The Experts below are selected from a list of 225 Experts worldwide ranked by ideXlab platform
Lawrence S Chan - One of the best experts on this subject based on the ideXlab platform.
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defective in vivo expression and apparently normal in vitro expression of a newly identified 105 kda lower Lamina Lucida protein in dystrophic epidermolysis bullosa
British Journal of Dermatology, 2006Co-Authors: Lawrence S Chan, J D Fine, C Hammerberg, E A Bauer, Kevin D CooperAbstract:Summary We have previously identified a novel 105-kDa lower Lamina Lucida protein detected by theautoantibodies from a group of patients who developed a unique immune-mediated subepidenna!bullous dermatosis. We sought to determine if this novel basement membrane zone (BMZ) protein isnormally expressed in the skin of patients with various subsets of epidermolysis bullosa (EB). Indirectimraunofluorescence microscopy performed on non-iesional skin sections from patients with threemajor EB subsets revealed absence or significantly reduced expression of this novel BMZ protein in 20out of 23 skin sections from patients with generalized dominant and recessive dystrophic EB.However, immunoblot analyses with the autoantibodies on Western-blotted proteins revealed that acomigrating U)5-kDa protein is present in both cytosol extracts (n ^ 6) and conditioned media (n =3) of cultured dermal fibroblasts derived from patients with dystrophic EB, as well as those culturedfrom two healthy individuals. Although the reason for such disparate findings is not known, thedefective in vivo expression of this novel 105-kDa protein in dystrophic EB is presumably not due to afailure of fibroblasts to synthesize or secrete the protein. It is possible, however, that the 105-kDaprotein may be unable to incorporate into the BMZ because it is produced in a dysfunctional form, orits BMZ binding site is missing. It is also possible that other structural alterations in skin BMZ. whichoccur in dystrophic EB, result in masking of the antigenic binding by the autoantibody when intactBMZ is probed. In any case, the reduced in vivo expression of the 105-kDa protein representsadditional evidence for a defect in BMZ composition in dystrophic EB which extends to a number ofmolecular components.Hereditary epidermolysis bullosa (EB) consists of a het-erogeneous group of skin diseases manifested by thedevelopment of bUsters at sites of minor mechanicaltrauma.^ EB subtypes are classified by clinical pheno-typic features, mode of inheritance, level of skin cleavage,and the presence or absence of additional ultrastructuralor immunohistochemical findings.^ Because sufficientphenotypic overlap exists among aC three major sub-types of EB clinically, precise classification usuallyrequires the performance of either transmission electronmicroscopy or immunofiuorescence
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Linear IgA bullous dermatosis. Characterization of a subset of patients with concurrent IgA and IgG anti-basement membrane autoantibodies.
Archives of Dermatology, 1995Co-Authors: Lawrence S Chan, Ted B Taylor, David T Woodley, Thomas N. Traczyk, Lynne R. Eramo, John J ZoneAbstract:Background and Design: The term linear IgA bullous dermatosis defines an immune-mediated blistering skin disease characterized by pruritic blisters, subepidermal separation with neutrophilic infiltration, and linear IgA antibody deposition at the basement membrane zone (BMZ). However, some patients with linear IgA bullous dermatosis demonstrate both IgA and IgG anti-BMZ autoantibodies on immunofluorescence. We describe four such patients and attempt to define this group of patients by studying their clinical, histopathologic, immunopathologic, immunoultrastructural, and immunochemical characteristics. Results: Clinically, all four patients had a generalized pruritic blistering skin disease consistent with linear IgA bullous dermatosis. Histopathologically, all four cases demonstrated subepidermal blister formation with a predominantly neutrophilic dermal infiltrate. Immunopathologically, tissue-bound IgA (in four of four patients) and IgG (in three of four patients) antibodies were detected at the BMZ. Circulating IgA (in four of four patients) and IgG (in four of four patients) labeled the epidermal BMZ of normal human skin fractured at the Lamina Lucida and did not label the dermal BMZ. By immunoblot analyses, IgA (in three of three patients) and IgG (in one of three patients) circulating antibodies labeled the 97-kd linear IgA bullous dermatosis antigen. By direct immunoelectron microscopy, IgA and IgG antibodies were localized (in two of two patients) exclusively within the upper Lamina Lucida of the BMZ. Conclusions: Four cases of an immune-mediated blistering skin disease typical for linear IgA bullous dermatosis demonstrated both IgA and IgG anti-BMZ autoantibodies against the linear IgA bullous dermatosis antigen within the upper Lamina Lucida. We conclude that linear IgA bullous dermatosis should include a subgroup of patients with both IgA and IgG anti-BMZ autoantibodies. (Arch Dermatol. 1995;131:1432-1437)
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a newly identified 105 kd lower Lamina Lucida autoantigen is an acidic protein distinct from the 105 kd γ2 chain of laminin 5
