The CCL2-CCR4 Axis Promotes Regulatory T Cell Trafficking to Canine Glioma Tissues.

Purpose: Spontaneously occurring glioma in pet dogs is increasingly recognized as a valuable translational model for human glioblastoma. Canine high grade glioma and human glioblastomas share many molecular similarities, including accumulation of immunosuppressive regulatory T cells (Tregs) that inhibit anti-tumor immune responses. Identifying in dog mechanisms responsible for Treg recruitment may afford targeting the cellular population driving immunosuppression, the results providing a rationale for translational clinical studies in human patients. Our group has previously identified C-C motif chemokine 2 (CCL2) as a glioma-derived T-reg chemoattractant acting on chemokine receptor 4 (CCR4) in a murine orthotopic model of glioma. Recently, we demonstrated a robust increase of CCL2 in the brain tissue of canine patients bearing high-grade glioma. Methods: We performed a series of in vitro experiments using canine Tregs and patient-derived canine glioma cell lines (GSC 1110, GSC 0514, J3T-Bg, G06A) to interrogate the CCL2-CCR4 signaling axis in the canine. Results: We established a flow cytometry gating strategy for identification and isolation of FOXP3+ Tregs in dogs. The canine CD4 + CD25high T-cell population was highly enriched in FOXP3 and CCR4 expression, indicating they are bona fide Tregs. Canine Treg migration was enhanced by CCL2 or by glioma cell line-derived supernatant. Blockade of the CCL2-CCR4 axis significantly reduced migration of canine Tregs. CCL2 mRNA was expressed in all glioma cell lines and expression increased when exposed to Tregs but not to CD4 + helper T-cells. Conclusion: Our study validates CCL2-CCR4 as a bi-directional Treg-glioma immunosuppressive and tumor-promoting axis in canine high-grade glioma.

The CCL2-CCR4 axis promotes Regulatory T cell trafficking to canine glioma tissues.

PURPOSE: Spontaneously occurring glioma in pet dogs is increasingly recognized as a valuable translational model for human glioblastoma. Canine high-grade glioma and human glioblastomas share many molecular similarities, including the accumulation of immunosuppressive regulatory T cells (Tregs) that inhibit anti-tumor immune responses. Identifying in dog mechanisms responsible for Treg recruitment may afford to target the cellular population driving immunosuppression, the results providing a rationale for translational clinical studies in human patients. Our group has previously identified C-C motif chemokine 2 (CCL2) as a glioma-derived T-reg chemoattractant acting on chemokine receptor 4 (CCR4) in a murine orthotopic glioma model. Recently, we demonstrated a robust increase of CCL2 in the brain tissue of canine patients bearing high-grade glioma. METHODS: We performed a series of in vitro experiments using canine Tregs and patient-derived canine glioma cell lines (GSC 1110, GSC 0514, J3T-Bg, G06A) to interrogate the CCL2-CCR4 signaling axis in the canine. RESULTS: We established a flow cytometry gating strategy for identifying and isolating FOXP3+ Tregs in dogs. The canine CD4 + CD25high T-cell population was highly enriched in FOXP3 and CCR4 expression, indicating they are bona fide Tregs. Canine Treg migration was enhanced by CCL2 or by glioma cell line-derived supernatant. Blockade of the CCL2-CCR4 axis significantly reduced migration of canine Tregs. CCL2 mRNA was expressed in all glioma cell lines, and expression increased when exposed to Tregs but not CD4 + helper T-cells. CONCLUSION: Our study validates CCL2-CCR4 as a bi-directional Treg-glioma immunosuppressive and tumor-promoting axis in canine high-grade glioma.

Electrophysiologic confirmation of heterogenous motor polyneuropathy in young cats.

