Feline Tissues Research Articles

Abstract 1265: Tissue is alive: Preserving phosphoproteins and tissue morphology in clinical trial samples

Abstract An urgent clinical goal is to identify subpopulations of cancer patients that may respond to targeted kinase inhibitors and/or their phosphorylated substrates. Consequently, phosphorylated signal pathway proteins have emerged as a critical class of analytes for therapeutic stratification. Technologies now exist that can measure protein phosphorylations and map cell signaling pathways in a single core-needle biopsy, thus relying on the inhibition of any kinase/phosphatase activity within the sample immediately following excision. Unfortunately applying these technologies to clinical samples has been hindered because phosphoprotein epitopes are not adequately preserved by formalin fixation and paraffin embedding, while freezing tissue samples may not be feasible in multi-center clinical trial sites and cannot adequately preserve morphology. To facilitate clinical trial molecular profiling, where immediate snap-freezing of tumor biopsies is not feasible, we have created a novel, one-step, room temperature preservative that stabilizes proteins/phosphoproteins equivalent to snap-freezing and tissue/cell morphology equivalent to neutral buffered formalin fixation. In addition, our preservative solution simultaneously fixes and decalcifies bony tissue, thus permitting molecular profiling of bony tissues that was never before possible. We have utilized our Biomarker and Histology Preservative (BHP-Cell and/or BHP-Tissue) in a breast cancer multi-site clinical trial (US Oncology 05-074/GSK LPT109096) and a clinical research multiple myeloma trial. BHP has been validated to a) function as a transport medium while preserving histomorphology, b) maintain full antigenicity for clinical immunohistochemistry (such as Ki-67, ER, PR, Her2, p63, and phosphorylated epitopes), c) preserve phosphoprotein epitopes for cell signaling pathway profiling by reverse phase protein microarray (RPMA), d) be compatible with frozen sections or paraffin embedding, and e) obviate the need for additional decalcification. Preservation of tissue morphology has been demonstrated in 25 mouse, feline, and human tissues with enhanced immunohistochemical properties for >20 antigens using standard IHC protocols. RPMA data from LPT10906 shows differentially deranged signaling networks in the pre-treatment biopsies for patients that did not have a pathologic complete response compared to responders. Our multiple myeloma trial quantifies cell signaling pathway expression pre/post kinase inhibitor treatment, in an ex vivo treatment model, on both myeloma and bone marrow cells, preserved in BHP. We are currently evaluating the integrity of nucleic acid preservation in parallel with proteins to provide a universal one step biospecimen tissue fixative for clinical trials. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1265. doi:1538-7445.AM2012-1265

Preliminary investigation of the influence of long-term dietary isoflavone intake on reproductive tract histology and sex steroid receptor expression in female domestic cats (Felis catus)

SummaryGenistein and daidzein are isoflavones which are reported to influence the reproductive system in a variety of mammalian species. This pilot study aimed to determine if dietary isoflavones could potentially influence reproductive tract histology or morphology in domestic cats, when consumed during the postnatal development period. Cats were maintained on either treatment (150 µg/g DM genistein and 150 µg/g DM daidzein, n = 4) or control (isoflavone free, n = 8) diets from weaning, up to 414 (±17.2) days post-weaning. Reproductive tissues were collected during routine ovario-hysterectomy and examined for histology and sex steroid receptor expression. Findings indicate that these dietary isoflavones influenced the expression of oestrogen receptor α (ERα) and oestrogen receptor β (ERβ), and progesterone receptor in feline reproductive tissues. One cat in the treatment group developed suppurative endometritis, but no evidence of uterotrophic or histological changes were found in any other cats. The potential to alter expression of hormone receptors in the reproductive tract of domestic cats exposed to genistein and daidzein warrants further investigation.