Journal of Investigative Dermatology, 1995Co-Authors: Lawrence S Chan, Xuesong Wang, Jean Christophe Lapiere, Peter M Marinkovich, Jonathan C R Jones, David T WoodleyAbstract:A 105-kD lower Lamina Lucida antigen (p105) has been detected by autoantibodies (anti-p105) from patients with a novel immunobullous disease. To distinguish p105 from other known Lamina Lucida components, we performed comparative irnmunoblotting on purified human amniotic laminin-5 (kalinin), 804G matrix (enriched in laminin-5), and keratinocyte and fibroblast proteins using anti-804G matrix antibody 0-18) and anti-p105. J-18 labeled the truncated laminin-5 γ2 chain in amniotic laminin-5, 804G matrix, and keratinocyte conditioned medium, but did not label fibroblast cytosol. Conversely, anti-p105 did not label amniotic laminin-5 or 804G matrix, but did label p105 in both keratinocyte conditioned medium and fibroblast cytosol. J-18 labeled the 105-kD laminin-5 γ2 chain in reduced keratinocyte proteins and a 400-kD laminin-5 complex under non-reducing conditions. In contrast, anti-p105 labeled p105 under both reducing and non-reducing conditions but did not label a 400-kD protein complex. Similarly, comparative immunoblotting on keratinocyte proteins using anti-p105 and anti-laminin-1 revealed no commonly labeled protein bands. Electrophoretic fractionations by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of these fractions revealed that the peak fractions of keratinocyte proteins reactive with anti-p105 are different from those reactive with J-18. Furthermore, keratinocyte proteins fractionated by Mono Q anionexchange chromatography revealed fractions immunoreactive with anti-p105, whereas J-18 showed no reactivity with these fractions. Two-dimensional gel electrophoresis and immunoblotting with anti-p105 revealed p105 to be an acidic protein with isoelectric points between 5.7 and 6.3, distinct from the isoelectric points of laminin-5 γ2 chain. We conclude that p105 is an acidic protein located in the Lamina Lucida and distinct from the truncated laminin-5 γ2 chain and the laminin-1 family.
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a novel 105 kda Lamina Lucida autoantigen association with bullous pemphigoid
Journal of Investigative Dermatology, 1994Co-Authors: Stephanie L Cotell, Lawrence S Chan, Jean Christophe Lapiere, John D Chen, Toshiroh Iwasaki, Paul A Krusinski, David T WoodleyAbstract:Abstract Several cases have been reported of patients with immunemediated subepidermal blistering disorders whose autoantibodies react to antigens present on both the dermal and epidermal side of 1 M NaCl-split skin. In this report, we identify, localize, and characterize the basement membrane zone antigen corresponding to the dermal staining in a patient whose serum stains both the dermal and epidermal side of 1 M NaCl-split skin. This patient's serum contains autoantibodies directed against a 105-kilodalton(kDa) dermal antigen and the 230-kDa epidermal (bullous permphigoid) antigen. This novel 105-kDa protein was previously identified as the sole antigen in another patient with a unique bullous disease whose autoantibodies were directed against only the dermal side of 1 M NaCl-split skin. This 105-kDa antigen was identical by one- and two-dimensional immunoblot analysis in these two patients. By immunoblot analysis, autoantibodies from our patient labeled a 105-kDa protein within various extracts of human skin basement membrane. Immunoblot analyses using epitope-selected autoantibodies directed against the 105-kDa protein demonstrated that this antigen is independent and distinct from other known basement membrane antigens. The 105-kDa antigen is an extracellular matrix component of the basement membrane, which is synthesized and secreted by both keratinocytes and fibroblasts. Identical electrophoretic migration of cellular and secreted forms of the protein suggested there is no major post-translational modification of the protein. Immunomapping of normal human skin fractured through the dermal-epidermal junction by incubation in 1 M NaCl or by suction blistering demonstrated that the location of the 105-kDa antigen within the basement membrane zone is between the bullous pemphigoid antigens and two other Lamina Lucida components, laminin and nicein. These data demonstrate clearly that a subepidermal autoimmune bullous disease may have autoantibodies directed against two distinct components of the dermal-epidermal junction.
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a novel immune mediated subepidermal bullous dermatosis characterized by igg autoantibodies to a lower Lamina Lucida component
Archives of Dermatology, 1994Co-Authors: Lawrence S Chan, Kevin D CooperAbstract:Background: Immune-mediated subepidermal bullous dermatoses characterized by in vivo—bound linear IgG deposition at the cutaneous basement membrane zone include bullous pemphigoid, ocular cicatricial pemphigoid, anti—bullous pemphigoid antigen mucosal pemphigoid, anti—epiligrin mucosal pemphigoid, epidermolysis bullosa acquisita, and the bullous eruption of systemic lupus erythematosus. In this article, we describe a novel IgG-mediated bullous dermatosis. Observations: Clinically, a unique nonscarring dermatosis was characterized by the sudden onset of extensive bullae and erosions on mucous membrane and skin, resembling toxic epidermal necrolysis or pemphigus vulgaris. Histologically, the patient's skin lesion demonstrated neutrophilic papillary dermal infiltration and subepidermal blister formation, resembling dermatitis herpetiformis. Immunopathologically, there was linear IgG and C3 deposition at the skin basement membrane zone. The patient responded well to prednisone and azathioprine immunosuppression and has achieved a lasting remission without further therapy. Further immunologic investigations revealed that this unique dermatosis is distinct from all other known IgG-mediated subepidermal bullous dermatoses. Conclusions: This novel deep Lamina Lucida pemphigoid can be distinctly termed anti—p105 pemphigoid on the basis of antigenic specificity of the autoantibodies. Although this novel dermatosis resembles toxic epidermal necrolysis clinically, prudent use of diagnostic immunofluorescence studies can clearly delineate its immunologic nature. Prompt recognition of this unique dermatosis and timely initiation of appropriate immunosuppressive therapy could be life-saving for those patients suffering from this dermatosis. ( Arch Dermatol. 1994;130:343-347 )
Kevin D Cooper - One of the best experts on this subject based on the ideXlab platform.
