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Neoplastic cells, through immunoediting mechanisms, can establish a state of immunosuppression to evade host immune defenses. The aims of this study were: (1) to validate a standard method for assessing tumor infiltrating lymphocytes (TILs) in canine mammary carcinoma by applying international human breast cancer guidelines; (2) to investigate if the TILs population was composed of a subset of regulatory T lymphocytes (Tregs); and (3) to evaluate the relationship between the number of TILs and Tregs and the biological behavior of the tumors. One hundred and twenty-nine canine mammary tumors were retrospectively selected for this study. Histological diagnosis, grading and histological evaluation of TILs was performed on hematoxylin and eosin-stained sections. TILs were evaluated using a three-tier semiquantitative method, previously validated in human medicine, based on the percentage of TILs (0-10%, 11-40% and 41-90%). Lymphocyte immunophenotype was confirmed by CD3 and CD79, while an anti-FoxP3 antibody was used to determine the presence of Tregs. The number of stromal TILs and invasive front TILs significantly correlated with each other (P < 0.0001) and increased with increasing histological grade (P = 0.002 and P = 0.004, respectively). A subset of TILs was composed of FOXP3 + Tregs. Stromal Tregs and invasive front Tregs were associated with stromal TILs and invasive front TILs (P = 0.03; P = 0.01 and P = 0.003; P = 0.007, respectively). In conclusion, in canine mammary carcinomas, an increased number of stromal and invasive front TILs is associated with increased malignancy and significant increase of Tregs that could lead to immunosuppression and evasion of the host immune system., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Papillomaviruses are among the most widespread animal viruses, with many hosts harbouring multiple virus types. The present study aimed to evaluate the haematological and immunophenotypic profile of cattle infected with bovine papillomavirus (BPV). Blood samples were collected from 10 animals with clinical cutaneous BPV and without clinical papillomatosis (control). Haematological analysis demonstrated a significant reduction in haemoglobin and haematocrit for BPV-infected animals. The results also showed an increase of natural killer cells and a decrease of γδ + T-cells and the CD4 + /CD8 + ratio for the BPV group when compared to the control group. The infection was also found to stimulate a pro-inflammatory profile with the participation of CD8 + T cells producing elevated IFN-γ and IL-17. These findings, although preliminary, provide a better understanding of the immune response of cattle infected with BPV., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Mast cell tumors (MCT) are among the most frequent tumors in dogs, but studies regarding canine mast cell immunophenotype are lacking. The aim of this study was to assess the feasibility of flow cytometric analysis of MCTs, to describe canine MCTs immunophenotype(s), and to evaluate the ability of flow cytometry to detect mast cells in lymph node aspirates. Thirty-four primary canine MCTs and 12 draining lymph nodes were evaluated regarding the expression of CD117, IgE, CD11b, CD18, CD44, CD34, CD25 and CD45. Distinct populations attributable to mast cells and eosinophils were recognized based on light scatters and CD117 positivity. Common antigens (CD18, CD45, CD44) and CD117 were detected in all cases; positivity for IgE and CD11b was found in 28 (82%) and 23 (68%) cases respectively, while CD34 and CD25 were occasionally expressed. A single multicolor tube (IgE/CD117/CD11b/CD21/CD5) allowed the identification of mast cells in lymph nodes, showing a high correlation with cytology in quantifying mast cells infiltration. In conclusion, flow cytometric analysis can be applied to characterize canine MCTs and can be used to detect the presence of mast cells in lymph nodes. The immunophenotype abnormalities observed may be useful to confirm the neoplastic nature of such mast cells but the diagnostic usefulness of atypical antigen expression remains to be clarified., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Multiparametric flow cytometry was used to compare peripheral blood lymphocyte subset distribution between healthy working police Labrador Retrievers (LRs; n=12) and German Shepherd dogs (GS; n=11) living in the same environment. The CD4/CD8 ratio was significantly higher in LR than in GS because of the lower percentage of CD8+ T lymphocytes in LR. GS showed the highest relative percentage of CD3-/CD21- lymphocytes, whereas LR had the highest percentages of MHC II+ lymphocytes. Because age, sex, environmental and housing conditions, dietary patterns, and training or working routines were similar in both breeds in the study, differences in peripheral blood lymphocyte subset distribution could be