Does the tumour microenvironment alter tumorigenesis and clinical response in transmissible venereal tumour in dogs?

The canine transmissible venereal tumour (CTVT) is a transmissible cancer that is spread naturally between dogs, with the ability to develop and evade the immune system, despite strict immune surveillance of the host. Furthermore, molecular signalling between cells of the immune system and the tumour microenvironment appear to influence the behaviour and development of the tumour. Thus, this study aimed to quantify the expression of genes related to the immune system such as IL‐6, IFN‐γ, and TGF‐β, as well as angiogenic factors (VEGF, CXCR4), in CTVT cells in vivo and in vitro (primary culture), correlating with the clinical response of the animals treated with vincristine. As expected, the most prevalent subtype was plasmacytoid cells, although lymphocytic cells were also found, indicating the possibility of polyclonality. When we compared the gene expressions of IFN‐γ and IL‐6, we mostly found low expression, concluding that MHC expression was probably not occurring in tumour cells, and no activation of immune cells to eliminate the tumour. The TGF‐β gene was normal in the majority of animals but demonstrated decreased expression in vincristine resistant animals, leading to the hypothesis that the concentration of tumour‐derived TGF‐β was affecting and even suppressing the real TGF‐β expression, favouring tumour proliferation and progression in these cases. VEGF expression was extremely high, demonstrating its angiogenic role in tumour growth, while CXCR4 was decreased, possibly because of CTVT’s low metastatic potential. Thus, we concluded that the tumour microenvironment, together with the immune system of the host, influences CTVT, presumably altering its tumorigenesis and the animal’s clinical response to treatment.     

Transfer of tumour necrosis factor-α via colostrum to foals

This study aimed to determine whether TNF-α is transferred to equine neonates via colostrum and the relationship between TNF-α and IgG concentrations in the equine neonate. Colostrum, presuckle and postsuckle foal serum samples were collected from healthy mares and their foals. Equine TNF-α ELISA and IgG SRID kits were used to determine the concentrations of TNF-α and IgG, respectively. Statistical analysis was performed using the Spearman rank correlation. TNF-α concentrations in all presuckle foal serum were below the limit of detection in 15/16 foals and increased in postsuckle foal serum to a mean concentration of 7.7 x 10(4) pg/ml. TNF-α concentrations in postsuckle foal serum and colostrum showed significant correlation (rho=0.668; P=0.005). However, TNF-α and IgG concentrations in colostrum or postsuckle foal serum did not correlate (rho<-0.016; P>0.05). Ratios of TNF-α/IgG in colostrum or postsuckle foal serum showed significant correlation (rho=0.750; P=0.0008). These results indicate that TNF-α is transferred to the foal via colostrum absorption and may play a role in early immunity.     

Clinical factors determining the efficacy of urinary bladder tumour treatments in dogs: surgery, chemotherapy or both?

In a study of 44 canine patients suffering from histopathologically proven urinary bladder tumour with a high incidence of transitional cell carcinoma (TCC) (n = 35), a close relationship was found either between the disease-free period and the age (r = -0.40) of animals or between the survival times and the age (r = -0.62) of animals after treatment. In addition to the dog breeds known to be prone to have urinary bladder tumour, we found an additional potentially sensitive breed, the Hungarian Vizsla. The median survival times obtained by the applied treatment types were as follow: 'surgery and chemotherapy' (n = 8/44) 475 days, 'surgery alone' (n = 19/44) 240 days, 'chemotherapy alone' (n = 7/44) 31 days, and 'no treatment' (n = 10/44) 7 days (P < 0.001). According to the findings, chemotherapy combined with surgery completed in time is the most effective protocol in the treatment of urinary bladder tumour cases in dogs. A rational and more effective procedure for the assessment and treatment of urinary bladder tumour cases is presented.     

Immunoexpression of cytokine tumour necrosis factor-α suggesting its role in formation and regression of corpus luteum in Indian buffalo

