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. 2008 May 15;3(5):304–315. doi: 10.1016/j.chom.2008.03.009

Figure 1.

Figure 1

α-PDC-E2 IgG and IgA Responses in Mice Infected with N. aromaticivorans

(A and B) Mice were inoculated intravenously or orally with N. aromaticivorans, E. coli, or PBS (uninfected) at week 0 and week 2, and serum IgG and IgA antibodies to recombinant PDC-E2 were measured by ELISA. Results show average and SD values for NOD 1101 mice (three to four mice per group) inoculated with N. aromaticivorans or E. coli DH5α (5 × 107 cfu intravenously [A] or 1 × 109 cfu orally [B]) and examined at week 10 (left panel) or week 5 (right panels). Data are representative of 20 NOD 1101 mice per group examined in six independent experiments between week 4 and week 12. Similar results were observed when E. coli ATCC 25922 was used instead of E. coli DH5α. Statistical significance was calculated using a Student's t test: ∗p < 0.05 and ∗∗p < 0.01. Error bars represent the standard error of the mean for each group.

(C) Long-term persistence of anti-PDC-E2 IgG antibodies after infection. Sera of NOD 1101 mice (three per group) were analyzed for the presence of anti-PDC-E2 IgG responses (average and SD) at weeks 10, 16, and 25 as indicated. Error bars represent the standard error of the mean for each group.

(D) Western blot analysis of N. aromaticivorans (N) or liver (L) lysates using week 5 and week 10 sera from NOD 1101 mice uninfected (uninf.) or inoculated with N. aromaticivorans (N. aro.) or E. coli (E.c.), as indicated. Bound antibodies were revealed with mouse IgG-specific secondary antibodies. Note the 47 kDa and 50 kDa bands corresponding to bacterial lipoylated PDC-E2 protein and the crossreactivity to a 70 kDa band in liver extracts corresponding to the molecular weight of mammalian PDC-E2. Results were based on pools of five sera and confirmed in five independent experiments.

(E) Western blot analysis of sera from uninfected or N. aromaticivorans-infected mice against recombinant PDC-E2. Control (rightmost lane) shows the reactivity of a monoclonal antibody raised against human PDC-E2 (Migliaccio et al., 1998).

(F) Presence of antibodies against mouse or human recombinant PDC-E2 in sera of individual B6 and NOD 1101 mice infected with N. aromaticivorans (week 7), as detected by western blot (serum dilution 1:500).