Critical Role of Perforin-dependent CD8+ T Cell Immunity for Rapid Protective Vaccination in a Murine Model for Human Smallpox

Vaccination is highly effective in preventing various infectious diseases, whereas the constant threat of new emerging pathogens necessitates the development of innovative vaccination principles that also confer rapid protection in a case of emergency. Although increasing evidence points to T cell immunity playing a critical role in vaccination against viral diseases, vaccine efficacy is mostly associated with the induction of antibody responses. Here we analyze the immunological mechanism(s) of rapidly protective vaccinia virus immunization using mousepox as surrogate model for human smallpox. We found that fast protection against lethal systemic poxvirus disease solely depended on CD4 and CD8 T cell responses induced by vaccination with highly attenuated modified vaccinia virus Ankara (MVA) or conventional vaccinia virus. Of note, CD4 T cells were critically required to allow for MVA induced CD8 T cell expansion and perforin-mediated cytotoxicity was a key mechanism of MVA induced protection. In contrast, selected components of the innate immune system and B cell-mediated responses were fully dispensable for prevention of fatal disease by immunization given two days before challenge. In conclusion, our data clearly demonstrate that perforin-dependent CD8 T cell immunity plays a key role in MVA conferred short term protection against lethal mousepox. Rapid induction of T cell immunity might serve as a new paradigm for treatments that need to fit into a scenario of protective emergency vaccination

Cutaneous apocrine gland neoplasia in 16 captive African wild dogs (Lycaon pictus)

AVAILABILITY OF DATA AND MATERIAL : Part of the case material included in the present study that derived from France was included in a poster presentation on familial occurrence of CATs in AWDs (cases 2, 9, 10 and 11) [19]. These cases and cases 3 and 8 were also included in a poster presentation as part of a retrospective study on neoplasms in AWDs in French zoological collections [7].This retrospective study documents the occurrence of single and multiple cutaneous apocrine gland tumours (CATs) on the dorsal midline of 16 captive African wild dogs (AWDs, Lycaon pictus) derived from 161 submissions to diagnostic laboratories in South Africa, France and Germany between 1997 and 2022. Animals included in the study came from zoological institutions in South Africa (n ¼ 2), France (n ¼ 5) and Germany (n ¼ 1) and ranged from 5 to 14 years of age. Fifteen affected animals were female (94%) and one was male. CATs presented as raised, hairless, multilobular, grey firm masses, consistently located along the dorsal midline. Apart from a single cutaneous apocrine adenoma and a cystadenoma occurring concurrently with two non-cystic adenocarcinomas, neoplasms were consistent with malignant cutaneous apocrine adenocarcinomas with lymphatic spread and visceral metastases. Advanced age and female sex were identified as risk factors. A genetic component or association with the increasing use of GnRH agonist contraceptives was suspected but could not be established. This study highlights the need for close clinical monitoring of AWDs over the age of 5 years for the development of CATs along the dorsal midline and supports early surgical intervention. More research is needed to determine the role of inbreeding, endocrine changes and husbandry factors that may play a role in the development of CATs on the dorsal midline of AWDs.http://www.elsevier.com/locate/jcpaam2024Centre for Veterinary Wildlife StudiesParaclinical SciencesSDG-03:Good heatlh and well-bein

Isolation of canine Anaplasma phagocytophilum strains from clinical blood samples using the Ixodes ricinus cell line IRE/CTVM20

