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Fig 1.

Flow diagram.

CONSORT flow diagram as applies for human challenge studies. In total 22 adult volunteers were assessed for eligibility, and 1 volunteer did not meet the inclusion criteria, resulting in a total of 21 volunteers allocated to experimental infection with different doses of ETEC: 1×106 (n = 3), 1×107 (n = 3), 1×108 (n = 3), 1×109 (n = 3) and 1×1010 (n = 9) CFU. Clinical data were analyzed for all volunteers, as well as the immunological data on day 10 and day 28 after experimental infection. There were none lost to follow-up, however, immunological data from 6 months, 1 year and 2 years after experimental infection were obtained from subgroups of the study population due to limited availability of personnel to perform the analyses.

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Table 1.

Description of diarrheal episodes among 21 volunteers experimentally infected with ETEC strain TW10722.

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Table 2.

Symptoms and signs other than diarrhea in 21 volunteers experimentally infected with ETEC strain TW10722.

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Table 3.

Mean disease severity scores among 21 volunteers experimentally infected with ETEC strain TW10722.

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Fig 2.

Antigen-specific CD4+ T cell responses after infection with STh-only ETEC strain TW10722.

The graph displays the mean percentage of CD4+ T cells that co-express CS25 and CD134 for each sampling timepoint and virulence factor after incubating whole blood with ETEC proteins CS6A, CS5 and YghJ. Grey circles represent volunteers in the 1×106 to 1×109 CFU dose groups, red circles represent volunteers in the 1×1010 CFU dose group. Error bars represent the 95% confidence intervals. Due to limited availibility of purified CS6 antigen in the initial parts of the study, CS6-specific CD4+ T cell responses were only measured for a subgroup of the volunteers (n = 12). Also, due to the stepwise inclusion of volunteers to the study, the long-term follow-up timepoints of each volunteer varied according to time of enrollment, with the first subgroup of volunteers (n = 9) having their follow-up at 2 years, while the last subgroup of volunteers (n = 12) had their follow-up at 6 months and 1 year after experimental infection. Abbreviations: d0: day 0, d10: day 10, d28: day 28, m6: 6 months, y1: 1 year, y2: 2 years, N: Number of volunteers, Stim: Antigen used for stimulation. a Antigen preparation differed from other timepoints.

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Fig 3.

Antigen-specific serum IgG and IgA responses after infection with STh-only ETEC strain TW10722.

The graphs display the median fluorescence intensity (MFI) value of CS5-, CS6AB-, and YghJ-specific IgG and IgA at each sampling time point for each volunteer. Grey circles represent volunteers in the 1×106 to 1×109 CFU dose groups, red circles represent volunteers in the 1×1010 CFU dose group. Error bars represent the 95% confidence intervals. Number of volunteers indicated in parentheses. Abbreviations: d0: day 0, d10: day 10, d28: day 28, m6: 6 months, y2: 2 years.

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