scholarly journals 1876. A Hepacivirus-Like Protein Is Targeted by the Antibody Response to Kawasaki Disease (KD)

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S48-S48
Author(s):  
Anne Rowley ◽  
Susan Baker ◽  
David Arrollo ◽  
Leah Gruen ◽  
Bodnar Tetyana ◽  
...  
Keyword(s):  
Inclusion Bodies ◽  
Nonstructural Protein ◽  
Infectious Agent ◽  
Bronchial Epithelium ◽  
Fourth Generation ◽  
Single Patient ◽  
Glutathione S Transferase ◽  
Coronary Aneurysms ◽  
Intracytoplasmic Inclusion Bodies ◽  
Portal Of Entry

Abstract Background Clinical and epidemiologic data support a viral cause of KD, but the etiology has eluded 50 years of study. We previously identified virus-like intracytoplasmic inclusion bodies (ICI) in ciliated bronchial epithelium of KD children but not infant controls, but the antigens within the ICI were unknown. At 1–2 weeks following infection, 75% of peripheral blood plasmablasts (PB) specifically target the infectious agent. We cloned the PB response to KD to identify KD-specific antibodies and their target antigens. Methods We isolated single PB from children with KD 1–3 weeks after fever onset by flow cytometry, and amplified immunoglobulin VDJ and VJ genes from each PB by RT-PCR. We sequenced the products and made monoclonal antibodies (Mab) from clonally expanded PB in individual patients. Mab were tested for binding to KD tissues and to a viral peptide array containing 29,939 peptides from known B cell epitopes of animal viruses (www.iedb.org). Results We sequenced 1156 PB from 11 KD patients, and identified 44 clonally expanded sets of PB. We prepared 61 Mab from clonally expanded and highly mutated IgA PB, and found that 33/61 bind to KD ICI, 10 strongly and 23 weakly. Of 10 Mab that strongly bind, 2 were VH3-33 (single patient), 2 VH3-23 (single patient), 1 VH3-15, 1 VH3-74, 3 VH1-46 (2 patients), and 1 VH4-59. These Mab CDR3s varied from 11 to 20 aacids, with 4–28 aacid mutations. Mab KD4-2H4 recognized multiple similar peptides from nonstructural protein 4A of hepacivirus C; pt KD4 sera was negative for hepatitis C by fourth-generation ELISA. Amino acid substitution analysis yielded an optimized peptide, and 6 KD Mab recognized this peptide by ELISA. These 6 Mab derived from 3 KD patients, all of whom had coronary aneurysms, and were VH3-74 (n = 1), VH3-33 (n = 2, single patient), VH1-45 (n = 1), and VH3-72 (n = 2, single patient). Strong binding of KD Mab KD4-2H4 and KD6-2B2 to ICI was totally blocked by pre-incubation with optimized peptide. KD but not control sera react with optimized peptide expressed as a glutathione S-transferase fusion protein by western blot. Conclusion Children with KD make antibodies to a hepacivirus-like protein, and KD ICI contain this protein. These results strongly suggest that a previously unidentified hepacivirus with a respiratory portal of entry is etiologically related to KD. Disclosures All Authors: No reported Disclosures.

scholarly journals A Protein Epitope Targeted by the Antibody Response to Kawasaki Disease

2020 ◽  
Vol 222 (1) ◽  
pp. 158-168 ◽  
Author(s):  
Anne H Rowley ◽  
Susan C Baker ◽  
David Arrollo ◽  
Leah J Gruen ◽  
Tetyana Bodnar ◽  
...  
Keyword(s):  
Kawasaki Disease ◽  
Antibody Response ◽  
Peripheral Blood ◽  
Inclusion Bodies ◽  
Specific Antigen ◽  
Illness Onset ◽  
Coronary Aneurysms ◽  
Acquired Heart Disease ◽  
Intracytoplasmic Inclusion Bodies ◽  
Developed Nations