AbstractAnaplasma phagocytophilum is an intracellular tick-borne rickettsial pathogen, which causes granulocytic anaplasmosis in various species of livestock and companion animals and also in humans. Previously A. phagocytophilum has been isolated and propagated in cell lines derived from the tick Ixodes scapularis and in the human promyelocytic cell line HL60. In this study we used the Ixodes ricinus-derived cell line IRE/CTVM20 to isolate and propagate two new canine strains of A. phagocytophilum.Blood samples were collected by veterinarians from two dogs, one from Germany and the other from Austria. Suspicion of clinical canine granulocytic anaplasmosis was raised by the treating veterinarians and after confirmation of A. phagocytophilum infection by real-time PCR, buffy coat cells were isolated and co-cultivated with IRE/CTVM20 cells maintained at 28°C in L15/L15B medium.In the tick cells, rickettsial inclusions were first recognised after 86 days of incubation. Electron microscopic examination of tick cells infected with one of the isolates revealed cytoplasmic vacuoles containing pleomorphic organisms with individual bacteria enveloped by a bilayer membrane. Sequencing of 16S rRNA genes confirmed the isolation of A. phagocytophilum and showed the highest identity to the A. phagocytophilum human HZ strain. The two A. phagocytophilum isolates were passaged several times in IRE/CTVM20 cells and transferred to the I. scapularis cell line ISE6. This confirms for the first time the successful establishment and continuous cultivation of this pathogen in I. ricinus cells as well as infectivity of these canine strains for I. scapularis cells

Immobility-associated thromboprotection is conserved across mammalian species from bear to human

Venous thromboembolism (VTE) comprising deep venous thrombosis and pulmonary embolism is a major cause of morbidity and mortality. Short-term immobility-related conditions are a major risk factor for the development of VTE. Paradoxically, long-term immobilized free-ranging hibernating brown bears and paralyzed spinal cord injury (SCI) patients are protected from VTE. Here we aimed to identify mechanisms of immobility-associated VTE protection in a cross-species approach. Mass spectrometry-based proteomics revealed an antithrombotic signature in platelets of hibernating brown bears with heat shock protein 47 (HSP47) as most substantially reduced protein. HSP47 downregulation or ablation attenuated immune cell activation and NET formation, contributing to thromboprotection in bears, SCI patients and mice. This cross-species conserved platelet signature may give rise to antithrombotic therapeutics and prognostic markers beyond immobility-associated VTE

Histomorphology and Immunohistochemistry of a Congenital Nephromegaly Demonstrate Concurrent Features of Heritable and Acquired Cystic Nephropathies in a Girgentana Goat (Capra falconeri)

Polycystic kidney diseases (PKD) represent frequent congenital and adult nephropathies in humans and domestic animals. This report illustrates an uncommon state of congenital PKD in a girgentana goat (Capra falconeri). A stillborn female goat kid was submitted for postmortem examination and underwent macroscopic and microscopic examination. The kidneys showed a bilateral nephromegaly and a perpendicular polycystic altered texture of the renal parenchyma. Renal tissue sections were comprehensively investigated by histopathology (overview and special stains), immunohistochemistry (CD10, CD117, pan-cytokeratin, cytokeratin 7, E-cadherin, Pax2, Pax8, and vimentin), and electron microscopy (SEM, TEM). Histopathology of renal tissue sections revealed polycystic alterations of the renal parenchyma as well as conspicuous polypoid proliferates/projections of the renal tubular epithelium, which showed clear cell characteristics. Furthermore, epithelial projections were indicative for epithelio-mesenchymal-transition, cellular depolarization, and strong expression of differentiation markers Pax2, Pax8, and CD10. Ultrastructural morphology of the projections was characterized by numerous diffusely distributed, demarked round cytoplasmic structures and several apico-lateral differentiations. Additionally, hepatic malformations comprising biliary duct proliferation with saccular dilation and bridging fibrosis were observed. Notably, this report describes the first case of a congenital cystic nephropathy with overlapping features of heritable and acquired nephropathies in any species. Epithelio-mesenchymal-transition and altered cadherin expression seem to be crucial components of a suspected pathomechanism during cystogenesis

Papillomavirus Antijenlerinin Kedi Oral Ve Kutano¨z Skuamo¨z Hu¨creli Karsinomalarda Tespiti

