Claude Carozzo

Natural killer cell trafficking in vivo requires a dedicated sphingosine 1-phosphate receptor

Consistent with their function in immune surveillance, natural killer (NK) cells are distributed throughout lymphoid and nonlymphoid tissues. However, the mechanisms governing the steady-state trafficking of NK cells remain unknown. The lysophospholipid sphingosine 1-phosphate (S1P), by binding to its receptor S1P1, regulates the recirculation of T and B lymphocytes. In contrast, S1P5 is detected in the brain and regulates oligodendrocyte migration and survival in vitro. Here we show that S1P5 was also expressed in NK cells in mice and humans and that S1P5-deficient mice had aberrant NK cell homing during steady-state conditions. In addition, we found that S1P5 was required for the mobilization of NK cells to inflamed organs. Our data emphasize distinct mechanisms regulating the circulation of various lymphocyte subsets and raise the possibility that NK cell trafficking may be manipulated by therapies specifically targeting S1P5.

Correction of Dog Dystrophic Epidermolysis Bullosa by Transplantation of Genetically Modified Epidermal Autografts

Recessive dystrophic epidermolysis bullosa (RDEB) is a severe skin blistering condition caused by mutations in the gene coding for collagen type VII. Genetically engineered RDEB dog keratinocytes were used to generate autologous epidermal sheets subsequently grafted on two RDEB dogs carrying a homozygous missense mutation in the col7a1 gene and expressing baseline amounts of the aberrant protein. Transplanted cells regenerated a differentiated and vascularized auto-renewing epidermis progressively repopulated by dendritic cells and melanocytes. No adverse immune reaction was detected in either dog. In dog 1, the grafted epidermis firmly adhered to the dermis throughout the 24-month follow-up, which correlated with efficient transduction (100%) of highly clonogenic epithelial cells and sustained transgene expression. In dog 2, less efficient (65%) transduction of primary keratinocytes resulted in a loss of the transplanted epidermis and graft blistering 5 months after transplantation. These data provide the proof of principle for ex vivo gene therapy of RDEB patients with missense mutations in collagen type VII by engraftment of the reconstructed epidermis, and demonstrate that highly efficient transduction of epidermal stem cells is crucial for successful gene therapy of inherited skin diseases in which correction of the genetic defect confers no major selective advantage in cell culture.

Prevalence of hip dysplasia according to official radiographic screening, among 31 breeds of dogs in France - A retrospective study

In order to determine the prevalence of hip dysplasia (HD) from radiographs that were submitted for authorative grading, and its changes over the time in 31 breeds of dogs in France, a 14 year-retrospective study was conducted. Significant differences were observed between breeds, with HD prevalence ranging from 59.7% (Cane Corso) to 3.9% (Siberian Husky). When comparing the 1993-1999 with the 2000-2006 period in 15 breeds, a significant decrease in HD prevalence was detected in Berger Picard, Bernese Mountain dog, Briard, Gordon Setter, White Swiss Sheepdog and Rottweiler. Modifications observed in the other breeds were not statistically significant.

Transiliac Approach for Exposure of Lumbosacral Intervertebral Disk and Foramen: Technique Description

OBJECTIVE To describe and evaluate a transiliac approach to the L7-S1 disk and intervertebral foramen in dogs. STUDY DESIGN Cadaver study. ANIMALS Fresh canine cadavers (n=10). METHODS A craniolateral approach was made to each iliac wing of 10 fresh canine mixed breed cadavers. An 18 mm hole was drilled in a standardized position through the iliac wing. The musculature connected to the cranial aspects of the sacral wing was dissected and retracted cranially through this iliac window. Endoscopic exploration of the area was performed. RESULTS The foramen and intervertebral disk were clearly observed in all specimens without iatrogenic injury of the L7 nerve branch. Access to the foramen was possible in 16 of 20 specimens without excision of the sacral wing; however, it was always partially excised to observe the intervertebral disk which lies more caudally and ventrally. CONCLUSION Transiliac approach to the lumbosacral joint allows direct exposure of the intervertebral disk and foramen through an iliac window. Endoscopic exploration provided good observation of the intervertebral disk and/or foramen. CLINICAL RELEVANCE Transiliac approach could be used for lateral corpectomy and foraminotomy in dogs with degenerative lumbosacral stenosis caused by ventral or ventrolateral disk protrusion, foramen stenosis, or OCD of the L7-S1 joint. Clinical study will be necessary to evaluate the efficacy of this approach.

Evaluation of intraoperative fluorescence imaging–guided surgery in cancer-bearing dogs: a prospective proof-of-concept phase II study in 9 cases

The objective was to prospectively evaluate the application of intraoperative fluorescence imaging (IOFI) in the surgical excision of malignant masses in dogs, using a novel lipid nanoparticle contrast agent. Dogs presenting with spontaneous soft-tissue sarcoma or subcutaneous tumors were prospectively enrolled. Clinical staging and whole-body computed tomography (CT) were performed. All the dogs received an intravenous injection of dye-loaded lipid nanoparticles, LipImage 815. Wide or radical resection was realized after CT examination. Real-time IOFI was performed before skin incision and after tumor excision. In cases of radical resection, the lymph nodes (LNs) were imaged. The margin/healthy tissues fluorescence ratio or LN/healthy tissues fluorescence ratio was measured and compared with the histologic margins or LN status. Nine dogs were included. Limb amputation was performed in 3 dogs, and wide resection in 6. No adverse effect was noted. Fluorescence was observed in all 9 of the tumors. The margins were clean in 5 of 6 dogs after wide surgical resection, and the margin/healthy tissues fluorescence ratio was close to 1.0 in all these dogs. Infiltrated margins were observed in 1 case, with a margin/healthy tissues fluorescence ratio of 3.2. Metastasis was confirmed in 2 of 3 LNs, associated with LN/healthy tissues fluorescence ratios of 2.1 and 4.2, whereas nonmetastatic LN was associated with a ratio of 1.0. LipImage 815 used as a contrast agent during IOFI seemed to allow for good discrimination between tumoral and healthy tissues. Future studies are scheduled to evaluate the sensitivity and specificity of IOFI using LipImage 815 as a tracer.