P-Cadherin Expression in Feline Mammary Tissues

The search for molecular markers in the feline mammary gland, namely, the adhesion molecules belonging to the cadherin family, is useful in the understanding of the development of mammary carcinomas in felines and humans. To study P-cadherin expression in the feline mammary gland, 61 samples of normal (n = 4), hyperplastic (n = 12), and neoplastic (n = 45) feline mammary tissues were examined. In both normal and hyperplastic mammary tissues as well as in benign tumours, P-cadherin immunolabelling was restricted to myoepithelial cells. In malignant tumours, however, there was an aberrant epithelial P-cadherin immunoexpression in 64.1% (n = 25) of cases, with a membranous and/or cytoplasmic pattern of distribution. A statistically significant relationship was seen between epithelial P-cadherin expression and malignant mammary lesions (P = 0.0001). In malignant mammary tumours, there was likewise a statistically significant relationship between aberrant P-cadherin immunoexpression and histological grade (P = 0.0132). Aberrant epithelial P-cadherin expression seems to be related to malignancy in the feline mammary gland. To confirm the results of this investigation, further studies with larger samples and follow-up studies are warranted.

Molecular characterization of the feline T-cell receptor γ alternate reading frame protein (TARP) ortholog

T-cell receptor γ alternate reading frame protein (TARP) is expressed by human prostate epithelial, prostate cancer, and mammary cancer cells, but is not found in normal mammary tissue. To date, this protein has only been described in humans. Additionally, no animal model has been established to investigate the potential merits of TARP as tumor marker or a target for adoptive tumor immunotherapy. In this study conducted to characterize feline T-cell receptor γ sequences, constructs very similar to human TARP transcripts were obtained by RACE from the spleen and prostate gland of cats. Transcription of TARP in normal, hyperplastic, and neoplastic feline mammary tissues was evaluated by conventional RT-PCR. In felines similarly to the situation reported in humans, a C-region encoding two open reading frames is spliced to a J-region gene. In contrast to humans, the feline J-region gene was found to be a pseudogene containing a deletion within its recombination signal sequence. Our findings demonstrated that the feline TARP ortholog is transcribed in the prostate gland and mammary tumors but not normal mammary tissues as is the case with human TARP.

Suppression of immunodeficiency virus-associated neural damage by the p75 neurotrophin receptor ligand, LM11A-31, in an in vitro feline model.

Feline immunodeficiency virus (FIV) infection like human immunodeficiency virus (HIV), produces systemic and central nervous system disease in its natural host, the domestic cat, that parallels the pathogenesis seen in HIV-infected humans. The ability to culture feline nervous system tissue affords the unique opportunity to directly examine interactions of infectious virus with CNS cells for the development of models and treatments that can then be translated to a natural infectious model. To explore the therapeutic potential of a new p75 neurotrophin receptor ligand, LM11A-31, we evaluated neuronal survival, neuronal damage and calcium homeostasis in cultured feline neurons following inoculation with FIV. FIV resulted in the gradual appearance of dendritic beading, pruning of processes and shrinkage of neuronal perikarya in the neurons. Astrocytes developed a more activated appearance and there was an enhanced accumulation of microglia, particularly at longer times post-inoculation. Addition of 10nM LM11A-31, to the cultures greatly reduced or eliminated the neuronal pathology as well as the FIV effects on astrocytes and microglia. LM11A-31 also, prevented the development of delayed calcium deregulation in feline neurons exposed to conditioned medium from FIV treated macrophages. The suppression of calcium accumulation prevented the development of foci of calcium accumulation and beading in the dendrites. FIV replication was unaffected by LM11A-31. The strong neuroprotection afforded by LM11A-31 in an infectious in vitro model indicates that LM11A-31 may have excellent potential for the treatment of HIV-associated neurodegeneration.

