Passive transfer of collagen XVII-specific antibodies induces sustained blistering disease in adult mice
© Chiriac et al.; licensee BioMed Central Ltd. 2013
Received: 31 July 2012
Accepted: 6 December 2012
Published: 29 January 2013
Bullous pemphigoid is a subepidermal blistering disorder associated with tissue-bound and circulating autoantibodies directed mainly to the hemidesmosomal component collagen XVII. While recapitulating the main immunopathological features of the human disease, frank skin blistering does not develop in the absence of skin rubbing in experimental pemphigoid models that have been established in neonatal mice. Moreover, due to their experimental design they only allow for short-term disease observation. In the present study we aimed to establish a model that reproduces the frank skin blistering seen in patients and allows for longer observation times.
Rabbit and sheep antibodies specific to several fragments of collagen XVII were generated and the purified antibodies were passively transferred into adult mice.
Collagen XVII-specific IgG bound to the basal membrane of the skin and mucous membranes activating murine complement in vivo. Mice injected with collagen XVII-specific antibodies, in contrast to mice receiving control antibodies, developed frank skin blistering disease, reproducing human bullous pemphigoid at the clinical, histological and immunopathological levels. Titres of circulating IgG in the serum of mice correlated with the extent of the clinical disease. Mice receiving sheep antibodies specific to murine collagen XVII showed an early onset and a more active disease when compared to litter mates receiving specific rabbit antibodies.
This novel animal model for bullous pemphigoid should facilitate further investigations of the pathogenesis of bullous pemphigoid and the development of innovative therapies for this disease.
KeywordsAutoimmunity Collagen XVII Inflammation Skin
Bullous pemphigoid (BP) is the most common autoimmune blistering disease in Western Europe and USA[1–4]. Immunopathologically, the disease is characterized by the presence of tissue-bound and circulating autoantibodies directed against the dermal-epidermal junction (DEJ). Antibodies in BP patients mainly recognize BP180 (bullous pemphigoid antigen of 180kDa) also known as collagen (C)XVII, a transmembrane protein with a type II orientation that spans the lamina lucida, and to a lesser extent BP230 (bullous pemphigoid antigen of 230kDa), an intracellular hemidesmosomal protein[5–10]. Since these proteins contribute to the tight anchoring of basal keratinocytes on the underlying basal membrane, disruption of their function either through genetic defects or immunopathological mechanisms results in a dermal-epidermal separation that ultra-structurally localizes within the lamina lucida[10–15].
The pathogenic relevance of autoantibodies in BP is supported by compelling evidence: 1) BP autoantibodies were shown to recruit and activate leukocytes resulting in dermal-epidermal separation in cryosections of human skin; 2) Antibodies against mouse and human BP180/ CXVII induce subepidermal blisters when passively transferred into wild-type or CXVII-humanized mice, respectively; in addition, antibodies to human type XVII collagen cross the placenta of immunized mice and induce disease in type XVII collagen-humanized neonates; 3) Transfer of splenocytes from mice immunized against human type XVII collagen into Rag2−/−/ CXVII-humanized mice results in sustained immunoglobulin G (IgG) production and BP phenotype; 4) Autoantibodies in the majority of BP patients recognize the non-collagenous 16th (NC16)A domain of BP180/ CXVII and pre-adsorption of pathogenic sera with NC16A abolished their pathogenic potential both in vivo[15, 18] and ex vivo.