BACKGROUND: Reports of motor polyneuropathies in young cats are scarce. Further, in-depth electrophysiologic evaluation to confirm a motor polyneuropathy in young cats of various breeds other than 2 Bengal cats is lacking. HYPOTHESIS/OBJECTIVES: To confirm a motor polyneuropathy in young cats of various breeds. ANIMALS: Five young cats with heterogenous chronic or relapsing episodes of weakness. METHODS: Retrospective case series. Cats were presented for evaluation of generalized neuromuscular disease and underwent electrophysiologic examination including electromyography, nerve conduction, and repetitive nerve stimulation. Minimum database and muscle and nerve biopsy analyses were carried out. Descriptive statistics were performed. RESULTS: Disease onset was at 3 months to 1 year of age and in 5 breeds. The most common clinical sign (5 of 5 cats) was weakness. Additional neurologic deficits consisted of palmigrade and plantigrade posture (4/4), low carriage of the head and tail (4/4), and variable segmental reflex deficits (5/5). Motor nerve conduction studies were abnormal for the ulnar (4/4), peroneal (5/5), and tibial (2/2) nerves (increased latencies, reduced amplitudes, slow velocities). A marked decrement was observed on repetitive nerve stimulation of the peroneal nerve in 3 cats for which autoimmune myasthenia gravis was ruled out. All sensory nerve conduction studies were normal. Histologic evaluation of muscle and nerve biopsies supported heterogenous alterations consistent with motor polyneuropathy with distal nerve fiber loss. CONCLUSIONS AND CLINICAL IMPORTANCE: Heterogenous motor polyneuropathies should be considered in young cats of any breed and sex that are presented with relapsing or progressive generalized neuromuscular disease.

Bilaterally symmetric focal cortical dysplasia in a golden retriever dog.

A 10-year-old golden retriever dog was referred with a 24-h history of generalized seizures. Magnetic resonance imaging of the brain found no abnormalities on 3 mm transverse sections and the dog was subsequently humanely destroyed. Microscopically there was bilaterally symmetrical focal disorganization of cortical grey matter within the tips of the right and left suprasylvian gyri of the temporal cortex. The focal abnormal cortical lamination was characterized by loss of pyramidal neurons with abnormal, irregular, angular, remaining neurons occasionally forming clusters, surrounded by fibrillary astrogliosis and microgliosis and vascular proliferation. These histological findings are consistent with focal cortical dysplasia, a cerebral cortical malformation that causes seizures in people, but not reported previously in the dog.

Clear cell ependymoma in a dog.

A 13-year-old, mixed breed dog presented with a 1-month history of seizures. Magnetic resonance imaging of the brain revealed a 2.2 × 1.0 × 0.9 cm ovoid and elongate cystic mass within the white matter of the left frontal lobe extending caudally from the cribriform plate to the rostral left lateral ventricle. Three fractions of stereotactic radiotherapy were administered and resulted in reduction of the volume of the tumour; however, the clinical signs failed to improve. On post-mortem examination, a single mass 1.5 × 0.3 × 1 cm was found within the left frontal lobe. It consisted of gelatinous, grey, friable tissue bordering a central empty cavity. Microscopical evaluation revealed polygonal neoplastic cells with distinct cytoplasmic borders and one or more intracytoplasmic solid, brightly eosinophilic, sharply defined globules. Immunohistochemically, the neoplastic cells expressed glial fibrillary acidic protein and S100 but were negative for pan cytokeratin, vimentin, olig-2 and synaptophysin. Ultrastructurally, neoplastic cells had dense whorls of intracytoplasmic intermediate filaments and were connected by multiple intermittent long zonula adherens-type junctions. Based on these findings, a diagnosis of clear cell ependymoma was made. This is the first report of this subtype in the dog.

Clinical signs, imaging features, neuropathology, and outcome in cats and dogs with central nervous system cryptococcosis from California.