PAPI·LLOMAVI·RUS ANTI·JENLERI·NI·N KEDI· ORAL VE KUTANO¨Z SKUAMO¨Z HU¨CRELI· KARSI·NOMALARDA TESPI·TI· 1Veteriner Patolojisi Enstitu¨su¨, Mu¨nih Ludwig Maximilian U¨niversitesi, 80539, Mu¨nih, Almanya 2Patoloji Anabilim Dalı, Veteriner Faku¨ltesi, I·stanbul U¨niversitesi-Cerrahpas¸a, 34320, I·stanbul, Tu¨rkiye Skuamo¨z hu¨creli karsinom (SCC), kedilerin oldukc¸a yaygın ve malign seyreden neoplazmlarıdır. C¸es¸itli fakto¨rler kedilerde SCC olus¸umuna yol ac¸maktadır. Bunlardan biri de, insanlarda da go¨ru¨ldu¨gˆu¨ gibi, papillomaviru¨s (PV) olarak belirtilmis¸tir. Bu c¸alıs¸mada, kedilerde bas¸ bo¨lgesinde bulunan oral ve kutano¨z SCC`lerin histopatolojik olarak degˆerlendirilmesi ve papillomaviru¨s ile arasındaki ilis¸kisinin ortaya konulması amac¸lanmıs¸tır. C¸alıs¸mada, Ludwig Maximilian U¨niversitesi Mu¨nih Veteriner Faku¨ltesi Patoloji Enstitu¨su¨nu¨n veri tabanından kırk kediden kırk u¨c¸ doku o¨rnegˆi kullanıldı. Is¸ık mikroskobu ile parafin doku kesitleri; hematoksilen eozin (H&E) ve immu¨nohistokimyasal (IHC) boya metotları kullanılarak degˆerlendirildi. Ayrıca PV, epoksi resin doku kesitleri kullanılarak transmisyon elektron mikroskobu (TEM) ile incelendi. Histopatolojik incelemede, 23/43 (%53,4) iyi diferansiye, 14/43 (%32,5) ko¨tu¨ diferansiye ve 6/43 (%13,9) iyi ve ko¨tu¨ diferansiye SCC bir arada go¨zlemlendi. Kedilerin yas¸ ortalaması 11,4 (dagˆılım 7 - 19 yas¸ arası) olarak belirlendi. Neoplastik lezyonlarda IHC ile PV antijeni; 6/43 doku o¨rnegˆinde pozitif olarak saptandı. Ancak, TEM ile viral parc¸acık bulunmadı. Ayrıca farklı diferansiyasyona sahip SCC’ler ile IHC pozitifligˆi arasında korelasyon go¨zlenmedi. C¸alıs¸mada, kedilerde bas¸ bo¨lgesindeki SCC histopatolojik olarak Du¨nya Sagˆlık O¨rgu¨tu¨`nu¨n (WHO) kriterleri go¨z o¨nu¨ne alınarak iyi ve ko¨tu¨ diferansiye olarak sınıflandırıldı. Bununla birlikte, SCC olus¸umunda PV`lerin olası rolu¨ IHC ve transmisyon elektron mikroskobisi kullanılarak aras¸tırıldı. Her ne kadar TEM de viral partiku¨lere rastlanılmasa da immunohistokimya sonuc¸larında go¨zlenen pozitif reaksiyonlar, bu konu ile ilgili digˆer c¸alıs¸malarla uyumlu bulundu. Sonuc¸ta IHC ile elde edilen bulgular, PV`ların SCC olus¸umunda indu¨kleyici bir role sahip olabilecegˆini du¨s¸u¨ndu¨ru¨r niteliktedir. Anahtar so¨zcu¨kler: I·mmunohistokimya, kedi, papillomavirus, skuamo¨z hu¨creli karsinom, transmisyon elektron mikroskobu

