Feline Tissues Research Articles

Feline injection-site sarcoma: Past, present and future perspectives

Feline injection-site sarcomas (FISS) have been known since the early 1990s. After an initial correlation with rabies and feline leukaemia virus vaccination, subsequent studies have demonstrated that an abnormal reaction of feline tissues to chronic inflammation was mainly responsible for the disease. The low incidence of FISS in the population is explained by its multifactorial aetiology, since individual genetic characteristics are also implicated. FISS is an infiltrative tumour with low metastatic potential but local recurrence is common. Multimodal treatment (extensive surgery, radiotherapy, chemotherapy) is recommended. The use of sophisticated imaging techniques can improve diagnosis and help in surgical planning. After the initial enthusiasm in understanding the disease, only few advances have been made in the last few years. New promising therapies may arise from a better knowledge of the molecular pathogenesis of FISS and the successful development of drugs modulating the immune system.

CONCOMITANT LIPOSOMAL DOXORUBICIN AND DAILY PALLIATIVE RADIOTHERAPY IN ADVANCED FELINE SOFT TISSUE SARCOMAS

Local recurrence of feline soft tissue sarcomas is common despite aggressive treatment. Liposomal doxorubicin might serve as a depot radiosensitizer if administered concomitantly with daily radiotherapy and thus improve tumor control. In this pilot study, the feasibility of concomitant liposomal radiochemotherapy was evaluated in a palliative setting in 10 cats with advanced soft tissue sarcomas. Cats were treated with median number of 5 (range 5-7) daily fractions of radiotherapy and a median total dose of 20 Gy (range 20-31.5 Gy). One dose of liposomal doxorubicin was administered at the beginning of radiotherapy. Seven cats received further free or liposomal doxorubicin after completion of the liposomal doxorubicin/radiation protocol. Seven of the treated 10 cats (70%) achieved a partial (n=5) or complete (n=2) response with a median response duration of 237 days. The median progression free interval in all 10 cats was 117 days and the median overall survival time was 324 days. Concomitant liposomal radiochemotherapy was tolerated well in nine cats, one cat experienced temporary anorexia. Although the number of patients is too small to make definitive conclusions, results appear promising enough to investigate the role of liposomal doxorubicin as a radiosensitizer further.

Quantitative TaqMan® real-time PCR assays for gene expression normalisation in feline tissues

BackgroundGene expression analysis is an important tool in contemporary research, with real-time PCR as the method of choice for quantifying transcription levels. Co-analysis of suitable reference genes is crucial for accurate expression normalisation. Reference gene expression may vary, e.g., among species or tissues; thus, candidate genes must be tested prior to use in expression studies. The domestic cat is an important study subject in both medical research and veterinary medicine. The aim of the present study was to develop TaqMan® real-time PCR assays for eight potential reference genes and to test their applicability for feline samples, including blood, lymphoid, endocrine, and gastrointestinal tissues from healthy cats, and neoplastic tissues from FeLV-infected cats.ResultsRNA extraction from tissues was optimised for minimal genomic DNA (gDNA) contamination without use of a DNase treatment. Real-time PCR assays were established and optimised for v-abl Abelson murine leukaemia viral oncogene homolog (ABL), β-actin (ACTB), β-2-microglobulin (B2M), β-glucuronidase (GUSB), hydroxymethyl-bilane synthase (HMBS), hypoxanthine phosphoribosyltransferase (HPRT), ribosomal protein S7 (RPS7), and tryptophan 5-monooxygenase activation protein, zeta polypeptide (YWHAZ). The presence of pseudogenes was confirmed for four of the eight investigated genes (ACTB, HPRT, RPS7, and YWHAZ). The assays were tested together with previously developed TaqMan® assays for feline glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the universal 18S rRNA gene. Significant differences were found among the expression levels of the ten candidate reference genes, with a ~106-fold expression difference between the most abundant (18S rRNA) and the least abundant genes (ABL, GUSB, and HMBS). The expression stability determined by the geNorm and NormFinder programs differed significantly. Using the ANOVA-based NormFinder program, RPS7 was the most stable gene in the tissues studied, followed by ACTB and ABL; B2M, HPRT, and the 18S rRNA genes were the least stable ones.ConclusionThe reference gene expression stability varied considerably among the feline tissues investigated. No tested gene was optimal for normalisation in all tissues. For the majority of the tissues, two to three reference genes were necessary for accurate normalisation. The present study yields essential information on the correct choice of feline reference genes depending on the tissues analysed.