Tumour necrosis factor-α (TNF-α) is a cytokine that plays multiple important roles in corpus luteum (CL). Immunolocalization of expression of TNF-α in CL of buffalo was studied in different stages of its development and regression. Corpus luteum of healthy buffaloes (24) was collected from local slaughterhouses and categorized into early (stage I, 1–5 days, n = 6), mid (stage II, 6–11 days, n = 6), late luteal (stage III, 12–16 days, n = 6) and regressing phase (stage IV, 17–20 days, n = 6). In earliest phase of cyclic CL, per cent immunoexpression of TNF-α was significantly (p < .05) lower as compared to all phases with its expression being restricted to few developing luteal cells, usually in neutrophils. A significantly (p < .05) higher number of neutrophils with TNF-α immunoexpression were observed as compared to mid-luteal phase that indicated its role in initiation of angiogenesis at this stage. TNF-α immunoexpression almost doubled in mid-luteal phase, but the number of neutrophils exhibiting TNF-α was significantly (p < .05) lower with respect to all phases of CL. Immunoexpression percentage in late luteal phase increased sharply being significantly (p < .05) higher than earlier two phases of CL. In regressing phase, per cent immunostaining was maximum with highly significant (p < .05) difference as compared to all other stages, observed in all degrading luteal cells, abundant immune cells, that is neutrophils and macrophages which finally led to apoptosis and phagocytosis. Immunoexpression of TNF-α in early luteal phases served its role in initiation of angiogenesis, and its intense expression in regressing phase of CL suggested a shift in its role to apoptosis and structural luteal regression signifying both luteotropic and luteolytic roles in buffalo. This is probably the first study of its kind in buffaloes.     

Primary cutaneous extragenital canine transmissible venereal tumour with Leishmania‐laden neoplastic cells: a further suggestion of histiocytic origin?

The clinical signs and histopathological features of a primary extragenital canine transmissible venereal tumour (TVT) are described. Three subcutaneous round alopecic nodules were located on the anterior and caudal dorsal region and in the ventral area of the neck. Cytologically, tumour cells were intermediate in size with a moderate amount of cytoplasm, and the nuclei were immature with finely reticular chromatin. The cytoplasm was lightly to heavily basophilic and contained distinct small vacuoles at the periphery. On the basis of these characteristics, a diagnosis of TVT was made and confirmed by histological and ultrastructural investigations. Leishmania amastigotes were detected in the cytoplasm of macrophages and neoplastic cells of the tumoral mass. The presence of the parasite within neoplastic cells is consistent with a histiocytic origin of TVT.     

Concentrations of interleukin-6, -8, -10 and tumour necrosis factor-α in the faeces of dogs with acute diarrhoea

Aim: To compare the concentration of faecal cytokines interleukin (IL)-6, -8, -10, and tumour necrosis factor (TNF)-α in dogs with acute diarrhoea with clinically normal (non-diarrhoeic) dogs.

Methods: A total of 14 dogs presenting with acute diarrhoea, and 25 dogs with no history of gastrointestinal signs in the 2 months prior to enrolment, were recruited from two veterinary hospitals in Melbourne, Australia. Concentrations of IL-6, -8, -10, and TNF-α were measured in faecal samples using canine-specific ELISA.

Results: The diarrhoeic dogs were diagnosed with or managed for acute gastroenteritis (n?=?6), extra-intestinal neoplasia (n?=?2), parvoviral enteritis (n?=?1), hepatopathy (n?=?1), acute pancreatitis (n?=?1), hypoadrenocorticism (n?=?1), gastric dilatation volvulus (n?=?1) and myelopathy (n?=?1). IL-6 was detectable in the faeces of 10/14 (71%) diarrhoeic and 7/25 (28%) non-diarrhoeic dogs, and median concentrations were 10.8 (min 0.0, max?54.0) and 2.0 (min 0.0, max15.0) pg/mL, respectively (p?=?0.01). IL-8 was detectable in the faeces of all diarrhoeic and 11 non-diarrhoeic dogs, and median concentrations were 149.7 (min 3.72, max?730.1) and 3.4 (min 0.0, max?22.5)?pg/mL, respectively (p?<?0.001). TNF-α was detected in the faeces of two of the diarrhoeic dogs (3.4 and 15.6?pg/mL) and none of the non-diarrhoeic dogs. IL-10 was not detected in the faeces of any dog.

Conclusions: Faecal concentrations of IL-6 and -8 were higher in diarrhoeic compared to non-diarrhoeic dogs, and are therefore potential candidates for non-invasive biomarkers to assess the severity and resolution of acute intestinal disease in dogs. However their correlation with disease progression and severity needs to be further investigated before their full clinical application can be determined.     

Dominant expression of interleukin-8 vs interleukin-1β and tumour necrosis factor alpha in lungs of lambs experimentally infected with Mannheimia haemolytica

Abstract

AIMS: To quantify the number of cells infected with Mannheimia haemolytica and expressing interleukin (IL)-1β, tumour necrosis factor alpha (TNFα) and IL-8 using immunohistochemistry, and to measure the immunoreactivity of cytokines in pulmonary tissue extracts using ELISA, in the lung of lambs experimentally infected with M. haemolytica, and to compare the patterns of expression of cytokines in airways at different times post-infection (p.i.).

