Fig 1: Substance P increases formation of antibody-secreting cells and their release of IgE.Splenic B cells were isolated from naive C57BL6 mice and cultured with or without LPS (10 µg/mL) and IL-4 (20 ng/mL) and exposed to vehicle, substance P (1 µM), and CGRP (300 nM). (A) After 4 days, the cells were immunophenotyped by flow cytometry, and immunoglobulin release was assessed by ELISA. LPS and IL-4 triggered the formation of (B) total, (C and E) IgG1+ and (D and F) IgE+ CD95+ (germinal center; C and D) and CD138+ (antibody-secreting cells; E and F) B cells. Additionally, LPS+IL-4 enhanced B cell release of (G) IgG1 and (H) IgE. These effects were augmented when B cells were coexposed to the neuropeptide substance P (1 µM). CGRP (300 nM) exposure had minimal impact on these parameters. Graphs show median and range. P values determined using 1-way ANOVA and Tukey’s multiple comparisons test. P < 0.05 is indicated by *; P < 0.01 is indicated by **. Representative experiment is shown; n = 3/group. Experiments were replicated at least 2 independent times.