Table 1.
Expression and surface density of MHC II products, CD80, and CD86 by DCs and B blasts
| Antibody | Mouse strain | Molecules per cell* |
Molecules per μm2 |
||
|---|---|---|---|---|---|
| DC† | B† | DC | B | ||
| I-Ab AF6 120.1 | C57/B6 | 111 ± 36 | 28 ± 9.0 | 77.9 | 115.7 |
| I-Ab TIB120‡ | C57/B6 | 191 ± 47 | 71 ± 43 | 134.1 | 290.5 |
| I-Ek TIB120‡ | B10.BR | 80 ± 21 | 16 ± 1.5 | 56.4 | 63.8 |
| I-Ek 14.4.4S | B10.BR | 124 ± 21 | 30 ± 4.2 | 87.3 | 124.6 |
| I-Ak 11-5.2 | B10.BR | 188 ± 77 | 39 ± 3.4 | 132.1 | 159.7 |
| CD86 GL1 | C57/B6, B10.BR | 208 ± 43 | 16 ± 1.6 | 146.3 | 67.3 |
| CD80 16-10A1 | C57/B6, B10.BR | 132 ± 20 | 1.6 ± 0.7 | 93.1 | 6.5 |
| CD40 3/23 | C57/B6, B10.BR | 17 ± 1.8 | 2.1 ± 1.0 | 11.9 | 8.5 |
| CD11a | C57/B6, B10.BR | 27 ± 7.6 | 9.7 ± 4.8 | 19.2 | 39.7 |
| Isotype control | C57/B6, B10.BR | 3.0 ± 1.6 | 0.9 ± 0.3 | 2.1 | 3.8 |
| Cell volume | 965 μm3 | 254 μm3 | |||
| Cell surface area | 1,421 μm2 | 243 μm2 | |||
Mature DCs or 24-hr B blasts were labeled at saturating concentrations with the indicated PE-conjugated Ab and processed for flow cytometry. Fluorescence standards were used to estimate the number of molecules per cell on mature DCs or fully blasted B cells by using beads with known quantities of fluors to perform a linear regression analysis. Numbers of molecules ± standard deviation (× 103) are listed for each Ab and mouse strain (columns 3 and 4). Pellets of mature DC or 24-hr B blasts were prepared for electron microscopy, serial sections were cut, and cell volume and surface area were calculated using the disector method. Numbers of molecules per square micron were obtained by dividing total number of molecules per cell by surface area in square microns (columns 5 and 6).
*Values are stated as numbers of molecules × 103.
†Gated on CD11c + or B220 + populations, respectively.
‡The mAb TIB 120 (M5/114.15.2) is specific for I-Ab, I-Aq, I-Ad, I-Ed, and I-Ek.