{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T19:48:33Z","timestamp":1768506513967,"version":"3.49.0"},"posted":{"date-parts":[[2023,6,7]]},"group-title":"Microbiology","reference-count":50,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,6,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  <jats:italic>Salmonella enterica<\/jats:italic>\n                  serovar Typhi (\n                  <jats:italic>S.<\/jats:italic>\n                  Typhi) is a human-restricted pathogen that replicates in macrophages. In this study, we investigated the roles of the\n                  <jats:italic>S.<\/jats:italic>\n                  Typhi Type 3 secretion systems (T3SSs) encoded on\n                  <jats:italic>Salmonella<\/jats:italic>\n                  Pathogenicity Islands (SPI) -1 (T3SS-1) and -2 (T3SS-2) during human macrophage infection. We found that mutants of\n                  <jats:italic>S<\/jats:italic>\n                  . Typhi deficient for both T3SSs were defective for intramacrophage replication as measured by flow cytometry, viable bacterial counts, and live time-lapse microscopy. T3SS-secreted proteins PipB2 and SifA contributed to\n                  <jats:italic>S.<\/jats:italic>\n                  Typhi replication and were translocated into the cytosol of human macrophages through both T3SS-1 and -2, demonstrating functional redundancy for these secretion systems. Importantly, an\n                  <jats:italic>S<\/jats:italic>\n                  . Typhi mutant strain that is deficient for both T3SS-1 and -2 was severely attenuated in the ability to colonize systemic tissues in a humanized mouse model of typhoid fever. Overall, this study establishes a critical role for\n                  <jats:italic>S.<\/jats:italic>\n                  Typhi T3SSs during its replication within human macrophages and during systemic infection of humanized mice.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Importance<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Salmonella enterica<\/jats:italic>\n                    serovar Typhi is a human-restricted pathogen that causes typhoid fever. Understanding the key virulence mechanisms that facilitate\n                    <jats:italic>S.<\/jats:italic>\n                    Typhi replication in human phagocytes will enable rational vaccine and antibiotic development to limit spread of this pathogen. While\n                    <jats:italic>S.<\/jats:italic>\n                    Typhimurium replication in murine models has been studied extensively, there is limited information available about\n                    <jats:italic>S.<\/jats:italic>\n                    Typhi replication in human macrophages, some of which directly conflicts with findings from\n                    <jats:italic>S.<\/jats:italic>\n                    Typhimurium murine models. This study establishes that both of\n                    <jats:italic>S.<\/jats:italic>\n                    Typhi\u2019s two Type 3 Secretion Systems (T3SS-1 and -2) contribute to intramacrophage replication and virulence.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2023.06.06.543980","type":"posted-content","created":{"date-parts":[[2023,6,7]],"date-time":"2023-06-07T09:45:20Z","timestamp":1686131120000},"source":"Crossref","is-referenced-by-count":0,"title":["<i>Salmonella enterica<\/i>\n                  serovar Typhi uses two type 3 secretion systems to replicate in human macrophages and to colonize humanized mice"],"prefix":"10.64898","author":[{"given":"Meagan","family":"Hamblin","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruth","family":"Schade","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ramya","family":"Narasimhan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Denise M.","family":"Monack","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023061008050510000_2023.06.06.543980v1.1","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1001\/archinte.1946.00220060002001","article-title":"Typhoid; clinical analysis of 360 cases","volume":"78","year":"1946","journal-title":"Arch Intern Med (Chic"},{"key":"2023061008050510000_2023.06.06.543980v1.2","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.20-0258"},{"key":"2023061008050510000_2023.06.06.543980v1.3","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.ijid.2022.03.036","article-title":"Toward Typhoid Fever Elimination","volume":"119","year":"2022","journal-title":"Int J Infect Dis"},{"key":"2023061008050510000_2023.06.06.543980v1.4","doi-asserted-by":"publisher","DOI":"10.3855\/jidc.219"},{"key":"2023061008050510000_2023.06.06.543980v1.5","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.83.14.5189"},{"key":"2023061008050510000_2023.06.06.543980v1.6","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2010.01904.x"},{"key":"2023061008050510000_2023.06.06.543980v1.7","doi-asserted-by":"crossref","first-page":"e12939","DOI":"10.1111\/cmi.12939","article-title":"Typhoidal Salmonella: Distinctive virulence factors and pathogenesis","volume":"20","year":"2018","journal-title":"Cell Microbiol"},{"key":"2023061008050510000_2023.06.06.543980v1.8","doi-asserted-by":"crossref","first-page":"102262","DOI":"10.1016\/j.mib.2022.102262","article-title":"One species, different diseases: the unique molecular mechanisms that underlie the pathogenesis of typhoidal Salmonella infections","volume":"72","year":"2023","journal-title":"Curr Opin Microbiol"},{"key":"2023061008050510000_2023.06.06.543980v1.9","doi-asserted-by":"crossref","first-page":"24","DOI":"10.15698\/mic2022.02.769","article-title":"Endomembrane remodeling and dynamics in Salmonella infection","volume":"9","year":"2022","journal-title":"Microb Cell"},{"key":"2023061008050510000_2023.06.06.543980v1.10","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1998.01048.x"},{"key":"2023061008050510000_2023.06.06.543980v1.11","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00065-13"},{"key":"2023061008050510000_2023.06.06.543980v1.12","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.041624-0"},{"key":"2023061008050510000_2023.06.06.543980v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2019.08.001"},{"key":"2023061008050510000_2023.06.06.543980v1.14","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0509183103"},{"key":"2023061008050510000_2023.06.06.543980v1.15","doi-asserted-by":"crossref","first-page":"e1009319","DOI":"10.1371\/journal.ppat.1009319","article-title":"Single cell analyses reveal distinct adaptation of typhoidal and non-typhoidal Salmonella enterica serovars to intracellular lifestyle","volume":"17","year":"2021","journal-title":"PLoS Pathog"},{"key":"2023061008050510000_2023.06.06.543980v1.16","doi-asserted-by":"crossref","unstructured":"Baldassarre M , Solano-Collado V , Balci A , Colamarino RA , Dambuza IM , Reid DM , Wilson HM , Brown GD , Mukhopadhyay S , Dougan G , Spano S . 2021. 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