-
Journal of Biomedical Science Jan 2011Dystonia musculorum (dt) is an autosomal recessive hereditary neuropathy with a characteristic uncoordinated movement and is caused by a defect in the bullous pemphigoid...
BACKGROUND
Dystonia musculorum (dt) is an autosomal recessive hereditary neuropathy with a characteristic uncoordinated movement and is caused by a defect in the bullous pemphigoid antigen 1 (BPAG1) gene. The neural isoform of BPAG1 is expressed in various neurons, including those in the central and peripheral nerve systems of mice. However, most previous studies on neuronal degeneration in BPAG1-deficient mice focused on peripheral sensory neurons and only limited investigation of the autonomic system has been conducted.
METHODS
In this study, patterns of nerve innervation in cutaneous and iridial tissues were examined using general neuronal marker protein gene product 9.5 via immunohistochemistry. To perform quantitative analysis of the autonomic neuronal number, neurons within the lumbar sympathetic and parasympathetic ciliary ganglia were calculated. In addition, autonomic neurons were cultured from embryonic dt/dt mutants to elucidate degenerative patterns in vitro. Distribution patterns of neuronal intermediate filaments in cultured autonomic neurons were thoroughly studied under immunocytochemistry and conventional electron microscopy.
RESULTS
Our immunohistochemistry results indicate that peripheral sensory nerves and autonomic innervation of sweat glands and irises dominated degeneration in dt/dt mice. Quantitative results confirmed that the number of neurons was significantly decreased in the lumbar sympathetic ganglia as well as in the parasympathetic ciliary ganglia of dt/dt mice compared with those of wild-type mice. We also observed that the neuronal intermediate filaments were aggregated abnormally in cultured autonomic neurons from dt/dt embryos.
CONCLUSIONS
These results suggest that a deficiency in the cytoskeletal linker BPAG1 is responsible for dominant sensory nerve degeneration and severe autonomic degeneration in dt/dt mice. Additionally, abnormally aggregated neuronal intermediate filaments may participate in neuronal death of cultured autonomic neurons from dt/dt mutants.
Topics: Animals; Autonomic Nervous System; Carrier Proteins; Cells, Cultured; Cytoskeletal Proteins; Dystonia Musculorum Deformans; Dystonin; Embryo, Mammalian; Humans; Mice; Mice, Mutant Strains; Nerve Tissue Proteins; Sensory Receptor Cells
PubMed: 21272373
DOI: 10.1186/1423-0127-18-9 -
Clinical and Experimental Immunology Nov 2010In this report,we present 15 patients with histological and immunopathologically proven pemphigus vulgaris (PV). After a mean of 80 months since the onset of disease,...
In this report,we present 15 patients with histological and immunopathologically proven pemphigus vulgaris (PV). After a mean of 80 months since the onset of disease, when evaluated serologically, they had antibodies typical of PV and pemphigoid (Pg). Similarly, 18 patients with bullous pemphigoid (BP) and mucous membrane pemphigoid (MMP) were diagnosed on the basis of histology and immunopathology.After a mean of 60 months since the onset of disease, when their sera were evaluated they were found to have Pg and PV autoantibodies. In both groups of patients the diseases were characterized by a chronic course, which included several relapses and recurrences and were non-responsive to conventional therapy. The major histocompatibility complex class II (MHC II) genes were studied in both groups of patients and phenotypes associated typically with them were observed. Hence, in 33 patients, two different pathogenic autoantibodies were detected simultaneously. The authors provide a computer model to show that each MHC II gene has relevant epitopes that recognize the antigens associated with both diseases. Using the databases in these computer models, the authors present the hypothesis that these two autoantibodies are produced simultaneously due to the phenomena of epitope spreading.
