In silico prediction and expression analysis of vaccine candidate genes of Campylobacter jejuni

Sabin Poudel; Linan Jia; Mark A Arick 2nd; Chuan-Yu Hsu; Adam Thrash; Anuraj T Sukumaran; Pratima Adhikari; Aaron S Kiess; Li Zhang

doi:10.1016/j.psj.2023.102592

In silico prediction and expression analysis of vaccine candidate genes of Campylobacter jejuni

Poult Sci. 2023 May;102(5):102592. doi: 10.1016/j.psj.2023.102592. Epub 2023 Feb 15.

Authors

Sabin Poudel¹, Linan Jia¹, Mark A Arick 2nd², Chuan-Yu Hsu², Adam Thrash², Anuraj T Sukumaran¹, Pratima Adhikari¹, Aaron S Kiess³, Li Zhang⁴

Affiliations

¹ Department of Poultry Science, Mississippi State University, Mississippi State, MS 39762, USA.
² Institute for Genomics, Biocomputing, and Biotechnology, Mississippi State University, Mississippi State, MS 39762, USA.
³ Prestage Department of Poultry Science, North Carolina State University, Raleigh, NC 27695, USA.
⁴ Department of Poultry Science, Mississippi State University, Mississippi State, MS 39762, USA. Electronic address: l.zhang@msstate.edu.

Abstract

Campylobacter jejuni (C. jejuni) is the most common food-borne pathogen that causes human gastroenteritis in the United States. Consumption of contaminated poultry products is considered as the major source of human Campylobacter infection. An effective vaccine would be a promising alternative to antibiotic supplements to curb C. jejuni colonization in poultry gastrointestinal (GI) tract. However, the genetic diversity among the C. jejuni isolates makes vaccine production more challenging. Despite many attempts, an effective Campylobacter vaccine is not yet available. This study aimed to identify suitable candidates to develop a subunit vaccine against C. jejuni, which could reduce colonization in the GI tract of the poultry. In the current study, 4 C. jejuni strains were isolated from retail chicken meat and poultry litter samples and their genomes were sequenced utilizing next-generation sequencing technology. The genomic sequences of C. jejuni strains were screened to identify potential antigens utilizing the reverse vaccinology approach. In silico genome analysis predicted 3 conserved potential vaccine candidates (phospholipase A [PldA], TonB dependent vitamin B12 transporter [BtuB], and cytolethal distending toxin subunit B [CdtB]) suitable for the development of a vaccine. Furthermore, the expression of predicted genes during host-pathogen interaction was analyzed by an infection study using an avian macrophage-like immortalized cell line (HD11). The HD11 was infected with C. jejuni strains, and the RT-qPCR assay was performed to determine the expression of the predicted genes. The expression difference was analyzed using ΔΔCt methods. The results indicate that all 3 predicted genes, PldA, BtuB, and CdtB, were upregulated in 4 tested C. jejuni strains irrespective of their sources of isolation. In conclusion, in silico prediction and gene expression analysis during host-pathogen interactions identified 3 potential vaccine candidates for C. jejuni.

Keywords: RT-qPCR; host-pathogen interaction; poultry; reverse vaccinology, Campylobacter jejuni.

MeSH terms

Animals
Campylobacter Infections* / genetics
Campylobacter Infections* / prevention & control
Campylobacter Infections* / veterinary
Campylobacter jejuni* / genetics
Campylobacter*
Chickens / genetics
Genes, Bacterial
Humans
Poultry
Vaccines*

Substances

Vaccines