Maria Elena Gelain

The dog as a possible animal model for human non-Hodgkin lymphoma: a review

Lymphoma represents the most frequent hematopoietic cancer in dogs, and it shows significant overlap with the human disease. Several environmental factors have been associated with canine lymphoma, suggesting that they may contribute to lymphomagenesis. Canine lymphoma often presents in advanced stage (III–V) at diagnosis and, most commonly, has an aggressive clinical course requiring prompt treatment, which relies on the use of polychemotherapy. In this review, we will summarize the state‐of‐the‐art of canine lymphoma epidemiology, pathobiology, diagnostic work‐up and therapy, and will highlight the links to the corresponding human disease, providing evidence for the use of dog as an animal model of spontaneous disease. Copyright

Immunophenotype predicts survival time in dogs with chronic lymphocytic leukemia.

BACKGROUND Chronic lymphocytic leukemia (CLL) is a hematologic disorder in dogs, but studies on prognostic factors and clinical outcome are lacking. In people, several prognostic factors have been identified and currently are used to manage patients and determine therapy. OBJECTIVES The aim of the study was to determine if the immunophenotype of neoplastic cells predicts survival in canine CLL. DESIGN Retrospective study. ANIMALS Forty-three dogs with CLL. PROCEDURES Records of dogs with a final diagnosis of CLL were reviewed. For each included dog, a CBC, blood smear for microscopic reevaluation, and immunophenotyping data had to be available. Data on signalment, history, clinical findings, therapy, follow-up, as well as date and cause of death were retrieved. RESULTS Seventeen dogs had B-CLL (CD21+), 19 had T-CLL (CD3+ CD8+), and 7 had atypical CLL (3 CD3- CD8+, 2 CD3+ CD4- CD8-, 1 CD3+ CD4+ CD8+, and 1 CD3+ CD21+). Among the variables considered, only immunophenotype was associated with survival. Dogs with T-CLL had approximately 3-fold and 19-fold higher probability of surviving than dogs with B-CLL and atypical CLL, respectively. Old dogs with B-CLL survived significantly longer than did young dogs, and anemic dogs with T-CLL survived a significantly shorter time than dogs without anemia. CONCLUSIONS Although preliminary, results suggested that immunophenotype is useful to predict survival in dogs with CLL. Young age and anemia are associated with shorter survival in dogs with B-CLL and T-CLL, respectively.

Identification of suitable endogenous controls and differentially expressed microRNAs in canine fresh-frozen and FFPE lymphoma samples

The elucidation of microRNA (miRNA) expression pattern in canine lymphoma is attractive for veterinary and comparative oncology due to similar genetics, physiology and exposure to environment in dogs and humans. In this work, the expression of a panel of mature miRNAs was quantitated in fresh-frozen and formalin-fixed paraffin-embedded (FFPE) lymph nodes from canine lymphoma. The major findings were: the detection of a panel of miRNAs expressed in canine lymph node; the identification of three suitable endogenous controls (let-7a, miR-16, and miR-26b) by NormFinder and geNorm analysis; the concordance between results obtained from fresh-frozen and FFPE samples; the detection of upregulation of miR-17-5p and miR-181a in B- and T-cell lymphomas respectively. This is the first study aimed to the application of miRNAs analysis in canine lymphoma.

Whole blood cytokine profiles in cats infected by feline coronavirus and healthy non-FCoV infected specific pathogen-free cats.

In this study, the cytokine profiles of clinically healthy cats naturally infected with feline coronavirus (FCoV), of cats with feline infectious peritonitis (FIP) and of specific pathogen-free (SPF) cats were investigated in whole blood using a traditional reverse-transcriptase polymerase chain reaction (RT-PCR) assay and a semi-quantitative method of analysis based on computerised quantification of positive bands. The low inter-assay coefficient of variation recorded demonstrated that this method is highly repeatable. Compared with SPF cats, cytokine production was upregulated in most of the samples from FCoV-positive non-symptomatic cats. The appearance of a case of FIP in the cattery was associated with an increased expression of cytokines, in particular there was an increased production of IL-1β and IFN-γ, suggesting that these cytokines might protect infected cats from the disease. This hypothesis was also supported by the low levels of IFN-γ recorded in blood from cats with FIP.

Flow‐cytometric detection of phenotypic aberrancies in canine small clear cell lymphoma

Histopathology and immunohistochemistry are mandatory to solve the differential between canine low-grade lymphoma and reactive hyperplasia. However, clinicians and owners often show reluctance toward these invasive tests. However, molecular biology techniques are still not sensitive and specific enough to be regarded as a reliable tool for final diagnosis. In humans, flow cytometry (FC) allows a definitive diagnosis of T-cell lymphoma based on high prevalence of antigen aberrancies. We describe here the immunophenotype of 26 cases of suspect canine small-clear cell lymphoma, determined by multi-colour FC. All cases showed antigen aberrancies and therefore neoplasia was always confirmed. As a consequence, we argue that the combined use of cytology and FC allows solving the differential diagnosis between small clear cell lymphoma and non-neoplastic reactive conditions when histopathology is not available. Further studies are needed to establish if any aberrancy can be considered indicative of specific histotypes.

