Figure 3

Influence of antigen-driven bystander effect on the threshold of epicutaneous sensitization with a new protein allergen. (A) Groups of mice (four per group) were similarly pretreated and immunized as in Fig. 1 (A). On day 11, draining LNs were obtained and OVA-specific proliferation assay was performed. (B) Groups of mice (four per group) were subcutaneously pretreated with BSA (sc) or not (no) on day -21. They received patch co-administration of serial dilutions of OVA (100, 10, 1 mg/ml) and BSA (100 mg/ml) on day 1–5. On day 11, draining LNs were obtained and OVA-specific proliferation assay was performed. (C) Groups of mice (four per group) were non-pretreated (no) or subcutaneously (sc) pretreated on day -21. One day before patch application, all mice received i.v. transfer of CFSE-labeled CD4 T cells prepared from DO11.10 transgenic mice. Patch application with serial dilutions of OVA (100, 10, 1 mg/ml) and BSA (100 mg/ml) were performed on day 1–3. Regional LNs were obtained on day 4 for staining and flow cytometric analysis. All experiments were performed independently for at least three times. One representative results are shown.