Topics: Adult; Aged; Aged, 80 and over; Amino Acid Sequence; Antibody Formation; Antigens, Surface; Autoantibodies; Autoantigens; Carrier Proteins; Cytoskeletal Proteins; Desmoglein 1; Desmoglein 3; Dystonin; Epitopes, T-Lymphocyte; Female; Genes, MHC Class II; HLA-DQ Antigens; HLA-DQ beta-Chains; HLA-DR Antigens; HLA-DRB1 Chains; Humans; Integrin alpha6; Integrin beta4; Keratinocytes; Male; Middle Aged; Molecular Sequence Data; Nerve Tissue Proteins; Non-Fibrillar Collagens; Pemphigoid, Benign Mucous Membrane; Pemphigoid, Bullous; Pemphigus; Software; Young Adult; Collagen Type XVII
PubMed: 21069937
DOI: 10.1111/j.1365-2249.2010.04239.x -
PloS One May 2010Malignant melanoma is one of the most aggressive types of tumor. Because malignant melanoma is difficult to treat once it has metastasized, early detection and treatment...
Malignant melanoma is one of the most aggressive types of tumor. Because malignant melanoma is difficult to treat once it has metastasized, early detection and treatment are essential. The search for reliable biomarkers of early-stage melanoma, therefore, has received much attention. By using a novel method of screening tumor antigens and their auto-antibodies, we identified bullous pemphigoid antigen 1 (BPAG1) as a melanoma antigen recognized by its auto-antibody. BPAG1 is an auto-antigen in the skin disease bullous pemphigoid (BP) and anti-BPAG1 auto-antibodies are detectable in sera from BP patients and are used for BP diagnosis. However, BPAG1 has been viewed as predominantly a keratinocyte-associated protein and a relationship between BPAG1 expression and melanoma has not been previously reported. In the present study, we show that bpag1 is expressed in the mouse F10 melanoma cell line in vitro and F10 melanoma tumors in vivo and that BPAG1 is expressed in human melanoma cell lines (A375 and G361) and normal human melanocytes. Moreover, the levels of anti-BPAG1 auto-antibodies in the sera of melanoma patients were significantly higher than in the sera of healthy volunteers (p<0.01). Furthermore, anti-BPAG1 auto-antibodies were detected in melanoma patients at both early and advanced stages of disease. Here, we report anti-BPAG1 auto-antibodies as a promising marker for the diagnosis of melanoma, and we discuss the significance of the detection of such auto-antibodies in cancer biology and patients.
Topics: Animals; Antibodies, Anti-Idiotypic; Antigens, Neoplasm; Autoantigens; Biomarkers, Tumor; Carrier Proteins; Cell Line, Tumor; Cytoskeletal Proteins; Dystonin; Female; Gene Expression Regulation, Neoplastic; Humans; Melanocytes; Melanoma; Mice; Mice, Inbred C57BL; Nerve Tissue Proteins; Non-Fibrillar Collagens; Peptide Library; Collagen Type XVII
PubMed: 20479946
DOI: 10.1371/journal.pone.0010566 -
PloS One Mar 2010Dystonin is a giant cytoskeletal protein belonging to the plakin protein family and is believed to crosslink the major filament systems in contractile cells. Previous...
Dystonin is a giant cytoskeletal protein belonging to the plakin protein family and is believed to crosslink the major filament systems in contractile cells. Previous work has demonstrated skeletal muscle defects in dystonin-deficient dystonia musculorum (dt) mice. In this study, we show that the dystonin muscle isoform is localized at the Z-disc, the H zone, the sarcolemma and intercalated discs in cardiac tissue. Based on this localization pattern, we tested whether dystonin-deficiency leads to structural defects in cardiac muscle. Desmin intermediate filament, microfilament, and microtubule subcellular organization appeared normal in dt hearts. Nevertheless, increased transcript levels of atrial natriuretic factor (ANF, 66%) beta-myosin heavy chain (beta-MHC, 95%) and decreased levels of sarcoplasmic reticulum calcium pump isoform 2A (SERCA2a, 26%), all signs of cardiac muscle stress, were noted in dt hearts. Hearts from two-week old dt mice were assessed for the presence of morphological and histological alterations. Heart to body weight ratios as well as left ventricular wall thickness and left chamber volume measurements were similar between dt and wild-type control mice. Hearts from dt mice also displayed no signs of fibrosis or calcification. Taken together, our data provide new insights into the intricate structure of the sarcomere by situating dystonin in cardiac muscle fibers and suggest that dystonin does not significantly influence the structural organization of cardiac muscle fibers during early postnatal development.