BACKGROUND: Cryptococcus spp. is a fungal pathogen with a predilection for the central nervous system (CNS). OBJECTIVES: To compare the clinical, advanced imaging, and neuropathologic findings in dogs and cats with CNS cryptococcosis, and to evaluate outcome of treatment in these animals. ANIMALS: Twenty-six cats and 21 dogs with CNS cryptococcosis. METHODS: Medical records were reviewed for clinical findings and results of CNS imaging. Archived cerebrospinal fluid and CNS tissue specimens were reviewed for pathology. Findings in cats were compared with those in dogs and the effects of variables on survival were determined by survival curve analysis. RESULTS: When present, pain was localized to the cervical region in dogs and was generalized or localized to the thoracolumbar spine or pelvic limbs in cats. Magnetic resonance imaging (MRI) findings were variable but correlated with CNS histopathological findings of meningitis, meningitis with gelatinous pseudocyst formation, and granulomatous mass lesions. Peripherally enhancing brain lesions were seen only in cats. Histopathologically, the inflammatory response was milder in cats compared with dogs. Remissions of ≥1 year occurred in 32% of treated animals. Altered mentation was associated with negative outcome. Glucocorticoid use after diagnosis was associated with improved survival in the first 10 days. CONCLUSIONS AND CLINICAL IMPORTANCE: Lesions seen on MRI reflected neuropathological findings and were similar to those reported in human patients. The immune response to infection may differ between cats and dogs, or relate to the infecting cryptococcal species. Long-term (>6 month median survival time) survival may be possible in animals surviving ≥4 days after diagnosis.

Cerebrospinal fluid eosinophilia in dogs.

BACKGROUND: Marked eosinophilic meningitis or meningoencephalomyelitis (EME) is rarely reported in dogs and the cause is usually undetermined. Long-term prognosis for dogs with cerebrospinal fluid (CSF) eosinophilia is variable. ANIMALS: Twenty-three client-owned dogs. METHODS: Retrospective case series. Dogs with eosinophilic CSF, defined as total nucleated cell count (TNCC) >3 cells/microL with >20% eosinophils, were identified by a computerized search of all dogs having cisternal and/or lumbar CSF analyzed as part of the diagnostic workup between 1992 and 2007. RESULTS: TNCC in CSF ranged from 4 to 4,740 cells/microL (median 84 cells/microL, reference range

Choroid plexus tumors in 56 dogs (1985-2007).

BACKGROUND: Choroid plexus tumors (CPTs) comprise approximately 10% of all primary brain tumors in dogs. The clinical utility of magnetic resonance imaging (MRI), cerebrospinal fluid (CSF) analysis, or both in the presumptive diagnosis of CPTs has not been determined. OBJECTIVES: To report MRI and CSF findings in dogs with CPT and determine if there are distinguishing features that allow clinical discrimination between the tumor grades. ANIMALS: Fifty-six client-owned dogs with naturally occurring CPT. METHODS: Retrospective case series. The inclusion criterion was histologically confirmed CPT. Blinded review of cranial MRI and cisternal CSF analysis was performed. RESULTS: Thirty-six of 56 dogs had a choroid plexus carcinoma (CPC) and 20 had a choroid plexus papilloma (CPP). Golden Retrievers were overrepresented compared with the hospital population (frequency 3.7 times that expected, confidence interval 95%= 2.0-6.7, P< .0002). Median CSF protein concentration in CPCs (108 mg/dL, range 27-380 mg/dL) was significantly higher than in CPPs (34 mg/dL, range 32-80 mg/dL) (P= .002). Only dogs with CPCs had a CSF protein concentration >80 mg/dL. Cytological evidence of malignancy in CSF was seen in 7 of 15 CPCs. Only CPCs had evidence of intraventricular or subarachnoid metastases on MRI. CONCLUSIONS AND CLINICAL IMPORTANCE: MRI, CSF analysis or both can help to differentiate between CPPs and CPCs, and may provide valuable prognostic and pretreatment information.

Necrotizing meningoencephalitis in five Chihuahua dogs.

