TY - JOUR
T1 - Demonstration of pemphigus antibodies on the cell surface of murine epidermal cell monolayers and their internalization
AU - Patel, H. P.
AU - Diaz, L. A.
AU - Anhalt, G. J.
AU - Labib, R. S.
AU - Takahashi, Y.
N1 - Funding Information:
Manuscript received March 5, 1984; accepted 11, 1984. Supported in part by U.S. Public Service Grants R01-AM 32081, R01-AM 32599, R23-AM 32079, and R23-AM 32490 from the National Institutes of Health, and a gift from the Estee Lauder Corporation. Reprint requests to: Harish P. Patel, M.D., Department of Dermatology, The Johns Hopkins University, Blalock Building-Room 909E, 600 N. W o lfe Street, Baltimore, Maryland 21205. Abbreviations: ABCPx: avidin-biotin peroxidase complex D A B: 3,3' -diaminobenzidine E M: electron microscopy FCS: fetal calf serum G B S : glutaraldehyde-borohydride-saponin ICS: intercellular spaces IF: immunofluorescence Immuno-EM: immunoelectron microscopy M-199: Medium 199 PBS: phosphate-buffered saline P V: pemphigus vulgaris P&S: penicillin and streptomycin
PY - 1984
Y1 - 1984
N2 - The pathogenic effects of pemphigus vulgaris (PV) antibodies on epidermal cells can be demonstrated both in vitro using skin organ culture or primary epidermal cell cultures (PECC) and in vivo by passive transfer of PV antibodies into neonatal BALB/c mice. Although PV antibodies have been localized on the epidermal cell surface by several techniques, little is known about the fate of these autoantibodies subsequent to their surface binding. We have examined this, using murine PECC which express pemphigus antigen on their surface, and followed the fate of the bound antibody molecules. Forty-eight-hour PECC were incubated at 27°C with PV antibodies for 30 min and then with horseradish peroxidase-labelled antihuman IgG. This was considered time 0. The monolayers were fixed with glutaraldehyde after 0, 0.5, 1, 3, 6, 18, and 24 h incubation at 37°C and then processed for electron microscopy. At time 0 hour, PV antibodies is detected bound evenly along the surface of keratinocytes. Within 20 min, the bound PV antibodies becomes clustered, internalized into submembranous vesicles via surface pits, and eventually fused with lysosomes. Widening of the intercellular spaces was also seen in PECC treated with PV antibodies within the first 24 h. PECC treated with normal human IgG in parallel cultures showed no such surface binding, internalization, or cell-cell detachment. Treatment with cytochalasin-D and/or colchicine did not affect the internalization of the PV antibodies, but fusion with lysosomes was not seen in treated cultures. These findings suggest that PV antibodies binds a surface antigen and the complex is internalized and fused with lysosomes in a process that may have pathophysiologic relevance.
AB - The pathogenic effects of pemphigus vulgaris (PV) antibodies on epidermal cells can be demonstrated both in vitro using skin organ culture or primary epidermal cell cultures (PECC) and in vivo by passive transfer of PV antibodies into neonatal BALB/c mice. Although PV antibodies have been localized on the epidermal cell surface by several techniques, little is known about the fate of these autoantibodies subsequent to their surface binding. We have examined this, using murine PECC which express pemphigus antigen on their surface, and followed the fate of the bound antibody molecules. Forty-eight-hour PECC were incubated at 27°C with PV antibodies for 30 min and then with horseradish peroxidase-labelled antihuman IgG. This was considered time 0. The monolayers were fixed with glutaraldehyde after 0, 0.5, 1, 3, 6, 18, and 24 h incubation at 37°C and then processed for electron microscopy. At time 0 hour, PV antibodies is detected bound evenly along the surface of keratinocytes. Within 20 min, the bound PV antibodies becomes clustered, internalized into submembranous vesicles via surface pits, and eventually fused with lysosomes. Widening of the intercellular spaces was also seen in PECC treated with PV antibodies within the first 24 h. PECC treated with normal human IgG in parallel cultures showed no such surface binding, internalization, or cell-cell detachment. Treatment with cytochalasin-D and/or colchicine did not affect the internalization of the PV antibodies, but fusion with lysosomes was not seen in treated cultures. These findings suggest that PV antibodies binds a surface antigen and the complex is internalized and fused with lysosomes in a process that may have pathophysiologic relevance.
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U2 - 10.1111/1523-1747.ep12273480
DO - 10.1111/1523-1747.ep12273480
M3 - Article
C2 - 6389716
AN - SCOPUS:0021714389
SN - 0022-202X
VL - 83
SP - 409
EP - 415
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 6
ER -