To evaluate delayed type hypersensitivity (DTH), 20 μg of the total antigen (in 20 μl) was inoculated into the plantar cushion of non-infected paw. After 48 h from the inoculation, the extent of swelling was measured using a caliper rule (Mitutoyo). The DTH was expressed as the difference between the paw thickness before and after the antigen inoculation.
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