Pharmacological analysis
The ASM contractile force in response to histamine, EFS, and QS was reported as mg normalized for 100 mg of bronchial tissue (mg/100mg). The protective effect of benralizumab and mepolizumab against AHR induced by contractile stimuli in passively sensitized airways was expressed as the percentage of the maximal AHR inhibition, that in turn was identified as the delta between the AHR induced in C+ airways and the normal contractile response elicited in C- airways. Since the effect of each single concentration of the mAbs investigated in this research was tested after overnight incubation during the sensitizing procedure, the CRCs to benralizumab and mepolizumab on the contractile plateau to specific histamine concentrations and EFS frequencies were generated by plotting in the same graph the data originated from different bronchial rings treated with different concentrations of the mAbs. Appropriate curve-fitting to sigmoidal models were used to calculate the effect (E), Emax, EC50, EC70, EC90, EF50, EF70, EF90, and the half maximal inhibitory concentration (IC50). The equations used to describe the models used in this study were: Y=Bottom + (Top-Bottom)/(1+10^((LogEC50_or_EF50-X))) and Y=Bottom + (Top-Bottom)/(1+10^((X-LogIC50))). EC50, EF50, and IC50were transformed in pEC50 (-LogEC50), pEF50 (-LogEF50), and pIC50 (-LogIC50) to perform the statistical analysis of the potency .
Data originated from ELISA kits were elaborated after normalization for the wet weight (100 mg) of isolated airways.