Differential Interleukin-8 Response of Intestinal Epithelial Cell Line to Reactogenic and Nonreactogenic Candidate Vaccine Strains of
            <i>Vibrio cholerae</i>

Rodrı́guez, Boris L.; Rojas, Armando; Campos, Javier; Ledón, Talena; Valle, Edgar; Toledo, William; Fando, Rafael

doi:10.1128/iai.69.1.613-616.2001

Cited by 33 publications

(42 citation statements)

References 24 publications

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“…5). This observation is consistent with the findings of a separate study, which demonstrated that strain CVD103-HgR induced less transcription and synthesis of IL-8 than other attenuated strains of V. cholerae in another intestinal epithelial cell line (47). These findings correlate with volunteer studies with these two classical strains, which found CVD101 to be reactogenic, while CVD103-HgR was nonreactogenic.…”

Section: Discussionsupporting

confidence: 91%

“…For example, CVD101 activated greater expression of the genes for IL-8 and MIP-3␣ than CVD103-HgR (Table 2). These results are consistent with previous studies that have shown CVD103-HgR to be nonreactogenic and well tolerated in human volunteers (30) and also to induce less IL-8 expression in the intestinal epithelial cell line HT29-18N2 (47). The nonreactogenic nature of CVD103-HgR may be due to it being derived from a strain (569B) with reduced ability to colonize the intestine (30).…”

Section: Effects Of V Cholerae Infection On Global Gene Expression Psupporting

confidence: 91%

“…Thus, increased transcription of these genes after infection with V. cholerae was anticipated. Previously it has been demonstrated that attenuated strains of V. cholerae induce IL-8 expression in the intestinal epithelial cell line HT29-18N2 (47). The findings of this investigation indicate that this response is also observed with the T84 intestinal epithelial cell line.…”

Section: Discussionsupporting

confidence: 70%

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Transcriptional Responses of Intestinal Epithelial Cells to Infection with Vibrio cholerae

et al. 2004

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Vibrio cholerae is a noninvasive enteric bacterium that causes the severe diarrheal disease cholera. Candidate cholera vaccines have been engineered by deleting genes encoding known virulence factors in V. cholerae; however, many of these attenuated strains were still reactogenic in human volunteers. In this study, DNA arrays were utilized to monitor the transcriptional responses of human intestinal epithelial cells (T84) to eight strains of V. cholerae, including attenuated, toxigenic, and environmental isolates. cDNA probes generated from host RNA samples were hybridized against low-and high-density gene arrays. V. cholerae induced the transcription of a variety of host genes and repressed the expression of a lower number of genes. Expression patterns were confirmed for certain genes by reverse transcriptase PCR and enzyme-linked immunosorbent assays. A core subset of genes was found to be differentially regulated in all experiments. These genes included genes involved in innate mucosal immunity, intracellular signaling, and cellular proliferation. Reactogenic vaccine strains induced greater expression of genes for certain proinflammatory cytokines than nonreactogenic strains. Wild-type and attenuated derivatives induced and repressed many genes in common, although there were differences in the transcription profiles. These results indicate that the types of host genes modulated by attenuated V. cholerae, and the extent of their induction, may mediate the symptoms seen with reactogenic cholera vaccine strains.The human diarrheal disease cholera results from infection with pathogenic strains of the gram-negative enteric bacterium Vibrio cholerae. Many serogroups of V. cholerae have been identified, but only two of these (O1 and O139) are important for epidemic disease. The O1 serogroup is further subdivided into the classical and El Tor biotypes, as well as the Inaba, Ogawa, and Hikojima serotypes. After ingestion, pathogenic V. cholerae bacteria colonize the small intestine and produce a number of virulence factors, notably cholera toxin (CT). This toxin targets and activates the adenylate cyclase within host epithelial cells, provoking the net secretion of chloride ions and water into the intestinal lumen, resulting in the extensive diarrhea that is characteristic of this disease (25). As such, the pathology of cholera has traditionally been considered noninflammatory (5). However, several reports provide evidence for an inflammatory response in cholera disease. Lymphocytes and mononuclear cells have been observed in the intestinal lamina propria in biopsy specimens from cholera patients (16, 40), and increased levels of lactoferrin, myeloperoxidase, and prostaglandins have been measured in stool samples from infected humans (43, 50). Leukocytes and erythrocytes were also detected in the stools (48), and elevated concentrations of nitric oxide metabolites were observed in urine and serum samples (22,43,44) from cholera patients. Finally, and of most relevance for this study, V. cholerae vaccine strains caused...

