A regulatory role for the C5a anaphylatoxin in type 2 immunity in asthma
J. Clin. Invest. Jörg Köhl, et al. 116:783 doi:10.1172/JCI26582 [Go to this article.]

Figure 4
Generation and characterization of C5aRA Tg mice. (A) Generation of C5aR Tg mice (see Methods). rCCSP, rat clara cell–specific promoter; hGHpA, human growth hormone polyadenylation signal; bGHpA, bovine growth hormone polyadenylation signal. (B, upper left panel) Ccsp-rtTa sTg mice (545 bp, lane 1) were bred to (tetO)₇-CMV-C5aRA sTg mice (228 bp, lane 2) to generate C5aRA-Cssp dTg progeny (C5aRA-Ccsp dTg, lane 3). M, marker (200–900 bp). (Upper right panel) Kinetic of induction of pulmonary C5aRA mRNA in C5aRA-Ccsp dTg mice. RT-PCR was performed from whole-lung samples of C5aRA-Ccsp dTg mice that received drinking water without dox (day 0) or dox-supplemented water (0.5 mg/ml) for 1 day, 3 days, or 7 days. Control reactions with GAPDH primers are shown in the lower panel. (Lower panel) Organ specificity of Tg activation in C5aRA-Ccsp dTg mice treated with dox for 7 days. Lane 1, lung from C5aRA sTg mouse; lanes 2–7, liver, spleen, kidney, heart, brain, and lung from C5aRA-Ccsp dTg mouse; lane 8, genomic DNA from C5aRA-Ccsp dTg (positive control). (C) C5aRA protein expression in lung epithelial cells (indicated by arrows) from C5aRA-Ccsp dTg mice treated for 7 days with dox. Immunohistochemical staining using anti-C5a mAb 561: Alexa⁴⁸⁸ (left panel); ISO-Ctrl (right panel). We found no C5aRA protein expression in lung epithelial cells of C5aRA sTg mice treated with dox (data not shown). (D, left panel) Total and differential cell counts in BAL. (Right panel) Cytokine profile of pulmonary cells harvested from BALB/c mice 72 hours after final in vivo HDM exposure. Cytokine profiles and BAL cell counts obtained from HDM-exposed non-Tg littermates were indistinguishable from those of HDM-exposed C5aRA sTg mice (data not shown). *P < 0.05; **P < 0.001.