Identification of in vivo-induced bacterial proteins during human infection with Salmonella enterica serotype paratyphi a

Mohammad Murshid Alam, Lillian L. Tsai, Sean M. Rollins, Alaullah Sheikh, Farhana Khanam, Meagan Kelly Bufano, Yanan Yu, Ying Wu-Freeman, Anuj Kalsy, Tania Sultana, M. Abu Sayeed, Nusrat Jahan, Regina C. LaRocque, Jason B. Harris, Daniel T. Leung, W. Abdullah Brooks, Stephen B. Calderwood, Richelle C. Charles, Firdausi Qadri, Edward T. Ryan

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Salmonella enterica serotype Paratyphi A is a human-restricted pathogen and the cause of paratyphoid A fever. Using a high-throughput immunoscreening technique, in vivo-induced antigen technology (IVIAT), we identified 20 immunogenic bacterial proteins expressed in humans who were bacteremic with S. Paratyphi A but not those expressed in S. Paratyphi A grown under standard laboratory conditions. The majority of these proteins have known or potential roles in the pathogenesis of S. enterica. These include proteins implicated in cell adhesion, fimbrial structure, adaptation to atypical conditions, oxidoreductase activity, proteolysis, antimicrobial resistance, and ion transport. Of particular interest among these in vivo-expressed proteins were S. Paratyphi A (SPA)2397, SPA2612, and SPA1604. SPA2397 and SPA2612 are prophage related, and SPA1604 is in Salmonella pathogenicity island 11 (SPI-11). Using real-time quantitative PCR (RT-qPCR), we confirmed increased levels of mRNA expressed by genes identified by IVIAT in a comparison of mRNA levels in organisms in the blood of bacteremic patients to those in in vitro cultures. Comparing convalescent- to acute-phase samples, we also detected a significant increase in the reaction of convalescent-phase antibodies with two proteins identified by IVIAT: SPA2397 and SPA0489. SPA2397 is a phage-related lysozyme, Gp19, and SPA0489 encodes a protein containing NlpC/P60 and cysteine, histidine-dependent amidohydrolase/peptidase (CHAP) domains. In a previous study utilizing a different approach, we found that transcripts for 11 and 7 of the genes identified by IVIAT were detectable in organisms in the blood of humans in Bangladesh who were bacteremic with S. Paratyphi A and Salmonella enterica serovar Typhi, respectively. S. Paratyphi A antigens identified by IVIAT warrant further evaluation for their contributions to pathogenesis and might have diagnostic, therapeutic, or preventive relevance.

Original languageEnglish
Pages (from-to)712-719
Number of pages8
JournalClinical and Vaccine Immunology
Volume20
Issue number5
DOIs
StatePublished - May 2013

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