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. 2021 Jul 21;10(8):917.
doi: 10.3390/pathogens10080917.

Immunogenic Peptides from Pap31 and SCS-α of Bartonella bacilliformis: One Step Closer to a Rapid Diagnostic Tool for Carrion's Disease

Cláudia Gomes  1 , Maria J Pons  2   3 , Juana Del Valle-Mendoza  3   4 , Mayumi Matsuoka  5 , Joaquim Ruiz  1   2
Affiliations

Immunogenic Peptides from Pap31 and SCS-α of Bartonella bacilliformis: One Step Closer to a Rapid Diagnostic Tool for Carrion's Disease

Cláudia Gomes et al. Pathogens. .

Abstract

Bartonella bacilliformis is the causal agent of Carrion's disease, an overlooked illness endemic in the Andean Mountains with Peru being the most affected country. The diagnostic of this illness is a challenge due to the limited resources and the common symptomatology with other infectious diseases. The goal of this study was to identify immunogenic peptides from Pap31 and succinyl-CoA synthetase α (SCS-α) of B. bacilliformis that might be suitable for developing a serologic tool. The immunodominant character of Pap31 and SCS-α was determined by Western blotting and in-silico analysis. Subsequently, 35 peptides were selected for epitope mapping and their immunoreactivity was tested by enzyme-linked immunosorbent assay (ELISA). A total of 30 sera were tested including pre-exposed people with high IgM levels for Pap31/SCS-α (23 sera), patients (2 sera) as well as 5 sera with no reactivity to Pap31/SCS-α. The results indicate that Pap31-8 (187QAIGSAILKGTKDTGT202) and SCS-α-12 (59IFASVAEGKEKTGANA74) are the most immunogenic peptides, with Pap31-8 showing potential to discriminate between B. bacilliformis and the remaining Bartonella spp., and SCS-α-12 differentiating Bartonella spp. from other microorganisms.

Keywords: Bartonella bacilliformis: diagnostic; Carrion’s disease; ELISA; immunogenic tools; peptides.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Expression and detection of Pap31 and SCS- α and their respective truncated constructs; SDS-PAGE showing the expression of the proteins of interest, full-length Pap31 and SCS- α and respective clones A, B, and C, corresponding to the three truncated constructs. The proteins of interest are marked with a dot. (a); Western blotting performed with the anti-Xpress antibody signalizing the proteins of interest. (b); Example of Western blotting performed with serum from exposed people for Pap31 and SCS-α and respective clones A, B, and C (c).
Figure 2
Figure 2
In-silico analysis of the immunogenic regions of (a) Pap31 protein; (b) SCS-α protein.
Figure 3
Figure 3
Absorbance levels of IgM ELISAs for all the antigenic peptides studied. (a) results for Pap31, and (b) results for SCS-α. Squares are the mean results for non-reactive samples; circles are the mean results of recently exposed sera, and triangles are the mean results for the positive samples.
Figure 4
Figure 4
Differences in the immunogenicity among peptides calculated with the Friedmann test. (a) Pap31, (b) SCS-α. p < 0.0001 (****); p < 0.0005 (***); p < 0.005 (**); p < 0.05 (*).
Figure 5
Figure 5
Immunogenicity levels for each peptide of (a) Pap31 and (b) SCS-α. The immunogenicity level of each peptide corresponds to the difference between the mean values of pre-exposed and non-reactive sera.
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