Disseminated histiocytic sarcoma with peripheral blood involvement in a Bernese Mountain dog

A 6-year-old Bernese Mountain dog was presented with a history of lethargy and weight loss of 2 weeks duration. On physical examination the dog had pale mucous membranes and tachypnea. Ultrasound examination revealed hepatomegaly, splenomegaly, and mesenteric lymphadenomegaly. Results of a CBC included marked normocytic normochromic nonregenerative anemia, marked thrombocytopenia, and moderate leukocytosis with mild neutrophilia and a large population of unclassified round cells (6.2 x 10(3)/microL). The unclassified cells occasionally were bi- or multinucleated and had variably abundant pale basophilic cytoplasm that contained multiple irregular clear vacuoles and occasionally erythrocytes. Fine needle aspirate specimens of the mesenteric lymph nodes and spleen were composed of a population of round pleomorphic cells with the same features as the circulating cells. On flow cytometric analysis of peripheral blood, the unclassified cells expressed CD18, CD45, CD11c, CD1c, and CD14; immunocytochemical analysis of blood smears also indicated the cells were positive for CD1c, CD1a, and CD11c. The dog died a few hours after referral. The histologic interpretation of samples collected from spleen, liver, and lymph nodes was malignant neoplasia of histiocytic origin. Immunohistochemical staining yielded negative results for CD11d, a marker of red-pulp macrophages, ruling out hemophagocytic histiocytic sarcoma. Based on clinical and pathologic findings, the final diagnosis was disseminated histiocytic sarcoma (DHS) with peripheral blood involvement. To our knowledge, DHS in a dog with evidence and immunophenotyping of neoplastic cells in peripheral blood has been reported only rarely.

Serum α1-acid glycoprotein (AGP) concentration in non-symptomatic cats with feline coronavirus (FCoV) infection

Previous studies have demonstrated that the concentration of α1-acid glycoprotein (AGP) transiently increases in asymptomatic cats infected with feline coronavirus (FCoV). In order to establish whether these fluctuations depend on the FCoV status, the serum concentration of AGP and anti-FCoV antibody titres and/or faecal shedding of FCoVs in clinically healthy cats from catteries with different levels of prevalence of FCoV infection were monitored over time. Serum AGP concentrations fluctuated over time in clinically healthy cats from the cattery with the highest prevalence of feline infectious peritonitis (FIP) and significantly increased just before an outbreak of FIP. Further studies are required to clarify whether the observed increase of AGP concentration is a consequence of the increased viral burden or a protective response against mutated viral strains. Nevertheless, the results of the present study suggest that AGP might be useful in monitoring FCoV–host interactions in FCoV-endemic catteries.

Immunophenotype-related microRNA expression in canine chronic lymphocytic leukemia.

MicroRNAs (miRNAs) are posttranscriptional regulatory noncoding RNAs used to profile human hematopoietic tumors. In this study, some mature miRNAs was quantitated in peripheral blood from dogs with chronic lymphocytic leukemia (CLL). Relative expression data were normalised against four endogenous controls (let-7a, miR-17-5p, miR-26b, and miR-223) selected by geNorm analysis. The results revealed distinct miRNA patterns in CLL depending on the immunophenotype. Also in dogs, the different miRNAs expression could reflect developmental lineage and tumor differentiation. The similar genetics, physiology and exposure to environment in dogs and humans make the miRNA expression study in canine CLL attractive for comparative oncology.

Long-term study of MRSA ST1, t127 mastitis in a dairy cow

METHICILLIN-resistant Staphylococcus aureus (MRSA) has been reported in human medicine as a cause of nosocomial and community-associated infections (Otter and French 2010). In veterinary medicine, MRSA strains have been identified in a wide range of animals and diseases (Leonard and Markey 2008, Fessler and others 2009, Huber and others 2010, Turkyilmaz and others 2010), thus it is considered an emerging threat with a high zoonotic potential (Juhasz-Kaszanyitzky and others 2007). MRSA sequence type (ST) 1, spa type (t) 127 has been mostly isolated from community-associated infections, but t127 has seldom been identified in bovine and swine (Hasman and others 2010). The present report regards a dairy cow with an intramammary infection by MRSA ST1, t127 that was investigated over an entire lactation and submitted to histological evaluation of mammary tissue to explore both bacterial molecular features and host immune response in the milk and mammary tissue. The study was performed because only a few mammary infections of the dairy cow by this particular strain have been reported and no information is available on the interaction between the bacteria and the mammary gland. A five-year-old Holstein-Friesian dairy cow from a closed herd located in Milano province, Italy, consisting of 25 lactating cows, showed an intramammary infection with S aureus in both right quarters. The isolate revealed a multidrug resistance profile by the agar-plate method, and PCR analysis (Costa and Palladini 2005) confirmed the presence of mecA genes in both isolates, which were consequently classified as MRSA. The farmer decided to antibiotic treat and dry off the animal, …

Association between faecal shedding of feline coronavirus and serum α1-acid glycoprotein sialylation

The sialylation pattern of serum α1-acid glycoprotein (AGP) in non-symptomatic cats infected by feline coronavirus (FCoV) and its possible relationship with the amount of FCoVs shed in faeces were investigated. Blood from three specific pathogen-free cats (group A) and from 10 non-symptomatic FCoV-positive cats from catteries with low (group B, three cats) or high (group C, seven cats) levels of faecal shedding were collected monthly. AGP was purified from serum and Western blotting followed by lectin-staining of α(2,3)-linked and α(2,6)-linked sialic acid. Faecal shedding was quantified in group C by quantitative polymerase chain reaction. Variations of AGP sialylation were recorded only in cats from group C, on which viral shedding peaked before the occurrence of feline infectious peritonitis (FIP) in the cattery, and decreased 1 month later, when serum AGP had an increase of α(2,3)-linked sialic acid. These results suggest that hypersialylation of AGP may be involved in host–virus interactions.