Table 3.
Functional capacity of hIgG exposed RF B cells
| Time after hIgG injection, days | Proliferation (cpm) in response to
|
HIgM secreted (ng/ml) in response to
|
||||
|---|---|---|---|---|---|---|
| Aggregated hIgG
|
F(ab′)2 anti-hIgM
|
agg. hIgG + B6 spleen | agg. hIgG + bm12 spleen | |||
| Day 1-2 | Day 2-3 | Day 1-2 | Day 2-3 | |||
| Response per 2 × 105 spleen cells | ||||||
| 34,867 (6,632) | 25,112 (5,705) | 77,514 (14,652) | 65,440 (3,854) | 1120 (140) | 2,692 (782) | |
| 1 | 21,603* (7,777) | 15,427* (4,276) | 67,711 (46,194) | 56,247 (31,433) | 1226 (855) | 1,706 (641) |
| 3 | 8,037* (2,280) | 7,451* (2,273) | 16,236* (17,999) | 30,446* (2,865) | 529 (45) | 1,093* (324) |
| Adjusted response per 2 × 105 hIgM+ spleen cells | ||||||
| 147,241 (30,298) | 106,859 (30,740) | 323,993 (43,054) | 276,697 (35,303) | 4520 (1084) | 11,157 (5,273) | |
| 1 | 81,028* (26,232) | 59,413* (23,316) | 257,116 (174,305) | 216,553 (132,944) | 4744 (3381) | 6,639 (2,765) |
| 3 | 64,422* (22,055) | 61,450* (29,741) | 131,298* (113,719) | 241,340 (32,903) | 3856 (709) | 8,077 (3,014) |
Mice were injected i.p. with 2 mg DHGG and killed at various times after treatment. Proliferation of splenocytes was measured in response to either aggregated hIgG at 50 ng/ml or F(ab′)2 fragments of affinity-purified goat anti-hIgM at 10 μg/ml or medium as control. Data are expressed as mean cpm incorporated for groups of three or four mice (after the subtraction of individual medium background cpm) and standard deviations from the mean (in parentheses) per 2 × 105 input spleen cells or as an adjusted value calculated for 2 × 105 hIgM+ B cells. Secretion of human IgM in vitro by 2 × 105 splenocytes stimulated with aggregated (agg) hIgG at 100 ng/ml final concentration or medium ± either 5 × 105 C57Bl/6bm12 splenocytes (T cell help) or 5 × 105 C57Bl/6 splenocytes (filler cells) as control. Triplicate samples of supernatants were harvested 5 days after the initiation of culture and pooled, and their hIgM levels were measured by standard ELISA. Data are shown as means and standard deviations (in parentheses) of hIgM produced (ng/ml) by 2 × 105 input spleen cells or as an adjusted value calculated for 2 × 105 input hIgM+ B cells. Data are shown for groups of three or four mice.
P < 0.05.