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Table I.

Increase in HDC Activity Induced by IgE or Various Activators of the Ca2+–PKC Pathway




HDC activity
 pmol/min/mg protein 
None 3.13 ± 1.68 
Anti-DNP IgE 652 ± 54.8a 
Anti-DNP IgG 5.80 ± 1.95 
Polyclonal IgG 1.43 ± 1.00 
Anti-DNP IgE + 2.4G2 626 ± 46.2a 
IgE/antigen 389 ± 10.3a,b 
Thapsigargin 618 ± 162a 
TPA 96.3 ± 23.3a 
A23187
 
1,240 ± 118a
 



HDC activity
 pmol/min/mg protein 
None 3.13 ± 1.68 
Anti-DNP IgE 652 ± 54.8a 
Anti-DNP IgG 5.80 ± 1.95 
Polyclonal IgG 1.43 ± 1.00 
Anti-DNP IgE + 2.4G2 626 ± 46.2a 
IgE/antigen 389 ± 10.3a,b 
Thapsigargin 618 ± 162a 
TPA 96.3 ± 23.3a 
A23187
 
1,240 ± 118a
 
a

P < 0.01 is regarded as significant by the Student's t test (vs. None).

b

P < 0.05 is regarded as significant by the Student's t test (vs. Anti-DNP IgE).

BMMCs were cultured in the presence of an anti-DNP IgE (3 μg/ml), an anti-DNP IgG (3 μg/ml), purified polyclonal IgG (3 μg/ml), thapsigargin (100 nM), TPA (10 nM), and A23187 (0.3 μM) for 6 h at 37°C. Blocking of FcγRII and FcγRIII was performed by pretreatment with the 2.4G2 antibody (10 μg/ml) for 10 min. In the experiment of antigen stimulation (IgE/antigen), BMMCs were incubated in the presence of 1 μg/ml anti-DNP IgE for 24 h at 37°C, washed, and then incubated with 30 ng/ml DNP-human serum albumin for 6 h. The concentration of each reagent was optimized in preliminary experiments to obtain its maximal effect. The values obtained are represented as the means ± SEM (n = 3).

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