Equine sarcoids, part 1: clinical presentation and epidemiology

Equine sarcoids are the most common skin tumors in horses and other equids. In their pathogenesis, the bovine papillomavirus (BPV) plays a major role. Many clinical manifestations have been described, ranging from small single lesions to multiple aggressively growing masses. Histopathologically, it is considered as a biphasic tumor with epidermal hyperplasia and subepidermal proliferation of transformed fibroblasts. The diagnosis can be made clinically, histopathologically and/or by detection of BPV DNA. Sarcoids can appear on any part of the body, but they are mostly localized on the ventral abdomen, the paragenital region, head and limbs. Sarcoids occur independent of breed, coat color, sex or age, but they develop more commonly in young adults and certain families and breeds are more vulnerable than others. Transmission of BPV is supposed to happen from cattle to horse or from horse to horse, possibly via insects

Equine sarcoids, part 3: association with bovine papillomavirus

The genetic material of the bovine papillomavirus (BPV) can be detected in virtually all equine sarcoids. Eight different types have been described, all inducing benign proliferation of epithelium in cattle. BPV-1 and -2 are less strictly species-specific and can induce equine sarcoids in horses. Historically, association between BPV and equine sarcoids has been demonstrated using inoculation studies and detection of BPV DNA and BPV gene expression. The BPV genome is composed of 6 early and 2 late genes, with E5 and E6 being the most important transforming genes. Specific BPV-1 variants associated with equine sarcoids have been reported, suggesting circulation of the virus between horses. In horses, a non-productive BPV infection occurs, with only transcription of early genes, responsible for genome maintenance, regulation of cell growth and cell transformation. There is no formation of new infectious virus particles as is the case in the natural host

Equine sarcoids, part 2: current treatment modalities

Treatment of sarcoids is often challenging, due to the variable clinical presentation of lesions and the frequent local recurrences. In this article, both the surgical and the non-surgical treatment of equine sarcoids are reviewed. It is generally accepted that the prognosis is worse if unsuccessful attempts have been made previously. Therefore, the best available treatment option should always be used at the first attempt of treatment. Different surgical approaches have been reported, including conventional excision, cryosurgery and CO2 laser surgery. Success rates are high if a non-touch approach, wide surgical margins and general anesthesia can be applied. Local chemotherapy is a valuable addition in the treatment of sarcoids and can be combined with surgery. Radiotherapy is a very successful treatment, but safety precautions prevent routine application. Local immunotherapy including Bacillus Calmette-Guerin vaccination and imiquimod cream are commonly applied treatments which induce rather effective tumour regression

Selection of a set of reliable reference genes for quantitative real-time PCR in normal equine skin and in equine sarcoids

BACKGROUND: Real-time quantitative PCR can be a very powerful and accurate technique to examine gene transcription patterns in different biological conditions. One of the critical steps in comparing transcription profiles is accurate normalisation. In most of the studies published on real-time PCR in horses, normalisation occurred against only one reference gene, usually GAPDH or ACTB, without validation of its expression stability. This might result in unreliable conclusions, because it has been demonstrated that the expression levels of so called "housekeeping genes" may vary considerably in different tissues, cell types or disease stages, particularly in clinical samples associated with malignant disease. The goal of this study was to establish a reliable set of reference genes for studies concerning normal equine skin and equine sarcoids, which are the most common skin tumour in horses. RESULTS: In the present study the gene transcription levels of 6 commonly used reference genes (ACTB, B2M, HPRT1, UBB, TUBA1 and RPL32) were determined in normal equine skin and in equine sarcoids. After applying the geNorm applet to this set of genes, TUBA1, ACTB and UBB were found to be most stable in normal skin and B2M, ACTB and UBB in equine sarcoids. CONCLUSION: Based on these results, TUBA1, ACTB and UBB, respectively B2M, ACTB and UBB can be proposed as reference gene panels for accurate normalisation of quantitative data for normal equine skin, respectively equine sarcoids. When normal skin and equine sarcoids are compared, the use of the geometric mean of UBB, ACTB and B2M can be recommended as a reliable and accurate normalisation factor

A Highly Stable Prefusion Rsv F Vaccine Derived from Structural Analysis of the Fusion Mechanism

Respiratory syncytial virus (RSV) causes acute lower respiratory tract infections and is the leading cause of infant hospitalizations. Recently, a promising vaccine antigen based on the RSV fusion protein (RSV F) stabilized in the native prefusion conformation has been described. Here we report alternative strategies to arrest RSV F in the prefusion conformation based on the prevention of hinge movements in the first refolding region and the elimination of proteolytic exposure of the fusion peptide. A limited number of unique mutations are identified that stabilize the prefusion conformation of RSV F and dramatically increase expression levels. This highly stable prefusion RSV F elicits neutralizing antibodies in cotton rats and induces complete protection against viral challenge. Moreover, the structural and biochemical analysis of the prefusion variants suggests a function for p27, the excised segment that precedes the fusion peptide in the polypeptide chain

Recombinant low-seroprevalent adenoviral vectors Ad26 and Ad35 expressing the respiratory syncytial virus (RSV) fusion protein induce protective immunity against RSV infection in cotton rats

AbstractRSV is an important cause of lower respiratory tract infections in children, the elderly and in those with underlying medical conditions. Although the high disease burden indicates an urgent need for a vaccine against RSV, no licensed RSV vaccine is currently available. We developed an RSV vaccine candidate based on the low-seroprevalent human adenovirus serotypes 26 and 35 (Ad26 and Ad35) encoding the RSV fusion (F) gene. Single immunization of mice with either one of these vectors induced high titers of RSV neutralizing antibodies and high levels of F specific interferon-gamma-producing T cells. A Th1-type immune response was indicated by a high IgG2a/IgG1 ratio of RSV-specific antibodies, strong induction of RSV-specific interferon-gamma and tumor necrosis factor-alpha cytokine producing CD8 Tcells, and low RSV-specific CD4 T-cell induction. Both humoral and cellular responses were increased upon a boost with RSV-F expressing heterologous adenovirus vector (Ad35 boost after Ad26 prime or vice versa). Both single immunization and prime-boost immunization of cotton rats induced high and long-lasting RSV neutralizing antibody titers and protective immunity against lung and nasal RSV A2 virus load up to at least 30 weeks after immunization. Cotton rats were also completely protected against challenge with a RSV B strain (B15/97) after heterologous prime-boost immunization. Lungs from vaccinated animals showed minimal damage or inflammatory infiltrates post-challenge, in contrast to animals vaccinated with formalin-inactivated virus. Our results suggest that recombinant human adenoviral Ad26 and Ad35 vectors encoding the RSV F gene have the potential to provide broad and durable protection against RSV in humans, and appear safe to be investigated in infants

Exploring Immune Development in Infants With Moderate to Severe Atopic Dermatitis

Background: Atopic dermatitis (AD) is the most common chronic inflammatory skin disease in infancy with a complex pathology. In adults, the clinical severity of AD has been associated with increases in T helper cell type (Th) 2, Th22, and Th17 serum markers, including high levels of CC chemokine ligand (CCL) 17 and CCL22 chemokines. Objective: To explore the possible association between serum chemokine levels and AD severity in infants with moderate-to-severe AD and elevated immunoglobulin E (IgE). Subjects and methods: Serum samples (n = 41) obtained from a randomized, double-blind, and clinical dietary intervention study were used to study biomarkers in infants with AD. Baseline- and post-intervention samples (4 months) were used, six chemokines and nine ratios thereof were analyzed using Luminex and correlated to AD severity. In the initial study, the infants were randomized to receive extensively hydrolyzed whey-based formula without (control) or with short-chain galacto-oligosaccharides/long-chain fructo-oligosaccharides (9:1) and Bifidobacterium breve M-16V (active). Results: 31 Infants up to 11 months of age, with an objective-SCORAD score (oSCORAD) ≥ 20 and elevated total-IgE and/or specific-IgE levels were included. In time, the median oSCORAD decreased in both groups by -8 (control, p < 0.05; active, p < 0.01). Irrespective of dietary intervention, several changes in Th2 chemokines (CCL17 and CCL22), inflammatory chemokine (CCL20), and the Th1 chemokine, CXC chemokine ligand (CXCL) 9, were detected over time. Overall CCL17 correlated to oSCORAD (r = 0.446, p < 0.01). After 4 months of dietary intervention, CXCL9 was higher (p < 0.01) in the active group compared with control [active, 2.33 (1.99-2.89); controls, 1.95 (1.77-2.43) log 10 median (range)]. In addition, a reduction in Th2/Th1 chemokine ratios for CCL17/CXCL9, CCL22/CXCL9, CCL20/CXCL10, and CCL20/CXCL11 was detected associated with the active intervention. Conclusion: While this study is small and exploratory in nature, these data contribute to immune biomarker profiling and understanding of AD in infants

Clinical and histological description of penile intraepithelial neoplasia in horses and possible association with papillomaviral infection.

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