A 45-year-old woman with epidermolysis bullosa aquisita is presented. The clinical, histological, and immunopathological features were in keeping with the previous reports of this disease. The patient also had anti-basal cell cytoplasmic antibodies at a significant titer, which is considered an unusual finding associated with this disorder. Treatment with a moderate dose of corticosteroid was effective in controlling the bullous lesions
Autoantibodies/*analysis ; Complement C3/analysis ; Cytoplasm/*immunology ; Epidermis/*immunology ; Epidermolysis Bullosa Acquisita/diagnosis/*immunology ; Female ; Humans ; Middle Aged

To explore the role of dentritic epidermal T lymphocytes ( DETCs) in immune rejection of skin allograft in mice and its related mechanism. Methods (1) Full-thickness skin was harvested from back of one male wild type (WT) C57BL/6 mouse. Epithelial cells were isolated for detection of the expression of DETCs and their phenotype with flow cytometer. Another male WT C57BL/6 mouse was used to harvest full-thickness skin from the back. Epidermis was isolated for observation of the morphological characteristics of DETCs with immunofluorescence technology. (2) Four male green fluorescence protein (GFP)-marked C57BL/6 mice, 7 female WT C57BL/6 mice (group WT), and 7 female ybT lymphocytes 8 gene knock-out (GK) C57BL/6 mice (group GK) were used. Full-thickness skin in the size of 1.4 cm x 1.4 cm on the back of mice in groups WT and GK were excised, and the wounds were transplanted with full-thickness skin in the size of 1.2 cm x 1.2 cm obtained from male GFP-marked C57BL/6 mice. The survival time of skin grafts was affirmed with small animal in vivo imager and naked eyes and recorded. (3) Two male WT C57BL/6 mice were used to isolate epithelial cells. Cells were inoculated into 48-well plate and divided into activation group (A) and control group (C) according to the random number table, with 4 wells in each group. Cells in group A were treated with 10 pL concanavalin A in the concentration of 2 microg/mL for 24 hours, while those in group C with PBS in the same volume as that in group A. The expression of interferon y in DETCs was detected with flow cytometer. (4) Four male GFP-marked C57BL/6 mice were used as donors. Fourteen female WT C57BL/6 mice were used as receptors and divided into interferon gamma neutralizing group (IN) and control group (C) according to the random number table, with 7 mice in each group. The skin transplantation model of C57BL/6 male to C57BL/6 female was established as in part (2). Before surgery and 72 hours after, mice in group IN were intraperitoneally injected with 200 pL interferon y neutralizing antibody in the concentration of 1 mg/mL, and those in group C with normal saline in the same volume as that in group IN. The survival time of skin grafts was observed and recorded using the methods in part (2), and the result of group IN was compared with that of group GK in part (2). The survival curve of skin grafts was processed with Log-rank ( Mantel-Cox) test. Results (1) The positive expression rate of DETCs in epithelial cells of skin in mouse was 7.27%, and they were all CD3 cells. DETCs were found to be scattered in the epidermis of skin in mouse with dendritic morphology. (2) The survival time of skin grafts of mice in group GK was 22-35 d, obviously longer than that in group WT (12-16 d, y2 = 14. 10 , P < 0.001). (3) Expression of interferon gamma was detected in 22. 70% DETCs in group A, which was obviously higher than that in group C (0.51%). (4) The survival time of skin grafts of mice in group IN was 19-24 d, which was obviously longer than that in group C (12-16 d, chi 2 = 13.60, P < 0.001) but close to that in group GK as in part (2) (chi2 = 0.06, P = 0.810). Conclusions DETCs are involved in promotion of immune rejection of skin allograft probably by secretinf interferon gamma.
Allografts ; Animals ; Epidermis ; Female ; Graft Survival ; immunology ; physiology ; Interferon-gamma ; immunology ; metabolism ; Lymphocytes ; Male ; Mice ; Mice, Inbred C57BL ; Skin ; Skin Transplantation ; T-Lymphocytes ; immunology

OBJECTIVETo explore the role of indirect antigen presentation pathway on the immunogenecity of epidermal cells.