An acute to chronic idiopathic necrotizing meningoencephalitis was diagnosed in 5 Chihuahua dogs aged between 1.5 and 10 years. Presenting neurologic signs included seizures, blindness, mentation changes, and postural deficits occurring from 5 days to 5.5 months prior to presentation. Cerebrospinal fluid analyses from 2 of 3 dogs sampled were consistent with an inflammatory disease. Magnetic resonance imaging of the brain of 2 dogs demonstrated multifocal loss or collapse of cortical gray/white matter demarcation hypointense on T1-weighted images, with T2-weighted hyperintensity and slight postcontrast enhancement. Multifocal asymmetrical areas of necrosis or collapse in both gray and white matter of the cerebral hemispheres was seen grossly in 4 brains. Microscopically in all dogs, there was a severe, asymmetrical, intensely cellular, nonsuppurative meningoencephalitis usually with cystic necrosis in subcortical white matter. There were no lesions in the mesencephalon or metencephalon except in 1 dog. Immunophenotyping defined populations of CD3, CD11d, CD18, CD20, CD45, CD45 RA, and CD79a immunoreactive inflammatory cells varying in density and location but common to acute and chronic lesions. In fresh frozen lesions, both CD1b,c and CD11c immunoreactive dendritic antigen-presenting cells were also identified. Immunoreactivity for canine distemper viral (CDV) antigen was negative in all dogs. The clinical signs, distribution pattern, and histologic type of lesions bear close similarities to necrotizing meningoencephalitis as described in series of both Pug and Maltese breed dogs and less commonly in other breeds.

Magnetic resonance imaging and histological classification of intracranial meningiomas in 112 dogs.

BACKGROUND: Intracranial meningiomas are the most common primary brain tumors in dogs. Classification of meningiomas by tumor grade and subtype has not been reported, and the value of magnetic resonance imaging (MRI) characteristics for predicting tumor subtype and grade has not been investigated. HYPOTHESIS: Canine intracranial meningiomas are a heterogenous group of tumors with differing histological subtypes and grades. Prediction of histopathological classification is possible based on MRI characteristics. ANIMALS: One hundred and twelve dogs with a histological diagnosis of intracranial meningioma. METHODS: Retrospective observational study. RESULTS: Meningiomas were overrepresented in the Golden Retriever and Boxer breeds with no sex predilection. The incidence of specific tumor grades was 56% benign (Grade I), 43% atypical (Grade II), and 1% malignant (Grade III). Grade I histological subtypes included meningothelial (43%), transitional (40%), microcystic (8%), psammomatous (6%), and angiomatous (3%). No statistically significant (P < .05) associations were found among tumor subtype or grade and any of the MRI features studied. CONCLUSIONS AND CLINICAL IMPORTANCE: Meningiomas in dogs differ from their counterparts in humans mainly in their higher incidence of atypical (Grade II) tumors observed. MRI characteristics do not allow for prediction of meningioma subtype or grade, emphasizing the necessity of histopathology for antemortem diagnosis. The higher incidence of atypical tumors in dogs may contribute to the poorer therapeutic response in dogs with meningiomas as compared with the response in humans with meningiomas.

Canine intraspinal meningiomas: imaging features, histopathologic classification, and long-term outcome in 34 dogs.

