TABLE 2.
Properties of MAbs to S. flexneri 2a, S. flexneri 3a, and S. sonnei
| Immunizing LPS serotype | MAb name | Isotype | Specificity by ELISA plates coated with LPS froma |
Specificity by flow cytometry with formalin-fixed target bacteriab |
|||||
|---|---|---|---|---|---|---|---|---|---|
| S. flexneri 2a | S. flexneri 3a | S. sonnei | E. coli | S. flexneri 2a | S. flexneri 3a | S. sonnei | |||
| S. flexneri 2a | Hflex2a1 | IgG3 | + | − | − | − | + | − | − |
| Hflex2a2 | IgG2b | + | + | − | − | + | − | − | |
| Hflex2a3 | IgM | + | ± | − | ± | ± | − | − | |
| Hflex2a4 | IgM | + | − | − | − | + | − | − | |
| Hflex2a5 | IgM | + | − | − | − | + | − | − | |
| Hflex2a6 | IgM | + | − | − | − | + | − | − | |
| Hflex2a7 | IgM | + | − | − | − | + | − | − | |
| Hflex2a8 | IgM | + | − | − | − | + | − | − | |
| Hflex2a9 | IgM | + | − | − | − | + | − | − | |
| Hflex2a10 | IgM | + | + | − | − | − | − | − | |
| S. flexneri 3a | Hflex3a1 | IgG2b | − | + | − | − | − | + | − |
| Hflex3a2 | IgG3 | − | + | − | − | − | + | − | |
| Hflex3a3 | IgG2b | + | + | − | − | − | − | − | |
| Hflex3a4 | IgG2b | + | + | − | ± | − | − | − | |
| Hflex3a5 | IgM | − | + | − | − | − | + | − | |
| Hflex3a6 | IgM | − | + | − | − | − | + | − | |
| Hflex3a7 | IgM | − | + | − | − | − | + | − | |
| Hflex3a8 | IgM | − | + | − | − | − | + | − | |
| Hflex3a9 | IgM | − | + | − | − | − | + | − | |
| Hflex3a10 | IgM | − | + | − | − | − | + | − | |
| Hflex3a11 | IgM | − | + | − | − | − | + | − | |
| Hflex3a12 | IgM | − | + | − | − | − | + | − | |
| Hflex3a13 | IgM | − | + | − | − | − | + | − | |
| Hflex3a14 | IgM | − | + | − | − | − | + | − | |
| Hflex3a15 | IgM | − | + | − | − | − | + | − | |
| Hflex3a16 | IgM | − | + | − | − | − | + | − | |
| Hflex3a17 | IgM | − | + | − | − | − | + | − | |
| Hflex3a18 | IgM | − | + | − | − | − | + | − | |
| S. sonnei | Hsoni1 | IgG3 | − | − | + | − | + | − | + |
| Hsoni2 | IgM | − | − | + | − | − | − | + | |
| Hsoni3 | IgM | − | − | + | − | − | − | + | |
| Hsoni4 | IgM | − | − | + | − | − | − | + | |
| Hsoni5 | IgM | − | − | + | − | − | − | + | |
| Hsoni6 | IgM | − | − | + | − | − | − | + | |
| Hsoni7 | IgM | − | − | + | − | − | − | + | |
| Hsoni8 | IgM | − | − | + | − | − | − | + | |
ELISA results were scored as “−” when the OD405 was less than 0.2, “±” when OD405 was greater than or equal to 0.2 but less than 0.5, and “+” when OD405 was greater than or equal to 0.5. The OD405 of wells with no supernatant was ∼0.1. Undiluted hybridoma supernatants were used for ELISA.
Target strains used for flow cytometric studies were ATCC 700930, J17B, and ATCC 9290, respectively, for S. flexneri 2a, S. flexneri 3a, and S. sonnei. The flow cytometric results are scored as “−” when real signal/background signal (S/N) is less than 2, “±” when S/N is greater than or equal to 2 but less than 5, and “+” when S/N is greater than or equal to 5. Background is the geometric mean fluorescence obtained with no MAb, and the signals are 20, 17, and 16 mean fluorescence units for S. flexneri 2a, S. flexneri 3a, and S. sonnei, respectively. Optimally diluted (ranging from 1:2 to 1:250) supernatants were used for flow cytometry.