Abstract Background Kawasaki disease (KD) is the leading cause of childhood acquired heart disease in developed nations and can result in coronary artery aneurysms and death. Clinical and epidemiologic features implicate an infectious cause but specific antigenic targets of the disease are unknown. Peripheral blood plasmablasts are normally highly clonally diverse but the antibodies they encode are approximately 70% antigen-specific 1–2 weeks after infection. Methods We isolated single peripheral blood plasmablasts from children with KD 1–3 weeks after onset and prepared 60 monoclonal antibodies (mAbs). We used the mAbs to identify their target antigens and assessed serologic response among KD patients and controls to specific antigen. Results Thirty-two mAbs from 9 of 11 patients recognize antigen within intracytoplasmic inclusion bodies in ciliated bronchial epithelial cells of fatal cases. Five of these mAbs, from 3 patients with coronary aneurysms, recognize a specific peptide, which blocks binding to inclusion bodies. Sera from 5/8 KD patients day ≥ 8 after illness onset, compared with 0/17 infant controls (P < .01), recognized the KD peptide antigen. Conclusions These results identify a protein epitope targeted by the antibody response to KD and provide a means to elucidate the pathogenesis of this important worldwide pediatric problem.

Round Table Discussion

PEDIATRICS ◽  
1948 ◽  
Vol 2 (4) ◽  
pp. 469-479
Author(s):  
RUSSELL J. BLATTNER
Keyword(s):  
Inclusion Bodies ◽  
Cellular Response ◽  
Infectious Agent ◽  
Round Table ◽  
Chemotherapeutic Agents ◽  
Tissue Response ◽  
Round Table Discussion ◽  
Obligate Intracellular ◽  
Form Of Life ◽  
Intracellular Parasites

Chairman Blattner: During recent years, there has been increasing interest shown in diseases caused by filterable viruses, and significant work has been accomplished in this comparatively new and absorbing field of endeavor. With the advent of chemotherapeutic agents and antibiotics, the presence and action of these infectious agents has become more apparent. Viral diseases, therefore, have assumed increasing importance in medical literature in general and in pediatric literature in particular. By way of review, it is well to bear in mind that viruses are filter-passing agents, obligate intracellular parasites, capable of reproducing themselves and of producing disease in plants and animals, including man. While these agents cannot be seen except by the most elaborate methods, their presence can be detected by their injurious effects. The pathologic picture produced by viral agents is rather characteristic and can be recognized readily by experienced observers acquainted with tissue response. In some instances, inclusion bodies are produced which may be intranuclear or intracytoplasmic, and represent cytologic changes which are considered typical of the pathologic response to viral invasion. When inclusion bodies are present they may serve as sign posts for the recognition of the type of infectious agent. The nature of a filterable virus is as yet unknown. Viruses may be a form of life similar to bacteria, but infinitely smaller in size. It is conceivable that viruses are enzymes capable of reproducing themselves and capable of producing cellular response. They may be non-living, crystallizable substances, such as the Stanley tobacco-mosaic virus; or a form of life, the definite nature of which is as yet unrecognized. Dr. Thomas M. Rivers has stated : "Viruses are a heterogeneous collection of diverse agents which happen to induce a state of broad similarity." He points out that the reaction of the tissues in general, and of the cells in particular, determines the nature of the pathologic process about as much as the infectious agent itself.

Expression, purification, and characterization of the carboxyl-terminal region of the Na+/H+ exchanger

1998 ◽  
Vol 76 (5) ◽  
pp. 837-842 ◽  
Author(s):  
Daniel Gebreselassie ◽  
Krishna Rajarathnam ◽  
Larry Fliegel
Keyword(s):  
Circular Dichroism ◽  
Membrane Protein ◽  
Fusion Protein ◽  
Gel Electrophoresis ◽  
Inclusion Bodies ◽  
Cytoplasmic Domain ◽  
Random Coil ◽  
Glutathione S Transferase ◽  
Carboxyl Terminal ◽  
Circular Dichroïsm