BACKGROUND: Meningioma is the most common primary intraspinal nervous system tumor in dogs. Clinical findings, clinicopathologic data, and treatment of these tumors have been reported sporadically, but little information is available regarding cerebrospinal fluid (CSF) analysis, histologic tumor grade, or efficacy of radiation therapy as an adjunct to cytoreductive surgery. ANIMALS: Dogs with histologically confirmed intraspinal meningiomas (n = 34). METHODS: A retrospective study of dogs with intraspinal meningiomas between 1984 and 2006 was carried out. Signalment, historical information, physical examination, clinicopathologic data, radiation therapy protocols, surgery reports, and all available images were reviewed. All tumors were histologically classified and graded as defined by the international World Health Organization classification scheme for central nervous system tumors. RESULTS: Intraspinal mengiomas in dogs are most common in the cervical spinal cord but can be found throughout the neuraxis. Location is correlated with histologic grade, with grade I tumors more likely to be in the cervical region than grade II tumors. Myelography generally shows an intradural extramedullary compressive lesion. On magnetic resonance imaging, the masses are strongly and uniformly contrast enhancing and a dural tail often is present. CSF analysis usually shows increased protein concentration with mild to moderate mixed pleocytosis. Surgical resection is an effective means of improving neurologic status, and adjunctive radiation therapy may lead to an improved outcome. CONCLUSIONS AND CLINICAL IMPORTANCE: Biopsy is necessary for definitive diagnosis, but imaging and CSF analysis can suggest a diagnosis of meningioma. Treatment of meningiomas with surgery and radiation therapy can result in a fair to excellent prognosis.

Middle ear tumours with brainstem extension treated by ventral bulla osteotomy and craniectomy in two cats.

Two geriatric domestic shorthaired cats (DSH) were treated surgically with a ventral bulla osteotomy and craniectomy for middle ear tumours that invaded the calvarium. Both cats had a history of vestibular disease. One cat had a normal neurological examination. Both cats underwent computed tomographic imaging of the head. After intravenous injection of an iodinated contrast material, one cat had a ring-enhancing intracranial lesion and the other had a uniform contrast-enhancing intracranial lesion, which either communicated with the bulla or was associated with bulla osseous lysis/production. One cat had a papillary adenoma and the other had an adenocarcinoma. Both cats had prolonged survivals (630 days; alive and lost to follow-up at 840 days), which is longer than reported in cats with middle ear neoplasia. Craniectomy, in addition to ventral bulla osteotomy, may be part of the treatment plan for middle ear tumours that invade the calvarium.

Granular cell tumor of the canine central nervous system: two cases.

Although pure granular cell tumors have been reported in various sites in the dog, only one tumor has been reported in the central nervous system. Two dogs presented with neurologic signs had brain lesions detected by magnetic resonance imaging in the area of the olfactory bulbs and frontal cortex. In both dogs, a clinical diagnosis of a granular cell tumor was made from tissues obtained from stereotactic biopsies guided by computed tomography. Surgical removal of the tumors was followed by histopathologic, ultrastructural, and immunocytochemical characterization. Although not conclusive, these studies indicated that the granular cells were not of leucocyte origin but may have been derived from the meninges. One dog died 12 months after surgery, and the other was alive 4 months later.

Extradural spinal synovial cysts in nine dogs.

Nine dogs presenting for investigation of cervical or thoracolumbar myelopathies were diagnosed with extradural spinal synovial cysts. Degenerative disease affecting the articular facets or intervertebral discs was present on plain spinal radiographs in all cases. Myelography was consistent with dorsolateral, extradural spinal cord compression. Two groups of dogs were identified: (1) young, giant breed dogs with multiple cysts involving one or more levels of the cervical spinal cord; and (2) older, large breed dogs with solitary cysts involving the thoracolumbar spinal cord. The synovial cysts constituted the major compressive lesions in four of the dogs. Analysis of lumbar cerebrospinal fluid demonstrated albuminocytological dissociation, consistent with chronic compressive myelopathy, in six dogs. All dogs underwent decompressive surgery and the diagnosis of synovial cysts was confirmed histologically. The mean follow-up period was 17 months (range four to 36 months). At the time of follow-up, all dogs were fully ambulatory with improved neurological function compared with that at initial presentation.

Primary canine and feline nervous system tumors: intraoperative diagnosis using the smear technique.