A nonsense mutation in PLD4

Background: Bovine hereditary zinc deficiency (BHZD) is an autosomal recessive disorder of cattle, first described in Holstein-Friesian animals. Affected calves suffer from severe skin lesions and show a poor general health status. Recently, eight calves with the phenotypic appearance of BHZD have been reported in the Fleckvieh cattle population. Results: In spite of the similar disease phenotypes, SLC39A4, the gene responsible for BHZD in Holstein-Friesian was excluded as underlying gene for the disorder in the affected Fleckvieh calves. In order to identify the disease-associated region, genotypes of eight affected calves obtained with the Illumina BovineHD BeadChip comprising 777,962 SNPs were contrasted with the genotypes of 1,339 unaffected animals. A strong association signal was observed on chromosome 21 (P = 5.87 x 10(-89)). Autozygosity mapping in the eight affected animals revealed a common segment of extended homozygosity encompassing 1,023 kb (BTA 21: 70,550,045 - 71,573,501). This region contains 17 genes/transcripts, among them two genes encoding gastro-intestinal zinc transporters (CRIP1, CRIP2). However, no mutation that was compatible with recessive inheritance could be detected in these candidate genes. One of the affected calves was re-sequenced together with 42 unaffected Fleckvieh animals. Analysis of the sequencing data revealed a nonsense mutation (p.W215X) in a phospholipase encoding gene (PLD4) as candidate causal polymorphism. To confirm the causality, genotypes of the p.W215X-mutation were obtained from 3,650 animals representing three different breeds. None of the unaffected animals was homozygous for the defect allele, while all eight affected calves were homozygous. The deleterious effect of the mutation is manifested in a significantly lower survival rate of descendants from risk matings when compared with the survival rate of descendants from non-risk matings. The deleterious allele has an estimated frequency of 1.1\% in the Fleckvieh population. Conclusion: Our results provide strong evidence that a newly identified recessive disorder in the Fleckvieh population is caused by a nonsense mutation in PLD4, most likely resulting in an impaired function of the encoded protein. Although the phenotype of affected calves strongly resembles BHZD, a zinc deficiency resulting from malabsorption is unlikely to be responsible for the diseased Fleckvieh calves

Hair follicle dystrophy in a litter of domestic cats resembling lanceolate hair mutant mice

Background: A new congenital hair-shaft abnormality resembling the lanceolate hair phenotype of rodents is described in a litter of four domestic short hair (DSH) cats. Data relating to hair shaft and follicle disorders remain scarce in veterinary medicine. Objectives: To describe and compare structural abnormalities in these cats with other hair dystrophies in cats and other mammals. Animals: A DSH cat litter with progressive noninflammatory alopecia. Methods and materials: Histopathological evaluation, scanning and transmission electron microscopy, and X-ray based element analysis defined the hair and skin changes in cats born with alopecia. Findings were compared to archival data from normal cats and lanceolate hair (Dsg4lahJ ) and Keratin 75 (Krt75tm1Der ) mutant mice. Results: Light and scanning electron microscopy of the hairs revealed lance- or spear-head shaped defects of the hair tip. Histological findings were swollen hair shafts, initially above the hair bulb matrix and later found in the distal parts of the telogen hair follicles, similar to those observed in Dsg4lahJ Krt75tm1Der mutant mice. Transmission electron microscopy of the hair shaft and hair follicles showed a loss in the normal structure of the guard hairs in the alopecic cats. There was a statistically significant decrease in sulfur content just below the defects in the hair shafts (trichothiodystrophy). Conclusion and clinical importance: A rare form of congenital alopecia resulting in follicular dystrophy is described in cats which is similar to hair follicle and hair-shaft changes reported in several mutant mouse strains with single gene mutations in adhesion molecules or keratin genes

Hair follicle dystrophy in a litter of domestic cats resembling lanceolate hair mutant mice.

BACKGROUND: A new congenital hair-shaft abnormality resembling the lanceolate hair phenotype of rodents is described in a litter of four domestic short hair (DSH) cats. Data relating to hair shaft and follicle disorders remain scarce in veterinary medicine. OBJECTIVES: To describe and compare structural abnormalities in these cats with other hair dystrophies in cats and other mammals. ANIMALS: A DSH cat litter with progressive noninflammatory alopecia. METHODS AND MATERIALS: Histopathological evaluation, scanning and transmission electron microscopy, and X-ray based element analysis defined the hair and skin changes in cats born with alopecia. Findings were compared to archival data from normal cats and lanceolate hair (Dsg4 RESULTS: Light and scanning electron microscopy of the hairs revealed lance- or spear-head shaped defects of the hair tip. Histological findings were swollen hair shafts, initially above the hair bulb matrix and later found in the distal parts of the telogen hair follicles, similar to those observed in Dsg4 CONCLUSION AND CLINICAL IMPORTANCE: A rare form of congenital alopecia resulting in follicular dystrophy is described in cats which is similar to hair follicle and hair-shaft changes reported in several mutant mouse strains with single gene mutations in adhesion molecules or keratin genes