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Section: Discussionsupporting

confidence: 91%

Section: Effects Of V Cholerae Infection On Global Gene Expression Psupporting

confidence: 91%

Section: Discussionsupporting

confidence: 70%

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Transcriptional Responses of Intestinal Epithelial Cells to Infection with Vibrio cholerae

et al. 2004

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“…The hap gene is positively regulated by HapR at the transcription level (20). It was reported that a CTX and hap-defective vaccine strain, 638, was not reactogenic in human volunteers and this strain induced lower levels of IL-8 than its parent wild-type strain in HT29 cells (38). The protease activity in V. cholerae vaccine strains has been associated with decreases in the transcellular epithelial resistance of polarized T84 intestinal epithelial cells (31).…”

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confidence: 99%

Induction of Interleukin-8 in T84 Cells byVibrio cholerae

et al. 2004

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The induction of interleukin-8 (IL-8) in vitro has been suggested to correlate with the reactogenicity of Vibrio cholerae vaccine candidates. V. cholerae vaccine candidate 638, a hemagglutinin protease/hap-defective strain, was recently reported to be well tolerated in human volunteers, suggesting a role for Hap in reactogenicity. We examined the role of hap in the induction of IL-8 in intestinal epithelial T84 cells. Wild-type V. cholerae strains 3038 and C7258 and a vaccine candidate strain, JBK70, induced levels of IL-8 similar to those of their isogenic hap mutants. Supernatant containing Hap did not stimulate IL-8 production at a variety of concentrations tested, suggesting that Hap itself does not induce IL-8 production. Furthermore, supernatant from CVD115, which had deletions of hap and rtxA (encoding repeats in toxin) and was derived from a reactogenic strain, CVD110, induced IL-8 production in T84 cells in a dose-dependent manner. The IL-8-stimulating activity of CVD115 culture supernatants was growth phase dependent and was strongest in stationary phase cultures. This IL-8 stimulator(s) was resistant to heat treatment but sensitive to proteinase. Protease activity in vitro did not correlate with the reactogenicity of V. cholerae vaccine candidates. Our data suggest that Hap is not an IL-8 inducer in T84 cells and that the IL-8 stimulator in the supernatant of V. cholerae culture may play a role in reactogenicity.The acute diarrheal disease cholera remains a significant public health problem, causing more than five million cases and 200,000 deaths annually in the world (47). Vibrio cholerae, the etiologic agent of cholera, is transmitted by contaminated water and food. It colonizes the surface of the small intestine, where it secretes cholera toxin (CT), which is largely responsible for the acute diarrhea characteristic of cholera. Although there are more than 200 O-antigen serogroups of V. cholerae, only the O1 and O139 serogroups are associated with epidemic cholera (22). The O1 serogroup can be divided further into two biotypes, classical and El Tor, based on various biochemical and phenotypic differences (for a review, see reference 22). One important difference in particular is that V. cholerae strains of the classical biotype do not have rtx genes, encoding repeats in toxin (27), and the majority of classical strains do not produce hemagglutinin/protease (Hap) or hemolysin (22). Several groups of investigators have constructed a variety of live attenuated V. cholerae O1 and O139 vaccine candidates by deleting the ctx genes encoding cholera toxin (21,26,30,46,48). However, in clinical trials most V. cholerae vaccine candidates still exhibit reactogenicity, which includes mild diarrhea, malaise, nausea, vomiting, abdominal cramps, low-grade fever, and headache (8,21,45,46,48). The only licensed oral live cholera vaccine, CVD103-HgR, is well tolerated and highly protective in North American volunteers (26, 43, 46); however, it was poorly protective in a field trial involving 67,500 people in Indones...

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“…The inflammatory response to V. cholerae is characterized by neutrophil infiltration in the intestinal epithelium and the underlying lamina propria [7]. We and others previously demonstrated elicitation of IL-8 in epithelial cells exposed to V. cholerae laboratory as well as vaccine strains [13,14]. A number of attempts have been made to identify the V. cholerae components contributing to IL-8 expression.…”

Section: Introductionmentioning

confidence: 99%

Interleukin‐8 gene regulation in epithelial cells by Vibrio cholerae: role of multiple promoter elements, adherence and motility of bacteria and host MAPKs

et al. 2012

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Interleukin (IL)-8 is an important mediator in neutrophil-mediated acute inflammation. We previously demonstrated that incubation of intestinal epithelial cells with Vibrio cholerae O395 resulted in increased IL-8 mRNA expression and IL-8 secretion, which was associated with the adherence and motility of bacteria. However, the mechanisms responsible for transcriptional regulation of the IL-8 gene in epithelial cells during V. cholerae infections were not explored. Transient transfection analysis of 5¢ deletions and mutations of the IL-8 promoter driving expression of the luciferase reporter gene indicates that multiple binding sites contribute to IL-8 promoter induction in response to V. cholerae and that cooperation among these different sites is required for full activation of the promoter. Stimulation with V. cholerae O395 insertional mutants, defective in adherence and motility, significantly reduced IL-8 promoter activity compared with the wild-type strain. We further demonstrate maximal involvement of extracellular signal-regulated kinase 1 ⁄ 2 ⁄ mitogen-activated protein kinase pathways to regulate IL-8 gene transcription. This study will help to design agents which could reduce the V. cholerae-induced inflammatory response and in the generation of safe vaccines.

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Differential Interleukin-8 Response of Intestinal Epithelial Cell Line to Reactogenic and Nonreactogenic Candidate Vaccine Strains of Vibrio cholerae

Cited by 33 publications

References 24 publications

Transcriptional Responses of Intestinal Epithelial Cells to Infection with Vibrio cholerae

Transcriptional Responses of Intestinal Epithelial Cells to Infection with Vibrio cholerae

Induction of Interleukin-8 in T84 Cells byVibrio cholerae

Interleukin‐8 gene regulation in epithelial cells by Vibrio cholerae: role of multiple promoter elements, adherence and motility of bacteria and host MAPKs

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