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defective in vivo expression and apparently normal in vitro expression of a newly identified 105 kda lower Lamina Lucida protein in dystrophic epidermolysis bullosa
British Journal of Dermatology, 2006Co-Authors: Lawrence S Chan, J D Fine, C Hammerberg, E A Bauer, Kevin D CooperAbstract:Summary We have previously identified a novel 105-kDa lower Lamina Lucida protein detected by theautoantibodies from a group of patients who developed a unique immune-mediated subepidenna!bullous dermatosis. We sought to determine if this novel basement membrane zone (BMZ) protein isnormally expressed in the skin of patients with various subsets of epidermolysis bullosa (EB). Indirectimraunofluorescence microscopy performed on non-iesional skin sections from patients with threemajor EB subsets revealed absence or significantly reduced expression of this novel BMZ protein in 20out of 23 skin sections from patients with generalized dominant and recessive dystrophic EB.However, immunoblot analyses with the autoantibodies on Western-blotted proteins revealed that acomigrating U)5-kDa protein is present in both cytosol extracts (n ^ 6) and conditioned media (n =3) of cultured dermal fibroblasts derived from patients with dystrophic EB, as well as those culturedfrom two healthy individuals. Although the reason for such disparate findings is not known, thedefective in vivo expression of this novel 105-kDa protein in dystrophic EB is presumably not due to afailure of fibroblasts to synthesize or secrete the protein. It is possible, however, that the 105-kDaprotein may be unable to incorporate into the BMZ because it is produced in a dysfunctional form, orits BMZ binding site is missing. It is also possible that other structural alterations in skin BMZ. whichoccur in dystrophic EB, result in masking of the antigenic binding by the autoantibody when intactBMZ is probed. In any case, the reduced in vivo expression of the 105-kDa protein representsadditional evidence for a defect in BMZ composition in dystrophic EB which extends to a number ofmolecular components.Hereditary epidermolysis bullosa (EB) consists of a het-erogeneous group of skin diseases manifested by thedevelopment of bUsters at sites of minor mechanicaltrauma.^ EB subtypes are classified by clinical pheno-typic features, mode of inheritance, level of skin cleavage,and the presence or absence of additional ultrastructuralor immunohistochemical findings.^ Because sufficientphenotypic overlap exists among aC three major sub-types of EB clinically, precise classification usuallyrequires the performance of either transmission electronmicroscopy or immunofiuorescence
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a novel immune mediated subepidermal bullous dermatosis characterized by igg autoantibodies to a lower Lamina Lucida component
Archives of Dermatology, 1994Co-Authors: Lawrence S Chan, Kevin D CooperAbstract:Background: Immune-mediated subepidermal bullous dermatoses characterized by in vivo—bound linear IgG deposition at the cutaneous basement membrane zone include bullous pemphigoid, ocular cicatricial pemphigoid, anti—bullous pemphigoid antigen mucosal pemphigoid, anti—epiligrin mucosal pemphigoid, epidermolysis bullosa acquisita, and the bullous eruption of systemic lupus erythematosus. In this article, we describe a novel IgG-mediated bullous dermatosis. Observations: Clinically, a unique nonscarring dermatosis was characterized by the sudden onset of extensive bullae and erosions on mucous membrane and skin, resembling toxic epidermal necrolysis or pemphigus vulgaris. Histologically, the patient's skin lesion demonstrated neutrophilic papillary dermal infiltration and subepidermal blister formation, resembling dermatitis herpetiformis. Immunopathologically, there was linear IgG and C3 deposition at the skin basement membrane zone. The patient responded well to prednisone and azathioprine immunosuppression and has achieved a lasting remission without further therapy. Further immunologic investigations revealed that this unique dermatosis is distinct from all other known IgG-mediated subepidermal bullous dermatoses. Conclusions: This novel deep Lamina Lucida pemphigoid can be distinctly termed anti—p105 pemphigoid on the basis of antigenic specificity of the autoantibodies. Although this novel dermatosis resembles toxic epidermal necrolysis clinically, prudent use of diagnostic immunofluorescence studies can clearly delineate its immunologic nature. Prompt recognition of this unique dermatosis and timely initiation of appropriate immunosuppressive therapy could be life-saving for those patients suffering from this dermatosis. ( Arch Dermatol. 1994;130:343-347 )
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identification and partial characterization of a novel 105 kdalton lower Lamina Lucida autoantigen associated with a novel immune mediated subepidermal blistering disease
Journal of Investigative Dermatology, 1993Co-Authors: Robert A Briggaman, Lawrence S Chan, Jodavid Fine, David T Woodley, Craig Hammerberg, Rhett J Drugge, Kevin D CooperAbstract:Certain skin basement membrane components, such as bullous pemphigoid antigens and epidermolysis bullosa acquisita antigen, were discovered as a result of an autoimmune reaction. In this report, we describe a unique Lamina Lucida determinant associated with a novel immune-mediated subepidermal bullous dermatosis. This unique bullous dermatosis resembled severe toxic epidermal necrolysis clinically. The histologic findings resemble dermatitis herpetiformis. Direct immunofluorescence microscopy detected linear immunoglobulin G (lgG) and C3 deposition at the cutaneous basement membrane zone of lesional and perilesional skin. Direct and indirect immunoelectron microscopy localized the LgG deposits to the lowest portion of the Lamina Lucida. The patient's autoantibodies, belonging to the IgG1 subclass, labeled basement membrane zone of normal intact human skin, oral mucosa, and conjunctiva, and localized to the dermal side of salt-split normal adult and neonatal human skin, but failed to react with human fetal skin up to 142 gestational days. The patient's autoantibodies failed to react with bullous pemphigoid antigens or epidermolysis bullosa acquisita antigen (type VII collagen) by immunoblotting. Instead, the patient's autoantibodies unequivocally labeled a 105-kilodalton (kD) protein in cellular extracts and conditioned media of human cultured keratinocytes and dermal fibroblasts. The titer of the patient's antibody against the cutaneous basement membrane zone and the intensity of the antibody reactivity against the 1O5-kD protein paralleled the patient's disease activity. Thus, this 105-kD lower Lamina Lucida protein represents a novel autoantigen and this patient's disease represents a deep Lamina Lucida pemphigoid, distinguishable from all other known autoimmune bullous dermatoses.
