Abstract
Escherichia coli K1 is an important neonatal pathogen that is usually transferred from maternal to infant gastrointestinal tract at the time of parturition. Approximately 20% of neonates are colonized, and a proportion of colonized infants goes on to have systemic infection. Entry into the bloodstream from the gastrointestinal tract is hypothesized to occur via epithelial cell invasion. Invasion of multiple epithelial cell lines was studied using gentamicin protection assays and transcytosis of polarized monolayers. Electron microscopy was used to confirm cellular invasion. Cell lines used include two human gastrointestinal lines, Caco-2 and T84; a human respiratory cell line, A549; a human laryngeal cell line, HEp-2; and a canine kidney cell line, MDCK. A virulent E. coli K1 strain, RS218, readily invaded HEp-2, A549, and T84 cell lines in gentamicin protection assays, but was less invasive into MDCK and Caco-2 cells. RS218 also demonstrated transcytosis of both T84 and Caco-2 cells. Four clinical isolates of E. coli K1 demonstrated levels of transcytosis of T84 cells similar to RS218. Caco-2 invasiveness correlated with length of time in tissue culture with maximum invasiveness demonstrated at 11 d in culture, when cells were polarized and differentiated.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- K:
-
capsular antigen
- ATCC:
-
American Type Culture Collection
- MEM:
-
modified Eagle medium
- CFU:
-
colony forming units
- ETEC:
-
enterotoxigenic E. coli
References
Robbins JB, McCracken GH, Gotschlich EC, Orskov F, Orskov I, Hanson LA 1974 Escherichia coli K1 capsular polysaccharide associated with neonatal meningitis. N Engl J Med 290: 1216–1220
Sarff LD, McCracken GH 1975 Epidemiology of Escherichia coli K1 in healthy and diseased newborns. Lancet 1: 1099–1104
Huang SH, Wass C, Fu Q, Prasadarao NV, Stins M, Kim KS 1995 Escherichia coli invasion of brain microvascular endothelial cells in vitro and in vivo: molecular cloning and characterization of invasion gene ibe10. Infect Immun 63: 4470–4475
Meier C, Oelschlaeger TA, Merkert H, Korhonen TK, Hacker J 1996 Ability of Escherichia coli isolates that cause meningitis in newborns to invade epithelial and endothelial cells. Infect Immun 64: 2391–2399
Isberg RR, Falkow S 1985 A single genetic locus encoded by Yersinia pseudotuberculosis permits invasion of cultured animal cells by Escherichia coli K-12. Nature 317: 262–264
Finlay BB, Falkow S 1990 Salmonella interactions with polarized human intestinal Caco-2 epithelial cells. J Infect Dis 162: 1096–1106
Silver RP, Vimr ER 1990 Polysialic acid capsule of Escherichia coli K1. In: Iglewski BH, Clark VL(eds) Molecular Basis of Bacterial Pathogenesis. Academic Press, San Diego, 39–60.
Bloch CA, Orndorff PE 1990 Impaired colonization by and full invasiveness of Escherichia coli K1 bearing a site-directed mutation in the type 1 pilin gene. Infect Immun 58: 275–278
Galan JE, Curtiss R 1989 Cloning and molecular characterization of genes whose products allow Salmonella typhimurium to penetrate tissue culture cells. Proc Natl Acad Sci USA 86: 6383–6387
Sambrook J, Fritsch EF, Maniatis T 1989 Molecular Cloning, A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY pp A1
Rousset M, Laburthe M, Pinto M, Chevalier G, Rouyer-Fessard C, Dussaulx E, Trugnan G, Boige N, Brun JL, Zweibaum A 1985 Enterocytic differentiation and glucose utilization in the human colon tumor cell Caco-2: modulation by forskolin. J Cell Physiol 123: 377–385
Lowry DH, Rosebough NJ, Farr AL, Randall RJ 1951 Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275
Messer M, Dahlqvist A 1966 A one-step ultramicro method for the assay of intestinal disaccharidases. Anal Biochem 14: 376–392
Garen A, Levinthal C 1960 A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. Biochim Biophys Acta 38: 470–483
Burns JL, Jonas M, Chi EY, Clark DK, Berger A, Griffith A 1996 Invasion of respiratory epithelial cells by Burkholderia (Pseudomonas) cepacia. Infect Immun 64: 4054–4059
