A library of 16 AA peptides, each overlapping by 8 AA, spanning the entire sequence of LukE, LukS-PV, Hla, SplB, and SspB was synthesized (A&A Labs, San Diego). ELISpot assays for quantification of IL-17A or IFNγ responses were performed as described above, with the following modifications. Splenocytes from convalescent BALB/c or C57BL/6 mice (8 weeks after infection) were incubated with 1 of 14 peptide pools (12 to 16 peptides each, each peptide at 10 μg/ml), with a response considered positive if the value was greater than double the mean negative control wells and was above the threshold of 10 spot-forming colonies/5 × 105 splenocytes. For each positive pool, splenocytes were incubated with each of the peptides from the pool to determine which of the peptides was immunogenic. The same procedures were used to identify immunogenic epitopes in C57BL/6 mice following immunization with LukE.

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