Akira Ishiko - One of the best experts on this subject based on the ideXlab platform.
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immunoglobulin g deposition to nonhemidesmosomal Lamina Lucida and early neutrophil involvement are characteristic features in a case of anti p200 pemphigoid
British Journal of Dermatology, 2013Co-Authors: Akira Ishiko, Takashi Hashimoto, Atsushi Shimizu, Takeru Funakoshi, M Ishibashi, Tadashi Yoshida, H Koga, Masayuki AmagaiAbstract:Summary The ultrastructural characteristics and immunolocalization of in vivo bound immunoglobulin G (IgG) in skin affected by anti-p200 pemphigoid have not been eLucidated. To give insight into the mechanism of blister formation we report a new case of anti-p200 pemphigoid, studied with stage-oriented morphological analysis and immunoelectron microscopy. Skin biopsy specimens were evaluated ultrastructurally and histologically with immunohistochemistry. By observing the nonblister site, the blister edge and centre of the blister, we determined that neutrophil infiltration increases gradually at the dermoepidermal junction in association with the destruction of type IV collagen. Ultrastructurally, many neutrophils were observed under the Lamina densa, with vacuole formation in the dermis. At the periphery of the blister, the Lamina densa became fragmented and was observed either at the roof or the floor of the blister. At the centre of the blister, the Lamina densa was mainly observed at the blister floor. Postembedding immunoelectron microscopy demonstrated that the IgG, bound in vivo, localized at the Lamina Lucida, while the area beneath the hemidesmosomes was spared. Together with the early involvement of neutrophils and the destruction of the basal Lamina, we suggest that the binding of autoantibodies to the nonhemidesmosomal Lamina Lucida may induce inflammation with neutrophils, resulting in blister formation.
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97 kda linear iga bullous dermatosis antigen localizes in the Lamina Lucida between the nc16a and carboxyl terminal domains of the 180 kda bullous pemphigoid antigen
Journal of Investigative Dermatology, 1998Co-Authors: Akira Ishiko, John J Zone, Hiroshi Shimizu, Takuji Masunaga, Kim B Yancey, George J Giudice, Takeji NishikawaAbstract:Linear IgA bullous dermatosis is an autoimmune blistering disease characterized by circulating IgA anti-basement membrane autoantibodies. A 97 kDa protein (97-LAD), which localizes at the basement membrane zone of normal human skin, is one of the major autoantigens associated with this disease and possesses multiple regions of amino acid identity with the extracellular domain of the 180 kDa bullous pemphigoid antigen, BPAG2. To investigate further the relationship between 97-LAD and BPAG2, immunogold electron microscopy was performed on cryo-ultrathin sections of normal human skin using a series of polyclonal and monoclonal antibodies. Gold particles immunolabeling two newly developed monoclonal antibodies against 97-LAD were localized to the Lamina Lucida. This immunolabeling pattern was associated with hemidesmosomes and localized at a mean distance of 28 nm beneath the plasma membrane of basal keratinocytes. In contrast, polyclonal antibodies against a fusion protein containing the NC16A domain of BPAG2 immunolabeled the plasma membrane of the hemidesmosomal complex, whereas polyclonal antibodies against the carboxyl terminus mainly immunolabeled the lower Lamina Lucida with a mean distance of 42 nm beneath the plasma membrane. By double immunolabeling, 97-LAD was localized as if being sandwiched between the NC16A and the carboxyl terminal domains of BPAG2. These results clearly demonstrated the co-localization of 97-LAD and the extracellular portion of BPAG2 in the Lamina Lucida, and suggested that 97-LAD is closely related to, and/or forms a complex with, the extracellular domain of BPAG2.
