ELISAs were performed as previously described (48 (link)). Samples were diluted in TBS containing 1% BSA (TBS-BSA) and analyzed for total and PC-specific IgM, IgG, and IgA. Diluted serum samples were added to Costar plates coated with 0.625 μg/mL PC(4 (link))BSA (Biosearch Technologies) or 5 μg/mL anti-mouse Ig(H+L) (Southern Biotechnology Associates) or AB1.2 (anti-T15 Id) Ab and blocked with TBS-BSA. Alkaline phosphatase-conjugated polyclonal goat anti-mouse IgM, IgG, IgG3, and IgA Abs (Southern Biotech) and pNPP (Sigma) were used to detect Ab. A PC-specific IgM hybridoma was used for quantitation of the PC IgM response. IL-4, IL-5, IL-6, IL-10, IFNγ, TNFα, and GM-CSF ELISAs were performed according to manufacturer’s instructions (BioLegend). Malondialdehyde-specific ELISAs used plates coated with MDA-BSA (Cell Biolabs, Inc) at 5 ug/ml. Phosphatidylcholine ELISAs were performed on air-dried plates that had been coated with L-alpha-phosphatidylcholine (Sigma) at 30 ug/ml in hexane. ELISPOT was performed as previously described (33 (link)), using both Image Acquisition 4.5 and ImmunoSpot software for analysis of spot number and size.