Carcinoembryonic antigen (CEA)-related cell adhesion molecules are co-expressed in the human lung and their expression can be modulated in bronchial epithelial cells by non-typable Haemophilus influenzae, Moraxella catarrhalis, TLR3, and type I and II interferons

Klaile, Esther; Klassert, Tilman E; Scheffrahn, Inka; Müller, Mario M; Heinrich, Annina; Heyl, Kerstin A; Dienemann, Hendrik; Grünewald, Christiane; Bals, Robert; Singer, Bernhard B; Slevogt, Hortense

Immunohistochemical staining for CEACAM1, CEACAM5, CEACAM6 and CEACAM8 using 20 μg/ml of the monoclonal antibodies B3-17, 5C8C4, 1H7-4B and 6/40c, respectively, was performed on paraffin wax sections obtained from 19 different human lung sections. Sections were deparaffinized (Histoclear 3 × 5 min, ethanol 100% 5 min, ethanol 96% 5 min, ethanol 80% 5 min, ethanol 70% 5 min) and rehydrated (H 2 O 2 × 5 min). Rabbit serumblocking (2% in PBS) was performed before the sections were incubated with primary antibody over night at 4°C, and with HRP-coupled secondary rabbit antibody for 1 h at RT. Then the nickel-glucose oxidase development technique was performed to enhance the DAB chromogen of the peroxidase immunohistochemistry. The specimens were counterstained with Calcium Red for 1-2 minutes to visualize the tissue structure. The result was a distinct purple/black staining product. As negative control served an IgG1 istotype matched control antibody. Non-cancer tissues from the sections were used for analysis.