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97 kda linear iga bullous dermatosis lad antigen localizes to the Lamina Lucida of the epidermal basement membrane
Journal of Investigative Dermatology, 1996Co-Authors: Akira Ishiko, Hiroshi Shimizu, Takashi Hashimoto, Takuji Masunaga, Marian Dmochowski, F Wojnarowska, B S Bhogal, M M Black, Takeji NishikawaAbstract:Linear IgA bullous dermatosis (LAD) is an autoimmune blistering disease in which IgA autoantibodies develop against the epidermal basement membrane zone. Target antigens of the circulating autoantibodies are thought to be heterogeneous, and their ultra-structural localization has not been fully eLucidated. Previous studies with immunoblotting have demonstrated that the 97-kDa autoantigen is detected most frequently in patients' sera and is thought to be a major LAD antigen. Although a recent report suggests that the 97-kDa antigen localized to the hemidesmosomal plaques and the adjacent Lamina Lucida, discrepancies still exist among previous immunoelectron microscopic findings. To identify the precise localization of the 97-kDa LAD antigen, we used two different low-temperature immunoelectron microscopic techniques. For iinmunolabeling, we selected five LAD sera that had a high titer of autoantibodies against the 97-kDa LAD antigen. A post-embedding method with cryofixation and freeze substitution failed to immunolabel the 97-kDa LAD antigen. Cryoultramicrotomy with immunoelectron microscopy succeeded in preserving the antigenicity of the 97-kDa LAD antigen. In all cases, the majority of labeling occurred in the Lamina Lucida beneath the hemidesmosomes. No specific labeling was observed in the hemidesmosomal attachment plaques or the Lamina densa or subLamina densa region, including anchoring fibrils. These results indicate that the 97-kDa LAD antigen is a component of the Lamina Lucida.
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coexistence of psoriasis and an unusual igg mediated subepidermal bullous dermatosis identification of a novel 200 kda lower Lamina Lucida target antigen
British Journal of Dermatology, 1996Co-Authors: Koron Chen, S Shimizu, Shunichi Miyakawa, Akira Ishiko, Hiroshi Shimizu, Takashi HashimotoAbstract:Summary Bullous pemphigoid (BP) is characterized by autoantibodies against 230- and 180-kDa hemidesmosomal antigens located in the most superficial layers of the basement membrane zone (BMZ). Histologically. there is a predominance of eosinophils in the infiltrate. In a psoriatic patient, we identified an unusual autoimmune subepidermal bullous eruption which clinically resembled BP, but which was characterized by IgG autoantibodies against a novel 200-kDa lower Lamina Lucida component, Histologically there was a predominance of neutrophils in the infiltrate. Direct immunofluorescence showed linear immunoglobulin (Ig)G and C3 deposition at the BMZ. The patient's IgG autoantibodies bound exclusively to the dermal side of salt-split normal human skin. Indirect immunogold electron microscopy showed a marked deposition of IgG at the lower Lamina Lucida and minimal deposition at the hemidesmosomes. Immunoblot analysis identified a unique 200-kDa autoantigen in dermal extracts and a faint band of the 230-kDa BP antigen in epidermal extracts. The patient responded dramatically well to cyclosporin A. Although the patient's serum also reacted slightly with the 230-kDa BP antigen, there were significant findings different from the usual immunopathological changes of BP. These included finding a novel 200-kDa lower Lamina Lucida target antigen, the binding of IgG autoantibodies exclusively to the dermal side of the split skin and a predominance of neutrophils in blister infiltrate. The IgG autoantibodies against the 200-kDa Lamina Lucida target antigen seemed to play a major role in the pathogenesis of this unique autoimmune subepidermal dermatosis.
Takashi Hashimoto - One of the best experts on this subject based on the ideXlab platform.
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immunoglobulin g deposition to nonhemidesmosomal Lamina Lucida and early neutrophil involvement are characteristic features in a case of anti p200 pemphigoid
British Journal of Dermatology, 2013Co-Authors: Akira Ishiko, Takashi Hashimoto, Atsushi Shimizu, Takeru Funakoshi, M Ishibashi, Tadashi Yoshida, H Koga, Masayuki AmagaiAbstract:Summary The ultrastructural characteristics and immunolocalization of in vivo bound immunoglobulin G (IgG) in skin affected by anti-p200 pemphigoid have not been eLucidated. To give insight into the mechanism of blister formation we report a new case of anti-p200 pemphigoid, studied with stage-oriented morphological analysis and immunoelectron microscopy. Skin biopsy specimens were evaluated ultrastructurally and histologically with immunohistochemistry. By observing the nonblister site, the blister edge and centre of the blister, we determined that neutrophil infiltration increases gradually at the dermoepidermal junction in association with the destruction of type IV collagen. Ultrastructurally, many neutrophils were observed under the Lamina densa, with vacuole formation in the dermis. At the periphery of the blister, the Lamina densa became fragmented and was observed either at the roof or the floor of the blister. At the centre of the blister, the Lamina densa was mainly observed at the blister floor. Postembedding immunoelectron microscopy demonstrated that the IgG, bound in vivo, localized at the Lamina Lucida, while the area beneath the hemidesmosomes was spared. Together with the early involvement of neutrophils and the destruction of the basal Lamina, we suggest that the binding of autoantibodies to the nonhemidesmosomal Lamina Lucida may induce inflammation with neutrophils, resulting in blister formation.