Despite important advances in disease management over the last decades, BP is still a disease with one-year mortality rates of up to 40%. The existence of animal models reproducing the disease is essential for our understanding of pathomechanisms with major implications for afflicted individuals. The most direct approach for reproducing autoantibody-induced autoimmune diseases in vivo has been to inject patients’ serum or purified antibodies specific to culprit autoantigens into healthy individuals. The method pioneered in the 1950s by the Harrington–Hollingsworth Experiment wherein Dr. Harrington received blood from an idiopathic thrombocytopenic purpura patient was first used to test the hypothesis that platelet destruction was caused by a factor circulating in the patient’s blood[21, 22]. Future studies using the passive transfer of IgG into laboratory animals demonstrated the pathogenic effect of antibodies in several diseases, including myasthenia gravis, pemphigus vulgaris and pemphigus foliaceus. Previous attempts to reproduce BP by this “classical” transfer of disease through antibodies from patients into experimental animals were unsuccessful[26–30]. The failure to transfer the disease in mice has been explained by a lack of reactivity of patients’ autoantibodies with the murine BP180/ CXVII-specific due to the low degree of homology between the human and mouse type XVII collagen[14, 15, 17, 18, 31]. A further reason for the lack of pathogenicity of pemphigoid patient’s autoantibodies in mice is related to their significantly weaker capacity of activating mouse innate immune factors when compared to human complement and granulocytes. The “alternative” strategy of generating antibodies to the murine form of type XVII collagen by immunizing rabbits and then transferring rabbit antibodies into mice has been used successfully for developing in vivo models for several other autoimmune diseases such as pemphigus vulgaris, anti-epiligrin cicatricial pemphigoid, and epidermolysis bullosa acquisita. Among these, the neonatal BP models have some major shortcomings including the fact that frank skin blistering does not occur and the very short observation times that precludes adequately dissecting disease pathogenesis and developing therapeutic strategies. In the present study, we aimed at addressing these shortcomings by developing a novel passive transfer model for bullous pemphigoid in adult mice. We generated antibodies against the murine BP180/ CXVII by immunizing rabbit and sheep with recombinant forms of the murine antigen. After being passively injected into adult wild type mice, these antibodies bound to the DEJ, activated complement and recruited inflammatory cells resulting in tissue damage. The phenotype of the disease mimicked human BP at the clinical, immunological and histopathological levels. Titres of BP180/ CXVII-specific antibodies in the peripheral blood of injected animals correlated well with disease activity. Immune sheep sera showed higher BP180/ CXVII-specific levels compared to rabbit antibodies and induced more extensive disease after their passive transfer in mice. This model provides a solid basis for further pathogenetic studies in BP and for the development of new therapeutic approaches.
Materials and methods
Six- to eight-week-old BALB/c mice with a body weight of approximately 20 g were used. Mice were obtained from the Cantacuzino Institute (Bucharest, Romania) and housed at our animal facility. All injections and bleedings were performed on mice narcotized by administration of a mixture of ketamine (100 μg/g) and xylazine (15 μg/g). Mice received subcutaneously 10 mg of ammonium sulfate-precipitated BP180/ CXVII-specific antibodies from either rabbit (end-titre 12.800-25.600) or sheep (end-titre 102.400) every second day for two weeks. Control mice received the same amounts of normal preimmune rabbit or sheep antibodies, referred to hereafter as control antibodies. The experiments were approved by the Ethics Committee (Babes-Bolyai University Cluj-Napoca no. 1146/2009 and 31458/2010) and performed by qualified personnel.
Heterologous expression of murine BP180/ CXVII fragments
Three extracellular and one intracellular fragments of murine BP180/ CXVII were expressed as glutathione-S-transferase (GST) fusion proteins as described. These were designated GST-mCXVII-EC1, GST-mCXVII-EC3, GST-mCXVII-EC7 and GST-mCXVII-IC2, and contain murine collagen XVII sequences stretching from amino acid positions 498–580, 856–901, 1030–1134, and 186–475, respectively. The proteins were produced using the recombinant vectors pGEX-mCOL17-EC1, pGEX-mCOL17-EC3, pGEX-mCOL17-EC7 and pGEX-mCOL17-IC2 in Escherichia coli and purified by glutathione agarose chromatography.
Generation of BP180/ CXVII-specific rabbit and sheep antibodies
Three New Zealand White rabbits and one sheep were immunized subcutaneously with either 200 μg or 400 μg, respectively, of a mixture of the four purified recombinant fragments of murine type XVII collagen (GST-mCXVII-EC1, GST-mCXVII-EC3, GST-mCXVII-EC7 and GST-mCXVII-IC2 in a molar ratio of 2:1:1:1) mixed with Freund’s complete adjuvant. The animals were boosted twice with the same protein preparation in incomplete Freund’s adjuvant at two weeks intervals. Control antibodies were collected before the first immunization and immune sera were obtained at regular intervals and characterized by immunofluorescence (IF) microscopy on cryosections of murine skin. For the passive transfer experiments, sera from rabbits were pooled and immunoglobulins were isolated by ammonium sulfate precipitation. Sheep antibodies were also subjected to ammonium sulfate precipitation before passive transfer experiments. Protein concentration was measured spectrophotometrically at 280 nm.