METHODSHuman epidermal cells (HEC), allogeneic human peripheral blood lymphocytes (PBL) and mononuclear cells (PBM, including monocytes) were isolated and cultured in vitro. HECs were transfected by human-originated CTLA4Ig-adenovirus vector. The CTLA4Ig expression was observed. Allogeneic PBLs or PBMs were added to the transfected and non-transfected HECs with simple cultured PBLs and PBMs as the control. The proliferation of PBL and PBM was determined by (3)H-TdR incooperation.

RESULTSHECs could be successfully transfected by CTLA4Ig-adenovirus vector and expressed corresponding proteins. The non-transfected HECs could stimulate slight proliferation of allogeneic PBLs (P < 0.05) and stimulate remarkable proliferation of PBMs (including monocytes) (P < 0.05). The proliferation reaction of PBLs and PBMs decreased significantly (P < 0.05) after being stimulated by HEC which was modulated by CTLA4Ig genes.

CONCLUSIONIndirect antigen presentation pathway might play important roles in the HEC immunogenicity which could be evidently inhibited by CTLA4Ig.

Adenoviridae ; genetics ; Antigen Presentation ; immunology ; physiology ; Antigens, CD ; Antigens, Differentiation ; genetics ; immunology ; CTLA-4 Antigen ; Cell Division ; immunology ; Cells, Cultured ; Coculture Techniques ; Epidermis ; cytology ; immunology ; metabolism ; Genetic Vectors ; genetics ; Humans ; Leukocytes, Mononuclear ; cytology ; immunology ; Lymphocytes ; cytology ; immunology ; Signal Transduction ; Transfection

BACKGROUNDThe presence of autoantibodies against multiple epidermal proteins is an important feature in paraneoplastic pemphigus (PNP). Circulating anti-desmoglein 3 autoantibody, the major pathogenic autoantibody in pemphigus vulgaris (PV), has been proved pathogenic in PNP. Because of many clinical differences between PNP and PV, we speculate about the involvement of other autoantibodies in the pathogenesis of PNP. Envoplakin (EPL) and periplakin (PPL) are recognized by most PNP sera. Their linker subdomains are highly homologous and necessary for the association of intermediate filaments.

METHODSWe characterized the autoantibodies against the linker subdomains of EPL and PPL in PNP patients' sera and their associated tumors by enzyme-linked immunosorbent assay (ELISA) and immunofluorence. We also applied the purified autoantibodies against EPL and PPL from PNP sera to cultured human epidermal keratinocytes (HEK), to evaluate the changes of cell-cell adhesion.

RESULTSAutoantibodies against EPL and PPL were detected in most PNP patients by ELISA, and the decrease of these autoantibodies after removal of the tumors was roughly comparable to the improvement of clinical symptoms. Cultured tumor cells from PNP patients secreted these autoantibodies. Specific immunoglobulin receptors for EPL and PPL were found on B lymphocytes in tumors from PNP. Furthermore, purified anti-EPL and anti-PPL autoantibodies from PNP sera were capable of dissociating cultured human epidermal keratinocytes.

CONCLUSIONAutoantibodies against EPL and PPL may also be pathogenic in PNP.

Adolescent ; Adult ; Autoantibodies ; immunology ; pharmacology ; Cell Adhesion ; drug effects ; Cells, Cultured ; Desmoglein 3 ; immunology ; Enzyme-Linked Immunosorbent Assay ; Epidermis ; cytology ; Female ; Humans ; Keratinocytes ; cytology ; drug effects ; Male ; Membrane Proteins ; immunology ; Middle Aged ; Paraneoplastic Syndromes ; immunology ; metabolism ; Pemphigus ; immunology ; metabolism ; Plakins ; immunology ; Protein Precursors ; immunology ; Young Adult

OBJECTIVETo explore a feasible method to repair full-thickness skin defects utilizing tissue engineered techniques.

