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The prototype strains\n                      <jats:italic toggle=\"yes\">Salmonella enterica<\/jats:italic>\n                      serovar Typhimurium (\n                      <jats:italic toggle=\"yes\">Salmonella<\/jats:italic>\n                      ) and\n                      <jats:italic toggle=\"yes\">Shigella flexneri<\/jats:italic>\n                      (\n                      <jats:italic toggle=\"yes\">Shigella<\/jats:italic>\n                      ) use Type-3-secretion-systems (T3SSs) to colonize intestinal epithelial cells (IECs). However, they have evolved unique sets of T3SS effectors and accessory virulence factors. A synthesis of how these differences impact the temporal progression of infection in non-transformed human epithelia is missing. Here, we followed\n                      <jats:italic toggle=\"yes\">Salmonella<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">Shigella<\/jats:italic>\n                      infections of human enteroids and colonoids by time-lapse imaging to pinpoint virulence factor modules that shape the divergent epithelial colonization strategies. By an apical targeting module that integrates flagella and the SPI-4-encoded adhesin system with T3SS-1,\n                      <jats:italic toggle=\"yes\">Salmonella<\/jats:italic>\n                      accomplishes appreciable numbers of apical IEC invasion events. These are promptly counteracted by IEC death, thus fostering a polyclonal iterative epithelial colonization strategy. The lack of a corresponding apical targeting module in\n                      <jats:italic toggle=\"yes\">Shigella<\/jats:italic>\n                      makes this pathogen reliant on external factors, for example, preexisting damage, for rare apical access to the intraepithelial environment. However,\n                      <jats:italic toggle=\"yes\">Shigella<\/jats:italic>\n                      compensates for this ineptness by an intraepithelial expansion module reliant on the tight coupling of OspC3-dependent temporal delay of cell death and IcsA-mediated mobility. This module enables intraepithelial\n                      <jats:italic toggle=\"yes\">Shigella<\/jats:italic>\n                      to evade the IEC death response just long enough to expand laterally, thus fostering an essentially monoclonal colonization strategy. Taken together, the study reveals how a small set of virulence factors shapes divergent epithelial colonization strategies by related enterobacteria.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Pathogenic bacteria employ partially overlapping sets of virulence factor functions to colonize host epithelia but can differ markedly in their pathogenesis and disease kinetics. This work combines bacterial genetics and real-time microscopy in enteroids and colonoids to decipher the divergent colonization strategies of\n                        <jats:italic toggle=\"yes\">Salmonella enterica<\/jats:italic>\n                        Typhimurium and\n                        <jats:italic toggle=\"yes\">Shigella flexneri<\/jats:italic>\n                        \u2014two major enteropathogens\u2014in non-transformed human intestinal epithelia. The results reveal how virulence factor modules enabling efficiency at either the invasion step (\n                        <jats:italic toggle=\"yes\">Salmonella<\/jats:italic>\n                        ) or the intraepithelial expansion stage (\n                        <jats:italic toggle=\"yes\">Shigella<\/jats:italic>\n                        ) drive the radically different infection cycles of these related enterobacteria. Moreover, our work emphasizes how real-time studies in infection models that retain primary host cell features are critical to understanding infectious disease progression.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.00911-25","type":"journal-article","created":{"date-parts":[[2025,5,30]],"date-time":"2025-05-30T05:45:27Z","timestamp":1748583927000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":10,"title":["Determinants of divergent\n                    <i>Salmonella<\/i>\n                    and\n                    <i>Shigella<\/i>\n                    epithelial colonization strategies resolved in human enteroids and colonoids"],"prefix":"10.1128","volume":"16","author":[{"given":"Petra","family":"Geiser","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/048a87296","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medical Biochemistry and Microbiology, Uppsala University","place":["Uppsala, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9491-4000","authenticated-orcid":true,"given":"Maria Letizia","family":"Di Martino","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/048a87296","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medical Biochemistry and Microbiology, Uppsala University","place":["Uppsala, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana C. 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