Characterization of murine BP180/ CXVII-specific antibodies
Frozen skin sections were prepared from tissue biopsies of mice and antibodies against murine BP180/ CXVII were analyzed by IF microscopy using 100-fold diluted antibodies specific to rabbit (DakoCytomation) or sheep IgG (Abcam). Complement-fixing activity of antibodies to the DEJ was determined as previously described. Recombinant proteins were fractionated by 12.5% and 5% SDS-PAGE, transferred to nitrocellulose, and analyzed by immunoblotting using HRP-conjugated secondary antibodies (DakoCytomation) and 3,3’ diaminobenzidine (Merck) as a chromogenic substrate.
Induction of disease in vivo and phenotype assessment
Mice were examined daily for their general condition and for evidence of lesions (i.e., erythema, blisters, erosions, crusts and alopecia). The extent of skin disease (disease activity) was scored as follows: 0, no lesions; 1, < 10 lesions or < 1% of the skin surface; 2, > 10 lesions or 1-5% of the skin surface; 3, 5-10%; 4, 10-20%; and 5, > 20% involvement of the skin surface. To evaluate the correlation of antibody titres with the extent of disease, sera were obtained from mice at three different time points (day 0, day 7 and day 14) and assayed by indirect IF microscopy on frozen sections of murine skin. Skin biopsies of perilesional and lesional skin and mucous membranes were obtained at the end of the observation period and assayed for immuno-reactants deposition by direct IF and hematoxylin and eosin staining, respectively. The staining intensity of immunoreactants in the skin of injected mice was assessed semi-quantitatively using a score comprising 0, for no staining; 1, focal, faint staining; 2, faint staining, 3, medium; and 4, intense staining.
To estimate the correlation between titres of circulating antibodies as detected by indirect IF microscopy, and disease activity, Spearman’s rank correlation test was applied. Differences in clinical disease activity among groups were calculated using the Chi-square test. All means are presented ± s.e.m.
Generation and characterization of BP180/ CXVII-specific antibodies
Antibodies to the dermal-epidermal junction (DEJ) produced for this study
BP180/ CXVII-specific antibodies induce frank skin blistering when passively transferred into adult mice
Mice injected with BP180/ CXVII-specific antibodies showed immunopathological features of human BP
BP180/ CXVII-specific antibodies induced subepidermal blistering in vivo
Lesional skin of mice was also obtained at the end of the observation period and examined for histological signs of disease by hematoxylin and eosin staining. Subepidermal blister formation accompanied by an inflammatory infiltrate was found in mice receiving either rabbit (Figure4g) or sheep (Figure4h) BP180/ CXVII-specific IgG. In contrast, histology from mice injected with control rabbit (not shown) or sheep antibodies (Figure4i) showed neither inflammatory cells nor dermal epidermal separation. Oesophagus specimens from both diseased or control mice showed no histopathological changes (not shown).
The extent of skin involvement correlates with levels of serum BP180/ CXVII-specific antibodies in diseased mice
Titres of antibodies correlate with the extent of disease in mice
R a-murine BP180/ CXVII
S a-murine BP180/ CXVII
Different models for bullous pemphigoid have been generated over the past two decades[14, 15, 17, 19, 27]. While the existing models demonstrated the pathogenicity of BP180/ CXVII-specific antibodies, they have several shortcomings. Neonatal mice injected with BP180/ CXVII-specific antibodies do not develop frank skin blistering; epidermal wrinkling and dermal-epidermal separation are induced by applying pressure (rubbing) on the mouse skin. In addition, due to their design, the short observation time of up to three days in the neonatal mice may not be optimal to reproduce the course of chronic disease and for pathogenetic studies and development of new therapeutic approaches. In the present study, to address these drawbacks, we explored whether the passive transfer of BP180/ CXVII-specific antibodies in adult mice results in blistering in the absence of skin rubbing and allows for longer observation times. In a first set of experiments, we generated several fragments of the BP180/ CXVII-specific antibodies, by immunization of rabbits and sheep with the recombinant autoantigen. Since only about 85% of the BP sera react with the immunodominant NC16A domain of BP180/ CXVII by ELISA and most BP sera also recognize other epitopes, we injected, antibodies directed against different fragments of murine BP180 in order to better reflect the broader autoimmune response against BP180 in patients. As shown in figure2, the polyclonal antibodies generated against recombinant fragments of the murine protein were able to recognize individual fragments by immunoblotting as well as the native form of the protein in cryosections of murine skin activating the complement system, a characteristic feature of several human and experimental autoimmune blistering diseases[15, 39–41]. Our present model provides a robust system for further studies dissecting the pathogenic potential of antibodies against different fragments outside the NC16A domain, which may arise in patients as a result of an epitope spreading.