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heterogeneity of brunsting perry type pemphigoid a case showing blister formation at the Lamina Lucida immune deposition beneath the Lamina densa and autoantibodies against the 290 kd polypeptide along the Lamina densa
Journal of Dermatology, 2011Co-Authors: Haruka Minato, Takashi Hashimoto, Norito Ishii, Shunpei Fukuda, Tomoko Wakasa, Kenichi Wakasa, Ryosuke Sogame, Yuji HoriguchiAbstract:: An otherwise healthy 31-year-old man presented with multiple, vesicular, subepidermal blistering on the head, face, chest and oral cavity, leaving shallow scar formation, typical of Brunsting-Perry type pemphigoid. Direct immunofluorescence showed linear deposition of immunoglobulin (Ig)G and C3 along the basement membrane zone (BMZ), and indirect showed anti-BMZ autoantibodies (IgG, >40×) reacting with the dermal side under the salt-split study. Immunofluorescence staining for type IV collagen and laminins, as well as routine electron microscopy, demonstrated that the cleavage level of the blister was intra-Lamina Lucida. The immunoperoxidase method applied to lesional skin demonstrated IgG deposits along the Lamina densa. The post-embedding immunogold method demonstrated that the autoantibodies against BMZ reacted with the Lamina densa and the dermis just beneath it. Immunoblot studies demonstrated that the autoantibodies reacted with the 290-kD polypeptide (suggesting type VII collagen) when dermal extract was used as the substrate. The patient was treated with combination therapy consisting of 30 mg prednisolone, 900 mg nicotinamide and 750 mg tetracycline, and the number of newly forming blisters decreased. We concluded that Brunsting-Perry type pemphigoid, a rare autoimmune blistering disease, includes cases showing characteristics of epidermolysis bullosa acquisita as well as bullous pemphigoid. This case showed discrepancy between the blistering level (intra-Lamina Lucida) and location of antigen (Lamina densa and sub-Lamina densa areas).
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97 kda linear iga bullous dermatosis lad antigen localizes to the Lamina Lucida of the epidermal basement membrane
Journal of Investigative Dermatology, 1996Co-Authors: Akira Ishiko, Hiroshi Shimizu, Takashi Hashimoto, Takuji Masunaga, Marian Dmochowski, F Wojnarowska, B S Bhogal, M M Black, Takeji NishikawaAbstract:Linear IgA bullous dermatosis (LAD) is an autoimmune blistering disease in which IgA autoantibodies develop against the epidermal basement membrane zone. Target antigens of the circulating autoantibodies are thought to be heterogeneous, and their ultra-structural localization has not been fully eLucidated. Previous studies with immunoblotting have demonstrated that the 97-kDa autoantigen is detected most frequently in patients' sera and is thought to be a major LAD antigen. Although a recent report suggests that the 97-kDa antigen localized to the hemidesmosomal plaques and the adjacent Lamina Lucida, discrepancies still exist among previous immunoelectron microscopic findings. To identify the precise localization of the 97-kDa LAD antigen, we used two different low-temperature immunoelectron microscopic techniques. For iinmunolabeling, we selected five LAD sera that had a high titer of autoantibodies against the 97-kDa LAD antigen. A post-embedding method with cryofixation and freeze substitution failed to immunolabel the 97-kDa LAD antigen. Cryoultramicrotomy with immunoelectron microscopy succeeded in preserving the antigenicity of the 97-kDa LAD antigen. In all cases, the majority of labeling occurred in the Lamina Lucida beneath the hemidesmosomes. No specific labeling was observed in the hemidesmosomal attachment plaques or the Lamina densa or subLamina densa region, including anchoring fibrils. These results indicate that the 97-kDa LAD antigen is a component of the Lamina Lucida.
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coexistence of psoriasis and an unusual igg mediated subepidermal bullous dermatosis identification of a novel 200 kda lower Lamina Lucida target antigen
British Journal of Dermatology, 1996Co-Authors: Koron Chen, S Shimizu, Shunichi Miyakawa, Akira Ishiko, Hiroshi Shimizu, Takashi HashimotoAbstract:Summary Bullous pemphigoid (BP) is characterized by autoantibodies against 230- and 180-kDa hemidesmosomal antigens located in the most superficial layers of the basement membrane zone (BMZ). Histologically. there is a predominance of eosinophils in the infiltrate. In a psoriatic patient, we identified an unusual autoimmune subepidermal bullous eruption which clinically resembled BP, but which was characterized by IgG autoantibodies against a novel 200-kDa lower Lamina Lucida component, Histologically there was a predominance of neutrophils in the infiltrate. Direct immunofluorescence showed linear immunoglobulin (Ig)G and C3 deposition at the BMZ. The patient's IgG autoantibodies bound exclusively to the dermal side of salt-split normal human skin. Indirect immunogold electron microscopy showed a marked deposition of IgG at the lower Lamina Lucida and minimal deposition at the hemidesmosomes. Immunoblot analysis identified a unique 200-kDa autoantigen in dermal extracts and a faint band of the 230-kDa BP antigen in epidermal extracts. The patient responded dramatically well to cyclosporin A. Although the patient's serum also reacted slightly with the 230-kDa BP antigen, there were significant findings different from the usual immunopathological changes of BP. These included finding a novel 200-kDa lower Lamina Lucida target antigen, the binding of IgG autoantibodies exclusively to the dermal side of the split skin and a predominance of neutrophils in blister infiltrate. The IgG autoantibodies against the 200-kDa Lamina Lucida target antigen seemed to play a major role in the pathogenesis of this unique autoimmune subepidermal dermatosis.
