Distribution of Anti-CD68 (EBM11) Immunoreactivity in Formalin-fixed, Paraffin-Embedded Bovine Tissues

Abstract

Abstract.A commercially acquired anti-human macrophage antibody (anti-CD68; EBM 1 1) was used in an immunocytochemical technique to detect macrophages in formalin-fixed, paraffin-embedded tissues from cattle, pigs, humans, rats, turkeys, dogs, and cats. In healthy cattle, the antibody labeled alveolar macrophages, pulmonary intravascular cells (presumably intravascular macrophages), and macrophage-like cells in other tissues. In bovine lungs infected with Pasteurellu haemolyticu, EBM 1 1 antibody labeled 95% of alveolar macrophages and macrophages within alveolar septa but only 0-2% of streaming or "oat" leukocytes. Alveolar macrophages were also stained by EBM 1 1 in pigs but not in rats, turkeys, dogs, and cats. The antibody also stained macrophage aggregates in the mesenteric lymph nodes and intestinal lamina propria of Mycobucterium purutuberculosisinfected cattle. This study shows that the anti-CD68 (EBM I 1) antibody is a useful marker of macrophages in normal bovine tissues or tissues from areas of acute or chronic inflammation that have been routinely processed. The study also adds strength to the growing evidence suggesting that streaming leukocytes seen in pneumonic pasteurellosis are neutrophils.

Examination of the Reticular Epithelium of the Bovine Pharyngeal Tonsil

“…In contrast to CD14, immunoreactivity for CD68 was noted among cells scattered within follicular and interfollicular regions with few CD68þ cells within either reticular or nonreticular epithelium. CD68 is a glycoprotein associated with lysosomes in macrophages, and has been previously documented in follicular and perifollicular regions of human palatine tonsil, similar to the distribution observed in the present study (Ackermann et al, 1994).…”

“…CD68 is another marker associated with human and bovine macrophages and multinucleated giant cells (Ackermann et al, 1994;Stent et al, 2002;Palmer et al, 2009;Ben-Yaakov et al, 2011). In contrast to CD14, immunoreactivity for CD68 was noted among cells scattered within follicular and interfollicular regions with few CD68þ cells within either reticular or nonreticular epithelium.…”

Immunophenotypic characterization and depletion of pulmonary intravascular macrophages of horses

“…Of these, ED-1 and CD68 recognize a 90-110 kDa protein in the lysosomes of rat and human macrophages, respectively, and have been used for immunohistochemistry in the respective species [1,24]. MAC-387 identifies another cytoplasmic, most probably lysosomal, protein called L1 in the lung and liver macrophages in humans [11,12].…”

“…These antibodies also show some reactivity against monocytes but very weak reaction with peripheral blood granulocytes. Bovine macrophages including PIMs are stained with CD68 antibody [1,22]. However, equine PIMs and alveolar macrophages did not stain with either CD68 or ED-1.…”

“…Mouse antihuman monocyte/macrophage antibody (MAC-387, Serotec Ltd., UK) recognizes the L1 cytoplasmic molecule and has been used in immunohistochemistry [11,12]. CD68 (DAKO A/S, Denmark) is a mouse anti-human macrophage antibody that reacts with a 110 kDa glycoprotein in tissue macrophages [1]. The sections were incubated with anti-human (MAC-387 and CD68, 1:75 each) and anti-rat macrophage (ED-1, 1:75) antibodies for 2 h followed by anti-mouse peroxidase-conjugated secondary antibodies (1:100, DAKO A/S Denmark) for 60 min.…”