization of immunodominant surface antigens of Haemophilus somnus

1991

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An immunodominant Haemophilus somnus outer membrane protein with an apparent molecular mass of 40 kDa on Western blots (immunoblots) of gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels was characterized because a monospecific antibody against this antigen was protective. This monospecific antibody was used for immunoaffinity purification of the antigen. The immunoaffinity-purified antigen reacted with a polyclonal antibody to the 40-kDa antigen but not with a monoclonal antibody (3G9) which reacted with the 40-kDa antigen in gradient gels. On 8 or 10% gels, the-40-kDa antigen was resolved as two bands, a 40-kDa band which reacted with the protective monospecific polyclonal antibody (p40) and a band of lower molecular mass which reacted with monoclonal antibody 3G9. The latter antigen was designated p39. Both antigens were conserved in all H. somnus isolates tested. The specific antibodies were also used to detect cross-reacting antigens in other gram-negative bacteria. Antibody to p40 reacted with proteins of 55 to 28 kDa, with the greatest intensity shown among proteins from other members of the family PasteureUlaceae. Antibody to p40 was reduced by absorption with live H. somnus or other members of the family Pasteurellaceae, so the antigen appears to be surface exposed. Antibody to p39 only cross-reacted with a broad band (38 to 40 kDa) in Haemophilus agni. Since H. agni is not a bovine pathogen and since convalescent-phase serum from H. somnus-infected animals did recognize p39, the latter may be a good immunodiagnostic antigen, if the lack of cross-reactivity with antigens in other gram-negative bacteria is confirmed with a polyclonal antibody to p39. The cross-reactivity of antiserum to p40 with antigens of members of the family Pasteurellaceae and the ability of this antiserum to protect against H. somnus pneumonia indicate that p40 may be a useful vaccine antigen for H. somnus disease and perhaps even diseases caused by other members of the family Pasteurellaceae.

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Characterization of a 78-kilodalton outer membrane protein of Haemophilus somnus

Cited by 24 publications

References 28 publications

Haemophilus somnus immunoglobulin binding proteins and surface fibrils

Haemophilus somnus immunoglobulin binding proteins and surface fibrils

Histophilosis as a Natural Disease

Characterization of immunodominant surface antigens of Haemophilus somnus

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