Tissue Distribution of P‐Glycoprotein in Cats

Permeability glycoprotein (P-gp) is a membrane-bound efflux pump that exports various substances out of the cell. Variations in P-gp expression play an important role in susceptibility to toxic substances, drug efficacy and disease risk. In the present study, the distribution of the MDR1-gene product P-gp was determined in normal tissues of domestic shorthair cats using immunohistochemistry. Two monoclonal antibodies C494 and C219 were used, recognizing a different epitope on the human P-gp molecule. A consistent positive immunolabelling was obtained. The tissue distribution and cellular locations with antibody C494 were similar to those in man and dogs; with liver, colon, adrenal cortex and brain capillaries being consistently and intensely labelled. However, the immunolabelling in the kidney was in contradiction to man and dogs. The C219 antibody seems to react with a specific form of P-gp, only expressed in feline tissues with a barrier function, i.e. endothelia of the brain, testes and ovaries, and intestinal epithelial cells in contact with the lumen.

Immunohistochemical Expression of c-KIT Protein in Feline Soft Tissue Fibrosarcomas

C-KIT is the cellular homolog of the feline sarcoma viral oncogene v-KIT, which encodes the tyrosine kinase receptor protein KIT. Mutations and varied expression of this gene have been demonstrated within multiple neoplasms in people and domestic animals. The purpose of this study was to determine if KIT protein is expressed in feline soft tissue fibrosarcomas (ST FSA) using immunohistochemistry (IHC). The computer database at the Oklahoma Animal Disease Diagnostic Laboratory was searched from January 1, 2006, to December 31, 2007, for any domestic cat with an ST FSA. Routinely stained slides from 46 feline ST FSAs were reviewed and graded based on the scale outlined by Kuntz et al. Immunohistochemistry for KIT protein was performed on one representative section from each cat. There were a total of 12/46 (26%) cats that were immunoreactive for KIT. Immunoreactivity was detected in greater than 80% of the neoplastic cells in 4/46 (9%) cats. Immunoreactivity was detected in less than 10% of the neoplastic cells in 8/46 (17%) cats. Immunoreactivity was characterized by evenly distributed cytoplasmic stippling within the neoplastic spindle-shaped cells and/or multinucleated giant cells. Based on these results, KIT immunoreactivity can be detected within feline ST FSAs using IHC. The results of this study also indicate that KIT immunoreactivity in feline ST FSA does not correlate with the histologic grade (P = .141, X(2) = 2.166), survivability (P = .241, X(2) = 1.373), or whether the neoplasm was a spontaneous or an injection site FSA (P = .074, X(2) = 3.184).

An Immunohistochemical Study of Feline Endometrial Adenocarcinoma

Feline endometrial adenocarcinomas are uncommon malignant neoplasms that have to date been poorly characterized. The present immunohistochemical study describes the expression of the pancytokeratins AE1 and AE3, cytokeratin-14, vimentin, alpha-actin, cyclo-oxygenase-2, E-cadherin, beta-catenin, the progesterone receptor, the oestrogen receptor and caveolin-1 within normal feline uterine tissue and tissue from six cats with endometrial adenocarcinoma. Synthesis of cyclo-oxygenase-2 and reduced expression of progesterone receptors may be involved in the neoplastic transformation of feline endometrium. The loss of cellular adhesion that occurs within these tumours does not require down-regulation of E-cadherin expression and nuclear translocation of beta-catenin is not a feature of these neoplasms.