To test whether mouse BP180/ CXVII-specific antibodies retain their capacity to recognize the basal membrane and to activate complement in vivo, we passively transferred antibodies from immunized animals or control antibodies into adult mice. The antibodies bound at the basement membrane and activated murine complement as shown by direct IF analysis of perilesional biopsies of the skin and oesophagus. In addition, they recruited murine leukocytes and induced skin blistering thus reproducing the human BP at the clinical, immunological and histopathological levels. These findings are in line with similar results of inflammatory blistering in another newly developed passive antibody transfer model of pemphigoid disease in which we pre-immunized adult mice with rabbit IgG followed by injection of collagen XVII-specific rabbit IgG. While having the advantage of an increased inflammatory reaction and more extensive disease in mice compared with the mouse model presented here, the model reported by Oswald et al., has some limitations due to the active immunization with the rabbit IgG, which pose constraints for designing studies addressing the pathophysiology of the pemphigoid disease in patients. Lesions developed at different sites, including the trunk, head, ears and the limbs. Extensive erosions were constantly accompanied by alopecia. Importantly, disease activity in animals receiving pathogenic antibodies correlated with the titres of these antibodies in the sera of injected mice, whereas control mice showed no clinical disease or circulating antibodies. These observations are in line with studies showing a correlation between BP180/ CXVII-specific antibody serum levels and clinical disease in patients[44–46] and their potential to induce tissue damage in an ex vivo model for BP. In the present study, in addition to rabbits we have also immunized a sheep in order to test for and compare the outcome of the passive transfer of these antibodies into mice as an alternate source of BP180/ CXVII-specific antibodies. An unexpected result of our present experiments was the fact that sheep antibodies were much more suitable for inducing the disease (Figure5). This may be due to the higher titre of injected sheep antibodies and/or to a better in vivo activation of murine innate immune players, including complement and inflammatory cells. We have obtained relatively high volumes of serum per bleeding and the immune sera showed high titres by indirect IF microscopy on murine skin sections. Since the serum yield of a single sheep can be equivalent to that of eight to ten rabbits, using antibodies generated in sheep may offer a cost-effective alternative for rabbit IgG. A further major advantage of the newly developed mouse model of BP is the fact that it allows for induction of sustained disease and longer observation times of weeks, which can be likely extended to months. This could be advantageous in addressing issues like the specific contribution of different immune players in disease initiation and progression. Probably the most striking difference between the neonatal models and our adult model is represented by the fact that the total IgG amount/g body weight received by adult mice is lower and yet they develop disease without the need to rub their skin. This might be explained at least in part by: 1) the fact that BP is a chronic disease with a complex pathogenesis in which different immune players act in a sequential manner. It is therefore imaginable that limited observation times preclude some immune players to become effectively activated for inducing a frank disease in the neonatal model; hence not only the dose but also the time frame of disease induction may play a critical role; 2) the fact that neonatal mice react differently in terms of activating immune factors in vivo. It is worth noting in this context, that neonatal mice injected with antibodies to type VII collagen in the experimental epidermolysis bullosa acquisita mouse model are resistant to blistering despite higher-doses of injected antibodies as compared to adult mice. Susceptibility was rendered in these neonatal mice only after treatment with IL-8 or C5a, two powerful attractants for granulocytes.
We describe here a model of pemphigoid disease in adult mice showing sustained blistering upon injection of BP180/ CXVII-specific antibodies. Pathogenic antibodies for this model may be produced in both sheep and rabbit. Finally, the present model allows for longer observation times. Collectively, these features will greatly facilitate further pathogenetic studies and should prove helpful for translating this information into novel therapeutic strategies for autoantibody-mediated inflammatory diseases.