Coconnier MH, Bernet-Camard MF, Servin AL 1994 How intestinal epithelial cell differentiation inhibits the cell-entry of Yersinia pseudotuberculosis in colon carcinoma Caco-2 cell line in culture. Differentiation 58: 87–94
Gaillard JL, Finlay BB 1996 Effect of cell polarization and differentiation on entry of Listeria monocytogenes into the enterocyte-like Caco-2 cell line. Infect Immun 64: 1299–1308
Pinto M, Robine-Leon S, Appay MD, Kedinger M, Triadou N, Dussaulx E, Lacroix B, Simon-Assmann P, Haffen K, Fogh J, Zweibaum A 1983 Enterocyte-like differentiation and polarization of the human colon carcinoma cell line Caco-2 in culture. Biol Cell 47: 323–330
Donnenberg MS, Donohue-Rolfe A, Keusch GT 1989 Epithelial cell invasion: an overlooked property of enteropathogenic Escherichia coli (EPEC) associated with the EPEC adherence factor. J Infect Dis 160: 452–459
Donnenberg MS, Kaper JB 1992 Enteropathogenic Escherichia coli. Infect Immun 60: 3953–3961
Elsinghorst EA, Kopecko DJ 1992 Molecular cloning of epithelial cell invasion determinants from enterotoxigenic Escherichia coli. Infect Immun 60: 2409–2417
Elsinghorst EA, Weitz JA 1994 Epithelial cell invasion and adherence directed by the enterotoxigenic Escherichia coli tib locus is associated with a 104-kilodalton outer membrane protein. Infect Immun 62: 3463–3471
Oelschlaeger TA, Barrett TJ, Kopecko DJ 1994 Some structures and processes of human epithelial cells involved in uptake of enterohemorrhagic Escherichia coli O157: H7 strains. Infect Immun 62: 5142–5150
Korth MJ, Lara JC, Moseley SL 1994 Epithelial cell invasion by bovine septicemic Escherichia coli. Infect Immun 62: 41–47
Prasadarao NV, Wass CA, Weiser JN, Stins MF, Huang SH, Kim KS 1996 Outer membrane protein A of Escherichia coli contributes to invasion of brain microvascular endothelial cells. Infect Immun 64: 146–153
Panigrahi P, Bamford P, Horvath K, Morris JG, Gewolb IH 1996 Escherichia coli transcytosis in a Caco-2 cell model: implications in neonatal necrotizing enterocolitis. Pediatr Res 40: 415–421
Kerneis S, Chauviere G, Darfeuille-Michaud A, Aubel D, Coconnier MH, Joly B, Servin AL 1992 Expression of receptors for enterotoxigenic Escherichia coli during enterocytic differentiation of human polarized intestinal epithelial cells in culture. Infect Immun 60: 2572–2580
Glode MP, Sutton A, Moxon ER, Robbins JB 1977 Pathogenesis of neonatal Escherichia coli meningitis: induction of bacteremia and meningitis in infant rats fed E. coli K1. Infect Immun 16: 75–80
Cohen MB, Guarino A, Shukla R, Giannella RA 1988 Age-related differences in receptors for Escherichia coli heat-stable enterotoxin in the small and large intestine of children. Gastroenterology 94: 367–373
Lencer WI, Chu SHW, Walker WA 1987 Differential binding kinetics of cholera toxin to intestinal microvillus membrane during development. Infect Immun 55: 3126–3130
Selander RK, Korkhonen TK, Vaisanen-Rhen V, Williams PH, Pattison PE, Caugant DA 1986 Genetic relationships and clonal structure of strains of Escherichia coli causing neonatal septicemia and meningitis. Infect Immun 52: 213–222
Acknowledgements
The authors thank Craig Rubens and Steve Moseley for technical advice and critical discussion during preparation of the manuscript.
Author information
Authors and Affiliations
Additional information
Supported by National Institutes of Health grant AI32577.
Rights and permissions
About this article
Cite this article
Burns, J., Griffith, A., Barry, J. et al. Transcytosis of Gastrointestinal Epithelial Cells by Escherichia coli K1. Pediatr Res 49, 30–37 (2001). https://doi.org/10.1203/00006450-200101000-00010
Received:
Accepted:
Issue date:
DOI: https://doi.org/10.1203/00006450-200101000-00010
This article is cited by
-
Engineered K1F bacteriophages kill intracellular Escherichia coli K1 in human epithelial cells
Scientific Reports (2018)
-
Response of the respiratory mucosal cells to mycobacterium avium subsp. Hominissuis microaggregate
Archives of Microbiology (2018)
-
Glucocorticoid receptor auto-upregulation and its relation with glucocorticoid sensitivity in idiopathic nephrotic syndrome
International Urology and Nephrology (2011)


