Immune System Cells in Healthy Ferrets

Abstract

The ferret has emerged as an excellent animal model to characterize several physiologic and pathologic conditions. The distribution and characterization of different types of immune system cells were studied in healthy ferret tissues. Eight primary antibodies were tested for immunohistochemistry in formalin-fixed tissues: anti-CD3, anti-CD79α, anti-CD20, anti-HLA-DR, anti-lysozyme, anti-CD163, anti-SWC3, and anti-Mac387. The anti-CD3 antibody labeled T cells mainly in interfollicular and paracortical areas of lymph nodes, cortex and thymic medulla, and periarteriolar lymphoid sheaths in the spleen. The anti-CD79α and anti-CD20 antibodies immunolabeled B cells located in lymphoid follicles at lymph nodes, spleen, and Peyer patches. The CD79α and CD20 antibodies also labeled cells with nonlymphoid morphology in atypical B-cell locations. The anti-HLA-DR antibody labeled macrophages, some populations of B and T lymphocytes, and different populations of dendritic cells in lymph nodes, Peyer patches, spleen, and thymus. The anti-lysozyme antibody immunolabeled macrophages in the liver, lymph nodes, spleen, and thymus. The Mac-387, CD163, and SWC3 antibodies did not show any positive reaction in formalin-fixed or frozen tissues. To elucidate the origin of the uncommon CD79α/CD20 positive cells, a double immunohistochemistry was carried out using the anti-HLA-DR + the anti-CD79α, the anti-HLA-DR + the anti-CD20, and the anti-lysozyme + the anti-CD79α antibodies. Double labeling was mainly observed when the anti-HLA-DR + the anti-CD79α antibodies were combined. The immunohistologic characterization and distribution of these immune system cells in healthy ferret tissues should be of value in future comparative studies of diseases in ferrets.

Heterogeneous pathological outcomes after experimental pH1N1 influenza infection in ferrets correlate with viral replication and host immune responses in the lung

“…Phenotyping of the different inflammatory cell lineages following IHC were performed on the lungs of the infected and control animals to determine which inflammatory cell types were associated with the different lung lesional patterns. Neutrophils and macrophages were detected by IHC using anti-lysozyme polyclonal antibodies (Dako, Polyclonal Rabbit Anti-Human Lysozyme EC 3.2.1.17, ref A0099), T and B cells were detected using anti-CD3 (Dako, Polyclonal Rabbit Anti-Human CD3, n°I S503) and anti-CD20 (Thermo Scientific, CD20 Rabbit Polyclonal Antibody, ref RB-9013-P) polyclonal antibodies as previously described [29,30].…”

Flow Cytometric and Cytokine ELISpot Approaches To Characterize the Cell-Mediated Immune Response in Ferrets following Influenza Virus Infection

“…Gene expression studies, using information gathered from the draft genome and cross-reactive probes for microarrays, have confirmed the biphasic kinetics of innate and adaptive immune responses following viral infection (42)(43)(44)(45). While limited, serological reagents (46,47) have been used in a number of applications, including immunohistochemical analyses to study pathological features of disease as it relates to the virus/host interface (48). Additionally, other studies have quantified antibody-secreting cells within the draining lymph node in live attenuated influenza virus vaccine (LAIV) recipients, finding that the abundance of these cells inversely correlated with virus replication during a heterologous virus challenge (49).…”

Improving immunological insights into the ferret model of human viral infectious disease

“…Markers targeting B‐cell antigens such as CD79a, CD20 and surface immunoglobulin also allow ferret B cells to be examined by immunohistochemistry and flow cytometry in ferret tissues . B‐cell frequencies are also transiently decreased 2 d.p.i after infection, with a corresponding increase in secondary lymphoid organs 2‐5 d.p.i .…”

“…Such non‐neutralising mechanisms have been shown to be important for broadly protective responses against antigenically distinct strains of influenza and development of universal influenza vaccines. Many of the current ADCC and related assays rely on the detection of Ab‐mediated activation of NK cells to express cytokines such as IFN‐γ or degranulation markers such as CD107a, but there are currently no reagents to differentiate NK cells from other cytotoxic lymphocyte populations such as CD8+ T cells . While a ferret‐specific T‐cell IFN‐γ expression assays to measure CD8 T‐lymphocyte activation have been developed and validated, the future elucidation of corresponding antibody/Fc receptor ferret orthologues will enable ferrets to be used to evaluate ADCC responses.…”