Identification and expression of the epithelial Muc1 mucin in normal feline tissues

Many pathogens require direct binding to, or penetration of, mucosal cells to cause pathology. Cell surface mucins are critical components of mucosal defense. Mucin 1, named MUC1 in humans and Muc1 in non-human species, is a transmembrane glycoprotein expressed in apical mammalian epithelial tissues. The aim of this study was to determine the Muc1 profile expression in healthy cat epithelial tissues. An extensive analysis of Muc1 expression was performed by immunohistochemistry (IHC), Western blot (WB) and RT-PCR. By IHC, the presence of Muc1 protein was observed in the epithelial cells of the esophagus, stomach, trachea, lung, small and large intestine, liver, pancreas, salivary glands, lactating mammary glands and bladder. The predominantly linear patterns of reaction as well as the ubiquitous expression of feline Muc1 were consistent with normal human tissues. By WB, a band of 35 kDa, corresponding to that predicted for the Muc1 cytoplasmic tail, was detected. The RT-PCR analysis showed a fragment of 115 bp, consistent with that found in MCF7 and T47D human cell lines. The results showed that the widest distribution of feline Muc1 expression is in the mucosal tissues most at risk of infections such as the gastrointestinal tract, respiratory tract and lactating mammary gland. This study provides a normal model of cat Muc1 pattern expression as a starting point to evaluate and compare the expression of this epithelial mucin in pathological feline tissues. We believe that the CT33 antibody and the universal primers designed could be valuable tools for veterinary pathologists involved in the diagnostic interpretation of alterations in Muc1 expression of infected tissues in cats.

Comparison of insulin signaling gene expression in insulin sensitive tissues between cats and dogs

Diabetes mellitus (DM) is a common endocrine disease in cats and dogs with increasing prevalence. Type 1 DM appears to be the most common form of diabetes in dogs whereas Type 2 DM prevails for cats. Since insulin resistance is more frequently encountered in cats than dogs, our laboratory was interested in determining whether differences at the insulin signaling pathway level and differences in glucose and lipid metabolism could be observed between cats and dogs. Insulin resistance has been positively correlated to insulin signaling pathway abnormalities. As such, this study measured insulin receptor substrate-1 (IRS-1), insulin receptor substrate-2 (IRS-2), and phosphatidylinositol 3-kinase (PI3-K) P-85alpha mRNA expression levels in classical insulin-responsive sensitive tissues (liver, skeletal muscle, and abdominal fat) and peripheral leukocytes between cats and dogs by qRT-PCR. Different tissues were sampled because it is currently unknown where insulin-resistance arises from. In addition, enzymes involved in glucose and lipid metabolism, malate dehydrogenase (MDH), glucose-6-phosphate dehydrogenase (G6PDH) and fatty acid synthase (FAS) were also assessed since glucose and lipid metabolism differs between cats and dogs. Overall, IRS-1, IRS-2, PI3-K, MDH, G6DPH, and FAS mRNA tissue expression profiles demonstrated different levels of expression, in various tissues for both canines and felines, which was expected. No distinct expression pattern emerged; however, differences were noted between canines and felines. In addition, IRS-1, IRS-2, PI3-K, MDH, G6DPH, and FAS mRNA expression was significantly higher in canine versus feline tissues, including peripheral leukocytes. Remarkable differences in insulin signaling gene expression between felines and canines indicate that cats may have an underlying low insulin sensitivity level due to low IRS-1, IRS-2, and PI3-K P-85alpha mRNA expression levels which would predispose cats to develop insulin resistance. Moreover, differences in glucose and lipid metabolism related gene expression (MDH, G6DPH, and FAS) demonstrate that felines have an overall lower metabolic rate in various tissues which may be attributed to overall lower insulin signaling gene expression and a lack of physical activity as compared to canines. Therefore, a combination of genetic and environmental factors appears to make felines more prone to suffer from insulin resistance and type 2 DM than canines.