METHODS: Twenty 3-month-old lambs of both sexes were randomly assigned to two groups, viz infected (n=15), and uninfected controls (n=5). Each lamb in the infected group was inoculated with 1.5 x 10⁹ cfu M. haemolytica in 5 mL sterile nutrient broth, control lambs were inoculated with 5 mL sterile nutrient broth and clinical signs were monitored. Infected and control animals were killed at 1, 3, 5, 7, and 15 days p.i. Histopathology and immunohistochemistry were conducted to determine the number of immunolabelled cells in pneumonic lungs, and study the pattern of expression of IL-1β, TNFα and IL-8 in lung extracts using ELISA.

RESULTS: Lesions in bronchi and bronchioles ranged from epithelial desquamation to bronchiolitis obliterans and necrosis. The alveoli had areas of seroproteinaceous fluid, fibrin and bacterial aggregates that evolved to foci of pyogranulomatous inflammation with clustered inflammatory cells, referred to as ‘oat cells’. M. haemolytica antigen was observed in the cytoplasm of inflammatory cells. Labelling of IL-1β, TNFα and IL-8 was observed in bronchial and bronchiolar epithelial cells, alveolar exudate, and in interstitial inflammatory infiltrate, with increased expression on 1 and 3 days p.i. for IL-1β and TNFα, and 1, 3, and 5 days p.i. for IL-8. In lung tissue extracts, peak concentrations of IL-1β (55 (SD 5) ng/mL), TNFα (92 (SD 6) pg/mL) and IL-8 (8 [SD 2] μg/mL) occurred at 3 days p.i.

CONCLUSIONS: The results of this study suggested that the inflammatory cytokines IL-1β, TNFα and IL-8 may play an important role in enhancing the biological response to M. haemolytica, and contribute to the development of lesions in the lung in pulmonary pasteurellosis in sheep. Given that the expression of IL-8 in lung was much greater than that of IL-1β and TNFα, anti-cytokine agents directed at this mediator could be useful in the prevention and treatment of this disease.     

Clinical and computed tomography tumour dimension assessments for planning wide excision of injection site sarcomas in cats: how strong is the agreement?

In injection site sarcoma (ISS) in cats lateral as well as deep margins should be correctly planned for a successful surgical outcome. The discrepancy between clinical and computed tomography (CT) measurements of dimension in resectable tumour has led to possible bias that affects the subsequent surgical dose. The aim of this study was to prospectively investigate the agreement between clinical and CT measurements of dimension in newly diagnosed ISS in cats. Fifty‐three client‐owned cats that underwent both clinical and CT measurements of the length and width of ISS were included. CT measurements showed a tendency towards being larger than clinical dimensions, and this difference increased with increasing tumour size. Based on our results, in further studies focusing on ISS in cats, the kind of assessment used to define tumour dimensions (CT versus clinic) should be declared and specified to properly consider surgical results and prognostic impact of this variable.     

Histologic margins and the residual tumour classification scheme: Is it time to use a validated scheme in human oncology to standardise margin assessment in veterinary oncology?

There is no consensus on the definition of a complete histologic excision in veterinary oncology; many definitions have been used in various studies, but these have been arbitrarily selected with no apparent justification. The residual tumour classification scheme, where a complete histologic excision is defined as a histologic tumour‐free margin >0 mm, has been used for >40 years in human oncology by all of the major clinical staging organizations and is considered highly prognostic for the vast majority of malignant tumours in people. Because of the widespread use of the residual tumour classification scheme both clinically and in research studies, this standardized approach permits better communication between clinicians, an evidence‐based decision‐making process for adjuvant treatment options following surgical resection, minimizes exposing patients to unnecessary adjuvant treatments and a better ability to compare local tumour control for specific tumours between different studies. The adoption of the residual tumour classification scheme in veterinary oncology would likely achieve similar outcomes and minimize the prevalent confusion within the veterinary community, amongst both general practitioners and specialists, regarding the definition of what constitutes a complete histologic excision.     

Ascites in a 13‐year‐old miniature horse mare with an ovarian granulosa theca‐cell tumour: A manifestation of Meigs' syndrome?

A 13‐year‐old miniature horse mare presented for evaluation of ascites. Abdominal ultrasound showed severe free peritoneal fluid accumulation as well as a caudal abdominal mass. The peritoneal fluid was characterised as a transudate with no evidence of sepsis. Surgical exploration of the abdomen was performed; copious free peritoneal fluid and a large right ovarian mass were discovered. The abnormal ovary was removed, and the mass was determined histologically to be a benign granulosa theca‐cell tumour. Although there was mild peritoneal fluid accumulation in the immediate post operative period, the ascites effectively resolved with removal of the tumour, and the mare returned to a normal lifestyle. This is similar to reports of Meigs' syndrome in human women, in which a benign ovarian mass of significant size causes ascites that resolves once the mass is removed.