Topics: Animals; Atrial Natriuretic Factor; Carrier Proteins; Cytoskeletal Proteins; Desmin; Dystonia Musculorum Deformans; Dystonin; Heart; Intermediate Filaments; Mice; Microtubules; Myocardial Contraction; Myocardium; Myosin Heavy Chains; Nerve Tissue Proteins; Protein Isoforms; Reverse Transcriptase Polymerase Chain Reaction; Sarcoplasmic Reticulum Calcium-Transporting ATPases
PubMed: 20209123
DOI: 10.1371/journal.pone.0009465 -
Acta Dermato-venereologica Mar 2010Bullous pemphigoid is associated with antibodies to a 230 kDa and a 180 kDa protein. In a literature review we investigated the role of auto-antibodies as detected by... (Review)
Review
Bullous pemphigoid is associated with antibodies to a 230 kDa and a 180 kDa protein. In a literature review we investigated the role of auto-antibodies as detected by different serological assays. Nine reports containing data on 143 patients were analyzed. Pre-treatment data showed that indirect immunofluorescence, enzyme-linked immunosorbent assay (ELISA) and immunoblotting offer an 82.2% or greater probability of being positive. At the end of the study period, all patients had clinically improved, whether or not they were on therapy. Auto-antibodies were present in 29% of patients evaluated by monkey esophagus immunofluorescence and 75% of those evaluated by human skin immunofluorescence. Positive titers were also reported in 67.6% of patients evaluated by ELISA. In 100% of patients in whom immunoblotting was performed the titers became negative. In 3 patients (5.3%) using human skin immunofluorescence and in one patient (1.4%) using ELISA the titers were increased at the end of the study period. The correlation between anti-basement membrane zone antibodies and the clinical course of bullous pemphigoid requires further and long-term studies.
Topics: Aged; Aged, 80 and over; Animals; Autoantibodies; Autoantigens; Biomarkers; Carrier Proteins; Cytoskeletal Proteins; Dystonin; Enzyme-Linked Immunosorbent Assay; Female; Fluorescent Antibody Technique; Haplorhini; Humans; Immunoblotting; Immunologic Tests; Male; Middle Aged; Nerve Tissue Proteins; Non-Fibrillar Collagens; Pemphigoid, Bullous; Predictive Value of Tests; Remission Induction; Sensitivity and Specificity; Time Factors; Treatment Outcome; Collagen Type XVII
PubMed: 20169293
DOI: 10.2340/00015555-0819 -
The Journal of Investigative Dermatology Jun 2010Epidermolysis bullosa (EB) is a group of autosomal dominant and recessive blistering skin diseases in which pathogenic mutations have been reported in 13 different genes...
A homozygous nonsense mutation within the dystonin gene coding for the coiled-coil domain of the epithelial isoform of BPAG1 underlies a new subtype of autosomal recessive epidermolysis bullosa simplex.
