Identification of Peptide Mimotopes of Trypanosoma brucei gambiense Variant Surface Glycoproteins

Nieuwenhove, Liesbeth Carolien Van; Rogé, Stijn; Balharbi, Fatima; Dieltjens, Tessa; Laurent, Thierry; Guisez, Yves; Büscher, Philippe; Lejon, Veerle

doi:10.1371/journal.pntd.0001189

Cited by 15 publications

(19 citation statements)

References 30 publications

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“…The ELISA was performed as previously described [16]. Briefly, the wells were incubated for 1 h at rT with 100 µL of phage dilution in PBS-Blotto (1/3 for culture plate supernatant or 1/20 for PEG-NaCl purified phage).…”

Section: Methodsmentioning

confidence: 99%

Identification of Mimotopes with Diagnostic Potential for Trypanosoma brucei gambiense Variant Surface Glycoproteins Using Human Antibody Fractions

et al. 2012

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Background At present, screening of the population at risk for gambiense human African trypanosomiasis (HAT) is based on detection of antibodies against native variant surface glycoproteins (VSGs) of Trypanosoma brucei ( T.b. ) gambiense . Drawbacks of these native VSGs include culture of infective T.b. gambiense trypanosomes in laboratory rodents, necessary for production, and the exposure of non-specific epitopes that may cause cross-reactions. We therefore aimed at identifying peptides that mimic epitopes, hence called “mimotopes,” specific to T.b. gambiense VSGs and that may replace the native proteins in antibody detection tests. Methodology/Principal Findings A Ph.D.-12 peptide phage display library was screened with polyclonal antibodies from patient sera, previously affinity purified on VSG LiTat 1.3 or LiTat 1.5. The peptide sequences were derived from the DNA sequence of the selected phages and synthesised as biotinylated peptides. Respectively, eighteen and twenty different mimotopes were identified for VSG LiTat 1.3 and LiTat 1.5, of which six and five were retained for assessment of their diagnostic performance. Based on alignment of the peptide sequences on the original protein sequence of VSG LiTat 1.3 and 1.5, three additional peptides were synthesised. We evaluated the diagnostic performance of the synthetic peptides in indirect ELISA with 102 sera from HAT patients and 102 endemic negative controls. All mimotopes had areas under the curve (AUCs) of ≥0.85, indicating their diagnostic potential. One peptide corresponding to the VSG LiTat 1.3 protein sequence also had an AUC of ≥0.85, while the peptide based on the sequence of VSG LiTat 1.5 had an AUC of only 0.79. Conclusions/Significance We delivered the proof of principle that mimotopes for T.b. gambiense VSGs, with diagnostic potential, can be selected by phage display using polyclonal human antibodies.

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Section: Methodsmentioning

confidence: 99%

Identification of Mimotopes with Diagnostic Potential for Trypanosoma brucei gambiense Variant Surface Glycoproteins Using Human Antibody Fractions

et al. 2012

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“…This observation led investigators to search for new ways of accurately identifying HAT-specific VSG epitopes (mimotopes) possibly recognized by the host immune system. Two studies illustrate well this idea (Van Nieuwenhove et al, 2011, 2012). In one study, two phage display libraries of random peptides (12-mer and cyclic 7-mer) were screened with mouse MAbs against the two predominant T.b.…”

Section: Phage Displaymentioning

confidence: 76%

“…gambiense VSGs proteins (LiTat 1.3 and LiTat 1.5). The result was the isolation of several peptides that mimicked epitopes on the native trypanosomal VSGs LiTat 1.5 and LiTat 1.3 (Van Nieuwenhove et al, 2011). Some of the selected peptides were then confirmed as mimotopes for the parasite VSGs since they were able to inhibit the binding of their homologous monoclonal to the corresponding VSG (Van Nieuwenhove et al, 2011).…”

Section: Phage Displaymentioning

confidence: 99%

Selection of binding targets in parasites using phage-display and aptamer libraries in vivo and in vitro

Tonelli¹,

Colli²,

Alves³

2013

Front. Immun.

