In vitro and in vivo evaluation of five low molecular weight proteins of Ehrlichia ruminantium as potential vaccine components

Sebatjane, S.I.; Pretorius, Ashley; Liebenberg, Junita; Steyn, H.C.; Kleef, Mirinda Van

doi:10.1016/j.vetimm.2010.05.011

Cited by 20 publications

(19 citation statements)

References 34 publications

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“…These genes were cloned in the pET102 TOPO vector as described previously [10]. For the present study, the proteins were freshly expressed in E.coli, precipitated using acetone and checked on SDS-PAGE and Western blot, as described previously [9]. Target cells were seeded in triplicate in a 48 well plate at 5 x 10 5 cells per well.…”

Section: Fatt-ctl Assaymentioning

confidence: 99%

“…Much effort has been spent on the identification of proteins that induce strong cell-mediated immune responses characterised by CD4 + or CD8 + T cell responses. To date in our laboratory several E. ruminantium proteins have been characterised [7,8,9,10,11]. These studies have shown that T cells induce IFN-γ production in immune sheep or bovine peripheral blood mononuclear cells (PBMC) but assays done have not included the cytotoxic T cell assay to show that CD8 + T cells induce lysis of target cells.…”

Section: Introductionmentioning

confidence: 99%

“…In previous E. ruminantium vaccine development studies in our laboratory, 14 genes were selected as vaccine candidates based mainly on their ability to induce IFN-γ [8,9,10,11].…”

Section: Introductionmentioning

confidence: 99%

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Ehrlichia ruminantium antigens and peptides induce cytotoxic T cell responses in vitro

Thema

Tshilwane

Sơn

et al. 2019

Veterinary Immunology and Immunopathology

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Since CD8 + T cells play an important role in resistance to infection with heartwater, effective vaccines against this disease will likely require identification of antigens that contain CD8 + T cell epitopes responsible for cytotoxic T lymphocyte (CTL) responses. With the use of the fluorescent antigen-transfected target cell (FATT)-CTL assay, IFN-γ ELISPOT and flow cytometry, peptides that induce CTL, proliferation of CD8+T cells and IFN-γ production were identified as possible target antigens for vaccine development. Of particular relevance was the finding that different peptides from different antigens were able to elicit varied cytotoxic activities by immune peripheral blood mononuclear cells (PBMC) from heartwater immune tick-infected sheep. Several peptides derived from Erum0660, Erum2330, Erum2540, Erum2580 and Erum5000 induced CTL in immune sheep PBMC. Peptide Erum2540-6 was the only peptide that induced significant CTL, CD8 + CD45RO + and CD8 + IFN-γ + by PBMC from all three sheep, and Erum2540 and p2540-20 induced the highest % CTL response in all three outbred sheep. These results suggest that these epitopes may be of major importance in heartwater recombinant vaccine development.

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Section: Fatt-ctl Assaymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Ehrlichia ruminantium antigens and peptides induce cytotoxic T cell responses in vitro

Thema

Tshilwane

Sơn

et al. 2019

Veterinary Immunology and Immunopathology

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“…This difference might be explained by same reasons which were just discussed for MHCI prediction. Also for B-cell epitope prediction, this is not the first report in literature of low epitope prediction performance [39]. …”

Section: Discussionmentioning

confidence: 99%

An assessment on epitope prediction methods for protozoa genomes

et al. 2012

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BackgroundEpitope prediction using computational methods represents one of the most promising approaches to vaccine development. Reduction of time, cost, and the availability of completely sequenced genomes are key points and highly motivating regarding the use of reverse vaccinology. Parasites of genus Leishmania are widely spread and they are the etiologic agents of leishmaniasis. Currently, there is no efficient vaccine against this pathogen and the drug treatment is highly toxic. The lack of sufficiently large datasets of experimentally validated parasites epitopes represents a serious limitation, especially for trypanomatids genomes. In this work we highlight the predictive performances of several algorithms that were evaluated through the development of a MySQL database built with the purpose of: a) evaluating individual algorithms prediction performances and their combination for CD8+ T cell epitopes, B-cell epitopes and subcellular localization by means of AUC (Area Under Curve) performance and a threshold dependent method that employs a confusion matrix; b) integrating data from experimentally validated and in silico predicted epitopes; and c) integrating the subcellular localization predictions and experimental data. NetCTL, NetMHC, BepiPred, BCPred12, and AAP12 algorithms were used for in silico epitope prediction and WoLF PSORT, Sigcleave and TargetP for in silico subcellular localization prediction against trypanosomatid genomes.ResultsA database-driven epitope prediction method was developed with built-in functions that were capable of: a) removing experimental data redundancy; b) parsing algorithms predictions and storage experimental validated and predict data; and c) evaluating algorithm performances. Results show that a better performance is achieved when the combined prediction is considered. This is particularly true for B cell epitope predictors, where the combined prediction of AAP12 and BCPred12 reached an AUC value of 0.77. For T CD8+ epitope predictors, the combined prediction of NetCTL and NetMHC reached an AUC value of 0.64. Finally, regarding the subcellular localization prediction, the best performance is achieved when the combined prediction of Sigcleave, TargetP and WoLF PSORT is used.ConclusionsOur study indicates that the combination of B cells epitope predictors is the best tool for predicting epitopes on protozoan parasites proteins. Regarding subcellular localization, the best result was obtained when the three algorithms predictions were combined. The developed pipeline is available upon request to authors.

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“…Nevertheless all these attempts failed. Nowadays, four vaccine strategies against heartwater have been developed; the "infection and treatment" method using live bacteria, infection with in vitro attenuated bacteria [14], immunization with inactivated in vitro grown bacteria [15] and recombinant vaccines [16,17] (Table 1). At the moment, the only commercially available vaccine is based on live Ball3 organisms (which are less virulent than the other strains) derived from the blood of infected sheep, and its use is limited to South Africa [18].…”

mentioning

confidence: 99%

Tick-borne diseases in cattle: Applications of proteomics to develop new generation vaccines

Marcelino

Almeida

Ventosa

et al. 2012

Journal of Proteomics

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In vitro and in vivo evaluation of five low molecular weight proteins of Ehrlichia ruminantium as potential vaccine components

Cited by 20 publications

References 34 publications

Ehrlichia ruminantium antigens and peptides induce cytotoxic T cell responses in vitro

Ehrlichia ruminantium antigens and peptides induce cytotoxic T cell responses in vitro

An assessment on epitope prediction methods for protozoa genomes

Tick-borne diseases in cattle: Applications of proteomics to develop new generation vaccines

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