Figure 3. FcɛRI-crosslinker, serotonin and histamine induce smooth muscle contractions in PCIS
(A) Example graphs showing PCIS filmed for 10min without stimulation to establish baseline movements, then an additional 10min following stimulation. FcɛRI-crosslinker, serotonin and histamine were used as positive controls. PCIS smooth muscle contraction was measured and quantified using movement per frame. X-axis indicates each frame of the video; y-axis indicates overall movement of individual pixels (n=1).(B) Total area under the curve (AUC) of muscle contraction responses to different stimulations (n=16). Unstimulated (MC); FcɛRI-crosslinker stimulation (α-FcɛRI); Serotonin stimulation (5-HT); Histamine stimulation (Hist). *P <.05, **P <.01, ****P <.0001, bars indicate mean, error bars represent ± SEM.
Figure 4. Passively sensitized PCIS show allergen-specific muscle contraction responses, mediator release and are affected by antihistamines
(A) Example graphs showing PCIS filmed for 10min without stimulation to establish baseline movements, then an additional 10min following stimulation. PCIS passively sensitized with peanut allergic plasma (PA) stimulated with peanut extract (PE, 1µg/ml) or irrelevant allergen (ovalbumin [OVA], 1µg/ml) (n=1) and PCIS passively sensitized with peanut sensitized non-allergic plasma (PS NA) stimulated with PE (1µg/ml) (n=1).(B) Total area under the curve (AUC) of muscle contraction responses to PE or irrelevant allergen (OVA) (n=16). (C)Tryptase concentration in culture supernatant from PCIS passively sensitized with PA stimulated with PE, FcɛRI-crosslinker (α-FcɛRI), irrelevant allergen (OVA) or unstimulated (MC), and PCIS passively sensitized with PS NA stimulated with PE (n=9).(D) Muscle contraction responses to histamine stimulation (10µg/ml) by passively sensitized PCIS untreated or pre-treated with antihistamines diphenhydramine hydrochloride (10µg/ml), cetirizine dihydrochloride (1µg/ml) or rupatadine (1µg/ml). Y-axis compares total AUC following histamine stimulation to the respective unstimulated (baseline) movements for each slice, represented by 100% on the y-axis and the horizontal dotted line (n=3). *P <.05, **P <.01, ****P <.0001, ns is not significant, bars indicate mean, error bars represent ± SEM.