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Parasite infections are largely dependent on interactions between pathogen and different host cell populations to guarantee a successful infectious process. This is particularly true for obligatory intracellular parasites as Plasmodium, Toxoplasma, and Leishmania, to name a few. Adhesion to and entry into the cell are essential steps requiring specific parasite and host cell molecules. The large amount of possible involved molecules poses additional difficulties for their identification by the classical biochemical approaches. In this respect, the search for alternative techniques should be pursued. Among them two powerful methodologies can be employed, both relying upon the construction of highly diverse combinatorial libraries of peptides or oligonucleotides that randomly bind with high affinity to targets on the cell surface and are selectively displaced by putative ligands. These are, respectively, the peptide-based phage display and the oligonucleotide-based aptamer techniques. The phage display technique has been extensively employed for the identification of novel ligands in vitro and in vivo in different areas such as cancer, vaccine development, and epitope mapping. Particularly, phage display has been employed in the investigation of pathogen–host interactions. Although this methodology has been used for some parasites with encouraging results, in trypanosomatids its use is, as yet, scanty. RNA and DNA aptamers, developed by the SELEX process (Systematic Evolution of Ligands by Exponential Enrichment), were described over two decades ago and since then contributed to a large number of structured nucleic acids for diagnostic or therapeutic purposes or for the understanding of the cell biology. Similarly to the phage display technique scarce use of the SELEX process has been used in the probing of parasite–host interaction. In this review, an overall survey on the use of both phage display and aptamer technologies in different pathogenic organisms will be discussed. Using these techniques, recent results on the interaction of Trypanosoma cruzi with the host will be highlighted focusing on members of the 85 kDa protein family, a subset of the gp85/TS superfamily.

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“…While evaluation of these recombinant antigens by ELISA gave very promising results, their diagnostic potential in an RDT format remains to be confirmed. Another approach has been to produce synthetic peptide mimotopes corresponding to epitopes of VSG LiTat 1.3 and VSG LiTat 1.5; however, the diagnostic accuracy of these peptides was found to be lower than that of full-length native antigens [63–65]. …”

Section: Discussionmentioning

confidence: 99%

Evaluation of Antigens for Development of a Serological Test for Human African Trypanosomiasis

Biéler

Waltenberger²,

Barrett

et al. 2016

PLoS ONE

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BackgroundControl and elimination of human African trypanosomiasis (HAT) can be accelerated through the use of diagnostic tests that are more accurate and easier to deploy. The goal of this work was to evaluate the immuno-reactivity of antigens and identify candidates to be considered for development of a simple serological test for the detection of Trypanosoma brucei gambiense or T. b. rhodesiense infections, ideally both.Methodology/Principal FindingsThe reactivity of 35 antigens was independently evaluated by slot blot and ELISA against sera from both T. b. gambiense and T. b. rhodesiense infected patients and controls. The antigens that were most reactive by both tests to T. b. gambiense sera were the membrane proteins VSG LiTat 1.3, VSG LiTat 1.5 and ISG64. Reactivity to T. b. rhodesiense sera was highest with VSG LiTat 1.3, VSG LiTat 1.5 and SRA, although much lower than with T. b. gambiense samples. The reactivity of all possible combinations of antigens was also calculated. When the slot blot results of 2 antigens were paired, a VSG LiTat 1.3- ISG75 combination performed best on T. b. gambiense sera, while a VSG LiTat 1.3-VSG LiTat 1.5 combination was the most reactive using ELISA. A combination of SRA and either VSG LiTat 1.3 or VSG LiTat 1.5 had the highest reactivity on T. b. rhodesiense sera according to slot blot, while in ELISA, pairing SRA with either GM6 or VSG LiTat 1.3 yielded the best results.ConclusionsThis study identified antigens that were highly reactive to T. b. gambiense sera, which could be considered for developing a serological test for gambiense HAT, either individually or in combination. Antigens with potential for inclusion in a test for T. b. rhodesiense HAT were also identified, but because their reactivity was comparatively lower, a search for additional antigens would be required before developing a test for this form of the disease.

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Identification of Peptide Mimotopes of Trypanosoma brucei gambiense Variant Surface Glycoproteins

Cited by 15 publications

References 30 publications

Identification of Mimotopes with Diagnostic Potential for Trypanosoma brucei gambiense Variant Surface Glycoproteins Using Human Antibody Fractions

Identification of Mimotopes with Diagnostic Potential for Trypanosoma brucei gambiense Variant Surface Glycoproteins Using Human Antibody Fractions

Selection of binding targets in parasites using phage-display and aptamer libraries in vivo and in vitro

Evaluation of Antigens for Development of a Serological Test for Human African Trypanosomiasis

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