- BP180/collagen XVII and BP230:
BP antigens of 180kDa and 230kDa respectively
16th non-collagenous domain of BP180/ CXVII.
The authors would like to thank Dr. Csaba Mate (Zoological Park Turda) for technical advice with sheep procedures. Funding sources: This work was supported by grants IDEI_1146/2009 and PCCE_ID_312/2008 and PCCE_ID_129/2008 by the Romanian National Council for Scientific Research in Higher Education (to MTC and OP, respectively) and by grants from the Deutsche Forschungsgemeinschaft SI-1281/2-1 and SI-1281/4-1 (to CS). MTC, IL and EL received financial support from the Sectoral Operational Programme for Human Resources Development 2007–2013, co-financed by the European Social Fund, under the project numbers POSDRU/21/1.5/G/36154 (Performant doctoral program for the development of highly qualified human resources in the interdisciplinary scientific research) and POSDRU 6/1.5/S/3 (Doctoral studies: through science towards society), respectively. MCT has been financially supported through the project POSDRU/89/1.5/S/61104, co-financed by the European Social Fund under the Sectoral Operational Program for Human Resources Development 2007–2013. The funders had no role in study design, data collection, analysis and interpretation, manuscript preparation and publication decisions.
- Zillikens D, Wever S, Roth A, Weidenthaler-Barth B, Hashimoto T, Bröcker EB: Incidence of autoimmune subepidermal blistering dermatoses in a region of central Germany. Arch Dermatol. 1995, 131: 957-958. 10.1001/archderm.1995.01690200097021.PubMedView Article
- Bernard P, Vaillant L, Labeille B, Bedane C, Arbeille B, Denoeux JP, Lorette G, Bonnetblanc JM, Prost C: Incidence and distribution of subepidermal autoimmune bullous skin diseases in three french regions. Bullous diseases french study group. Arch Dermatol. 1995, 131: 48-52. 10.1001/archderm.1995.01690130050009.PubMedView Article
- Langan SM, Smeeth L, Hubbard R, Fleming KM, Smith CJ, West J: Bullous pemphigoid and pemphigus vulgaris--incidence and mortality in the UK: population based cohort study. BMJ. 2008, 337: a180-10.1136/bmj.a180.PubMed CentralPubMedView Article
- Marazza G, Pham HC, Scharer L, Pedrazzetti PP, Hunziker T, Trueb RM, Hohl D, et al: Incidence of bullous pemphigoid and pemphigus in Switzerland: a 2-year prospective study. Br J Dermatol. 2009, 161: 861-868. 10.1111/j.1365-2133.2009.09300.x.PubMedView Article
- Stanley JR, Hawley-Nelson PYuspa SH, Shevach EM, Katz SI: Characterization of bullous pemphigoid antigen: a unique basement membrane protein of stratified squamous epithelia. Cell. 1981, 24: 897-903. 10.1016/0092-8674(81)90115-X.PubMedView Article
- Labib RS, Anhalt GJ, Patel HP, Mutasim DF, Diaz LA: Molecular heterogeneity of the bullous pemphigoid antigens as detected by immunoblotting. J Immunol. 1986, 136: 1231-1235.PubMed
- Stanley JR, Tanaka T, Mueller S, Klaus-Kovtun V, Roop D: Isolation of complementary DNA for bullous pemphigoid antigen by use of patients' autoantibodies. J Clin Invest. 1988, 82: 1864-1870. 10.1172/JCI113803.PubMed CentralPubMedView Article
- Diaz LA, Ratrie HD, Saunders WS, Futamura S, Squiquera HL, Anhalt G, et al: Isolation of a human epidermal cDNA corresponding to the 180-kD autoantigen recognized by bullous pemphigoid and herpes gestationis sera. Immunolocalization of this protein to the hemidesmosome. J Clin Invest. 1990, 86: 1088-1094. 10.1172/JCI114812.PubMed CentralPubMedView Article
