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.