Figure 1.
Passive systemic anaphylaxis (PSA) depends on mast cells and histamine.
PSA was induced by i.v. sensitization with 10 µg of anti-DNP IgE and challenge 24hrs later with 500 µg of DNP-HSA (black symbols) or PBS as control (white symbols) in MC-deficient mice homozygotes mutants (W/Wv Ho) (circles), heterozygotes (W/Wv He) (triangles) and their WT littermates (+/+) (squares) (A) and in Histamine receptor-1-deficient (HR1°/°) (triangles) and WT B6 mice (squares) mice (B). Body temperature was measured at various time points after challenge. Data are presented as mean (± SEM) of body temperature (n= 5 mice/group).
Figure 2.
Analysis of anaphylaxis in settings of CD4+CD25+ T cell deficiency.
DNP-specific PSA was induced A) in MHC class II-deficient mice (Aβ°/°) (triangles) and WT B6 (squares) mice and B) in B6 mice injected with either an anti-CD25 mAb (triangles), an anti-CD4 mAb (circles) or a control rat IgG mAb (squares). All mice were passively sensitized with anti-DNP IgE and challenge with DNP-HSA (black symbols) or PBS (open symbols). Data show mean (± SEM) values of body temperature at various times after challenge. Figure 2A shows 1 representative experiment out of 3 (n= 5-6 mice/group); p** = 0,0027 for DNP-challenged Aβ KO versus WT B6 (p** = 0,0074 and p = * 0,0286, for the other two experiments). Figure 2B data are from 2 pooled experiments (n= 5 mice/group); p* = 0,017 for DNP-challenged WT B6 versus anti-CD4-mAb treated mice and p* = 0,05 for DNP-challenged WT versus anti-CD25 mAb-treated B6 mice.
Figure 3.
Foxp3+ Treg control the severity and duration of anaphylaxis symptoms.
A: DEREG mice were injected with 1µg of DT (triangles) or with PBS (squares) on day -1, injected with anti-DNP IgE on day 0 and challenged with DNP-HSA (black symbols) or PBS (white symbols) on day +1. Body temperature was measured at various times after challenge. The data show pooled values from 3 experiments using 4-6 mice/group in each. p** = 0,0007 (2 way ANOVA) for DNP-challenged DEREG with and without DT (p values for each independent experiments are p= 0,032; p= 0,0176 and p=0,0105).
B: plasma histamine (left panel) and serum mMCP-1 (right panel) were titrated in IgE-sensitized DEREG mice that were DT-untreated (black bars) or DT-treated (grey bars). IgE-sensitized and PBS-challenged DT-untreated DEREG mice (white bars) were used as negative controls. Data show mean ± SEM values (n= 8-10 mice/group) of each factor expressed as ng/ml. Statistical analysis by 2 way ANOVA comparing DEREG with and without DT show significance with p** = 0,0085.
C: DT-untreated (white symbol) or DT-treated (black symbols) DEREG mice were either un-transferred (black triangles) or transferred with CD4+Foxp3+ Treg from either naïve mice (black diamond) or from day 6 DNFB-sensitized mice (black circles). Mice were passively sensitized with IgE anti-DNP, challenged with DNP-HSA as indicated and body temperature was measured at various times after challenge. Data show mean (± SEM) of body temperature using 5 mice/group.