Sulphido-leukotriene production from peripheral leukocytes and skin in clinically normal dogs and house dust mite positive atopic dogs

Pathogenesis of canine atopy has not been completely elucidated. In humans, sulphido-leukotrienes (s-LT) play a role in atopy, and increased production of s-LT occurs in the skin and peripheral leukocytes after allergen challenge. The study population included 16 clinically normal and 13 atopic dogs. All atopic dogs had in common a positive reaction (4+) to the intradermal injection of house dust mite (allergen of reference). Blood samples and skin biopsies were collected. Sulphido-LT synthesis by peripheral leukocytes after stimulation was measured, and no statistically significant difference was found between clinically normal and atopic dogs. Sulphido-LT concentrations in skin samples from stimulated and unstimulated sites were measured, and no statistically significant difference was detected between clinically normal and atopic dogs or between lesional and nonlesional skin within the atopic group. Clinical signs of atopic dogs were graded by owners and no correlation was found between their severity and cutaneous concentrations of s-LT. In this study there was no increase in s-LT synthesis in atopic dogs.     

Failure to induce mucosal disease in cattle persistently infected with bovine viral diarrhea virus by treatment with adrenocorticotropic hormone

Recent research has shown that cattle that develop mucosal disease (MD) often, if not always, have been persistently infected with bovine viral diarrhea virus (BVDV) since birth. The purpose of the present study was to determine whether MD could be induced by immunosuppression of persistently BVDV-infected cattle. For that purpose, adrenocorticotropic hormone (ACTH) was injected intramuscularly, twice daily for 5 consecutive days in 4 persistently BVDV-infected cattle and in 3 control cattle. Before the ACTH treatment, the numbers of leukocytes, neutrophils and mononuclear cells (MNC) per litre of blood in BVDV-infected cattle were in the same range as in the controls. Similarly, the proportions of B cells, T cells, monocytes and Fcγ⁺ cells (cells with receptor for the Fc part of IgG) were the same in the 2 groups of animals. On the other hand, the proliferative response to mitogen stimulation of MNC obtained from the control animals was twice as high as the corresponding value of the persistently BVDV-infected cattle.In all animals, ACTH treatment caused increased Cortisol concentrations, leukocytosis, neutrophilia and decreased mitogen-induced lymphocyte stimulation. However, the MNC count and the proportions of B cells, T cells, Fcγ⁺ cells and monocytes remained unaltered. In spite of the immunosuppression, indicated by the decrease in mitogen-induced lymphocyte stimulation. ACTH treatment did not provoke any clinical signs of MD in the persistently BVDV-infected cattle.     

Multinucleated giant cells with an osteoclast phenotype derived from caprine peripheral blood mononuclear cells

Formation of multinucleated giant cells (MGCs) by macrophage fusion is a typical cytopathic effect of lentiviral replication in caprine monocytes and MGC formation from cultured caprine peripheral blood mononuclear cells (PBMCs) has been considered to be diagnostic for small ruminant lentivirus (SRLV) infection. In this study, formation of MGCs was observed after 7–14 days when PBMCs were cultured from healthy goats free from SRLV infection. These MGCs expressed tartrate-resistant acid phosphatase, calcitonin receptor, integrin αVβ3, cathepsin K and matrix metalloproteinase 9 and were able to resorb bone in vitro in the absence of RANKL and macrophage colony stimulating factor, consistent with an osteoclast phenotype.     

Autologous point‐of‐care cellular therapies variably induce equine mesenchymal stem cell migration,proliferation and cytokine expression

Reasons for performing study: Autologous cellular therapy products including adipose‐derived stromal vascular fraction (SVF), bone marrow mononuclear cells (BMMNs), cord blood mononuclear cells (CBMNs) and platelet rich plasma are options for treatment of acute orthopaedic lesions while mesenchymal stem cells (MSCs) are culture expanded. These products may contribute to healing by secreting matrix proteins or growth factors, but they may also act on endogenous MSCs to facilitate healing. Objectives: To determine the effects of cell therapy products on MSCs function in vitro. The hypothesis was that cell therapy products promote MSCs functions including proliferation, migration and mediator release. Methods: Fat, bone marrow (BM), cord blood and platelets were obtained from 6 Quarter Horses. The BM‐MSCs and their autologous cell therapy products were co‐incubated in transwells. Mesenchymal stem cells proliferation, migration, gene expression and cytokine concentrations were determined. Results: All cell therapy products increased MSCs proliferation, but SVF induced significantly more proliferation than any other product. Also SVF elicited more MSCs chemotaxis and, along with BMMNs, significantly more MSCs chemoinvasion. Cord blood mononuclear cells stimulated MSCs to produce high concentrations of interleukin‐6 (IL‐6), transforming growth factor‐β1 (TGF‐β1), and prostaglandin E₂ (PGE₂). Stromal vascular fraction and platelet lysate did not stimulate MSCs but SVF and platelet lysate themselves contained high concentrations of PGE₂ and IL‐6 (SVF) and TGF‐β1 (platelet lysate). Conclusions: Autologous cell products variably stimulate MSCs functions with 2 primary patterns apparent. Products either contained preformed mediators that may have intrinsic healing function, or products stimulated MSCs to secrete mediators. Potential relevance: The specific clinical indications for these products may differ to include administration as a sole treatment modality prior to MSCs injection for intrinsic cell and cytokine activity (i.e. SVF) or administration concurrently with MSCs to activate MSCs for treatment of chronic lesions (i.e. CBMNs).     

