Effects of pemphigus antibody on the organization of microtubules and keratin-intermediate filaments in cultured human keratinocytes

Kitajima, Yasuo; Inoue, S; Yaoita, Hideo

doi:10.1111/j.1365-2133.1986.tb02795.x

Cited by 33 publications

(22 citation statements)

References 19 publications

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“…The first proposes that anti-Dsg3 antibody-dependent steric hindrance interferes with intercellular adhesive function(s) of Dsg3, leading directly to desmosomal dissociation (12)(13)(14). However, we have observed no inhibition of Ca 2ϩ -induced desmosome formation by PV-IgG binding to surface PV-antigens (15), suggesting that PV-IgG does not directly inhibit desmosome formation, even though the antibodies may cause steric hindrance between homophilic Dsg3 interactions.…”

mentioning

confidence: 63%

“…Cell Culture-DJM-1 cells, a cell line isolated from human skin squamous cell carcinoma (15) were seeded either in 35-mm dishes or on glass coverslips. Cells were cultured in Eagle's minimum essential medium (MEM) (Nissui Phama, Tokyo, Japan) containing 10% fetal calf serum, 0.4 g/ml hydrocortisone (Sigma), 20 ng/ml epidermal growth factor (TOYOBO, Osaka, Japan), 84 ng/ml cholera toxin (List Biological Laboratories, Campbell, CA), and 1.8 mM Ca 2ϩ .…”

Section: Methodsmentioning

confidence: 99%

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Anti-desmoglein 3 (Dsg3) Monoclonal Antibodies Deplete Desmosomes of Dsg3 and Differ in Their Dsg3-depleting Activities Related to Pathogenicity

Yamamoto

Aoyama

Shu

et al. 2007

Journal of Biological Chemistry

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Pemphigus vulgaris (PV) is an autoimmune blistering disease, characterized by the loss of cell-cell adhesion between epidermal keratinocytes and the presence of autoantibody against desmoglein 3 (Dsg3), which provides adhesive integrity to desmosomes between adjacent keratinocytes. We have previously shown that PV-IgG purified from patients depletes desmosomes of Dsg3. However, PV-IgG contains not only antibodies against a variety of different epitopes of Dsg3 but also against other unknown antigens. Therefore, we examined whether the Dsg3-depleting activity of PV-IgG is generated specifically by antiDsg3 activity in a human squamous cell carcinoma cell line (DJM-1) and normal human keratinocytes by using four different pathogenic and nonpathogenic monoclonal antibodies against Dsg3. We demonstrate that these monoclonal antibodies deplete cells and desmosomes of Dsg3, as PV-IgG does. Individual monoclonal anti-Dsg3 antibodies display characteristic limits to their Dsg3-depleting activity, which correlates with their pathogenic activities. In combination, these antibodies exert a cumulative or synergistic effect, which may explain the potent Dsg3-depleting capability of PV-IgG, which is polyclonal. Finally, although Dsg3-depletion activity correlated with AK-monoclonal antibody pathogenicity in mouse models, the residual level of Dsg3, when below ϳ50%, does not correlate with the adhesive strength index in the present study. This may suggest that although the Dsg3 depletion is not indicative for adhesive strength, the level of Dsg3 can be used as a read-out of pathogenic changes within the cell and that the Dsg3 depletion from desmosomes plays an important role in skin fragility or susceptibility to blister formation in PV patients.Pemphigus is a group of an autoimmune blistering diseases that includes two major types; i.e. pemphigus vulgaris (PV), 2 characterized by suprabasal acantholysis (the loss of cell-cell adhesion between keratinocytes) in the epidermis and autoantibodies against Dsg3 (1-4), and pemphigus foliaceus, characterized by superficial acantholysis in the granular cell layer of the epidermis and autoantibodies against Dsg1 (5, 6). Dsg3 and Dsg1 are members of the desmosomal cadherins that provide adhesive integrity to desmosomes between adjoining keratinocytes in the epidermis; the former distributes primarily in the deeper epidermis, whereas the latter is localized to the superficial epidermis. This distribution profile of these target antigens, Dsg3 and Dsg1, of PV and pemphigus foliaceus autoantibodies explains the characteristic clinical features and histopathological localization of acantholysis in their respective diseases. PV, but not pemphigus foliaceus, affects the mucous membrane, where Dsg3 is localized but Dsg1 is rarely expressed (7-10).Recent work has suggested that two principal hypotheses emerge as pathomechanisms underlying the generation of PV blisters (4, 11). The first proposes that anti-Dsg3 antibody-dependent steric hindrance interferes with intercellular adhesive function(s...