David T Woodley - One of the best experts on this subject based on the ideXlab platform.
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Linear IgA bullous dermatosis. Characterization of a subset of patients with concurrent IgA and IgG anti-basement membrane autoantibodies.
Archives of Dermatology, 1995Co-Authors: Lawrence S Chan, Ted B Taylor, David T Woodley, Thomas N. Traczyk, Lynne R. Eramo, John J ZoneAbstract:Background and Design: The term linear IgA bullous dermatosis defines an immune-mediated blistering skin disease characterized by pruritic blisters, subepidermal separation with neutrophilic infiltration, and linear IgA antibody deposition at the basement membrane zone (BMZ). However, some patients with linear IgA bullous dermatosis demonstrate both IgA and IgG anti-BMZ autoantibodies on immunofluorescence. We describe four such patients and attempt to define this group of patients by studying their clinical, histopathologic, immunopathologic, immunoultrastructural, and immunochemical characteristics. Results: Clinically, all four patients had a generalized pruritic blistering skin disease consistent with linear IgA bullous dermatosis. Histopathologically, all four cases demonstrated subepidermal blister formation with a predominantly neutrophilic dermal infiltrate. Immunopathologically, tissue-bound IgA (in four of four patients) and IgG (in three of four patients) antibodies were detected at the BMZ. Circulating IgA (in four of four patients) and IgG (in four of four patients) labeled the epidermal BMZ of normal human skin fractured at the Lamina Lucida and did not label the dermal BMZ. By immunoblot analyses, IgA (in three of three patients) and IgG (in one of three patients) circulating antibodies labeled the 97-kd linear IgA bullous dermatosis antigen. By direct immunoelectron microscopy, IgA and IgG antibodies were localized (in two of two patients) exclusively within the upper Lamina Lucida of the BMZ. Conclusions: Four cases of an immune-mediated blistering skin disease typical for linear IgA bullous dermatosis demonstrated both IgA and IgG anti-BMZ autoantibodies against the linear IgA bullous dermatosis antigen within the upper Lamina Lucida. We conclude that linear IgA bullous dermatosis should include a subgroup of patients with both IgA and IgG anti-BMZ autoantibodies. (Arch Dermatol. 1995;131:1432-1437)
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a newly identified 105 kd lower Lamina Lucida autoantigen is an acidic protein distinct from the 105 kd γ2 chain of laminin 5
Journal of Investigative Dermatology, 1995Co-Authors: Lawrence S Chan, Xuesong Wang, Jean Christophe Lapiere, Peter M Marinkovich, Jonathan C R Jones, David T WoodleyAbstract:A 105-kD lower Lamina Lucida antigen (p105) has been detected by autoantibodies (anti-p105) from patients with a novel immunobullous disease. To distinguish p105 from other known Lamina Lucida components, we performed comparative irnmunoblotting on purified human amniotic laminin-5 (kalinin), 804G matrix (enriched in laminin-5), and keratinocyte and fibroblast proteins using anti-804G matrix antibody 0-18) and anti-p105. J-18 labeled the truncated laminin-5 γ2 chain in amniotic laminin-5, 804G matrix, and keratinocyte conditioned medium, but did not label fibroblast cytosol. Conversely, anti-p105 did not label amniotic laminin-5 or 804G matrix, but did label p105 in both keratinocyte conditioned medium and fibroblast cytosol. J-18 labeled the 105-kD laminin-5 γ2 chain in reduced keratinocyte proteins and a 400-kD laminin-5 complex under non-reducing conditions. In contrast, anti-p105 labeled p105 under both reducing and non-reducing conditions but did not label a 400-kD protein complex. Similarly, comparative immunoblotting on keratinocyte proteins using anti-p105 and anti-laminin-1 revealed no commonly labeled protein bands. Electrophoretic fractionations by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of these fractions revealed that the peak fractions of keratinocyte proteins reactive with anti-p105 are different from those reactive with J-18. Furthermore, keratinocyte proteins fractionated by Mono Q anionexchange chromatography revealed fractions immunoreactive with anti-p105, whereas J-18 showed no reactivity with these fractions. Two-dimensional gel electrophoresis and immunoblotting with anti-p105 revealed p105 to be an acidic protein with isoelectric points between 5.7 and 6.3, distinct from the isoelectric points of laminin-5 γ2 chain. We conclude that p105 is an acidic protein located in the Lamina Lucida and distinct from the truncated laminin-5 γ2 chain and the laminin-1 family.