Detection of bovine herpesvirus type 4 antibodies and bovine lymphotropic herpesvirus in New Zealand dairy cows

AIM: To detect the presence of bovine herpesvirus (BoHV) type 4 in New Zealand dairy cows with clinical metritis.

METHODS: Serum samples taken from 92 dairy cows with clinical metritis, each from a different farm, were tested for the presence of antibodies against BoHV-4 using a commercially available, indirect ELISA. Peripheral blood mononuclear cells (PBMC) were collected from 10 BoHV-4 seropositive cows, and PBMC were examined by a pan-herpesvirus nested PCR to detect herpesvirus. PCR products were sequenced directly and a proportion of the PCR products were cloned and sequenced to identify the virus present.

RESULTS: Antibodies to BoHV-4 were detected in 23/92 (25%) serum samples. The pan-herpesvirus PCR was positive in 8/10 PBMC samples. Cloning and sequencing identified that all of the eight PCR-positive PBMC contained bovine lymphotropic herpesvirus (BLHV); no BoHV-4 DNA was detected.

CONCLUSIONS: This study reports the finding of the presence of apparent antibodies to BoHV-4, and BLHV DNA in New Zealand dairy cows affected by metritis.

CLINICAL RELEVANCE: Bovine herpesvirus type 4 and BLHV are reported to have the potential to cause reproduction failure in cows. This is the first report of apparent BoHV-4 antibodies, and BLHV in New Zealand. The importance and epidemiology of these viruses in cattle in New Zealand requires further investigation.     

The composition of the inflammatory infiltrate in three cases of polyneuritis equi

Polyneuritis equi (PNE) is a rare neurological disease in the horse. Because of the suspicion in PNE of a T-lymphocyte mediated immune response against the myelin, the objective of this study was to determine the composition of the inflammatory infiltrate in the involved nerves of 3 horses with PNE, studied retrospectively. T-lymphocytes were demonstrated in the lesions, which suggests a T-lymphocyte mediated immune response against myelin. In addition, the presence of B-lymphocytes indicated a local production of antibodies. More research, involving a prospective study, is needed to determine whether the T-lymphocytes are cytotoxic or T-helper lymphocytes.     

Surgical enucleation of unilateral mandibular radicular cysts in an 11-year-old Thoroughbred mare

An 11-year-old Thoroughbred broodmare was diagnosed with extensive radicular cysts that affected the left horizontal mandibular ramus. A left mandibular swelling was present clinically, and mandibular lesions appearing cystic in nature were identified with successive radiographic and ultrasonographic examinations. Surgical enucleation of the cysts was performed under a standing sedation protocol and the excised cystic structures submitted for histopathological analysis. Histological examination of the cystic structures confirmed a diagnosis of multiple radicular cysts.     

Identification of CD3+ T lymphocytes in the green turtle Chelonia mydas

To understand the role of the immune system with respect to disease in reptiles, there is the need to develop tools to assess the host's immune response. An important tool is the development of molecular markers to identify immune cells, and these are limited for reptiles. We developed a technique for the cryopreservation of peripheral blood mononuclear cells and showed that a commercially available anti-CD3 epsilon chain antibody detects a subpopulation of CD3 positive peripheral blood lymphocytes in the marine turtle Chelonia mydas. In the thymus and in skin inoculated with phytohemagglutinin, the same antibody showed the classical staining pattern observed in mammals and birds. For Western blot, the anti-CD3 antibodies identified a 17.6 kDa band in membrane proteins of peripheral blood mononuclear cell compatible in weight to previously described CD3 molecules. This is the first demostration of CD3+ cells in reptiles using specific antibodies.     

不同重悬细胞法在洗涤过程中对水牛外周血单个核细胞活率的影响

为了探讨在离心洗涤水牛外周血单个核细胞（peripheral blood mononuclear cell, PBMC）过程中以移液器吹打法和晃动离心管法重悬细胞对其活率的影响。对经密度梯度离心后初步收集到的PBMC,分别用移液器吹打和晃动离心管2种方法重悬细胞,离心洗涤后对细胞活率进行计数。结果发现,移液器吹打法得到的细胞活率为（74.9±2.6）%,而晃动离心管法使细胞活率达到（98.5±0.5）%,差异极显著（P<0.01）。说明在洗涤PBMC时,晃动离心管法重悬细胞能大大提高细胞活率,可为那些对细胞活率要求高的试验提供一个较为有效的方法。