Evaluation of a panel of antibodies for the immunohistochemical identification of immune cells in paraffin-embedded lymphoid tissues of new- and old-world camelids

Abstract

Different species of camelids play an important role in the epidemiology of various emerging infectious diseases such as Middle East respiratory syndrome. For precise investigations of the immunopathogenesis in these host species, appropriate immunohistochemical markers are highly needed in order to phenotype distinct immune cells populations in camelids. So far, specific immunohistochemical markers for camelid immune cells are rarely commercially available, and cross-reactivity studies are restricted to the use of frozen dromedary tissues. To bridge this gap, 14 commercially available primary antibodies were tested for their suitability to demonstrate immune cell populations on formalin fixed paraffin-embedded (FFPE) tissue sections of dromedaries, Bactrian camels, llamas, and alpacas in the present study. Out of these, 9 antibodies directed against CD3, CD20, CD79α, HLA-DR, Iba-1, myeloid/histiocyte antigen, CD204, CD208, and CD68 antigen exhibited distinct immunoreaction patterns to certain camelid immune cell subsets. The distribution of these antigens was comparatively evaluated in different anatomical compartments of thymus, spleen, mesenteric, and tracheobronchial lymph nodes. The presented results will provide a basis for further investigations in camelids, especially with respect to the role of the immune response in certain infectious diseases, which harbor a considerable risk to spill over to other species.

Vascular Inflammation Is Associated with Loss of Aquaporin 1 Expression on Endothelial Cells and Increased Fluid Leakage in SARS-CoV-2 Infected Golden Syrian Hamsters

“…Immunohistochemistry of lung sections was performed using the avidin-biotin-peroxidase complex (ABC; Vector Laboratories, Burlingame, CA, USA) method or the Dako EnVision+ polymer system (Dako Agilent Pathology Solutions) as previously described [ 26 , 27 , 28 ]. Primary antibodies were applied overnight at 4 °C.…”

Experimental infection of dromedaries with Middle East respiratory syndrome-Coronavirus is accompanied by massive ciliary loss and depletion of the cell surface receptor dipeptidyl peptidase 4

“…Immunohistochemistry was performed for the detection of CD3 + T cells (polyclonal rabbit anti-CD3, DakoCytomation GmbH), CD20 + B cells (polyclonal rabbit anti-CD20, Thermo Fisher scientific), and CD204 + macrophages (monoclonal mouse anti-human macrophage scavenger receptor, human CD204, BioLogo) as summarized in Table 1 . The used markers have been previously established for lymphoid dromedary tissue 28 . Additionally, epithelial cells were labeled by application of different antibodies (Table 1 ) directed against pan-cytokeratin (CK; monoclonal mouse anti-Human Cytokeratin, DakoCytomation GmbH), CK5/6 (monoclonal mouse anti-Cytokeratin 5/6, DakoCytomation GmbH), CK 14 (polyclonal rabbit anti-Keratin 14, Thermo Fisher Scientific) and CK18 (monoclonal mouse anti-Cytokeratin 18, Abcam).…”

“…Despite two recent animal trials 21 , 22 and the successful establishment of an orthopoxvirus-based vaccine against MERS-CoV in dromedaries 22 , the virus infection triggered immune response and the exact cell tropism have not been evaluated so far, since valuable markers for immune cell phenotyping in dromedaries were hardly established. In order to facilitate such studies and other investigations in camels, a panel of antibodies for identification of different immune cell subsets has been recently tested 28 . Since dromedaries play a key role in transmitting MERS-CoV to humans, it was the aim of the present study to describe MERS-CoV associated lesions in the nasal epithelium and viral cell tropism of experimentally infected dromedaries in closer detail.…”

Solitary tracheal B-cell lymphoma in an adult alpaca (Vicugna pacos)

“…Immunohistochemical staining using CD3, PAX-5, BLA36, CD 79 and CD20 [ 8 , 10 , 12 , 13 ] allows the differentiation of these tumors into T- or B-cell lymphomas. Although B-cell lymphomas appear to occur more frequently in SAC [ 5 , 6 , 8 ], the juvenile disseminated form observed in young alpacas is usually of T-cell origin [ 10 ].…”