The Na+/H+ exchanger is a pH regulatory protein that is responsible for removal of excess intracellular protons in exchange for extracellular Na+. It is a plasma membrane protein with a large cytoplasmic carboxyl terminal domain that regulates activity of the membrane domain. We overexpressed and purified the cytoplasmic domain that was produced in Escherichia coli. This region (516-815 amino acids) was under control of the tac promoter from the plasmid pGEX-KG and was fused with glutathione S-transferase. Upon induction, the fusion protein was principally found in inclusion bodies. Purified inclusion bodies were solubilized and fractionated using preparative SDS polyacrylamide gel electrophoresis. To obtain free Na+/H+ exchanger protein the fusion protein was dialyzed against cleavage buffer and cleaved at the thrombin cleavage site between glutathione S-transferase and the Na+/H+ exchanger domain. Free Na+/H+ exchanger protein was obtained by rerunning the sample on preparative gel electrophoresis. The final yield of the purified protein was 2.15 mg protein/L of cell culture. After exhaustive dialysis the secondary structure of the purified protein was assessed using circular dichroism spectroscopy. The results indicated that the protein was 35% alpha-helix, 17% beta-turn, and 48% random coil. They suggest that the cytoplasmic domain is structured and some regions may be compact in nature.Key words: Na+/H+ exchanger, pH regulation, membrane protein, circular dichroism.

Intracytoplasmic inclusion bodies in the chondrocytes of type I lethal achondrogenesis

1976 ◽  
Vol 7 (6) ◽  
pp. 667-673 ◽  
Author(s):  
Sheng-S. Yang ◽  
Kathleen P. Heidelberger ◽  
Jay Bernstein
Keyword(s):  
Inclusion Bodies ◽  
Type I ◽  
Intracytoplasmic Inclusion ◽  
Intracytoplasmic Inclusion Bodies

Naturally Occurring Infectious Fibroma in the Domestic Rabbit

1973 ◽  
Vol 10 (6) ◽  
pp. 509-519 ◽  
Author(s):  
L. T. Pulley ◽  
J. N. Shively
Keyword(s):  
Tumor Cells ◽  
Inclusion Bodies ◽  
Inflammatory Cells ◽  
Naturally Occurring ◽  
Intracytoplasmic Inclusion ◽  
Domestic Rabbit ◽  
Cutaneous Tumor ◽  
Intracytoplasmic Inclusion Bodies ◽  
Vascular Channels ◽  
Domestic Rabbits

Infectious fibromas developed in eight domestic rabbits that were all kept in outside cages. Each rabbit had a single cutaneous tumor. Six of these tumors, which were studied by light microscopy, consisted of tumor cells resembling fibroblasts and large areas of inflammation. The overlying epidermis was hyperplastic and contained easily recognizable intracytoplasmic inclusion bodies. Inclusion bodies were also in subepithelial fibroma cells. Electron microscopy of three of these tumors revealed typical pox-virus inclusions, sites of viral replication, maturation, and mature particles in epithelial cells of the epidermis and in subepithelial tumor cells. Virus was not seen in inflammatory cells or leukocytes within vascular channels.

scholarly journals Morbilliviral Dermatitis in Seals

2001 ◽  
Vol 38 (6) ◽  
pp. 724-726 ◽  
Author(s):  
T. P. Lipscomb ◽  
M. G. Mense ◽  
P. L. Habecker ◽  
J. K. Taubenberger ◽  
R. Schoelkopf
Keyword(s):  
Inclusion Bodies ◽  
Marine Mammals ◽  
Skin Lesions ◽  
Basal Cells ◽  
Hooded Seal ◽  
Cystophora Cristata ◽  
Phoca Groenlandica ◽  
Phocine Distemper Virus ◽  
Intracytoplasmic Inclusion Bodies ◽  
Polymerase Chain

A juvenile female hooded seal ( Cystophora cristata) and a juvenile male harp seal ( Phoca groenlandica) stranded separately on the New Jersey (USA) coast and were taken to a marine mammal rehabilitation center. Both were lethargic and emaciated, had dermatitis, and died. Histologic skin lesions in the seals were similar and consisted of epidermal and follicular epithelial hyperplasia, hyperkeratosis, degeneration, and necrosis. The most distinctive finding was extensive syncytial zones bounded superficially by hyperkeratosis and deeply by hyperplastic basal cells. Eosinophilic intracytoplasmic inclusion bodies were present in epithelial cells. Morbilliviral antigen was demonstrated in the skin lesions by immunohistochemistry. Phocine distemper virus was detected in the skin by reverse transcription polymerase chain reaction and a phocine distemper virus-specific probe using the Southern blot technique. This is the first report of morbilliviral dermatitis in marine mammals.