Effect of Sample Quality on the Sensitivity of Endoscopic Biopsy for Detecting Gastric and Duodenal Lesions in Dogs and Cats

The quality of histopathology slides of endoscopic biopsies from different laboratories varies, but the effect of biopsy quality on outcome is unknown. The ability to demonstrate a histologic lesion in the stomach or duodenum of a dog or cat is affected by the quality of endoscopic biopsy samples submitted. More endoscopic samples are needed to find a lesion in poor-quality tissue specimens. Tissues from 99 dogs and 51 cats were examined as clinical cases at 8 veterinary institutions or practices in 5 countries. Histopathology slides from sequential cases that underwent endoscopic biopsy were submitted by participating institutions. Quality of the histologic section of tissue (inadequate, marginal, adequate), type of lesion (lymphangiectasia, crypt lesion, villus blunting, cellular infiltrate), and severity of lesion (normal, mild, moderate, severe) were determined. Sensitivity of different quality tissue samples for finding different lesions was determined. Fewer samples were required from dogs for diagnosis as the quality of the sample improved from inadequate to marginal to adequate. Duodenal lesions in cats displayed the same trend except for moderate duodenal infiltrates for which quality of tissue sample made no difference. Gastric lesions in dogs and mild gastric lesions in cats had the same trend, whereas the number of tissue samples needed to diagnose moderately severe gastric lesions in cats was not affected by the quality of tissue sample. The quality of endoscopically obtained tissue samples has a profound effect on their sensitivity for identifying certain lesions, and there are differences between biopsies of canine and feline tissues.

Induction of the Human Heat Shock Promoter HSP70B by Nutritional Stress: Implications for Cancer Gene Therapy

Background: We designed and tested, in vitro, an adenoviral construct containing the feline interleukin-12 (IL-12) gene under control of the heat-inducible promoter HSP70B. This construct, AdhspfIL12, was used in a phase I trial in feline soft tissue sarcomas. During the course of our experiments, we noted that IL-12 was being produced in the transfected Crandell Feline Kidney (CrFK) cells under certain conditions even in the absence of hyperthermia. This observation was further explored to identify the cause of this unintended HSP70B induction. Materials and Methods: We used real-time PCR as a sensitive method to quantitatively detect the presence of even small amounts of IL-12 mRNA. This served as a surrogate indicator of HSP70B induction. Various conditions were tested to induce the heat shock promoter, including nutritional deprivation, radiation and changes in pH. Results: Nutritional stresses, specifically the absence of glucose and glutamine, could induce the heat shock promoter, thus, resulting in production of the downstream gene product. Other factors known to trigger the heat shock response, pH change, and reactive oxygen species production were also studied but were not found to contribute to heat shock promoter induction in our setting. Conclusions: The human heat shock promoter (HSP70B) is reported to be an efficient and tightly regulated promoter. We discovered, using sensitive real-time PCR techniques, that it can also be induced in response to cellular nutrient stresses. The pros and cons of this phenomenon and its implications for cancer gene therapy are discussed.

A validation of 10 feline reference genes for gene expression measurements in snap-frozen tissues

For a proper determination of relative mRNA expression levels with real-time quantitative PCR (Q-PCR) internal standards, such as the expression of reference genes, are of utmost importance. For cats, in contrast to dogs, no validation of reference genes has been published. Our goal was to evaluate frequently used reference genes for the analysis of relative mRNA levels from feline tissues in a SYBR Green-based Q-PCR protocol. First, primers were optimized on mRNA-derived cDNA from liver and kidney tissues of randomly chosen (healthy and diseased) cats. Then, the expression variation and stability of each reference gene within a specific tissue was determined. Dental roots and crowns, heart (left ventricle), renal, liver, lung, and mammary gland tissues from 3 to 11 cats of different breeds, sexes, ages, and disease status were included in this study. Averaging relative stabilities over these six tissues revealed the usefulness of each tested gene as reference gene. In order to compensate for the expression variation of a reference gene within a specific tissue, as much as six reference genes (e.g. RPL17, RPL30, RPS7, YWHAZ, and HPRT) were required to obtain highly reliable data in cat tissues. The optimal set of reference genes depended on the tissue analyzed and should, ideally, be selected and evaluated at the start of each experimental condition. A comparison with a similar evaluation in dogs revealed three issues: (i) most ribosomal genes are suitable in both species; (ii) good non-ribosomal reference genes differ; (iii) more feline than canine reference genes are required for proper analysis.

