Fibronectin-Facilitated Invasion of T84 Eukaryotic Cells by Campylobacter jejuni Occurs Preferentially at the Basolateral Cell Surface

Abstract

Previous studies have indicated that the ability to bind to fibronectin is a key feature in successful cell invasion by Campylobacter jejuni. Given the spatial distribution of fibronectin and the architecture of the epithelium, this suggests the possibility that C. jejuni cell invasion might preferentially occur at the basolateral cell surface. To test this hypothesis, we examined the interaction of C. jejuni with T84 human colonic cells. When grown under the appropriate conditions, T84 cells form a polarized cell monolayer. C. jejuni translocation of a T84 cell monolayer appeared to occur via a paracellular (extracellular) route as opposed to a transcellular (intracellular) route based on the finding that a C. jejuni noninvasive mutant translocated as efficiently as its isogenic parent. Additional studies revealed that two distinct C. jejuni wild-type isolates could compete with one another for host cell receptors, whereas a C. jejuni fibronectin-binding-deficient mutant could not compete with a wild-type isolate for host cell receptors. Further, C. jejuni adherence and internalization were significantly inhibited by antifibronectin antibodies but only when cells were first treated with EGTA to expose basolateral cell surfaces. Together, these results support the theory that C. jejuni invasion occurs preferentially at the basolateral surface of eukaryotic cells.

Enteric Campylobacter: Purging Its Secrets?

“…Although some isolates of C. jejuni, such as the well characterized 81-176 strain, are highly invasive in these experimental models, many isolates show low levels of host cell entry in vitro (65). It has been suggested that campylobacters may not efficiently enter the host cell via the apical membrane, and recent evidence supports the contention that C. jejuni preferentially enters polarized epithelial cells via the basolateral membrane (66). In support of this model, there exists evidence both for paracellular passage (66) and M-cell transcytosis (64, 65) of C. jejuni.…”

Campylobacter-Induced Interleukin-8 Secretion in Polarized Human Intestinal Epithelial Cells RequiresCampylobacter-Secreted Cytolethal Distending Toxin- and Toll-Like Receptor-Mediated Activation of NF-κB

“…The adherence, invasion, and transcytosis of Campylobacter in polarized epithelial cells were analyzed as previously described (42). Briefly, polarized T84 epithelial-cell monolayers were washed with invasion medium and incubated with Campylobacter (ϳ2 ϫ 10 6 CFU; MOI, ϳ10) apically at 37°C for 4 h. The transepithelial resistance of the epithelial-cell monolayers was checked before and after incubation with the bacteria, and no significant change was detected.…”

“…Previous studies have shown that the interaction of enteric pathogens, such as Salmonella enterica serovar Typhimurium (37), Shigella flexneri (5), and Yersinia enterocolitica (50), with T84 polarized human intestinal epithelial cells induced the secretion of IL-8 and the transmigration of subepithelial neutrophils to the apical surface (5,37,38). Like those bacteria, campylobacters colonize, invade, and transmigrate across polarized human intestinal epithelial cells (30,42) and induce IL-8 secretion in nonpolarized and polarized human intestinal epithelial cell lines (9,21,36).…”

Campylobacter jejuni response to human mucin MUC2: modulation of colonization and pathogenicity determinants

“…Adherence involves different types of lectin-like molecules attaching to their corresponding glycosylated receptors on the epithelial cells (Guerry et al, 2002). Major outermembrane proteins such as the putative rod-shapedetermining protein MreB, the putative periplasmic protein MreC and C. jejuni lipoprotein JlpA play important roles in bacterial adhesion (Ashgar et al, 2007;del Rocio Leon-Kempis et al, 2006;Jin et al, 2003;Kakuda & DiRita, 2006;Karlyshev et al, 2000;Konkel et al, 1997;Linton et al, 2000;Mamelli et al, 2006;McSweegan & Walker, 1986;Monteville & Konkel, 2002;Moser et al, 1997;Pei & Blaser, 1993). The genes encoding these three proteins were upregulated in response to MUC2.…”