scholarly journals A Patient with Proximal Myotonic Myopathy and Parkinsonism

2002 ◽  
Vol 29 (2) ◽  
pp. 188-190 ◽  
Author(s):  
Kon Chu ◽  
Jin-Whan Cho ◽  
Eun-Chol Song ◽  
Beom S. Jeon
Keyword(s):  
Case Report ◽  
Myotonic Dystrophy ◽  
Inclusion Bodies ◽  
Neurodegenerative Disorder ◽  
Case Reports ◽  
Proximal Myotonic Myopathy ◽  
Intracytoplasmic Inclusion Bodies ◽  
Gene Study ◽  
Stooped Posture

Abstract:Introduction:There are two case reports of patients who had proximal myotonic myopathy (PROMM) / myotonic dystrophy (DM) Type 1 and parkinsonism. The combination of myotonic myopathy and parkinsonism is so rare that it may appear to be just a coincidence. However, previous neuropathological examinations of patients who had myotonic dystrophy showed that there were intracytoplasmic inclusion bodies in the nigra and striatum, which raises the possibility that myotonic myopathy may be associated with parkinsonism. In this report we describe a patient with PROMM and a clinically definite parkinsonism to highlight this possibility.Case Report:A 65-year-old man developed proximal muscle weakness, myotonia and atrophy around the age of 55 and was diagnosed as having PROMM at the age of 62. Needle electromyography and muscle biopsy supported the diagnosis. A gene study of the DM Type 1 showed a normal CTG repeat length. At age 63, he developed rest tremor, bradykinesia, hypomimia, stooped posture, and gait disturbance. The postural instability worsened rapidly. The tremor and rigidity were much worse in his right side, where myotonia was more severe. Levodopa therapy was only partially effective.Conclusion:This is a case report of a patient with PROMM that shows an association with a rapidly progressive form of parkinsonism. We suggest that this may be a novel form of a neurodegenerative disorder, which we name ‘Parkinsonism- Myotonic Myopathy-Complex’.

scholarly journals RNA-Containing Cytoplasmic Inclusion Bodies in Ciliated Bronchial Epithelium Months to Years after Acute Kawasaki Disease

PLoS ONE ◽  
2008 ◽  
Vol 3 (2) ◽  
pp. e1582 ◽  
Author(s):  
Anne H. Rowley ◽  
Susan C. Baker ◽  
Stanford T. Shulman ◽  
Francesca L. Garcia ◽  
Linda M. Fox ◽  
...  
Keyword(s):  
Kawasaki Disease ◽  
Inclusion Bodies ◽  
Cytoplasmic Inclusion ◽  
Bronchial Epithelium ◽  
Cytoplasmic Inclusion Bodies ◽  
Acute Kawasaki Disease

scholarly journals A Case of Contagious Ecthyma (Orf Virus) in a Nonmanipulated Laboratory Dorset Sheep (Ovis aries)

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Gwynne E. Kinley ◽  
Connie W. Schmitt ◽  
Julie Stephens-Devalle
Keyword(s):  
Inclusion Bodies ◽  
Clinical Case ◽  
Ovis Aries ◽  
Orf Virus ◽  
Contagious Ecthyma ◽  
Medial Canthus ◽  
Presumptive Diagnosis ◽  
Intracytoplasmic Inclusion ◽  
Histopathological Evaluation ◽  
Intracytoplasmic Inclusion Bodies

An approximately 5-month-old laboratory wether, originating from a local vendor with a closed flock and maintained on a preventative medicine plan, presented with a continuum of lesions from hemorrhagic papules, vesicles, and pustules, to multifocal necrotic scabs at the commissure of the lips, medial canthus of the left eye, and distal prepuce. A presumptive diagnosis of Orf virus (ORFV) was made and the sheep was euthanized. A full necropsy was performed, and histopathological evaluation of affected tissues revealed multifocal-to-coalescing necrotizing and proliferative cheilitis and dermatitis with eosinophilic intracytoplasmic inclusion bodies. Electron microscopy findings revealed degenerate keratinocytes containing numerous typical 200–300 nm wide cytoplasmic parapoxvirus virions, confirming the diagnosis of ORFV. We believe that this animal developed a clinical case of ORFV either due to an adverse reaction to an ORFV vaccine, or this animal had a case of preexisting ORFV which manifested after arrival at our facility.

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