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mentioning

confidence: 63%

Section: Methodsmentioning

confidence: 99%

Anti-desmoglein 3 (Dsg3) Monoclonal Antibodies Deplete Desmosomes of Dsg3 and Differ in Their Dsg3-depleting Activities Related to Pathogenicity

Yamamoto

Aoyama

Shu

et al. 2007

Journal of Biological Chemistry

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“…The primary events after binding of pemphigus antibodies to keratinocytes are changes in the cytoskeleton of keratinocytes with retraction of tonofilaments [17][18][19] and breaking of the bond between tonofilaments and desmosomes culminating in shrinkage of the basal cells. 8,[20][21][22] The detachment of tonofilaments occurs before the desmosomes show any visible alteration 8,23 or loss in adhesion.…”

Section: Nondesmosomal Mechanisms Appear To Be Involved In Pv Acanthomentioning

confidence: 99%

A novel explanation for acantholysis in pemphigus vulgaris: The basal cell shrinkage hypothesis

Bystryn¹,

Grando²

2006

Journal of the American Academy of Dermatology

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“…DJM-1 cells, a human squamous cell carcinoma cell line (Kitajima et al, 1986(Kitajima et al, , 1987, were cultured in s-MEM (Life Technologies, Inc.) supplemented with 10% (v/v) Ca 2+ -chelated fetal bovine serum (IS-Japan, Tokyo, Japan), 84 ng/ml choleratoxin (Life Technologies, Inc.) and 0.4 mg/ ml hydrocortisone (Sigma) and CaCl2 (the final Ca 2+ concentration of the medium was adjusted to 0.09 mM). Cells were cultured on 24´24 mm glass coverslips (Matsunami Glass Co. Tokyo, Japan) for immunofluorescent microscopy in 35 mm cell culture dishes (Becton-Dickinson and Co., Franklin Lakes, NJ, USA) and 100 mm cell culture dishes (Corning Costar Corp., Cambridge, MA, USA) for biochemical study in 37°C CO2-incubator.…”

Section: Cellsmentioning

confidence: 99%

Scraped-Wounding Causes Activation and Association of C-Src Tyrosine Kinase with Microtubules in Cultured Keratinocytes.

Yamada¹,

Aoyama²,

Owada³

et al. 2000

Cell Struct. Funct.

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ABSTRACT. In order to elucidate the function of c-Src in keratinocytes, we studied the intracellular distribution of its active and inactive form in cultured normal human keratinocyte, using anti-c-Src monoclonal antibody clone 28, which recognizes the active form of c-Src (dephosphorylated at COOH-terminal residue Tyr 530), and monoclonal antibody clone 327 which recognizes both active and inactive forms. Since c-Src has been suggested to be involved in the control of cell adhesion in other cells, we produced a dynamic condition of cell migration by cutting culture cell colonies into squares to form a mesh pattern with a blade (culture wound model). Before cutting, the active form was expressed in cells located only at the periphery of colonies or isolated migrating cells, and was associated with microtubules. Wounding the colony generated a dramatic and rapid activation of c-Src in a few rows of cells along the cut edges, which were made even at the middle of colony, resulting in the association of the active form with microtubules. This increase of the active form was also detected by immunoblotting of cell extracts. These reactions were inhibited by 1 mM sodium orthovanadate, a protein-tyrosine phosphatase inhibitor. ST 638, a potent Src family tyrosine kinase inhibitor, inhibited the migration of keratinocytes in the culture wound healing model. These results suggest that wounding the culture causes activation of c-Src in keratinocytes, and thus activated c-Src may play a role in the function of microtubules during cell migration, especially at an early stage of wound healing.Key words: c-Src/keratinocyte/wound healing/microtubules During wound healing, keratinocytes (KCs) must dynamically regulate cell-cell contacts and cytoskeletons in order to migrate to and cover the surface of the wound. Major cell-cell contacts between KCs are adherens junctions and desmosomes, the former of which are specifically associated with actin microfilaments and the latter with keratin-intermediate filaments. The intercellular contacts of adherens junctions are mediated in a homotypic manner with Ecadherin (Takeichi, 1991), and linked to actin filaments via vinculin, =-actinin, =-catenin, >-catenin, plakoglobin and P120-Cas (Kam et al., 1995;Tsukita et al., 1992;Reynolds et al., 1992Reynolds et al., , 1994Reynolds et al., , 1996. Increase in tyrosine phosphorylation of >-catenin, plakoglobin and P120-Cas, which are known as substrate of c-Src, has been shown to be correlated with decrease of cell adhesion, which is one of the typical characteristics of neoplastic transformation (Matsuyoshi et al., 1992; Behrens et al., 1993;Hamaguchi et al., 1993;Papkoff, 1997). The regulation of cell-cell contacts is thought to be one of the most important events for KCs to cover the wound surface (Grinnell, 1990), and the following differentiation. Therefore, it is of potential importance to clarify the intracellular signaling mechanisms involved in wound healing in skin.Src is a non receptor type tyrosine kinase, which is widely expressed ...

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Effects of pemphigus antibody on the organization of microtubules and keratin-intermediate filaments in cultured human keratinocytes

Cited by 33 publications

References 19 publications

Anti-desmoglein 3 (Dsg3) Monoclonal Antibodies Deplete Desmosomes of Dsg3 and Differ in Their Dsg3-depleting Activities Related to Pathogenicity

Anti-desmoglein 3 (Dsg3) Monoclonal Antibodies Deplete Desmosomes of Dsg3 and Differ in Their Dsg3-depleting Activities Related to Pathogenicity

A novel explanation for acantholysis in pemphigus vulgaris: The basal cell shrinkage hypothesis

Scraped-Wounding Causes Activation and Association of C-Src Tyrosine Kinase with Microtubules in Cultured Keratinocytes.

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