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a novel 105 kda Lamina Lucida autoantigen association with bullous pemphigoid
Journal of Investigative Dermatology, 1994Co-Authors: Stephanie L Cotell, Lawrence S Chan, Jean Christophe Lapiere, John D Chen, Toshiroh Iwasaki, Paul A Krusinski, David T WoodleyAbstract:Abstract Several cases have been reported of patients with immunemediated subepidermal blistering disorders whose autoantibodies react to antigens present on both the dermal and epidermal side of 1 M NaCl-split skin. In this report, we identify, localize, and characterize the basement membrane zone antigen corresponding to the dermal staining in a patient whose serum stains both the dermal and epidermal side of 1 M NaCl-split skin. This patient's serum contains autoantibodies directed against a 105-kilodalton(kDa) dermal antigen and the 230-kDa epidermal (bullous permphigoid) antigen. This novel 105-kDa protein was previously identified as the sole antigen in another patient with a unique bullous disease whose autoantibodies were directed against only the dermal side of 1 M NaCl-split skin. This 105-kDa antigen was identical by one- and two-dimensional immunoblot analysis in these two patients. By immunoblot analysis, autoantibodies from our patient labeled a 105-kDa protein within various extracts of human skin basement membrane. Immunoblot analyses using epitope-selected autoantibodies directed against the 105-kDa protein demonstrated that this antigen is independent and distinct from other known basement membrane antigens. The 105-kDa antigen is an extracellular matrix component of the basement membrane, which is synthesized and secreted by both keratinocytes and fibroblasts. Identical electrophoretic migration of cellular and secreted forms of the protein suggested there is no major post-translational modification of the protein. Immunomapping of normal human skin fractured through the dermal-epidermal junction by incubation in 1 M NaCl or by suction blistering demonstrated that the location of the 105-kDa antigen within the basement membrane zone is between the bullous pemphigoid antigens and two other Lamina Lucida components, laminin and nicein. These data demonstrate clearly that a subepidermal autoimmune bullous disease may have autoantibodies directed against two distinct components of the dermal-epidermal junction.
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identification and partial characterization of a novel 105 kdalton lower Lamina Lucida autoantigen associated with a novel immune mediated subepidermal blistering disease
Journal of Investigative Dermatology, 1993Co-Authors: Robert A Briggaman, Lawrence S Chan, Jodavid Fine, David T Woodley, Craig Hammerberg, Rhett J Drugge, Kevin D CooperAbstract:Certain skin basement membrane components, such as bullous pemphigoid antigens and epidermolysis bullosa acquisita antigen, were discovered as a result of an autoimmune reaction. In this report, we describe a unique Lamina Lucida determinant associated with a novel immune-mediated subepidermal bullous dermatosis. This unique bullous dermatosis resembled severe toxic epidermal necrolysis clinically. The histologic findings resemble dermatitis herpetiformis. Direct immunofluorescence microscopy detected linear immunoglobulin G (lgG) and C3 deposition at the cutaneous basement membrane zone of lesional and perilesional skin. Direct and indirect immunoelectron microscopy localized the LgG deposits to the lowest portion of the Lamina Lucida. The patient's autoantibodies, belonging to the IgG1 subclass, labeled basement membrane zone of normal intact human skin, oral mucosa, and conjunctiva, and localized to the dermal side of salt-split normal adult and neonatal human skin, but failed to react with human fetal skin up to 142 gestational days. The patient's autoantibodies failed to react with bullous pemphigoid antigens or epidermolysis bullosa acquisita antigen (type VII collagen) by immunoblotting. Instead, the patient's autoantibodies unequivocally labeled a 105-kilodalton (kD) protein in cellular extracts and conditioned media of human cultured keratinocytes and dermal fibroblasts. The titer of the patient's antibody against the cutaneous basement membrane zone and the intensity of the antibody reactivity against the 1O5-kD protein paralleled the patient's disease activity. Thus, this 105-kD lower Lamina Lucida protein represents a novel autoantigen and this patient's disease represents a deep Lamina Lucida pemphigoid, distinguishable from all other known autoimmune bullous dermatoses.
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Labeling of fractured human skin with antibodies to BM 600/nicein, epiligrin, kalinin and other matrix components.
Journal of Dermatological Science, 1993Co-Authors: Elizabeth Ceilley, Robert A Briggaman, Eugene A. Bauer, Noriko Watanabe, Dorit Shapiro, Patrick Verrando, Robert E. Burgeson, David T WoodleyAbstract:Abstract A variety of methods were used to fracture the dermal-epidermal junction (DEJ) of human skin. These included warm and hot phosphate buffered saline, trypsin, cold 1 M salt, potassium bromide and proteolytic digestion with dispase. The localization and sensitivity of basement membrane components (bullous pemphigoid antigen, BM 600/nicein, epiligrin, kalinin, laminin, collagens IV and VII (EBA antigen) and linkin) were determined after the DEJ was fractured by each method. We found that the basement membrane zone proteins, BM 600/nicein, epiligrin and kalinin remained with the dermal side of the DEJ fractured through the Lamina Lucida by cold salt, phosphate buffered saline and potassium bromide. BM 600/nicein, epiligrin and kalinin were not detected after treatment with trypsin. In contrast, laminin, another glycoprotein in the Lamina Lucida, was insensitive to all of the procedures, but co-localized to the dermal side of DEJ-fractured skin. We also found that separation of the DEJ with brief exposure of skin to 56°C provided a useful substrate for testing the autoantibodies in the sera of patients with epidermolysis bullosa acquisita (EBA). Heat-separated skin can be prepared in a significantly shorter period of time than salt-separated skin.