Epidermolysis bullosa (EB) is a group of autosomal dominant and recessive blistering skin diseases in which pathogenic mutations have been reported in 13 different genes encoding structural proteins involved in keratinocyte integrity, as well as cell-matrix or cell-cell adhesion. We now report an inherited skin fragility disorder with a homozygous nonsense mutation in the dystonin gene (DST) that encodes the coiled-coil domain of the epithelial isoform of bullous pemphigoid antigen 1, BPAG1-e (also known as BP230). The mutation, p.Gln1124X, leads to the loss of hemidesmosomal inner plaques and a complete absence of skin immunostaining for BPAG1-e, as well as reduced labeling for plectin, the beta4 integrin subunit, and for type XVII collagen. The 38-year-old affected individual has lifelong generalized trauma-induced spontaneous blisters and erosions, particularly around the ankles. In addition, he experiences episodic numbness in his limbs, which started at the age of 37 years. These neurological symptoms may also be due to DST gene mutation, although he has a concomitant diagnosis of CADASIL (cerebral arteriopathy, autosomal dominant, with subcortical infarcts and leukoencephalopathy), a cerebral small-vessel arteriopathy, which thus complicates the genotype-phenotype interpretation. With regard to skin blistering, the clinicopathological findings expand the molecular basis of EB by identifying BPAG1-e pathology in a new form of autosomal recessive EB simplex.
Topics: Adult; Blister; Carrier Proteins; Codon, Nonsense; Collagen Type VII; Cytoskeletal Proteins; Dystonin; Epidermolysis Bullosa Simplex; Hemidesmosomes; Homozygote; Humans; Integrin beta4; Male; Nerve Tissue Proteins; Plectin; Protein Isoforms; Skin
PubMed: 20164846
DOI: 10.1038/jid.2010.19 -
Molecular Biology of the Cell Jun 2009alpha6beta4 integrin, a component of hemidesmosomes, also plays a role in keratinocyte migration via signaling through Rac1 to the actin-severing protein cofilin. Here,...
alpha6beta4 integrin, a component of hemidesmosomes, also plays a role in keratinocyte migration via signaling through Rac1 to the actin-severing protein cofilin. Here, we tested the hypothesis that the beta4 integrin-associated plakin protein, bullous pemphigoid antigen 1e (BPAG1e) functions as a scaffold for Rac1/cofilin signal transduction. We generated keratinocyte lines exhibiting a stable knockdown in BPAG1e expression. Knockdown of BPAG1e does not affect expression levels of other hemidesmosomal proteins, nor the amount of beta4 integrin expressed at the cell surface. However, the amount of Rac1 associating with beta4 integrin and the activity of both Rac1 and cofilin are significantly lower in BPAG1e-deficient cells compared with wild-type keratinocytes. In addition, keratinocytes deficient in BPAG1e exhibit loss of front-to-rear polarity and display aberrant motility. These defects are rescued by inducing expression of constitutively active Rac1 or active cofilin. These data indicate that the BPAG1e is required for efficient regulation of keratinocyte polarity and migration by determining the activation of Rac1.
Topics: Actin Depolymerizing Factors; Actins; Carrier Proteins; Cell Line; Cell Movement; Cell Polarity; Cytoskeletal Proteins; Cytoskeleton; Dystonin; Enzyme Activation; Epidermal Cells; Focal Adhesions; Gene Knockdown Techniques; Humans; Integrin beta4; Keratinocytes; Lentivirus; Nerve Tissue Proteins; Plectin; Protein Transport; Pseudopodia; RNA, Small Interfering; rac1 GTP-Binding Protein
PubMed: 19403692
DOI: 10.1091/mbc.e09-01-0051 -
Revista de NeurologiaBullous pemphigoid is the most frequent blistering disease and has been found associated to several neurological diseases, including amyotrophic lateral sclerosis.
INTRODUCTION
Bullous pemphigoid is the most frequent blistering disease and has been found associated to several neurological diseases, including amyotrophic lateral sclerosis.
CASE REPORT
A 63-year-old male with bulbar-onset amyotrophic lateral sclerosis who presented clinical and histological signs and symptoms of bullous pemphigoid.
CONCLUSIONS
The association does not seem to occur by chance and we suggest an autoimmune pathogenetic mechanism consisting in a crossed reaction between bullous pemphigoid antigen 1 and the protein dystonin, which is involved in the organisation/integrity of the neuronal cytoskeleton.
