Expression of CD68 in Non‐Myeloid Cell Types

CD68, the human homologue of macrosialin, is commonly regarded as a selective marker for human monocytes and macrophages. Its expression is thought to be regulated by a macrophage-specific promoter. However, several immunohistochemical studies have indicated that CD68 antibodies also react with other haematopoietic and non-haematopoietic cell types. We investigated the expression of CD68 in various primary cells and carcinoma cell lines using immunohistochemistry, flow cytometry, Western blot analysis and qRT-PCR. Weak but significant immunoreactivity was detected in lymphocytes and several tumour cell lines whereas staining of primary fibroblasts and endothelial cells was comparable to macrophages. The intensity of CD68 staining in individual cell types depended on the antibody clone and the fixation technique. Anti-CD68 mAb KP1 should be used with great caution for frozen tissue sections due to its reactivity with a wide variety of cell types. Also, care should be taken when distinguishing macrophages from fibroblasts/stromal cells in paraffin sections after formalin fixation since both cell types are stained highly positive for CD68. In accordance, mRNA expression of CD68 was not only detected in macrophages and monocytes but also in fibroblasts as well as endothelial cells and tumour cells, although with a varying intensity. Cloning of full length 5'-sequences and determination of transcription start sites shows that macrophages and fibroblasts initiate transcription within the known promoter region; however, from different start sites, indicating alternative promoter architecture in myeloid versus non-myeloid cells. We suggest that CD68 is not a selective macrophage marker but rather a lysosomal protein that is enriched in macrophages.