130 EXPRESSION OF STEROIDOGENIC ENZYMES IN THE CAT OVARY DURING FOLLICULAR GROWTH

This study was designed to describe the expression of steroidogenic enzymes in the various compartments of the feline ovary, from primordial to preovulatory follicles: P450 side-chain cleavage (P450scc), 17α-hydroxylase (17α-OH), 3β-hydroxysteroid dehydrogenase (3β-HSD). Nine female cats were ovariectomized during behavioural oestrus. After validation of their specificity by Western blot on feline tissue, 3 polyclonal antibodies obtained in the rabbit were used for immunohistochemistry (anti-bovine P450scc; anti-pig 17α-OH; anti-human 3β-HSD). Three successive ovarian cryosections were incubated with antibodies and 2 sections were used as controls (purified immunoglobulin G from a nonimmunized rabbit). Immunostaining was revealed by streptavidin-biotin system (LSAB kit, DAKO S.A., Trappes, France), with aminoethylcarbazole as a chromogen. The staining was evaluated according to its localization, intensity (from 1 to 4) and number of cell layers stained. Histological examination after haematoxylin-eosin-safran staining allowed follicle diameter measurement and evaluation of the atresia. Analysis of variance was used to compare follicles from various size classes. A total of 343 follicles from 140 to 3250 μm in diameter were observed (i.e. all follicles present on the slides), of which 15.2% were atretic. The observations were similar for the 3 enzymes. From the preantral stage onward (diameter >140 μm), more than 85% of the healthy follicles had the theca interna positive for P450scc, 3β-HSD and 17α-OH (respectively, 87.3, 86.0 and 99.0%). Approximately 20 to 25% were also positive in the theca externa. Granulosa were stained in 9.5% (P450scc), 0.8% (3β-HSD) and 1.8% (17α-OH) of the healthy follicles, but always with a low intensity (<2). The follicular size did not affect the proportion of stained follicles, but the staining intensity decreased progressively until 1200 μm and increased thereafter. The number of positive layers decreased continuously until 1500 μm in diameter. Between 90 and 95% of the preovulatory follicles (>2000 μm) were positive in the theca interna; the signal was more intense and the number of stained layers was higher than in smaller follicles. The 3 enzymes were detected in all the atretic follicles, with a similar intensity as in healthy ones; they were also present in the interstitial tissue, but with a higher intensity. The staining pattern in the various ovarian compartments suggests that the major steroidogenic tissues in the cat ovary are the theca interna of healthy follicles and the interstitial tissue. The lack of expression of the 3 key enzymes in the granulosa cells, even in preovulatory follicles, strongly suggests that these cells, shaping the future corpus luteum, are probably unable to produce progesterone.

Widespread Mismatch Repair Expression in Feline Small Intestinal Lymphomas

Small intestinal lymphoma is a common feline tumour that most often develops in older cats, but also occurs in younger animals. In man, germline defects in the mismatch repair (MMR) genes most commonly cause hereditary non-polyposis colorectal cancer (HNPCC), or Lynch syndrome, while MMR defects have also been implicated in the development of lymphoid tumours in mice and in people. It was hypothesized that inherited MMR defects predispose a proportion of younger cats to the development of small intestinal lymphoma. MMR expression in 10 small intestinal lymphomas from younger cats (group 1, mean age 4.5 years) was compared with MMR expression in 30 small intestinal lymphomas from older cats (group 2, mean age 12.6 years). The cross-reactivity of the antibodies specific for the human MMR proteins MLH1, MSH2 and MSH6 with the corresponding proteins in feline tissues was first confirmed by western blotting. MMR expression was then investigated immunohistochemically in feline lymphomas. MLH1, MSH2 and MSH6 were detected immunohistochemically within neoplastic lymphocytes from all tumours examined. There were no significant differences between the two groups in either the intensity of immunolabelling or the percentage of neoplastic cells within which MMR proteins were detected. These results confirm the cross-reactivity of the human MMR antibodies with the corresponding proteins in feline tissues, but do not support the hypothesis that inherited germline MMR defects are a significant cause of feline small intestinal lymphomas.