Topics: Amyotrophic Lateral Sclerosis; Humans; Male; Middle Aged; Pemphigoid, Bullous
PubMed: 19012256
DOI: No ID Found -
The Journal of Neuroscience : the... Jun 2008Schwann cells integrate signals deriving from the axon and the basal lamina to myelinate peripheral nerves. Integrin alpha6beta4 is a laminin receptor synthesized by...
Schwann cells integrate signals deriving from the axon and the basal lamina to myelinate peripheral nerves. Integrin alpha6beta4 is a laminin receptor synthesized by Schwann cells and displayed apposed to the basal lamina. alpha6beta4 integrin expression in Schwann cells is induced by axons at the onset of myelination, and rises in adulthood. The beta4 chain has a uniquely long cytoplasmic domain that interacts with intermediate filaments such as dystonin, important in peripheral myelination. Furthermore, alpha6beta4 integrin binds peripheral myelin protein 22, whose alteration causes the most common demyelinating hereditary neuropathy. All these data suggest a role for alpha6beta4 integrin in peripheral nerve myelination. Here we show that ablating alpha6beta4 integrin specifically in Schwann cells of transgenic mice does not affect peripheral nerve development, myelin formation, maturation, or regeneration. However, consistent with maximal expression in adult nerves, alpha6beta4 integrin-null myelin is more prone to abnormal folding with aging. When the laminin receptor dystroglycan is also ablated, major folding abnormalities occur, associated with acute demyelination in some peripheral nervous system districts. These data indicate that, similar to its role in skin, alpha6beta4 integrin confers stability to myelin in peripheral nerves.
Topics: Aging; Animals; Cell Differentiation; Dystroglycans; Integrin alpha6beta4; Mice; Mice, Inbred C57BL; Mice, Knockout; Myelin Sheath; Nerve Fibers, Myelinated; Peripheral Nerves; Protein Folding; Schwann Cells
PubMed: 18579745
DOI: 10.1523/JNEUROSCI.0326-08.2008 -
American Journal of Transplantation :... Mar 2008Donor-reactive memory T cells undermine the survival of transplanted organs through multiple pathways. We have previously reported that memory CD4 T cells resist...
Donor-reactive memory T cells undermine the survival of transplanted organs through multiple pathways. We have previously reported that memory CD4 T cells resist treatment with anti-CD154 antibody and donor-specific transfusion (DST/MR1) and promote cardiac allograft rejection via generation of effector CD4 T cells and alloantibody. We hypothesized that the helper functions of memory CD4 T cells are independent of T-cell costimulation through CD154 but instead are regulated by alternative costimulatory pathways. This study investigated how blocking ICOS/B7RP-1 interactions affects functions of donor-reactive memory CD4 T cells. Treatment with blocking anti-ICOS mAb synergized with DST/MR1 and prolonged mouse cardiac allograft survival despite the presence of donor-reactive memory CD4 T cells. While blocking ICOS did not diminish the expansion of preexisting memory CD4 T cells or the induction of allospecific effector T cells, it did inhibit recruitment of the activated memory and effector T cells into the graft. In addition, anti-ICOS mAb treatment in combination with DST/MR1 prevented help provided by memory CD4 T cells for production of donor-specific IgG antibody. These results demonstrate the potential efficacy of ICOS blockade in sensitized transplant patients and provide the foundation for rational use of ICOS blockade in combination with other graft-prolonging strategies.
Topics: Animals; Antibodies, Monoclonal; Antigens, Differentiation, T-Lymphocyte; B7-1 Antigen; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Carrier Proteins; Cytoskeletal Proteins; Dystonin; Female; Graft Rejection; Graft Survival; Heart Transplantation; Histocompatibility Antigens Class I; Immunoglobulin Switch Region; Immunologic Memory; Inducible T-Cell Co-Stimulator Ligand; Inducible T-Cell Co-Stimulator Protein; Male; Mice; Mice, Inbred Strains; Minor Histocompatibility Antigens; Nerve Tissue Proteins
PubMed: 18294146
DOI: 10.1111/j.1600-6143.2007.02096.x