Ocular immunology in equine recurrent uveitis

Equine recurrent uveitis (ERU) is a disease with high prevalence and relevance for the equine population, since it results in blindness. Over the last decade, important advancements have been made in our understanding of the underlying immune responses in this disease. ERU is mediated by an autoaggressive Th1 response directed against several retinal proteins. Interphotoreceptor-retinoid binding protein (IRBP) and cellular retinaldehyde-binding protein (CRALBP) are capable to induce ERU-like disease in experimental horses, with the unique possibility to activate relapses in a well-defined manner. Further, proteomic evidence now suggests that retinal Mueller glial cells (RMG) may play a fatal role in uveitic disease progression by directly triggering inflammation processes through the expression and secretion of interferon-γ. Ongoing relapses in blind eyes can be associated with stable expression of the major autoantigens in ERU retinas. This review briefly summarizes the most significant developments in uveitis immune response research.     

Detection of Leptospira interrogans DNA and antigen in fixed equine eyes affected with end-stage equine recurrent uveitis.

Equine recurrent uveitis (ERU) is the most frequent cause of blindness in horses worldwide. Leptospira has been implicated as an etiologic agent in some cases of ERU and has been detected in fresh ocular tissues of affected horses. The objective of this study was to determine the presence of Leptospira antigen and DNA in fixed equine ocular tissues affected with end-stage ERU. Sections of eyes from 30 horses were obtained. Controls included 1) 10 normal equine eyes and 2) 10 equine eyes with a nonrecurrent form of uveitis. The experimental group consisted of 10 eyes diagnosed with ERU based on clinical signs and histologic lesions. Sections were subjected to immunohistochemical staining with an array of rabbit anti-Leptospira polyclonal antibodies. DNA extractions were performed by using a commercial kit designed for fixed tissue. Real-time PCR analysis was completed on extracted DNA. The target sequence for PCR was designed from alignments of available Leptospira 16S rDNA partial sequences obtained from GenBank. Two of 10 test samples were positive for Leptospira antigen by immunohistochemical assay. Zero of 20 controls were positive for Leptospira antigen. All test samples and controls were negative for Leptospira DNA by real-time PCR analysis. Leptospira was detected at a lower frequency than that previously reported for fresh ERU-affected aqueous humor and vitreous samples. Leptospira is not frequently detectable in fixed ocular tissues of horses affected with ERU when using traditional immunohistochemical and real-time PCR techniques.     

Physiological and pathological expression of intermediate filaments in the equine endometrium

The aim of this study was to investigate the expression of the intermediate filaments cytokeratin, vimentin and desmin in the equine endometrium by immunohistological techniques. For this purpose, endometrial biopsies of 151 mares were examined to determine physiological cycle patterns and changes resulting from endometriosis. During the physiological cycle epithelial cells and mesenchymal cells express cytokeratin and vimentin, respectively, whilst desmin and vimentin were coexpressed by the smooth muscle cells. Epithelial coexpression of cytokeratin and vimentin was seen in numerous fibrotic glands and in the uterine glands of three mares with pathologically inactive endometria. Three different staining patterns (basal, perinuclear, diffuse) of vimentin were associated with typical morphological alterations of the affected epithelia. In addition, in 14 cases a stromal coexpression of vimentin and desmin was found, indicating an atypical stromal differentiation in inactive endometria of older mares, barren for several years.     

Immunological response of chickens to eastern equine encephalomyelitis virus

The dominant immunoglobulin against eastern equine encephalomyelitis (EEE) virus and its duration and the longevity of the EEE virus haemagglutination inhibition (HI) antibodies were determined in sentinel and 125 immunised and hyperimmunised domestic chickens by enzyme-linked immunosorbent assay and the HI test respectively. The chickens ranged in age from 10 weeks to 18 months, were of varied pedigrees and from different countries. Results show that the HI antibody (IgG) is short-lived. It peaks and disappears within 30 days. The secondary response is dominated by the IgM immunoglobulin which is relatively long-lasting. These results are contrary to classical expectations and were observed in all the chickens studied. If these observations are found to be characteristic of birds generally, the present standard method of EEE virus seroepidemiological surveillance must be modified to be effective.     

Immunological characterization of the equine airway epithelium and of a primary equine airway epithelial cell culture model

