ABSTRACT
OBJECTIVE: To determine the characteristic clinical and spinal MRI phenotypes of sarcoidosis-associated myelopathy (SAM), we analyzed a large cohort of patients with this disorder. METHODS: Patients diagnosed with SAM at a single center between 2000 and 2018 who met the established criteria for definite and probable neurosarcoidosis were included in a retrospective analysis to identify clinical profiles, CSF characteristics, and MRI lesion morphology. RESULTS: Of 62 included patients, 33 (53%) were male, and 30 (48%) were African American. SAM was the first clinical presentation of sarcoidosis in 49 patients (79%). Temporal profile of symptom evolution was chronic in 81%, with sensory symptoms most frequently reported (87%). CSF studies showed pleocytosis in 79% and CSF-restricted oligoclonal bands in 23% of samples tested. Four discrete patterns of lesion morphology were identified on spine MRI: longitudinally extensive myelitis (n = 28, 45%), short tumefactive myelitis (n = 14, 23%), spinal meningitis/meningoradiculitis (n = 14, 23%), and anterior myelitis associated with areas of disc degeneration (n = 6, 10%). Postgadolinium enhancement was seen in all but 1 patient during the acute phase. The most frequent enhancement pattern was dorsal subpial enhancement (n = 40), followed by meningeal/radicular enhancement (n = 23) and ventral subpial enhancement (n = 12). In 26 cases (42%), enhancement occurred at locations with coexisting structural changes (e.g., spondylosis). CONCLUSIONS: Recognition of the clinical features (chronically evolving myelopathy) and distinct MRI phenotypes (with enhancement in a subpial and/or meningeal pattern) seen in SAM can aid diagnosis of this disorder. Enhancement patterns suggest that SAM may have a predilection for areas of the spinal cord susceptible to mechanical stress.
Subject(s)
Central Nervous System Diseases , Sarcoidosis , Spinal Cord Diseases , Adult , Central Nervous System Diseases/cerebrospinal fluid , Central Nervous System Diseases/diagnostic imaging , Central Nervous System Diseases/pathology , Female , Humans , Leukocytosis/cerebrospinal fluid , Magnetic Resonance Imaging , Male , Meningitis/cerebrospinal fluid , Meningitis/diagnostic imaging , Meningitis/pathology , Middle Aged , Myelitis/cerebrospinal fluid , Myelitis/diagnostic imaging , Myelitis/pathology , Radiculopathy/cerebrospinal fluid , Radiculopathy/diagnostic imaging , Radiculopathy/pathology , Retrospective Studies , Sarcoidosis/cerebrospinal fluid , Sarcoidosis/diagnostic imaging , Sarcoidosis/pathology , Spinal Cord Diseases/cerebrospinal fluid , Spinal Cord Diseases/diagnostic imaging , Spinal Cord Diseases/pathologyABSTRACT
Geminiviruses are single-stranded DNA viruses that infect a wide variety of plants and cause severe crop losses worldwide. The geminivirus replication initiator protein (Rep) binds to the viral replication origin and catalyzes DNA cleavage and ligation to initiate rolling circle replication. In this study, we found that the Tomato golden mosaic virus (TGMV) Rep is phosphorylated at serine-97 by sucrose nonfermenting 1-related protein kinase 1 (SnRK1), a master regulator of plant energy homeostasis and metabolism. Phosphorylation of Rep or the phosphomimic S97D mutation impaired Rep binding to viral DNA. A TGMV DNA-A replicon containing the Rep S97D mutation replicated less efficiently in tobacco (Nicotiana tabacum) protoplasts than in wild-type or Rep phosphorylation-deficient replicons. The TGMV Rep-S97D mutant also was less infectious than the wild-type virus in Nicotiana benthamiana and was unable to infect tomato (Solanum lycopersicum). Nearly all geminivirus Rep proteins have a serine residue at the position equivalent to TGMV Rep serine-97. SnRK1 phosphorylated the equivalent serines in the Rep proteins of Tomato mottle virus and Tomato yellow leaf curl virus and reduced DNA binding, suggesting that SnRK1 plays a key role in combating geminivirus infection. These results established that SnRK1 phosphorylates Rep and interferes with geminivirus replication and infection, underscoring the emerging role for SnRK1 in the host defense response against plant pathogens.
Subject(s)
Begomovirus/metabolism , Plant Proteins/metabolism , Protein Serine-Threonine Kinases/metabolism , Viral Proteins/metabolism , Virus Replication , Amino Acid Sequence , Begomovirus/genetics , Begomovirus/physiology , Host-Pathogen Interactions , Solanum lycopersicum/enzymology , Solanum lycopersicum/virology , Mutation , Phosphorylation , Plant Diseases/virology , Plant Proteins/genetics , Protein Binding , Protein Domains , Protein Serine-Threonine Kinases/genetics , Sequence Homology, Amino Acid , Serine/genetics , Serine/metabolism , Viral Proteins/chemistry , Viral Proteins/geneticsABSTRACT
Coupled with the advantages afforded by the model plant Arabidopsis, virus-induced gene silencing (VIGS) offers a rapid means to assess gene function. The geminivirus vector based on Cabbage leaf curl virus described here has the benefits of small insert size and persistent silencing of the target gene through the life cycle of the plant. Here, we show that genetic variation in the vast collection of Arabidopsis accessions can be leveraged to ameliorate viral symptomology that accompanies the VIGS procedure. The plasticity of phenotypes under different day lengths or temperature conditions can be exploited to achieve maximum silencing efficacy in either vegetative or inflorescence tissue, according to the question being asked. Protocols and vectors for Agro-infiltration of primary leaves, subapical pricking in older plants, and microprojectile bombardment are described.