METHODSThe Changfeng hybrid swines were used and the skin specimens were cut from the posterior limb girdle region, from which the keratinocytes and fibroblasts were isolated and harvested by trypsin, EDTA, and type II collagenase. The cells were seeded in Petri dishes for primary culture. When the cells were in logarithmic growth phase, they were treated with trypsin to separate them from the floor of the tissue culture dishes. A biodegradable material, Pluronic F-127, was prefabricated and mixed with these cells, and then the cell-Pluronic compounds were seeded evenly into a polyglycolic acid (PGA). Then the constructs were replanted to the autologous animals to repair the full-thickness skin defects. Histology and immunohistochemistry of the neotissue were observed in 1, 2, 4, and 8 postoperative weeks.

RESULTSThe cell-Pluronic F-127-PGA compounds repaired autologous full-thickness skin defects 1 week after implantation. Histologically, the tissue engineered skin was similar to the normal skin with stratified epidermis overlying a moderately thick collageneous dermis. Three of the structural proteins in the epidermal basement membrane zone, type IV collagen, laminin, and type VII collagen were detected using immunohistochemical methods.

CONCLUSIONSBy studying the histology and immunohistochemistry of the neotissue, the bioengineered skin graft holds great promise for improving healing of the skin defects.

Animals ; Disease Models, Animal ; Epidermis ; pathology ; Skin ; immunology ; injuries ; pathology ; Skin Transplantation ; methods ; Swine ; Tissue Engineering ; methods ; Transplantation, Autologous ; Transplants ; Treatment Outcome ; Wounds and Injuries ; surgery

OBJECTIVETo study the mechanism of promoting wound healing with mixed grafting of autologous and allogeneic microskin in rats.

METHODSFifteen male Wistar rats served as alloskin donor. Forty-five female SD rats with full-thickness skin defect served as recipients in the study. In part one experiment, 27 SD rats were randomly divided into group I (n = 9, without allogeneic microskin), group II (n = 9, with mixed grafting of allogeneic microskin at area expansion rate of 10:1); group III (n = 9, with mixed grafting of allogeneic microskin at area expansion rate of 10:3) with grafting with the same amount of autologous microskin at area expansion rate of 10:1. In part two experiment, 18 SD rats were also divided into group I (n = 6, with autologous microskin only); group II (n = 6, with mixed grafting of autologous and allogeneic microskin with area expansion rate of 20:1 and 20:3 respectively); group III (n = 6, with mixed grafting of autologous and allogeneic microskin with area expansion rate of 20:1 and 20:6, respectively). Biopsy samples were obtained from healed wound area of SD rats in each group at different time points after operation. The histological changes, epidermal thickness, and immunohistochemical staining of Integrin beta1 were observed.

RESULTS(1) HE staining showed the thickness of epidermis in each group increased obviously, and various amounts of mononuclear cell infiltration and different degrees of vasodilation appeared in the dermal layer during 2 - 4 weeks. (2) Epidermal thickness in group II and III of part one experiment were significantly thicker than that in group I during 2 - 4 weeks after operation (P < 0.05), and the similar result was also seen in part two experiment on 3 and 4 weeks after operation (P < 0.05). (3) A positive staining pattern for Integrin beta1 was seen in the suprabasal layers (especially in the spinous and granular layers) in all groups. In part one experiment, the expression of Integrin beta1 in group II and III were obviously higher than that in group I on 2 week after operation (P < 0.01), and the expression of Integrin beta1 in group II (10 982 +/- 2169) was also higher than that in group III (4240 +/- 512, P < 0.01); the expression of Integrin beta1 in group II was still higher than that in group I and III (P < 0.01) 3 and 4 weeks after operation. In part two experiment, the expression of Integrin beta1 in group III (1618 +/- 171) was higher than that in group I 3 weeks after operation (1060 +/- 146, P < 0.05).

CONCLUSIONThe ectopic and increased expression of Integrin beta1 was closely associated with the proliferation and differentiation of epidermal cells, wound reepithelialization and thickened epidermis in mixed grafting of autologous and allogeneic microskin. Integrin beta1 may be responsible in promoting wound healing.

Animals ; Epidermis ; metabolism ; Epithelium ; metabolism ; Female ; Integrin beta1 ; immunology ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Skin Transplantation ; Transplantation, Homologous ; Wound Healing