Fig 1.
C1-INH plasma levels after i.v. or i.p. bolus injection.
Human plasma derived C1-INH antigen (Ag) and activity (Act) in plasma after bolus injection of 5U (white bars) or 15U (black bars) C1-INH either i.v. (A-B) or i.p. (C-D) at indicated time points up to 72 hours following injection. Data are presented as mean ± SEM, n = 3 per time point.
Fig 2.
C1-INH and C5a levels in plasma and BALF.
Human plasma derived C1-INH antigen (Ag) and activity (Act) in plasma (A-B) and BALF (C-D) of NaCl (white bars) and HDM (black bars) exposed C1-INH treated mice. Complement activation was determined in BALF (E) by C5a ELISA in vehicle (white bars) and C1-INH (black bars) treated NaCl and HDM exposed mice. Data are presented as mean ± SEM, n = 6 (NaCl) and n = 10 (HDM). *P<0.05, **P<0.01.
Fig 3.
Eosinophil influx in BALF and lung.
(A) Total cell count in BALF of NaCl or HDM challenged vehicle (white bars) or C1-INH (black bars) treated mice. (B) Differential cell count in BALF showing eosinophil, neutrophil, macrophage, and lymphocyte numbers in NaCl challenged vehicle (dotted bars) and C1-INH treated (blocked bars) mice and in HDM challenged vehicle (white bars) and C1-INH treated (black bars) mice. (C) Influx of eosinophils in the lung was determined by digital analysis of Major Basic Protein (MBP) stained lung sections in C1-INH (black bars) and vehicle (white bars) treated mice following challenge with NaCl or HDM. Representative pictures of each group are shown, magnification 4x. Data are presented as mean ± SEM, n = 6 (NaCl) and n = 10 (HDM).
Fig 4.
Lung damage and mucus production.
Lung sections were stained with HE (A) or PasD (B) to determine damage, depicted as total pathology, or mucus production, respectively in NaCl or HDM exposed mice treated with C1-INH (black bars) or vehicle (white bars). Representative pictures of each staining per group are shown, magnification 10x. Data are presented as mean ± SEM, n = 6 (NaCl) and n = 10 (HDM).
Fig 5.
Following NaCl or HDM exposure MLN were dissected from mice treated with C1-INH (black bars) or vehicle (white bars). Equal numbers of MLN cells from C1-INH (black bars) or vehicle (white bars) treated HDM exposed mice were restimulated ex vivo with HDM and afterwards production of IL-4 (A), IL-5 (B), and IL-13 (C) was determined in the medium. In BALF of NaCl or HDM exposed mice treated with C1-INH (black bars) or vehicle (white bars) the cytokines IL-4 (D), IL-5 (E) and IL-13 (F) were determined by a magnetic luminex screening assay. Data are presented as mean ± SEM, n = 6 (NaCl) and n = 10 (HDM). *P<0.05.
Fig 6.
Plasma and BALF protein levels.
In plasma of NaCl or HDM exposed mice treated with C1-INH (black bars) or vehicle (white bars) IgE (A), HDM specific IgG1 (B) and E-selectin (C) were determined. In BALF of C1-INH (black bars) or vehicle (white bars) treated NaCl or HDM exposed mice E-selectin (D), total protein (E), IgM (F) and TATc (G) were determined. Data are presented as mean ± SEM, n = 6 (NaCl) and n = 10 (HDM). **P<0.01.