Identification of the principal serological immunodeterminants of African swine fever virus by screening a virus cDNA library with antibody

Kollnberger, Simon; Gutiérrez-Castañeda, Benito; Foster-Cuevas, Mildred; Corteyn, A.; Parkhouse, R. M. E.

doi:10.1099/0022-1317-83-6-1331

Cited by 86 publications

(71 citation statements)

References 35 publications

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“…Thus, the only unarguable proof for an antigen to become a real vaccine candidate comes from its potential to clinically protect individuals (60). Confirming this theory, in vitro screening of both B-and T-cell epitopes identified ASFV determinants that, however, failed to induce any measurable protection (61,62). This is most probably the main reason why there are only a few reports identifying optimal vaccine candidates screened from successful ELI libraries obtained from large and complex pathogens (63).…”

Section: Fig 4 Kinetics Of the Detection Of Ifn-␣ (A) And Tnf-␣ (B) Imentioning

confidence: 87%

Expression Library Immunization Can Confer Protection against Lethal Challenge with African Swine Fever Virus

Lacasta

Ballester

Monteagudo

et al. 2014

J Virol

106

112

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African swine fever is one of the most devastating pig diseases, against which there is no vaccine available. Recent work from our laboratory has demonstrated the protective potential of DNA vaccines encoding three African swine fever viral antigens (p54, p30, and the hemagglutinin extracellular domain) fused to ubiquitin. Partial protection was afforded in the absence of detectable antibodies prior to virus challenge, and survival correlated with the presence of a large number of hemagglutinin-specific CD8 IMPORTANCEAfrican swine fever is a highly contagious disease of domestic and wild pigs that is endemic in many sub-Saharan countries, where it causes important economic losses and is currently in continuous expansion across Europe. Unfortunately, there is no treatment nor an available vaccine. Early attempts using attenuated vaccines demonstrated their potential to protect pigs against experimental infection. However, their use in the field remains controversial due to safety issues. Although inactive and subunit vaccines did not confer solid protection against experimental ASFV infection, our DNA vaccination results have generated new expectations, confirming the key role of T-cell responses in protection and the existence of multiple ASFV antigens with protective potential, more of which are currently being identified. Thus, the future might bring complex and safe formulations containing more than a single viral determinant to obtain broadly protective vaccines. We believe that obtaining the optimal vaccine formulation it is just a matter of time, investment, and willingness.

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Section: Fig 4 Kinetics Of the Detection Of Ifn-␣ (A) And Tnf-␣ (B) Imentioning

confidence: 87%

Expression Library Immunization Can Confer Protection against Lethal Challenge with African Swine Fever Virus

Lacasta

Ballester

Monteagudo

et al. 2014

J Virol

106

112

View full text Add to dashboard Cite

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“…Most probably, the assay takes advantage of specific biological attributes associated with the p30. On one hand, p30 as a main ASFV- induced protein which is incorporated in the virus particle carries strong immunogenic determinants and consequently represents a very efficient inducer of humoral responses (7,8,10). On the other hand, this protein is expressed and secreted at early times after infection (1,21), which would explain its rapid immune recognition after the infection.…”

Section: Discussionmentioning

confidence: 99%

Optimization and Validation of Recombinant Serological Tests for African Swine Fever Diagnosis Based on Detection of the p30 Protein Produced inTrichoplusia niLarvae