Porphyrins are not present in feline ocular tissues or corneal sequestra

To determine if feline lacrimal glands, glands of the third eyelid, corneas, and corneal sequestra contain porphyrins, which could be responsible for the brown/amber discoloration of corneal sequestra and tears in affected cats. Samples of grossly normal cornea, lacrimal gland, gland of the third eyelid, and sequestra obtained via keratectomy were collected. Porphyrin concentrations of the homogenate were determined by spectrofluorometry with protoporphyrin IX and coproporphyrin III dihydrochloride used as standards. A hamster harderian gland was used as a positive control. Normal tissues were harvested from one eye each of 14 nonclient owned, adult, mixed-breed, short-hair cats euthanized for reasons not associated with this study. Eighteen sequestra were acquired from cats undergoing unilateral lamellar keratectomies. Breeds of the affected cats included eight Himalayan, five domestic shorthair, and one each of four other breeds. Only the positive control and standards contained levels of porphyrins above background. All feline samples examined were histologically normal with no evidence of porphyrins. Porphyrins are absent in normal feline lacrimal glands, corneas, and corneal sequestra. Porphyrins do not appear to be the cause of the brown/amber color of feline corneal sequestra.

Quantification and molecular characterization of the feline leukemia virus A receptor

Virus receptors and their expression patterns on the cell surface determine the cell tropism of the virus, host susceptibility and the pathogenesis of the infection. Feline thiamine transport protein 1 (fTHTR1) has been identified as the receptor for feline leukemia virus (FeLV) A. The goal of the present study was to develop a quantitative, TaqMan real-time PCR assay to investigate fTHTR1 mRNA expression in tissues of uninfected and FeLV-infected cats, cats of different ages, in tumor tissues and leukocyte subsets. Moreover, the receptor was molecularly characterized in different feline species. fTHTR1 mRNA expression was detected in all 30 feline tissues investigated, oral mucosa scrapings and blood. Importantly, identification of significant differences in fTHTR1 expression relied on normalization with an appropriate reference gene. The lowest levels were found in the blood, whereas high levels were measured in the oral mucosa, salivary glands and the musculature. In the blood, T lymphocytes showed significantly higher fTHTR1 mRNA expression levels than neutrophil granulocytes. In vitro activation of peripheral blood mononuclear cells with concanavalin A alone or followed by interleukin-2 led to a transient increase of fTHTR1 mRNA expression. In the blood, but not in the examined tissues, FeLV-infected cats tended to have lower fTHTR1 mRNA levels than uninfected cats. The fTHTR1 mRNA levels were not significantly different between tissues with lymphomas and the corresponding non-neoplastic tissues. fTHTR1 was highly conserved among different feline species (Iberian lynx, Asiatic and Indian lion, European wildcat, jaguarundi, domestic cat). In conclusion, while ubiquitous fTHTR1 mRNA expression corresponded to the broad target tissue range of FeLV, particularly high fTHTR1 levels were found at sites of virus entry and shedding. The differential susceptibility of different species to FeLV could not be attributed to variations in the fTHTR1 sequence.

Detection of Tritrichomonas foetus and Pentatrichomonas hominis in intestinal tissue specimens of cats by chromogenic in situ hybridization

In this retrospective study 102 cats were analyzed for the presence of trichomonads in intestinal tissue sections using chromogenic in situ hybridization (CISH). Two intestinal trichomonad species are described in cats: Pentatrichomonas hominis and Tritrichomonas foetus. While P. hominis is considered a mere commensal, T. foetus has been found to be the causative agent of feline large-bowel diarrhea. For the detection of both agents within intestinal tissue CISH assays using three different probes were performed. In the first CISH run a probe specific for all relevant members of the order Trichomonadida (OT probe) was used. In a second CISH run all positive samples were further examined on three consecutive tissue sections using the OT probe, a probe specific for the family of Tritrichomonadidae (Tritri probe) and a newly designed probe specifically detecting P. hominis (Penta hom probe). In total, four of the 102 cats were found to be positive with the OT probe. Thereof, one cat gave a positive reaction with the P. hominis probe and three cats were positive with the T. foetus probe. All Trichomonas-positive cats were pure-bred and between 8 and 32 weeks of age. In one cat positive for T. foetus large amounts of parasites were found in the gut lumen and invading the intestinal mucosa. The species of the detected trichomonads were confirmed by polymerase chain reaction and nucleotide sequencing of a part of the 18S ribosomal RNA gene. In this study, the usefulness of CISH to detect intestinal trichomonads within feline tissue samples was shown. Additionally, the specific detection of P. hominis using CISH was established. Generally, it was shown that CISH is well suited for detection and differentiation of trichomonosis in retrospective studies using tissue samples.