Endoscope‐Assisted Thoracolumbar Lateral Corpectomy

OBJECTIVE To report an endoscope-assisted lateral approach to expose the intervertebral foramen and disk and perform lateral corpectomy of thoracolumbar disks in dogs. STUDY DESIGN Cadaver study. ANIMALS Fresh canine cadavers (n=6). METHODS A thoracic (T10-T11) and a lumbar (L3-L4) lateral approach were made on 6 fresh canine mixed breed cadavers. Through a limited skin incision, musculature was dissected and retracted using a neurosurgical self-retaining retractor and lateral corpectomy performed. The approach and bone removal, both performed under endoscopic control, were assessed. RESULTS The foramen and intervertebral disk were clearly observed in all specimens without any iatrogenic injury of the ventral and dorsal nerve branches. Access to the foramen was possible in all specimens; in the thoracic area the head of the rib was always partially excised to observe the intervertebral disk medially. Lateral corpectomy was easily performed in all cadavers and spinal cord observation was good. Extension to foraminotomy or mini hemilaminectomy could easily be performed through the same limited approach. CONCLUSION Endoscopic exploration provided a good viewing of the intervertebral disk and/or foramen. An endoscope-assisted thoracolumbar lateral corpectomy could effectively be performed through a limited approach to the thoracolumbar disks and allowed good ventral spinal cord assessment.

Canine hip dysplasia radiographic screening. Prevalence of rotation of the pelvis along its length axis in 7,012 conventional hip extended radiographs

The prevalence of rotation of the pelvis along its length axis was noted, as was the number of rotations towards the right or left hand side of the dog, on 7,012 conventional hip extended radiographs, which were sent for official screening. 29.8% of the radiographs showed a rotation the pelvis. The rotation was statistically more frequent towards the left hand side of the dog. The number of rejected radiographs for too important pelvis rotation was only 5.2%. The consequences of the pelvis rotation on the Norberg-Olsson angle, on the dorsal femoral head coverage, and in the aspect of cranial acetabular edge have to be taken into account when scoring the dog for hip dysplasia.

Risk of simultaneous phenotypic expression of hip and elbow dysplasia in dogs: a study of 1,411 radiographic examinations sent for official scoring.

In order to look for phenotypic correlation between hip (HD) and elbow (ED) dysplasia, we used radiographic scoring obtained from 1,411 dogs of different breeds, which were evaluated for authoritative grading of both conditions. In this population, we found that the risk ratio for an animal to be simultaneously affected by HD and ED is 1.67. For a dog with ED, the risk ratio to be affected by HD increases as the ED grade increases. Similarly, for a dog affected by HD, the risk ratio to be affected by ED increases as the HD grade increases. In a dog affected by HD or ED, the clinician should look for the second condition in the same animal. Due to the low, yet positive correlation, selection against one trait will not affect the other trait sufficiently. Therefore selection has to be conducted at reduction of HD as well as reduction of ED.

Two cases of surgically treated feline patellar fractures

A transverse patellar fracture in a six-month-old cat was successfully treated by figure-of-eight dorsal wiring of the patella. A longitudinal patellar fracture with luxation of a large medial fragment in a 2.5-year-old cat was treated by lateral marginal patellectomy with a positive outcome. While adding material to the few veterinary reports in that species, in this brief communication, the authors discuss the aetiology, diagnosis, and the treatment of the presented cases with regards to findings in previously published feline cases.

Phase-0/phase-I study of dye-loaded lipid nanoparticles for near-infrared fluorescence imaging in healthy dogs.

Near-infrared (NIR) fluorescence imaging using FDA-approved indocyanine green (ICG) has been the subject of numerous studies during the past few years. It could constitute a potentially exciting new paradigm shift in veterinary oncology, especially to develop in vivo fluorescence imaging diagnostics and surgery guidance methods. The objective of this study was to evaluate the pharmacologic and toxicological characteristics in healthy beagle dogs of LipImage™ 815, a formulation made of NIR-dye-loaded lipid nanoparticles. The initial dosage for the evaluation of biodistribution was extrapolated from data in mice and then adapted to define the more adapted dose (MAD) according to the fluorescence results obtained in 5 dogs using a Fluobeam® 800 imaging device (phase 0 study). A single dose acute toxicity study was then performed (3 dogs, phase I study). Before the systemic administration of LipImage™ 815, the dogs presented a very mild residual fluorescence, particularly in the liver and kidneys. After injection, the plasma fluorescence continuously decreased, and the signal was relatively homogeneously distributed throughout the different organs, though more pronounced in the liver and to a lesser extent in the steroid-rich organs (adrenal, ovaries), intestines, lymph nodes and kidneys. A MAD of 2.0μg/kg was found. No evidence of acute or delayed general, hepatic, renal or hematologic toxicity was observed at 1-fold, 5-fold or 10-fold MAD. The results of this phase-0/phase-I study showed that an optimal dosage of LipImage™ 815 of 2.0μg/kg allowed the achievement of a fluorescence signal suitable for surgery guidance application without any acute side effects.