title: ATR/TEM8 is highly expressed in epithelial cells lining Bacillus anthracis' three sites of entry: implications for the pathogenesis of anthrax infection.
abstract: Anthrax is a disease caused by infection with spores from the bacteria Bacillus anthracis. These spores enter the body, where they germinate into bacteria and secrete a tripartite toxin that causes local edema and, in systemic infections, death. Recent studies identified the cellular receptor for anthrax toxin (ATR), a type I membrane protein. ATR is one of the splice variants of the tumor endothelial marker 8 (TEM8) gene. ATR and TEM8 are identical throughout their extracellular and transmembrane sequence, and both proteins function as receptors for the toxin. ATR/TEM8 function and expression have been associated with development of the vascular system and with tumor angiogenesis. TEM8 is selectively upregulated in endothelial cells during blood vessel formation and tumorigenesis. However, selective expression of TEM8 in endothelial cells contradicts the presumably ubiquitous expression of the receptor. To resolve this controversial issue, we evaluated the distribution of ATR/TEM8 in a variety of tissues. For this purpose, we generated and characterized a novel anti-ATR/TEM8 polyclonal antibody. Here, we show that this novel antibody recognizes all three ATR/TEM8 isoforms, which are widely and differentially expressed in various tissue types. We found that ATR/TEM8 expression is not only associated with tumor endothelial cells, as previously described. Indeed, ATR/TEM8 is highly and selectively expressed in the epithelial cells lining those organs that constitute the anthrax toxin's sites of entry, i.e., the lung, the skin, and the intestine. In fact, we show that ATR/TEM8 is highly expressed in the respiratory epithelium of the bronchi of the lung and is particularly abundant in the ciliated epithelial cells coating the bronchi. Furthermore, immunostaining of skin biopsies revealed that ATR/TEM8 is highly expressed in the keratinocytes of the epidermis. Finally, we show that the epithelial cells lining the small intestine strongly express ATR/TEM8 isoforms. This is the first demonstration that the ATR/TEM8 protein is highly expressed in epithelial cells, which represent the primary location for bacterial invasion. These results suggest that the ATR/TEM8 expression pattern that we describe here is highly relevant for understanding the pathogenesis of anthrax infection.
authors.affiliation: Department of Molecular Pharmacology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461, USA.
authors.forename: Gloria
authors.initials: G
authors.lastname: Bonuccelli
authors.forename: Federica
authors.initials: F
authors.lastname: Sotgia
authors.forename: Philippe G
authors.initials: PG
authors.lastname: Frank
authors.forename: Terence M
authors.initials: TM
authors.lastname: Williams
authors.forename: Cecilia J
authors.initials: CJ
authors.lastname: de Almeida
authors.forename: Herbert B
authors.initials: HB
authors.lastname: Tanowitz
authors.forename: Philipp E
authors.initials: PE
authors.lastname: Scherer
authors.forename: Kylie A
authors.initials: KA
authors.lastname: Hotchkiss
authors.forename: Bruce I
authors.initials: BI
authors.lastname: Terman
authors.forename: Brent
authors.initials: B
authors.lastname: Rollman
authors.forename: Abdelkrim
authors.initials: A
authors.lastname: Alileche
authors.forename: Jürgen
authors.initials: J
authors.lastname: Brojatsch
authors.forename: Michael P
authors.initials: MP
authors.lastname: Lisanti
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000881:Anthrax / Q000503:physiopathology*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004847:Epithelial Cells / Q000502:physiology*; D015870:Gene Expression / Q000502:physiology*; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D008840:Microfilament Proteins; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D020033:Protein Isoforms; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000737:chemistry / Q000502:physiology*; D012867:Skin / Q000378:metabolism
chemical_list: C473287:ANTXR1 protein, human; D008565:Membrane Proteins; D008840:Microfilament Proteins; D009363:Neoplasm Proteins; D020033:Protein Isoforms; D011956:Receptors, Cell Surface