Serum IgA, IgG, and IgG subclass responses specific for (A) OMV, (B) Ptx, (C) Prn, (D) FHA, and (E) Fim2/3 were determined by using a MIA. Data were obtained in naive mice and protected mice prior to challenge (D0) and 14 days post infection (p.i.) or 14 days post-challenge (p.c.) (n = 3/time point). (F) Pulmonary IgA responses against these antigens were determined on the same time points. * = p<0.05 experimental group versus unprotected group (D0), + = p<0.05 protected group (D0) versus protected group (14 days p.c.). (G) The kinetics of the anti-OMV IgA antibody formation in lung lysates were analyzed at more time points (n = 3/time point) and expressed in fluorescence intensity (F.I.). **** = p<0.0001, challenged unprotected or protected group versus unprotected or protected group (day 0), ++ and +++ = p<0.01 and p<0.001 unprotected group versus protected group (for each time point). (H) Western blot on separated B. pertussis B1917 proteins was performed with pooled lung lysates (1:50) of unprotected and protected mice prior to challenge (D0), and of protected mice 14 days p.c. with IR800-labeled secondary antibody. Left panel shows whole protein range (260kDa-<10kDa) of B. pertussis lysate. Right panel shows more detailed separation of the 110-60kDa protein range. Antigen identification for Vag8 and LPS is depicted.