attributed to the influence of breed on the immune system., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The diagnosis of canine lymphoma is a multi-step process involving clinical examination, diagnostic imaging, cytology, haematology, biochemical profiling, histopathology and ancillary techniques such as flow cytometry (FC). In human medicine, FC (in addition to cytology) is reported to increase the accuracy of diagnosis of most lymphoma sub-types. In dogs, FC can add a number of useful diagnostic features to the morphological evaluation of lymphoma including the evaluation of B or T cell lineage, antigen quantification and evaluation of aberrant patterns, the assessment of clonality, staging and the evaluation of minimal residual disease. In comparison to other immunophenotyping techniques, FC is rapid and easy to perform and 'gating' techniques can resolve mixed cell populations although the use of fresh samples is required and the appropriate equipment and its maintenance are quite expensive. The use of FC to refine cytological diagnosis could be further enhanced by the use of a multi-parametric approach and by the development of a wider panel of standardised canine-specific antibodies., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The immunopathogenesis of cutaneous adverse food reactions (CAFRs) in dogs is unknown. Since the clinical manifestations in the skin are like those found in canine atopic dermatitis (AD), this study investigated the similarity in T cell phenotypes and gene expression of cytokines and transcription factors in CAFRs. In addition, the influence of an elimination diet on these parameters was tested. In the skin of canine CAFRs, a predominant presence of CD8(+) T cells and increased expression of the IL-4, IL-13, Foxp3 and SOCS-3 genes were observed. IFN-γ gene expression was increased in lesional compared to non-lesional skin. The predominance of CD8(+) T cells indicates that the immunopathogenesis of CAFRs is different from that of canine AD. The elimination diet relieved clinical signs, but did not influence T cell phenotypes or expression of the cytokine and transcription factor genes in the skin of dogs with CAFRs, indicating a continuously pre-activated immune status in dogs sensitised to food constituents., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Six cases of lymphosarcoma (LSA) in South American camelids (SACs) were selected from submissions to a diagnostic laboratory network servicing England and Wales. Immunophenotyping was carried out using anti-human CD3 and anti-human CD5 for T-cells; and anti-human CD79a and anti-human CD79b for B-cells/plasma cells. On the basis of labelling with mainly anti-CD3, four of the tumours were classified as T-cell tumours. One case was labelled with anti-CD79a and anti-CD79b, and was classified as a B-cell tumour. In the other case the majority of cells were labelled with anti-CD3, anti-CD79a and anti-CD79b, and was classified as a mixed T- and B-cell tumour. To the authors' knowledge this is the first reported attempt at immunophenotyping LSA in SACs on British premises and is only the second time that a presumptive mixed T- and B-cell LSA has been reported in alpacas and the veterinary literature in general.

A two-year-old female alpaca with multicentric lymphosarcoma presented because of progressive weakness and recumbency. The diagnosis of lymphosarcoma was based on the detection of immature and atypical lymphocytes in a lumbosacral CSF sample. Post mortem examination confirmed multicentric lymphosarcoma involving multiple organs. Immunophenotyping using cross reactive T- and B-cell antibodies characterized the tumour as a T-cell lymphosarcoma.

The aim of this study was to determine the response of different morphological subtypes of canine lymphoma to a standardized therapeutic protocol. Diagnosis of lymphoma was based on cytohistological analysis and immunophenotyping with antibodies against CD3 and CD79a of an enlarged lymph node or an extranodal mass. Fifty-seven cases were classified according to the updated Kiel classification adapted to the canine species, into 24 B-cell lymphomas (20 centroblastic polymorphic and four Burkitt-type subtypes), and 33 T-cell lymphomas (10 pleomorphic mixed, 10 lymphoblastic, eight unclassifiable high grade plasmacytoid, and five small clear-cell subtypes). All dogs were clinically staged at diagnosis. The protocol used l-asparaginase, vincristine, cyclophosphamide, doxorubicin, and prednisone. First remission duration and overall survival time were evaluated. Although the T-cell phenotype was associated, on the whole, with a poor prognosis, as previously reported in veterinary and human medicine, the study showed significant prognostic differences between the B- and the T-cell subtypes of canine lymphoma and suggests that clinico-morphological characterization of the disease is justified in dogs, as in humans.