Pharmacokinetic/pharmacodynamic modelling of robenacoxib in a feline tissue cage model of inflammation

Robenacoxib is a novel nonsteroidal anti-inflammatory drug developed for use in cats. It is a highly selective COX-2 inhibitor. Results from previous feline studies showed that, despite a short half-life in blood, the effect of robenacoxib persisted for 24 h in clinical studies. A tissue cage model of acute inflammation was used to determine robenacoxib's pharmacokinetics and its ex vivo and in vivo selectivity for COX-1 and COX-2 using serum TxB(2) and exudate PGE(2) as surrogate markers for enzyme activity, respectively. After intravenous, subcutaneous and oral administration (2 mg/kg), the clearance of robenacoxib from blood was rapid (0.54-0.71 L·h/kg). The mean residence time (MRT) in blood was short (0.4, 1.9 and 3.3 h after intravenous, subcutaneous and oral administration, respectively), but in exudate MRT was approximately 24 h regardless of the route of administration. Robenacoxib inhibition of COX-1 in blood was transient, occurring only at high concentrations, but inhibition of COX-2 in exudate persisted to 24 h. The potency ratio (IC(50) COX-1: IC(50) COX-2) was 171:1, and slopes of the concentration-effect relationship were 1.36 and 1.12 for COX-1 and COX-2, respectively. These data highlight the enzymatic selectivity and inflamed tissue selectivity of robenacoxib and support the current recommendation of once-daily administration.

Localised tetanus in a cat

TETANUS is a neurological disorder caused by tetanospasmin, a neurotoxin of the anaerobic spore-forming bacillus Clostridium tetani (Caleo and Schiavo 2009). Tetanus spores are commonly introduced into tissues through wounds...

Development of a chromogenic in situ hybridization for Giardia duodenalis and its application in canine, feline, and porcine intestinal tissues samples

In the present study, a chromogenic in situ hybridization for the identification of Giardia duodenalis in paraffin-embedded tissue samples was developed. The sensitivity and specificity of the probe was validated by testing it on cultured reference samples of different assemblages of G. duodenalis as well as culture and tissue samples containing other protozoa and infectious agents. The probe gave a positive reaction with the Giardia samples and a negative reaction with all other samples. Further, the probe was used for screening of histological slides of intestine from different animal species (99 canine samples, 85 feline samples, and 202 porcine samples) for the presence of G. duodenalis trophozoites. With this assay, the parasites were detected in samples from 8 dogs (8.08%), 6 cats (7.06%), and zero pigs. The results clearly indicate that the described method is useful for detection of Giardia trophozoites in routinely processed intestinal tissue of different animal species.