The recent application of neuroimaging techniques in veterinary neurology has led to the accurate localization of many types of intracranial lesions but has also created a clinical need, particularly with brain tumors, for a specific intraoperative diagnosis. For human brain tumors, a smear technique has been used successfully for many years to provide an extremely rapid, highly accurate intraoperative diagnosis. In similar smear preparations of intracranial lesions, obtained either by computed tomography (CT)-guided stereobiopsy or from a craniotomy, we have described distinguishing cytologic features of some primary spontaneous nervous system tumors in 80 dogs and 13 cats. A final diagnosis was confirmed by evaluation of paraffin-embedded sections from the same sample and, when appropriate, by immunocytochemical staining. Preliminary findings indicate that, in dogs and cats, this procedure is useful for rapid, accurate intraoperative diagnosis of many primary nervous system tumors. The distinguishing features of the canine and feline tumors bear a remarkably close resemblance to their human counterparts.

Intervertebral disc extrusion in six cats.

Existing reports concerning intervertebral disc disease (IVDD) have focused almost exclusively on dogs, although a small number of individual case reports of IVDD of cats has been published. The medical records of six cats with IVDD were reviewed. Radiographic studies confirmed narrowed intervertebral disc spaces, mineralised intervertebral discs, and one or more extradural compressive lesions of the spinal cord in each cat. All disc extrusions were located in the thoracolumbar region. Surgical decompression of the spinal cord was achieved in all cats by means of hemilaminectomy and removal of compressive extradural material confirmed to be degenerative disc material. Good to excellent neurological recovery was noted in five of the six cats included in this report. Based on this review, it appears that IVDD of cats has many similarities to IVDD of dogs, and that healthy cats with acute intervertebral disc extrusion(s) respond favourably to surgical decompression of the spinal cord.

CT-guided brain biopsy using a modified Pelorus Mark III stereotactic system: experience with 50 dogs.

This report describes the results of CT-guided stereotactic brain biopsies performed on 50 consecutive dogs using a modified Pelorus Mark III Stereotactic System. Based on available histopathologic samples (stereotactic biopsy [n = 50], surgery [n = 17], necropsy [n = 9]) the patient population consisted of 34 dogs with primary brain tumors, 2 with invasive nasal adenocarcinomas, and 13 with non-neoplastic brain lesions. Brain tissue was not obtained from one dog. In 22 dogs a final diagnosis was made from tissue subsequently obtained from surgical resection or at necropsy. The final diagnosis was in agreement with the stereotactic biopsy diagnosis in 20 of these 22 dogs. In 17 other dogs without follow-up, stereotactic biopsy provided a diagnosis of a specific primary brain tumor subtype. Postoperative complications associated with the biopsy procedure were assessed in 41 dogs. The other 9 dogs either went directly to surgery (n = 7) or were killed (n = 2) immediately after the biopsy procedure. Thirty-six dogs recovered without apparent clinical complications. Postoperative clinical complications in the remaining 5 dogs included transient epistaxis (1 dog), transient exacerbation of cerebellar signs (1 dog), obtundation progressing to coma (1 dog), and medically uncontrollable seizures (2 dogs). The latter 3 dogs with severe neurologic complications all had large primary brain tumors and had been receiving high doses of phenobarbital and glucocorticoids to control seizures at the time of biopsy. These results suggest that this CT-guided biopsy procedure can provide an accurate pathologic diagnosis of brain lesions detected by CT and MR neuroimaging. Further refinement of both technique and case selection is expected to reduce the rate of clinical complications and to improve the accuracy of the procedure.

Feline vestibular disorders. Part I: anatomy and clinical signs.

An understanding of the anatomy of the feline vestibular system is essential for interpretation of the clinical signs associated with vestibular dysfunction, for precise lesion localisation, and for accurate interpretation of results of diagnostic imaging. Appropriate recognition and interpretation of the clinical signs of vestibular disease is also an essential aspect of the precise diagnosis of the cause of vestibular dysfunction in cats. The objectives of this review are to provide an overview of the anatomy of the feline vestibular system, and to review the clinical signs of peripheral and central vestibular dysfunction of cats.