Sustained generation of tissue dendritic cells from cats using organ stromal cell cultures

Currently most dendritic cells (DC) for in vitro study are generated from bone marrow or peripheral blood by culture in high concentrations of GM-CSF and other cytokines. However, in mice it is also possible to generate DC from spleen cells using long-term stromal cell cultures. To determine whether tissue DC could be also be generated from cats, we established stromal cell cultures from a number of different tissues of newborn cats. We found that stromal cell cultures from spleen, lung, liver, kidney, brain, and lymph node tissues were all capable of spontaneously generating DC over long periods of time (months), without requiring the addition of exogenous cytokines. The tissue DC generated from these stromal cell cultures could be readily isolated at high purity by simple mechanical detachment. The feline tissue DC expressed high levels of CD11c, CD11b, and MHC Class II and variable levels of CD80 and CD14 and exhibited high levels of spontaneous macropinocytosis. Moreover, DC from spleen stromal cell cultures, but not DC from lung or liver stromal cell cultures, stimulated mixed-lymphocyte reactions. The DC generated from the stromal cell cultures were relatively independent of GM-CSF for survival and proliferation, indicative of a dependence on other growth factors produced by the stromal cells. These results suggest that tissues of young cats contain a population of resident DC progenitor cells that under appropriate conditions are capable of spontaneous proliferation and generation of immature DC.

Effect of donor age on success of spermatogenesis in feline testis xenografts

Ectopic xenografting of 'donor' feline testicular tissue into a 'recipient' immunodeficient mouse is a promising tool to preserve the male genome from genetically valuable felids. To define parameters under which the technique can succeed, we compared the effect of donor age on xenograft spermatogenesis among four age groups of domestic cats (Felis catus; age range 8 weeks to 15 months). In all cases, fresh tissue was grafted into castrated mice and collected 10, 30 and 50 weeks later. The percentage of xenografts recovered decreased as donor age increased. Mature testicular spermatozoa were observed in xenografts from the 8 and 9-16 week age groups; only a single 7-month-old donor produced elongating spermatids and xenografts from donors >/= 8 months of age degenerated. Seminal vesicle weight, an indicator of bioactive testosterone, was not significantly different between donors aged 8 weeks to 7 months and controls, suggesting that xenograft Leydig cells were ultimately functional even in the 5-7 month age group. Regardless of donor age, production of mature spermatozoa from xenografts was markedly delayed compared with controls. Comparison of xenografts that produced sperm with normal controls revealed a decrease in tubule cross-sections having post-meiotic germ cells. Together, these results indicate that the maximum practical donor age was just before the onset of puberty and that even successful xenografts had abnormalities in spermatogenesis.

A phase I trial of hyperthermia-induced interleukin-12 gene therapy in spontaneously arising feline soft tissue sarcomas