An Immunohistochemical Assay to Detect Trophoblasts in Frozen Feline Placenta

The placenta, a fetal endocrine organ, is composed of subpopulations of trophoblasts, including cytotrophoblasts, and syncytiotrophoblasts. Trophoblastic populations can be distinguished based upon their expression of cytokeratin. The purpose of the current study was to develop an immunohistochemistry (IHC) method to identify trophoblasts selectively in frozen feline placental tissue using antibodies specific for cytokeratin. The mouse monoclonal antibody anti-human pan-cytokeratin AE1/AE3 and a commercial detection system were used. Nonspecific immunoreactivity was encountered that could not be eliminated with altered blocking methods. The nonspecific reactivity was attributed to the goat anti-mouse/rabbit immunoglobulin G (IgG) peroxidase polymer included in the commercial kit. Alternatively, a polyclonal rabbit anti-cow cytokeratin wide spectrum screening antibody with goat anti-rabbit IgG polyclonal secondary antibody was used to detect cytokeratin in feline placental tissue. The IHC procedure eliminated nonspecific immunoreactivity while specifically labeling cytokeratin. This new approach provides an IHC method to identify trophoblasts specifically in feline placenta.

Assessment of feline abdominal adipose tissue using computed tomography.

Obesity is a common nutritional disorder in cats and it increases the risk factors for various diseases. The aim of this study is to suggest a method for the evaluation of feline obesity using computed tomography. The attenuation range from -156 to -106 was determined as the range of feline abdominal adipose tissue. With this range, total (TAT), visceral (VAT) and subcutaneous (SAT) adipose tissues were measured. The best correlation between the adipose tissue in cross-sectional image and entire abdomen volume was obtained at the L3 and L5 levels. The mean VAT/SAT ratio was 1.18±0.32, which was much higher than in humans. The cats with an overweight body condition had a significantly lower VAT/SAT ratio than cats with an ideal body condition. This technique may contribute to both the clinical diagnosis and the experimental study of feline obesity.

Preliminary Immunohistochemical Study of Natriuretic Peptide Receptor Localization in Canine and Feline Heart

We examined the immunohistochemical distributions of natriuretic peptide receptor (NPR)-A, -B and -C that bind with natriuretic peptide hormones A, B and C in four healthy crossbreed young canine and feline cardiac tissues using specific antibodies against human antigens. Cross-immunoreactivities between antigens and antibodies were confirmed using western blot analysis. NPR-A and -C were expressed more strongly in dogs than cats. In both species, these expressions were stronger in the atria than the ventricles, with stronger expression in the left ventricles than the right. NPR-B was largely very weekly or undetected. In canine and feline cardiac tissues, the expressional distribution of NPR-A, -B, and -C closely matched with that of atrial natriuretic peptide, brain natriuretic peptide, and C-type natriuretic peptide as the ligands for corresponding receptors.

Expression of Multidrug Resistance-Associated P-Glycoprotein in Feline Tumours

Permeability glycoprotein (P-gp) is a membrane-bound efflux pump that mediates the active transmembrane transport of a variety of substrates. Several studies in human and canine normal and neoplastic tissues indicate that P-gp is involved in resistance to chemotherapy and in the development of multidrug resistance (MDR). The purpose of this study was to evaluate P-gp expression with the monoclonal antibody C494 in common feline tumours from 88 patients not pretreated with chemotherapy. Tumours arising from tissues with intrinsic P-gp expression consistently showed positive labelling for P-gp in a cellular pattern identical to that described for the normal feline tissues. Both P-gp positive and negative tumour cells, however, were found in mammary gland tumours, lymphomas, mastocytomas and squamous cell carcinomas. Feline mammary gland tumours in particular showed strong membranous P-gp labelling, especially in areas with infiltrative growth and atypical cells, although this was not correlated with the grade of malignancy. These findings might have implications for future response to chemotherapy.

