Table 1.
Mouse # | Globo-H Ceramide | GM2 Ceramide | OSM for sTn | dPSM for TF | dOSM for Tn | |||||
---|---|---|---|---|---|---|---|---|---|---|
IgG | IgM | IgG | IgM | IgG | IgM | IgG | IgM | IgG | IgM | |
Pre-Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 160 | 0 | 0 |
| ||||||||||
1 | 160 | 160 | 160 | 0 | 400 | 320 | 400 | 400 | 800 | 0 |
2 | 640 | 160 | 640 | 0 | 1600 | 40 | 400 | 400 | 1600 | 0 |
3 | 640 | 160 | 640 | 0 | 800 | 320 | 200 | 100 | 6400 | 200 |
4 | 640 | 640 | 2560 | 0 | 1600 | 2560 | 1600 | 800 | 3200 | 400 |
5 | 640 | 160 | 640 | 0 | 1600 | 20 | 1600 | 800 | 1600 | 400 |
| ||||||||||
Median | 640 | 160 | 640 | 0 | 1600 | 320 | 800 | 400 | 1600 | 200 |
IgM or IgG antibody reciprocal titers of pre- and post-vaccination sera tested for the five antigens in the pentavalent vaccines. ELISA assays were performed to determine IgM and IgG serum antibody titers as previously described.9 In brief, Globo-H ceramide, GM2 ceramide, ovine submaxillary mucin (OSM expressing sTn), desialylated ovine submaxillary mucin (dOSM expressing Tn), or desialylated porcine submaxillary mucin (dPSM expressing TF) was coated on ELISA plates at an antigen dose of 0.1 μg/well and incubated overnight at 4 °C. Nonspecific sites were blocked with 1% human serum albumin (HSA) for 2 h, and serially diluted antiserum was added to each well. After 1 h of incubation, the plates were washed, and alkaline phosphatase labeled goat anti-mouse IgM or IgG was added at 1:400 dilution (Southern Biotechnology Associates Inc., Birmingham, AL). The antibody titer was defined as the highest dilution with absorbance of 0.1 or greater over that of normal control mouse sera.