Figure 5. Neu5Gc expressed on NTHi induces anti-Neu5Gc antibodies in Cmah−/− mice. (A and B) Cmah−/− mice were injected intraperitoneally with heat-killed NTHi (without adjuvant) which had been grown in sialic-acid free media (Sia-free NTHi, n = 16) or with sialic-acid free media with 1 mM Neu5Gc (Gc-NTHi, n = 17). All mice were injected a total of three times at 2-wk intervals. Sera collected after the third injection were analyzed by ELISA for IgM (A) and IgG (B) antibodies against Neu5Gcα-PAA. The highest and lowest value from each group was removed before graphing. Horizontal lines represent the mean values. Statistical analysis was performed using an unpaired one-tailed Student’s t test. (C) Pooled mouse serum from mice injected intraperitoneally with heat-killed NTHi grown in sialic-acid free media with 1 mM Neu5Gc was analyzed in an ELISA for levels of anti-Neu5Gc IgG antibodies against Neu5Gcα2–3Galβ1–4Glcβ-HSA (black bar), Neu5Gcα2–6Galβ1–4Glcβ-HSA (gray bar), Neu5Gcα2–3Galβ1–4GlcNAcβ-HSA (white hashed bar), and Neu5Gcα2-6Galβ1-4GlcNAc-HSA (white bar). Values represent mean IgG levels, quantified according to a mouse IgG standard curve. Error bars represent SD. For comparison, serum from 12-mo-old infants (n = 15) was analyzed for levels of anti-Neu5Gc antibodies against Neu5Gcα2–3Galβ1–4Glcβ-HSA (black bar), Neu5Gcα2–6Galβ1–4Glcβ-HSA (gray bar; same data as shown in Fig. 1 B). Values represent mean IgG levels, quantified according to a human IgG standard curve. Error bars represent SEM. Data are representative of greater than three independent experiments.
Figure 5.

Neu5Gc expressed on NTHi induces anti-Neu5Gc antibodies in Cmah−/− mice. (A and B) Cmah−/− mice were injected intraperitoneally with heat-killed NTHi (without adjuvant) which had been grown in sialic-acid free media (Sia-free NTHi, n = 16) or with sialic-acid free media with 1 mM Neu5Gc (Gc-NTHi, n = 17). All mice were injected a total of three times at 2-wk intervals. Sera collected after the third injection were analyzed by ELISA for IgM (A) and IgG (B) antibodies against Neu5Gcα-PAA. The highest and lowest value from each group was removed before graphing. Horizontal lines represent the mean values. Statistical analysis was performed using an unpaired one-tailed Student’s t test. (C) Pooled mouse serum from mice injected intraperitoneally with heat-killed NTHi grown in sialic-acid free media with 1 mM Neu5Gc was analyzed in an ELISA for levels of anti-Neu5Gc IgG antibodies against Neu5Gcα2–3Galβ1–4Glcβ-HSA (black bar), Neu5Gcα2–6Galβ1–4Glcβ-HSA (gray bar), Neu5Gcα2–3Galβ1–4GlcNAcβ-HSA (white hashed bar), and Neu5Gcα2-6Galβ1-4GlcNAc-HSA (white bar). Values represent mean IgG levels, quantified according to a mouse IgG standard curve. Error bars represent SD. For comparison, serum from 12-mo-old infants (n = 15) was analyzed for levels of anti-Neu5Gc antibodies against Neu5Gcα2–3Galβ1–4Glcβ-HSA (black bar), Neu5Gcα2–6Galβ1–4Glcβ-HSA (gray bar; same data as shown in Fig. 1 B). Values represent mean IgG levels, quantified according to a human IgG standard curve. Error bars represent SEM. Data are representative of greater than three independent experiments.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal