RIPK3 deficiency or catalytically inactive RIPK1 provides greater benefit than MLKL deficiency in mouse models of inflammation and tissue injury.

Necroptosis is a caspase-independent form of cell death that is triggered by activation of the receptor interacting serine/threonine kinase 3 (RIPK3) and phosphorylation of its pseudokinase substrate mixed lineage kinase-like (MLKL), which then translocates to membranes and promotes cell lysis. Activation of RIPK3 is regulated by the kinase RIPK1. Here we analyze the contribution of RIPK1, RIPK3, or MLKL to several mouse disease models. Loss of RIPK3 had no effect on lipopolysaccharide-induced sepsis, dextran sodium sulfate-induced colitis, cerulein-induced pancreatitis, hypoxia-induced cerebral edema, or the major cerebral artery occlusion stroke model. However, kidney ischemia-reperfusion injury, myocardial infarction, and systemic inflammation associated with A20 deficiency or high-dose tumor necrosis factor (TNF) were ameliorated by RIPK3 deficiency. Catalytically inactive RIPK1 was also beneficial in the kidney ischemia-reperfusion injury model, the high-dose TNF model, and in A20(-/-) mice. Interestingly, MLKL deficiency offered less protection in the kidney ischemia-reperfusion injury model and no benefit in A20(-/-) mice, consistent with necroptosis-independent functions for RIPK1 and RIPK3. Combined loss of RIPK3 (or MLKL) and caspase-8 largely prevented the cytokine storm, hypothermia, and morbidity induced by TNF, suggesting that the triggering event in this model is a combination of apoptosis and necroptosis. Tissue-specific RIPK3 deletion identified intestinal epithelial cells as the major target organ. Together these data emphasize that MLKL deficiency rather than RIPK1 inactivation or RIPK3 deficiency must be examined to implicate a role for necroptosis in disease.

Mouse Models of Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a chronic debilitating autoimmune disorder characterized by synovitis that leads to cartilage and bone erosion by invading fibrovascular tissue. Mouse models of RA recapitulate many features of the human disease. Despite the availability of medicines that are highly effective in many patient populations, autoimmune diseases (including RA) remain an area of active biomedical research, and consequently mouse models of RA are still extensively used for mechanistic studies and validation of therapeutic targets. This review aims to integrate morphologic features with model biology and cover the key characteristics of the most commonly used induced and spontaneous mouse models of RA. Induced models emphasized in this review include collagen-induced arthritis and antibody-induced arthritis. Collagen-induced arthritis is an example of an active immunization strategy, whereas antibody- induced arthritis models, such as collagen antibody-induced arthritis and K/BxN antibody transfer arthritis, represent examples of passive immunization strategies. The coverage of spontaneous models in this review is focused on the TNFΔ (ARE) mouse, in which arthritis results from overexpression of TNF-α, a master proinflammatory cytokine that drives disease in many patients.

TNF superfamily member TL1A elicits type 2 innate lymphoid cells at mucosal barriers.

Immune responses at mucosal barriers are regulated by innate type 2 lymphoid cells (ILC2s) that elaborate effector cytokines interleukins 5 and 13 (IL5 and IL13). IL25 and IL33 are key cytokines that support ILC2s; however, mice deficient in these pathways retain some functional ILC2s. Analysis of human and murine cells revealed that ILC2s highly express tumor necrosis factor (TNF)-receptor superfamily member DR3 (TNFRSF25). Engagement of DR3 with cognate ligand TL1A promoted ILC2 expansion, survival, and function. Exogenous protein or genetic overexpression of TL1A activated ILC2s independent of IL25 or IL33. Dr3(-/-) mice failed to control gut helminthic infections, and failed to mount ILC2 responses in the lung after nasal challenge with papain. Our data demonstrate a key role for TL1A in promoting ILC2s at mucosal barriers.

Resistance to scrapie in PrP ARR/ARQ heterozygous sheep is not caused by preferential allelic use.

BACKGROUND: In sheep, susceptibility to scrapie, which is similar to human prion diseases such as Kuru and variant Creutzfeldt-Jakob disease (vCJD), is determined by prion protein (PrP) gene (Prnp) polymorphisms. Sheep with genotype ARQ/ARQ, denoting polymorphisms at codons 136, 154, and 171, are susceptible, whereas those with genotypes ARR/ARQ and ARR/ARR are resistant, indicating dominance of ARR over the ARQ allele. AIMS: Based on familial CJD E200K, 129V, where preferential use of the 200E allele in EK heterozygous individuals confers resistance, heterozygous ARR/ARQ sheep were used to test the hypothesis that resistance is caused by preferential use of the ARR allele. METHODS: After assessment of equivalent PrP expression across genotypes, allele use was analysed by sequencing reverse transcription polymerase chain reaction derived DNA clones containing the Prnp gene coding sequence. RESULTS: The ARR to ARQ ratio was 1.1 in 133 clones, representing Prnp mRNA from three ARR/ARQ sheep, indicating equal use of both alleles. CONCLUSIONS: Dominance of the resistant associated allele in sheep scrapie involves mechanisms other than the absence of PrP derived from the disease associated ARQ allele.

Encephalitozoon cuniculi placentitis and abortion in a quarterhorse mare.

Encephalitozoon cuniculi is a microsporidial parasite, which has rarely been reported to cause placentitis in animals. A late-term aborted fetus and placenta from a Quarterhorse were presented to the Livestock Disease Diagnostic Center, University of Kentucky, for diagnostic examination. There was a necrotizing placentitis, with distension of many chorionic epithelial cells by intracytoplasmic vacuoles containing 1-2-microm-diameter, elongated, gram-positive organisms. The organisms were identified as E. cuniculi by electron microscopy and by polymerase chain reaction using primers to microsporidial ribosomal DNA. Joints of the fetus were swollen, with gross and microscopic lesions of synovitis; however, E. cuniculi DNA was not detected.

[Borna disease in Switzerland and in the principality of Liechtenstein]. / Die "Bornasche Krankheit" in der Schweiz und im Fürstentum Liechtenstein.

Borna disease (BD) is a rare immunopathological disorder of the central nervous system (CNS) caused by infection with Borna disease virus (BDV) and histologically characterized by mononuclear encephalomyelitis. BD primarily affects equines and sheep in well defined endemic areas of central Europe, but BDV infections have also been reported in other host species including humans, as well as in non endemic regions. In this paper recent data on the pathogenesis of BD are reviewed and the current situation in Switzerland and the Principality of Liechtenstein is summarized.

Lack of association between Borna disease virus infection and neurological disorders among HIV-infected individuals.

Viruses have been proposed as etiologic cofactors in the pathogenesis of HIV-related neurological disease. To investigate the etiologic potential of Borna disease virus (BDV) in these disorders, two populations were studied: (1) 27 prospectively identified patients with various neurological disorders were evaluated with BDV RT-PCR (CSF, PBMC), and BDV serology, and (2) a separate group of 25 retrospectively studied patients with AIDS dementia complex was evaluated using BDV serology only. A novel, BDV p40 gene RT-PCR assay was developed: conserved primers were used in a non-nested amplification, detecting less than 100 BDV RNA copies and all of nine wild-type, confirmed animal BDV infections. BDV seroprevalences were 12.5% and 8.0%, respectively, which are similar to the general HIV-infected population. None of the prospectively studied patients had detectable BDV RNA in their CSF or PBMC. Our findings do not support the hypothesis that BDV infections are responsible for HIV-related neurological disorders.

[Investigations on the treatment of cerebrospinal nematode infections in goats]. / Untersuchungen zur Behandlung der zerebrospinalen Nematodiasis bei Ziegen.

In 17 goats the therapeutical outcome of cerebrospinal nematodiasis, a parasitic infection in the central nervous system with the nematode Elaphostrongylus cervi, is described. The diagnosis was made on the basis of physical findings and cerebrospinal fluid eosinophilia. The therapy was based on the administration of a non-steroidal antiinflammatory drug (flunixin meglumine) and two anthelmintics (fenbendazol, ivermectin) for five days. The response to therapy was documented immediately and 30 days after therapy and by owners report up to 30 months. Eleven goats (64.7%) showed an improvement of the neurological signs directly after therapy. Three had a complete recovery. Signs of a light and middle ataxie were still present in 6 and 2 goats, respectively. Six recumbent goats (35.3%) had to be euthanasied because of a non-therapeutical response. This study has shown a clinical impression of successful use of flunixin meglumine, fenbendazole and ivermectin in goats with light to middle neurological signs. The goats could accommodate the remaining ataxia without showing a reduced production. Goats with high neurological deficits could not be successful treated based on the high degree of the neurological damage.

Neuropathological and aetiological studies of sporadic non-suppurative meningoencephalomyelitis of cattle.

Sporadically occurring non-suppurative encephalitis appears to be a frequent condition of Swiss cattle. Fifty-one such cases diagnosed over a period of 10 years were examined retrospectively to investigate whether they constituted one or more distinct diseases, and to search for aetiological agents. Three cases were characterised by periventricular granulomatous encephalitis, and most probably represented a different disease, but the remaining 48 cases had disseminated non-suppurative encephalitis with widespread neuronal changes. Neuronal degeneration was very marked in the hippocampus of 10 cases and in the cerebellar Purkinje cells of 11. It was thought that the latter cases represented morphological variations of the same disease rather than a different disease because of their overlapping morphological features. The 48 cases had the following features in common: the disease had primarily neurological signs affecting mostly adult cattle, it was a sporadic condition, and there was a clear tendency for it to have a subacute to chronic course. Polymerase chain reaction (PCR) amplification for chlamydial DNA was negative except in one of 32 specimens, and immunohistochemistry did not demonstrate the presence of chlamydial antigens either in the one PCR-positive case or in the other cases examined. Immunohistochemistry for rabies virus, Borna disease virus, and central European tickborne encephalitis virus was negative. In four cases, immunolabelled cells were found in the lesions with antibodies against paramyxovirus antigens.

Borna disease in a dog with lethal meningoencephalitis.

A dog was euthanatized because of progressive neurological signs. Histologically, a nonsuppurative meningoencephalitis was found. By immunohistochemistry, in situ hybridization, and nested PCR procedures, Borna disease virus (BDV) antigen and BDV-specific RNA were demonstrated in brain tissues of the dog. The nucleotide sequence of the PCR product showed 94 to 98% homology to published BDV sequences. This is the first description of Borna disease in a dog.

[Pestivirus as causative agent of abortion and perinatal mortality in cattle and sheep in Switzerland]. / Ursächliche Beteiligung von Pestiviren an Aborten und perinatalen Todesfällen bei Rindern und Schafen in der Schweiz.

The causal involvement of bovine viral diarrhoea virus (BVDV) and border disease virus (BDV) infection in bovine and ovine abortion and perinatal mortality remain unclear. From 1992 until 1994, 213 bovine and 31 ovine foetuses as well as 36 calves and 25 lambs which had died perinatally were investigated. Tissue samples were tested for the presence of pestiviruses and serum or fluid from the body cavities were analysed for the presence of pestivirus antibodies. Detection of pestiviruses was performed by (i) cell culture isolation, (ii) antigen ELISA and (iii) immunohistochemical staining for viral antigen. For antibody-testing an indirect ELISA was used. In nine bovine foetuses and in two calves BVDV was isolated. Pestiviruses, most likely BDV were detected in one ovine foetus and three lambs. In 6% of the bovine and 11% of the ovine foetuses anti-pestivirus antibodies were detected. However, clinical features and history of bovine cases did not show a correlation with the diagnostic results, In contrast, the presence of central nervous system signs in neonatal lambs and the detection of BDV was correlated.

Spontaneous Borna disease in sheep and horses: immunophenotyping of inflammatory cells and detection of MHC-I and MHC-II antigen expression in Borna encephalitis lesions.

Borna disease (BD) has been recognized as a virally induced T-cell dependent immunopathological disorder of the central nervous system (CNS), as shown by experimental infection of rats with Borna disease virus (BDV). In contrast to the rat model, little is known about the pathogenesis of spontaneous BD in sheep and horses. The present study describes the brain lesions of 12 ovine and 11 equine cases of naturally occurring BD. A set of monoclonal and polyclonal antibodies was used in order to determine the cells operative in encephalitic lesions and to detect expression of MHC-I and MHC-II products in the brains of affected animals. In all cases investigated, a reaction pattern similar to that reported for the acute phase of BD in experimentally infected rats was noted. In brief, the majority of inflammatory cells in perivascular infiltrates (PVI) as well as parenchymal and meningeal infiltrates were CD3 +. CD4 + cells outnumbered CD8 + cells in PVI as well as in the parenchyma. Macrophages (defined by lysozyme immunoreactivity) were seen less often and B-cells or plasma cells (cells positive for lambda or kappa light chains) were demonstrated at lower numbers. TCR-1 + cells were found on very rare occasions in PVI of some sheep. MHC-I and MHC-II products were constantly expressed on inflammatory cells but inconsistently on astrocytes and neurons. Neuronal degeneration was not a major feature.

[Polycythemia caused by liver carcinoma in cattle and sheep]. / Polyglobulie infolge Leberkarzinom bei Rind und Schaf.

Polycythemia associated with hepatic carcinoma was diagnosed in a 10-year-old cow, an 8-month-old heifer and a 3-year-old sheep. The cow was referred to our clinic because of weight loss, reduced appetite, hematuria, marked reddening of the mucosa of the tear canal, oral cavity and vestibule and an increased hematocrit. Clinical examination also revealed injected scleral vessels. The erythrocyte count, the PCV, hemoglobin concentration and the activities of the hepatic enzymes were increased. Ultrasonographic examination revealed a very large liver with a focal echogenic lesion. Based on all findings, a diagnosis of polycythemia associated with a liver tumor, was made. The cow was slaughtered. Multifocal liver tumors were diagnosed histologically as hepatocellular carcinoma. The heifer and sheep had similar clinical and hematological findings. Hepatocellular carcinoma was diagnosed in the heifer and cholangiocellular carcinoma in the sheep. It was concluded that in all three patients, polycythemia was caused by hepatic carcinoma.

[Cerebrospinal nematodiasis in seven goats]. / Zerebrospinale Nematodiasis bei sieben Ziegen.

Clinical findings in seven goats affected with cerebrospinal nematodiasis are described. The animals originated from different parts of Switzerland. The disease occurred mainly in winter. The animals were admitted to the clinic because of progressive pelvic limb ataxia, recumbency, vestibular disease and circling. Clinical findings were complete or incomplete posterior paresis, pelvic limb ataxia, circling, reduced cutaneous sensation and proprioceptive deficits as well as cranial nerve reflexes deficits. The general condition was slightly reduced and the appetite was normal. In three goats predominance of mononuclear and eosinophilic cells in the cerebrospinal fluid was interpreted as typical findings for parasite infestation in the central nervous system. Histopathological changes and the finding of a nematode in cross sections in two affected animals confirmed the diagnosis. Infection with Elaphostrongylus cervi is discussed due to close contact with deer.

Echocardiographic diagnosis of a cardiac fibrosarcoma in the right atrium of a sheep.

This is a case report of a six-year-old female White Alpine sheep with a cardiac fibrosarcoma in the right atrium. Clinically, the sheep had right-sided cardiac insufficiency with tachycardia, engorgement of the jugular veins, brisket edema, and ascites. Chronic congestion of the liver resulted in increased hepatic enzyme activity. Based on clinical findings, a tentative diagnosis of endocarditis or pericarditis was made. Radiography of the thorax revealed hydrothorax. An echogenic mass was observed in the right atrium via echocardiography; it was interpreted as a tumor or thrombus. Ultrasonography of the abdomen revealed severe ascites and chronic congestion of the liver attributable to right-sided cardiac insufficiency. The clinical and sonographic findings were verified at post mortem. The mass in the right atrium was a pedunculated fibrosarcoma.

Borna disease in naturally infected cattle.

Based on the immunohistochemical demonstration of viral antigen and on the histological brain lesions, Borna disease was diagnosed in a cow and a bull which had suffered from a severe, subacute progressive disorder of the central nervous system. Virus-specific antigen was characteristically localized in neurons, predominantly in the perikaryon and dendrites. In a serum sample available from one of the animals a Borna disease virus antibody titre of 1 in 80 was demonstrated. This is the first report of the natural disease in cattle.