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In this study, we report a novel murine in vitro BBB model combining primary brain endothelial cells, Mycobacterium bovis bacillus Calmette-Gu\u00e9rin\u2013infected dendritic cells (DCs), PBMCs, and bacterial Ag-specific CD4+ T cells. We show that mycobacterial infection limits DC mobility and also induces cellular cluster formation that has a similar composition to pulmonary mycobacterial granulomas. Within the clusters, infection from DCs disseminates to the recruited monocytes, promoting bacterial expansion. Mycobacterium-induced in vitro granulomas have been described previously, but this report shows that they can form on brain endothelial cell monolayers. Cellular cluster formation leads to cluster-associated damage of the endothelial cell monolayer defined by mitochondrial stress, disorganization of the tight junction proteins ZO-1 and claudin-5, upregulation of the adhesion molecules VCAM-1 and ICAM-1, and increased transmigration of bacteria-infected cells across the BBB. TNF-\u03b1 inhibition reduces cluster formation on brain endothelial cells and mitigates cluster-associated damage. These data describe a model of bacterial dissemination across the BBB shedding light on a mechanism that might contribute to CNS tuberculosis infection and facilitate treatments.<\/jats:p>","DOI":"10.4049\/jimmunol.2001094","type":"journal-article","created":{"date-parts":[[2021,7,28]],"date-time":"2021-07-28T14:40:10Z","timestamp":1627483210000},"page":"1065-1077","source":"Crossref","is-referenced-by-count":7,"title":["<i>Mycobacterium bovis<\/i>\n                    Bacillus Calmette\u2013Gu\u00e9rin\u2013Infected Dendritic Cells Induce TNF-\u03b1\u2013Dependent Cell Cluster Formation That Promotes Bacterial Dissemination through an In Vitro Model of the Blood\u2013Brain Barrier"],"prefix":"10.1093","volume":"207","author":[{"given":"Trey E","family":"Gilpin","sequence":"first","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, School of Medicine and Public Health, University of Wisconsin-Madison , Madison, WI"},{"name":"Graduate Training Program of Cellular and Molecular Pathology, School of Medicine and Public Health, University of Wisconsin-Madison , Madison, WI"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8145-2823","authenticated-orcid":false,"given":"Fruzsina R","family":"Walter","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, School of Medicine and Public Health, University of Wisconsin-Madison , Madison, WI"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melinda","family":"Herbath","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, School of Medicine and Public Health, University of Wisconsin-Madison , Madison, WI"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matyas","family":"Sandor","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, School of Medicine and Public Health, University of Wisconsin-Madison , Madison, WI"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zsuzsanna","family":"Fabry","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, School of Medicine and Public Health, University of Wisconsin-Madison , Madison, WI"},{"name":"Graduate Training Program of Cellular and Molecular Pathology, School of Medicine and Public Health, University of Wisconsin-Madison , Madison, WI"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2021,8,15]]},"reference":[{"key":"2025022610315322100_r1","doi-asserted-by":"crossref","first-page":"111","DOI":"10.3329\/jhpn.v28i2.4879","article-title":"Tuberculosis: a global health problem","volume":"28","author":"Zaman","year":"2010","journal-title":"J. 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