Interleukin-12 (IL-12), a proinflammatory cytokine, shows anticancer properties. Systemically administered IL-12 causes dose-dependent toxicity. To achieve localized intratumoral gene expression, an adenoviral gene therapy vector with IL-12 controlled by a heat-inducible promoter (heat shock promoter 70B) was developed and tested in a phase I clinical trial in cats with spontaneously arising soft tissue sarcoma. A feasibility study was done in 16 cats with soft tissue sarcoma using murine IL-12 and/or enhanced green fluorescent protein adenoviral vectors under cytomegalovirus or heat shock promoter 70 control. Subsequently, we conducted a phase I clinical trial using an adenoviral feline IL-12 construct in 13 cats with soft tissue sarcoma. The soft tissue sarcomas were irradiated (48 Gy/16 fractions) followed by intratumoral injection of adenovirus. Twenty-four hours postinjection, tumors were heated (41 degrees C, 60 min). Tumor expression of feline IL-12 and IFN-gamma was determined. Cats were monitored for systemic toxicity. For the murine IL-12 construct, an association was noted between viral dose and murine IL-12 levels within tumor, whereas serum levels were minimal. Mild toxicity was noted at 10(11) plaque-forming units (pfu). With the feline IL-12 construct, high levels of feline IL-12 mRNA were detected in tumor biopsies with low or absent IFN-gamma mRNA following gene therapy. Hematologic and hepatic toxicities were noted at the highest viral doses and were associated with detection of IFN-gamma mRNA in tumor. It is possible to localize gene expression and limit systemic toxicity of IL-12 using the hyperthermia-induced gene therapy approach. The maximum tolerated dose of the feline IL-12 adenoviral vector was 10(10) pfu/tumor as dose-limiting toxicities were noted at the 4 x 10(10) pfu dose.

Immunohistochemical Study of Phospho-Stat3-ser727 Expression in Feline Mammary Gland Tumours

We describe the expression of pStat3-ser727 (signal transducer and activator of transcription 3 phosphorylated on serine 727) in normal, hyperplastic and neoplastic feline mammary gland tissue assessed by immunohistochemistry in 56 cats. The samples included 4 normal mammary non-lactating tissues, 13 hyperplastic lesions (9 lobular and 4 fibroepithelial) and 39 tumours (6 benign and 33 carcinomas). For immunohistochemistry, tissue sections were incubated with anti-pStat3-ser727 monoclonal antibodies and visualized with EnVision-DAB polymer. pStat3-ser727 positivity was quantified in a semi-quantitative manner, differentiating cytoplasmic and nuclear localization. Intense anti-pStat3-ser727 immunoreactivity was detected in epithelial neoplastic cells and in the fibro-component in two fibroepithelial hyperplastic lesions. The immunostaining was dot-like in the cytoplasm and homogeneous in the nuclei in both benign and malignant lesions. Statistically significant relations were observed between nuclear expression of pStat3-ser727 and the pleomorphism score (p = 0.006), mitotic activity (p < 0.0001), and histological grade (p < 0.0001). In contrast, no significant correlations were observed for cytoplasmic pStat3-ser727. These findings add new and interesting information on the potential role of the phosphorylated form of Stat3 in malignant lesions.

Immunohistochemical evaluation of STAT3-p-tyr705 expression in feline mammary gland tumours and correlation with histologic grade

STAT3 (signal transducer and activator of transcription 3) is a cytoplasmic transcription factor that plays a role in the G1 to S phase cell-cycle transition and is induced by cytokines and growth factors. The expression of STAT3 phosphorylated on tyrosine 705 (STAT3-p-tyr705) in normal, hyperplastic and neoplastic feline mammary gland tissue was assessed by immunohistochemistry in 45 cats. The samples included 4 normal mammary non-lactating tissues, 9 hyperplastic tissues (5 fibroepithelial hyperplasia and 4 lobular epithelial hyperplasia), 2 benign tumours (1 complex adenoma, and 1 simple adenoma), and 30 carcinomas (18 simple tubular papillary, 6 simple tubular, 2 simple solid, 3 cribriform, and 1 adenosquamous carcinoma). For immunohistochemistry, tissue sections were incubated with an anti-STAT3-p-tyr705 monoclonal antibody and visualized with EnVision-DAB polymer. STAT3-p-tyr705 positivity was quantified in a semi-quantitative manner. All positive samples showed cytoplasmic and/or nuclear positivity. Normal non-lactating mammary tissue showed a low number of positive cells, similar to hyperplastic tissue. In neoplastic tissues, a high number of positive cells with a moderate to intense reaction was observed. Moreover, a correlation was observed between nuclear positivity for STAT3-p-tyr705 and histologic grade ( P = 0.013; r = 0.447), tubular formation ( P = 0.043; r = 0.820), and mitotic activity ( P < 0.0001; r = 0.689). In contrast, no such correlations were observed for cytoplasmic reactivity of STAT3-p-tyr705. A significant difference was observed between malignant lesions and hyperplasia with regards to expression of STAT3-p-tyr 705 in the cytoplasm ( P = 0.008; U = 59.00) and nuclei ( P = 0.002; U = 47.00). These results confirm previous our data and reinforce the potential role of STAT3 in malignancy as reported for human breast cancer and other sporadic tumours.

