Cross-protection and Antigen Expression by Chicken Embryo-grown Pasteurella multocida in Chickens

Ibrahim, Roshita; Sawada, Takuo; Shahata, Mostafa A.; Ibrahim, Ass

doi:10.1053/jcpa.2000.0419

Cited by 10 publications

(4 citation statements)

References 18 publications

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“…Immunoblot of OMPs probed with hyperimmune sera against whole cell antigen of P. multocida P52 revealed a deeply immunostained band corresponding to a molecular weight of 39 kDa. The results clearly indicated that PlpE is most likely to be a 39 kDa cross protective immunodominant antigen, as described by Ibrahim et al [11] and Rimler [21]. It also indicates a satisfactory level of expression of PlpE grown in in-vitro conditions.…”

Section: Discussionsupporting

confidence: 79%

“…OMPs have been found to be potent immunogens and protective to mice, rabbits, chicken and calves [6,16,26]. A 39 kDa cross-protective antigen was identified by Rimler [20] in P. multocida A: 3 (strain P1059) which induced active and passive cross-protection in chickens and turkeys [11,21]. The monoclonal antibodies against 39 kDa protein of P. multocida P-1059 (serovar A: 3) were produced and characterized by Ali et al [1,2].…”

Section: Discussionmentioning

confidence: 99%

“…Among these, a 39 kDa protein has been reported to be highly immunogenic and cross protective, when challenged with heterologous strains [11,21]. The monoclonal antibodies to this protein were found to be protective in mice against lethal challenge with virulent strains [2].…”

Section: Introductionmentioning

confidence: 99%

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Molecular heterogeneity ofplpE gene in Indian isolates ofPasteurella multocidaand expression of recombinant PlpE in vaccine strain ofP. multocidaserotype B: 2

Singh

Ranjan

et al. 2010

J Vet Sci

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Outer membrane proteins of Pasteurella (P.) multocida have been known to be protective immunogens. Pasteurella lipoprotein E (PlpE) has been reported to be an important cross reactive outer membrane protein in P. multocida. The gene encoding the PlpE of P. multocida serotypes A: 3, B: 2 and D: 1 was amplified from the genomic DNA. The amplified products were cloned and the nucleotide sequence was determined. Sequence analysis of the recombinant clones revealed a single open reading frame of 1,011 bp, 1,008 bp and 1,017 bp encoding a protein with a calculated molecular mass of 37.829 kDa, 37.389 kDa and 37.965 kDa for serotypes A: 3, B: 2 and D: 1 respectively. The comparison of the plpE sequence in different capsular types revealed a high degree (>90%) of homology. Furthermore, the plpE gene of Haemorhhagic septicaemia causing serotype (B: 2) was expressed in E. coli and recombinant PlpE was strongly immunostained by antiserum against whole cell antigen, indicating that the protein is expressed in vivo.

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Section: Discussionsupporting

confidence: 79%

Section: Discussionmentioning

confidence: 99%

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Molecular heterogeneity ofplpE gene in Indian isolates ofPasteurella multocidaand expression of recombinant PlpE in vaccine strain ofP. multocidaserotype B: 2

Singh

Ranjan

et al. 2010

J Vet Sci

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“…These results indicates that the 39 kDa antigen of in vitro grown avian P. multocida is a cross-protective over serovars A:1 and A:3. Induction of a cross-protection in chickens was reportedly achieved by inoculation of inactivated vaccine prepared from in vivo grown P. multocida strain X-73 that expressed major protein bands at a 36-39 kDa position [8] or by a purified 39 kDa antigen of both in vivo and in vitro grown strain P-1059 [12]. SDS-PAGE analysis of strain P-1059 in our previous study [1] and the antigen purified in the present study showed that 39 kDa antigen occurred very near the molecular-mass region of H protein, which has been found at the outer surface of P. multocida [3,9].…”

mentioning

confidence: 99%

Protectivity of an Immunoaffinity-Purified 39kDa Capsular Protein of Avian Pasteurella multocida in Mice