et al. 2006

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We describe the validation of an enzyme-linked immunosorbent assay (ELISA) and confirmatory immunoblotting assays based on a recombinant p30 protein (p30r) produced in insect larvae using a baculovirus vector. Such validation included the following: (i) the scaling up and standardization of p30r production and the associated immunoassays, (ii) a broad immunological analysis using a large number of samples (a total of 672) from Spain and different African locations, and (iii) the detection of the ASF virus (ASFV)-antibody responses at different times after experimental infection. Yields of p30r reached up to 15% of the total protein recovered from the infected larvae at 3 days postinfection. Serological analysis of samples collected in Spain revealed that the p30r-based ELISA presented similar sensitivity to and higher specificity than the conventional Office International des Epizooties-approved ASFV ELISA. Moreover, the p30r ELISA was more sensitive than the conventional ELISA test in detecting ASFV-specific antibodies in experimentally infected animals at early times postinfection. Both the recombinant and conventional ELISAs presented variable rates of sensitivity and specificity with African samples, apparently related to their geographical origin. Comparative analyses performed on the sequences, predicted structures, and antigenicities of p30 proteins from different Spanish and African isolates suggested that variability among isolates might correlate with changes in antigenicity, thus affecting detection by the p30r ELISA. Our estimations indicate that more than 40,000 ELISA determinations and 2,000 confirmatory immunoblotting tests can be performed with the p30r protein obtained from a single infected larva, making this a feasible and inexpensive strategy for production of serological tests with application in developing countries.

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“…Indeed, previous studies have demonstrated that recombinant viral proteins can give improved specificity and sensitivity when they are applied to the analysis of European field sera (9,19,22). In previous studies, 12 serological immunodeterminants of ASFV were characterized by exhaustive screening of a representative lambda phage cDNA expression library of the tissue culture-adapted Ba71V isolate of ASFV for antibodies (12). These included four proteins encoded by previously unassigned open reading frames (ORFs) (B602L, C44L, CP312R, and K205R), as well as some of the more well studied structural proteins (pA104R, p10, p32, p54, and p73) and three enzymes (RNA reductase, DNA ligase, and thymidine kinase).…”

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confidence: 99%

Recombinant Antigen Targets for Serodiagnosis of African Swine Fever

Gallardo

Reis

Kalema‐Zikusoka

et al. 2009

Clin Vaccine Immunol

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African swine fever (ASF) is an infectious and economically important disease of domestic pigs. There is no vaccine, and so reliable diagnosis is essential for control strategies. The performance of four recombinant ASF virus (ASFV) protein (pK205R, pB602L, p104R, and p54)-based enzyme-linked immunosorbent assays (ELISAs) was evaluated with European porcine field sera that had been established by Office International des Epizooties (OIE)-approved tests to be ASFV negative (n ‫؍‬ 119) and ASFV positive (n ‫؍‬ 80). The values showed that there was almost perfect agreement between the results of the "gold standard" test (immunoblotting) and the results obtained by the p54-specific ELISA ( ‫؍‬ 0.95; 95% confidence interval [CI], 0.90 to 0.99) and the pK205R-specific ELISA or the pB602L-specific ELISA ( ‫؍‬ 0.92; 95% CI, 0.86 to 0.97). For the pA104R-specific ELISA, there was substantial to almost perfect agreement ( ‫؍‬ 0.81; 95% CI, 0.72 to 0.89). Similar results were observed by the OIE-approved ELISA ( ‫؍‬ 0.89; 95% CI, 0.82 to 0.95). Importantly, antibodies against these proteins were detectable early after infection of domestic pigs. Preliminary testing of 9 positive and 17 negative serum samples from pigs from West Africa showed identical results by the recombinant protein-based ELISA and the OIE-approved tests. In contrast, there was a high degree of specificity but a surprisingly a low level of sensitivity with 7 positive and 342 negative serum samples from pigs from East Africa. With poorly preserved sera, only the p104R-specific ELISA showed a significant reduction in sensitivity compared to that of the OIE-approved ELISA. Finally, these recombinant proteins also detected antibodies in the sera of the majority of infected warthogs. Thus, recombinant ASFV proteins p54, pB602L, and pK205R provide sensitive and specific targets for the detection of antibodies in European and West African domestic pigs and warthogs.

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Identification of the principal serological immunodeterminants of African swine fever virus by screening a virus cDNA library with antibody

Cited by 86 publications

References 35 publications

Expression Library Immunization Can Confer Protection against Lethal Challenge with African Swine Fever Virus

Expression Library Immunization Can Confer Protection against Lethal Challenge with African Swine Fever Virus

Optimization and Validation of Recombinant Serological Tests for African Swine Fever Diagnosis Based on Detection of the p30 Protein Produced inTrichoplusia niLarvae

Recombinant Antigen Targets for Serodiagnosis of African Swine Fever

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