Expression of Stat3 in Feline Mammary Gland Tumours and its Relation to Histological Grade

The expression of Stat3 (signal transducer and activator of transcription 3) in normal and neoplastic feline mammary gland tissue was assessed by immunohistochemistry in 72 cats. The samples included 3 normal nonlactating mammary tissues, 17 hyperplastic lesions (11 lobular and 6 fibroepithelial) and 52 neoplasms (5 benign and 47 malignant). For immunohistochemistry, tissue sections were incubated with anti-Stat3 monoclonal antibody and visualized with EnVision-DAB polymer. Stat3 positivity was assessed in a semiquantitative manner. Normal mammary tissue showed occasional cytoplasmic and nuclear positivity, although the number of positive cells was lower than in the hyperplastic specimens. In neoplastic tissue, high numbers of neoplastic cells with a moderate to intense cytoplasmic and nuclear positivity were observed. Selected variations in the staining pattern correlated with malignancy. Moreover, a positive association was seen between the histological grade and the Stat3 score (p < 0.0001; correlation coefficient 0.750). A linear regression model showed a positive association between mitotic count and Stat3 expression (p < 0.001). These results further support published data found in humans regarding to the relation between the expression of Stat3 and malignancy.

Steroid hormone receptors in normal, dysplastic and neoplastic feline mammary tissues and their prognostic significance

The expression of oestrogen-α and progesterone receptors was determined in 13 normal, 21 dysplastic and 53 neoplastic feline mammary tissues. Expression of the receptors was correlated with cell proliferation, as...

Ten Cases of Feline Mesothelioma: an Immunohistochemical and Ultrastructural Study

In the cat only 10 cases of mesothelioma, mainly of the peritoneum, have been previously reported. This paper describes a further 10 cases, eight pleural and two peritoneal, in males and females aged 1-17 years. Histologically, five tumours were epithelial, three fibrosarcomatous and two biphasic. Immunohistochemical markers used in human pathology for the identification of mesotheliomas include vimentin, cytokeratin (CK) AE1/AE3, HBME-1, CK 5/6, calretinin, thrombomodulin, carcinoembryonic antigen (CEA), CD15, E-cadherin and desmin. All 10 feline mesotheliomas were positive for vimentin and CK AE1/AE3, six were positive for HBME-1, two for CK5/6, three for CEA and four for E-cadherin. All were negative for desmin and calretinin. Antibodies to thrombomodulin and CD15 failed to cross-react with feline tissues. Electron microscopy, performed in four cases, revealed microvillar structures, desmosomes and intracytoplasmic lumina, confirming its value as a diagnostic tool. The study showed that mesothelial marker antibodies commonly used in human patients can be used for the diagnosis of feline mesothelioma, preferably as a panel of antibodies rather than only one.

Cloning of the Feline GADD45 cDNA and Analysis of its Mutation in Feline Lymphoma Cell Lines

Growth arrest and DNA damage-inducible protein 45 (GADD45) plays an important role in suppressing multistep carcinogenesis. In this report, we describe the isolation of the complete wild-type feline GADD45 cDNA from feline tissues. Expression of feline GADD45 mRNA was detected in the liver, spleen, kidney, lung, and testis. The predicted amino acid sequences encoded by the full-length feline GADD45 cDNA display sequence homology with those from other vertebrates, and as in the case of human GADD45, cell growth suppression was observed by ectopic expression of feline GADD45. However, no mutations were detected by sequence analysis of feline GADD45 in several feline lymphoma cell lines, indicating that the GADD45 mutation might be uncommon in feline oncogenesis.