Immunohistological staining of lymphoid tissue in four Australian marsupial species using species cross‐reactive antibodies

Abstract

Antibodies raised against evolutionarily conserved intracytoplasmic peptide sequences of human T and B lymphocyte antigens and an antibody raised specifically against koala serum IgG were assessed for their capacity for immunoperoxidase staining of formalin fixed lymphoid tissues of koalas (Phascolarctos cinereus), common brushtail possums (Trichosurus vulpecula), common ringtail possums (Pseudocheirus peregrinus) and tammar wallabies (Macropus eugenii). Utilizing microwave pretreatment and a streptavidin biotin-horseradish peroxidase method for histochemical staining, polyclonal anti-human CD3 (for T cells), monoclonal anti-human CD5 (for T cells), monoclonal anti-human CD79b cy (for B cells) and polyclonal anti-koala IgG (for plasma cells) consistently stained lymphoid elements in a wide range of tissues from all four marsupial species. Monoclonal anti-human CD79a cy (for B cells) stained lymphoid elements in only brushtail possums. This is the first report of immunohistological staining of lymphocytes in Australian marsupials, thereby vastly increasing the scope for immunological studies in these species.

Cellular Response in the Dermis of Common Wombats (Vombatus Ursinus) Infected With Sarcoptes Scabiei Var. Wombati

“…Immunohistochemical staining of tissues of the common wombat is similar to that of the koala, its closest relative, and other marsupials, the ringtail possum and the tammar wallaby, with monoclonal antibodies anti-CD5, anti-CD3, and antiCD79b differentially staining cells (Hemsley et al, 1995). Similarly, the monoclonal antibody anti-CD79a did not stain cells and the polyclonal antibody anti-CD3 stained cells more strongly than monoclonal anti-CD3 (Hemsley et al, 1995).…”

“…Similarly, the monoclonal antibody anti-CD79a did not stain cells and the polyclonal antibody anti-CD3 stained cells more strongly than monoclonal anti-CD3 (Hemsley et al, 1995). However, anti-koala IgG antibodies did not differentially stain wombat plasma cells although they stained plasma cells from koala, brushtail and ringtail possums, and tammar wallabies.…”

“…In this study, conventional staining methods as well as immunohistochemistry using antibodies raised against conserved antigens were used to identify cell types in the dermis of common wombats infected with S. scabiei. The immunohistochemical staining of T and B lymphocytes of Australian marsupials such as the koala (Phascolarctos cinereus), brushtail possum (Trichosurus vulpecula), tammar wallaby (Macropus eugenii), and ringtail possum (Pseudocheirus peregrinus) has been achieved using antibodies raised against intracytoplasmic regions of receptors such as CD3 and CD79b of human T and B lymphocytes (Hemsley et al, 1995;Wilkinson et al, 1995), suggesting that these antibodies may also identify T and B lymphocytes of FIGURE 1. Sites on lateral surfaces, along the dorsal midline and on the ventral surface of common wombats from which 7 mm diameter biopsies were taken.…”

Devil Facial Tumor Disease

“…It is unknown what gave rise to the first devil facial tumor (DFT), but genomic analysis demonstrated that this primary tumor appeared in a female devil <20 years ago. 49 DFTD was first observed in 1996 in the far northeast of Tasmania 19 and has since spread to affect the majority of the species' geographic range, up to 90% of individuals within certain locations. 41 It causes mortality in all affected animals, seemingly within 6 months of the tumor's appearance.…”

“…Detailed investigation and clarification of immune cell function, however, are significantly hampered by the lack of species-specific reagents. 19,54 For example, antibodies against CD4 and CD8 have only recently been developed and only for use in immunohistochemistry. 21 Results of T-cell proliferation assays revealed that devils have a relatively robust mitogen-induced response.…”