Ali

Sawada

Noda³

2004

The Journal of Veterinary Medical Science

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ABSTRACT. A 39 kDa protein of avian Pasteurella multocida strain P-1059 (serovar A:3) was purified from a crude capsular extract by immunoaffinity chromatography by using a ligand of purified mouse monoclonal antibody to the 39 kDa capsular protein of the strain. Protective activity of the purified 39 kDa protein antigen was determined by inoculation of ddY mice twice with 25 or 125 µg of the protein and challenge-exposure with 10 or 50 LD 50 of strains P-1059 or X-73 (serovar A:1). The results showed that the antigen gave high protection (60 to 100%). These results indicated that the 39 kDa protein of avian P. multocida is a cross-protective antigen over serovars A:1 and A:3. KEY WORDS: active cross-protection, avian Pasteurella multocida, 39 kDa capsular protein.J. Vet. Med. Sci. 66(12): 1603-1604, 2004 Fowl cholera, typically an acute septicemic disease of many avian species, is caused mainly by Pasteurella multocida serovars A:1, A:3 and A:3•4 [6]. Despite extensive vaccination efforts, the disease often causes marked economic loss in the poultry industry. Commercial vaccines for control of the disease contain P. multocida bacterins or liveattenuated bacteria. Neverthless, the failure of bacterins to protect the birds against heterologous challenge [11] and outbreaks of fowl cholera caused by vaccination with liveattenuated vaccine strains [6,7] have been hampering vaccine development. In our previous study [1], monoclonal antibodies (Mabs) to a 39 kDa capsular protein of avian P. multocida strain P-1059 (serovar A:3) were produced and characterized. Two representative Mabs (HAA1 and HAA3) showed high protective activity in mice immunized passively with the Mabs and challenge-exposed with the strains of serovars A:1 and A:3. In the present study we attempted to purify the 39 kDa antigen with Mab HAA3 and to determine the protective activity of the antigen in mice.A large amount of Mab HAA3 was produced in a growth medium and in pristane (2,6,10,14-tetramethylpentadecane; Sigma Chemicals Co., MO, U.S.A.) primed BALB/c mice. Antibodies from the growth medium were purified by ammonium sulfate precipitation and antibodies of ascitic fluids were collected and purified by affinity chromatography through Affi-gel protein A (MAPII kit; Bio-Rad Laboratories, CA, U.S.A.). Prior to the coupling procedure antibodies were dialyzed against 0.1 M HEPES (Sigma) buffer (pH 8.0) and then concentrated. Targeted antigen was purified by immunoaffinity chromatography. Affi-gel-10 (Bio-Rad) was used for matrix support. Purified Mab HAA3 was coupled with the gel as described by the manufacturer. A crude capsular extract (CCE) of strain P-1059 [13] was passed through the column at 4°C, overnight. The antigen was eluted with 0.1 M glycine (pH 2.2) and the pH was neutralized by addition of 0.1 M Tris-HCl buffer (pH 9.0) and dialyzed against 0.02 M PBS (pH 7.2). The antigen was concentrated by ultra-filtration with a diaflomembrane of PM 10 (M.W. cut-off 10,000; Millipore Corp., MA, U.S.A.) and verified by SDS-PAGE analysis. To collect...

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PasteurellaandMannheimia

Boyce¹,

Lo²,

Wilkie³

et al. 2004

Pathogenesis of Bacterial Infections in Animals

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Cross-protection and Antigen Expression by Chicken Embryo-grown Pasteurella multocida in Chickens

Cited by 10 publications

References 18 publications

Molecular heterogeneity ofplpE gene in Indian isolates ofPasteurella multocidaand expression of recombinant PlpE in vaccine strain ofP. multocidaserotype B: 2

Molecular heterogeneity ofplpE gene in Indian isolates ofPasteurella multocidaand expression of recombinant PlpE in vaccine strain ofP. multocidaserotype B: 2

Protectivity of an Immunoaffinity-Purified 39kDa Capsular Protein of Avian Pasteurella multocida in Mice

PasteurellaandMannheimia

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