P-cadherin expression in feline mammary tissues

Placental cadherin (P-cadherin) is a classical cadherin [1] expressed by myoepithelial cells of the human mammary gland [2,3]. Changes to P-cadherin expression have been observed and implicated in human breast carcinogenesis [4-6]. Feline mammary tumours show similarities with the women tumours concerning many histological characteristics and clinical evolution, being proposed as good animal model to study mammary carcinogenesis [7]. To study P-cadherin expression in feline mammary gland an immunohistochemistry assay was performed in 61 samples of normal (n = 4), hyperplastic (n = 12), benign (n = 6) and malignant (n = 39) feline mammary tissues and the immunostaining assessment was based on the estimated percentage of luminal epithelial cells labeling (aberrant expression). In normal mammary gland, mammary hyperplasia and benign tumours, P-cadherin immunolabelling was restricted to myoepithelial cells. Nevertheless, in malignant tumours there was an aberrant epithelial P-cadherin immunoexpression in 64,1% (n = 25) of the cases, with a membranar and/or cytoplasmic pattern of cellular distribution. Consequently, P-cadherin expression in feline mammary lesions was not exclusive of myoepithelial cells. It was possible to observe a significant statistical correlation between P-cadherin expression intensity and feline mammary lesions (p = 0.0001). In malignant mammary tumors a statistical correlation between P-cadherin immunoexpression intensity and histological grade was observed (p = 0.0132). Aberrant epithelial P-cadherin expression seems to be correlated with tumor malignancy in feline mammary gland.

Expression of Regulatory T cell (Treg) Activation Markers in Endometrial Tissues from Early and Late Pregnancy in the Feline Immunodeficiency Virus (FIV)-Infected Cat

Regulatory T cells (Tregs) support pregnancy maintenance by suppressing placental inflammation, while diminished Treg function may accompany reproductive failure. Experimental FIV infection frequently results in vertical transmission and increased pregnancy failure in the cat. The mechanism of reproductive compromise is unknown. We hypothesized that FIV infection alters endometrial Treg population dynamics and function, potentiating vertical transmission and reproductive failure. RNA collected from early and late gestation reproductive tissue and fetuses from FIV infected and control cats was probed for expression of FIV gag and Treg markers CD25, FOXP3, and CTLA4, using real time reverse-transcriptase (RT)-PCR. Frequent placental and fetal infection and reproductive failure were detected at early and late pregnancy. Expression of FOXP3 and CTLA4 was higher in early gestation tissues from control cats. FIV infection significantly reduced expression of FOXP3 and CTLA4 at early, but not late pregnancy. At late pregnancy, CTLA4 was expressed to higher levels in infected tissues. The number of tissues with decreased co-expression of FOXP3 and CTLA4 was significant in infected cats at early pregnancy. No significant changes in CD25 expression occurred between FIV-infected and control animals at early or late pregnancy. Differences in Treg marker expression were not significant between viable and non-viable pregnancies in infected cats. The detection of Treg markers in these feline tissues provides the first evidence of feline endometrial Tregs and suggests that such cells diminish as pregnancy progresses. These cells may be depleted or rendered less functional by viral infection, but understanding their role in pregnancy requires further study.

Distribution of the Feline Calicivirus Receptor Junctional Adhesion Molecule A in Feline Tissues

Junctional adhesion molecule A (JAM-A) is an immunoglobulin superfamily protein that plays an important role in the assembly and maintenance of tight junctions and the establishment of epithelial cell polarity. The feline JAM-A (fJAM-A) is a functional receptor for feline calicivirus (FCV). Among natural diseases associated with FCV infection, isolates that cause oral vesicular disease are detected in epithelial cells; however, isolates that cause systemic disease are detected in multiple cell types. The distribution of an FCV receptor or receptors in feline tissues is relevant to viral pathogenesis in that it should reflect the wide latitude of clinical sequelae associated with FCV infection. The authors examined the expression of feline JAM-A in the cat by using confocal immunofluorescence localization on normal tissues, with special regard to tissue targets of naturally occurring FCV. As described in the human and the mouse, fJAM-A was widely distributed in feline tissues, where it localized at cell-cell junctions of epithelial and endothelial cells. fJAM-A was highly expressed on feline platelets, with lower levels of expression on feline peripheral blood leukocytes. Additionally, FCV infection of a feline epithelial cell monolayer causes redistribution of fJAM-A to the cytosol of infected cells. It is reasonable to propose that the spectrum of lesions caused by FCV reflects disruption of intercellular junctions that rely on fJAM-A function and tight junctional integrity.