Our understanding of innate immunity within the equine respiratory tract is limited despite growing evidence for its key role in both the immediate defense and the shaping of downstream adaptive immune responses to respiratory disease. As the first interface to undergo pathogen invasion, the respiratory epithelium is a key player in these early events and our goal was to examine the innate immune characteristics of equine respiratory epithelia and compare them to an in vitro equine respiratory epithelial cell model cultured at the air-fluid interface (AFI). Respiratory epithelial tissues, isolated epithelial cells, and four-week old cultured differentiated airway epithelial cells collected from five locations of the equine respiratory tract were examined for the expression of toll-like receptors (TLRs) and host defense peptides (HDPs) using conventional polymerase chain reaction (PCR). Cultured, differentiated, respiratory epithelial cells and freshly isolated respiratory epithelial cells were also examined for the expression of TLR3, TLR9 and major histocompatibility complex (MHC) class I and class II using fluorescence-activated cell sorting (FACS) analysis. In addition, cytokine and chemokine profiles from respiratory epithelial tissues, freshly isolated respiratory epithelial cells, and cultured, differentiated, epithelial cells from the upper respiratory tract were examined using real-time PCR. We found that respiratory epithelial tissues and isolated epithelial cells expressed TLRs 1-4 and 6-10 as well as HDPs, MxA, 2'5' OAS, β-defensin-1, and lactoferrin. In contrast, epithelial cells cultured at the AFI expressed TLRs 1-4 and 6 and 7 as well as MxA, 2'5' OAS, β-defensin-1, but had lost expression of TLRs 8-10 and lactoferrin. In addition, MHC-I and MHC-II surface expression decreased in epithelial cells cultured at the AFI compared to isolated epithelial cells whereas TLR3 and TLR9 were expressed at similar levels. Lastly, we found that equine respiratory epithelial cells express an array of pro-inflammatory, antiviral and regulatory cytokines and that after four weeks of in vitro growth conditions, equine respiratory epithelial cells cultured at the AFI retained expression of GM-CSF, IL-10, IL-8, TGF-β, TNF-α, and IL-6. In summary, we describe the development of an in vitro equine respiratory epithelial cell culture model that is morphologically similar to the equine airway epithelium and retains several key immunological properties. In the future this model will be a used to study equine respiratory viral pathogenesis and cell-to-cell interactions.     

Strongylus equinus: development and pathological effects in the equine host.

The development and pathological effects of Strongylus equinus were studied in 17 pony foals and one horse foal raised in isolation and examined at necropsy from seven days to 40 wk postinfection (PI). Following inoculation of 15000 +/- 6% or 16000 +/- 6% infective larvae by stomach tube foals were monitored for clinical signs and selected blood changes. Larvae penetrated the wall of the ileum, cecum and colon. The molt to the fourth stage occurred mostly in the wall of the ventral colon before 2 wk PI and larvae attained the liver mainly via the peritoneal cavity as early as eight days PI and persisted in the liver until 17 wk PI. Following active migration within the liver, invasion of the pancreas was accomplished at least by 7 wk PI with maximum numbers at 17 wk. The fourth molt occurred about 15 wk PI and preadults were present in the wall of the ventral colon at 30 wk PI and in the lumen of the colon at 40 wk. Strongylus equinus tends to wander retroperitoneally to the flanks, perirenal fat, diaphragm, omentum and occasionally to the lungs. Between 1 and 4 wk PI small raised hemorrhagic areas were present on the serosa of the ileum and colon. Small white foci on the surface of the liver at 1 wk PI were followed by tortuous tracks 3 wk later. Pathological changes in the pancreas were evident at three months PI and more severe by four months. Granulomas containing larvae were common in the flanks, diaphragm, omentum and occasionally beneath the pleura of the lungs. Clinical signs were correlated with invasion of the pancreas, the fourth molt, maximum globulin values and high eosinophil counts.     

Relationship between spinal biomechanics and pathological changes in the equine thoracolumbar spine

The relationship between spinal biomechanics and pathological changes occurring in functionally normal equine thoracolumbar spines was studied in 23 horses. Ventrolateral vertebral body osteophytes occurred in 36 per cent of the spines. The majority occurred between the 10th and 17th thoracic vertebrae with the largest being found between the 11th and 13th thoracic vertebrae, the region of the thoracic spine where the greatest amount of lateral bending and axial rotation occurs. Impingement of the dorsal spinous processes was detected in 86 per cent of the spines with most lesions occurring between the 13th and 18th thoracic vertebrae. The severity of occurrence of impingement did not appear to be related to regional spinal mobility. Degeneration of intervertebral discs was observed in three of four specimens that were sectioned sagittally. It occurred in the first thoracic and the lumbosacral intervertebral discs and appeared to be related to the increased dorsoventral mobility and the increased disc thickness of these joints. The characteristic distribution of fractures of the thoracolumbar spine is discussed with respect to the biomechanics of the spine.     

Immunological and hematological response in experimental Toxocara canis-infected pigs

The relationship between the immunological and hematological response to infection was studied in pigs inoculated experimentally with Toxocara canis. Two groups of four pigs were infected with doses of 1000 and 2000 infective eggs, respectively. Two uninfected animals were used as negative controls. Blood samples were collected from each pig once a week. Serological examination by ELISA to determine antibody levels against T. canis L2/L3 excretory-secretory (ES) antigens showed values higher than the positive cut-off point (1:32) for both the infected groups. These values increased from day 7 p.i. and remained high during the experimental period until day 56. Significant differences were recorded for the two inoculating doses (p相似文献