- Giudice GJ, Emery DJ, Diaz LA: Cloning and primary structural analysis of the bullous pemphigoid autoantigen BP180. J Invest Dermatol. 1992, 99: 243-250. 10.1111/1523-1747.ep12616580.PubMedView Article
- Ishiko A, Shimizu H, Kikuchi A, Ebihara T, Hashimoto T, Nishikawa T: Human autoantibodies against the 230-kD bullous pemphigoid antigen (BPAG1) bind only to the intracellular domain of the hemidesmosome, whereas those against the 180-kD bullous pemphigoid antigen (BPAG2) bind along the plasma membrane of the hemidesmosome in normal human and swine skin. J Clin Invest. 1993, 91: 1608-1615. 10.1172/JCI116368.PubMed CentralPubMedView Article
- Schaumburg-Lever G, Rule A, Schmidt-Ullrich R, Lever WF: Ultrastructural localization of in vivo bound immunoglobulins in bullous pemphigoid--a preliminary report. J Invest Dermatol. 1975, 64: 47-49. 10.1111/1523-1747.ep12540902.PubMedView Article
- Karpati S, Stolz W, Meurer M, Braun-Falco O, Krieg T: Herpes gestationis: ultrastructural identification of the extracellular antigenic sites in diseased skin using immunogold techniques. Br J Dermatol. 1991, 125: 317-324. 10.1111/j.1365-2133.1991.tb14164.x.PubMedView Article
- McGrath JA, Gatalica B, Christiano AM, Li K, Owaribe K, McMillan JR, Eady R, et al: Mutations in the 180-kD bullous pemphigoid antigen (BPAG2), a hemidesmosomal transmembrane collagen (COL17A1), in generalized atrophic benign epidermolysis bullosa. Nat Genet. 1995, 11: 83-86. 10.1038/ng0995-83.PubMedView Article
- Nishie W, Sawamura D, Goto M, Ito K, Shibaki A, McMillan JR, Sakai K, Nakamura H, Olasz E, et al: Humanization of autoantigen. Nat Med. 2007, 13: 378-383. 10.1038/nm1496.PubMedView Article
- Liu Z, Sui W, Zhao M, Li Z, Li N, Thresher R, Giudice GJ, Fairley JA, et al: Subepidermal blistering induced by human autoantibodies to BP180 requires innate immune players in a humanized bullous pemphigoid mouse model. J Autoimmun. 2008, 31: 331-338. 10.1016/j.jaut.2008.08.009.PubMed CentralPubMedView Article
- Sitaru C, Schmidt E, Petermann S, Munteanu LS, Brocker EB, Zillikens D: Autoantibodies to bullous pemphigoid antigen 180 induce dermal-epidermal separation in cryosections of human skin. J Invest Dermatol. 2002, 118: 664-671. 10.1046/j.1523-1747.2002.01720.x.PubMedView Article
- Liu Z, Diaz LA, Troy JL, Taylor AF, Emery DJ, Fairley J, et al: A passive transfer model of the organ-specific autoimmune disease, bullous pemphigoid, using antibodies generated against the hemidesmosomal antigen, BP180. J Clin Invest. 1993, 92: 2480-2488. 10.1172/JCI116856.PubMed CentralPubMedView Article
- Nishie W, Sawamura D, Natsuga K, Shinkuma S, Goto M, Shibaki A, Ujiie H, Olasz E, Yancey KB, Shimizu H: A novel humanized neonatal autoimmune blistering skin disease model induced by maternally transferred antibodies. J Immunol. 2009, 183: 4088-4093. 10.4049/jimmunol.0800389.PubMedView Article
- Ujiie H, Shibaki A, Nishie W, Sawamura D, Wang G, Tateishi Y, Li Q, Moriuchi R, Qiao H, Nakamura H, et al: A novel active mouse model for bullous pemphigoid targeting humanized pathogenic antigen. J Immunol. 2010, 184: 2166-2174. 10.4049/jimmunol.0903101.PubMedView Article
- Joly P, Roujeau JC, Benichou J, Picard C, Dreno B, Delaporte E, Vaillant L, D'Incan M, Plantin P, et al: A comparison of oral and topical corticosteroids in patients with bullous pemphigoid. N Engl J Med. 2002, 346: 321-327. 10.1056/NEJMoa011592.PubMedView Article
- Harrington WJ, Minnich V, Hollingsworth JW, Moore CV: Demonstration of a thrombocytopenic factor in the blood of patients with thrombocytopenic purpura. J Lab Clin Med. 1951, 38: 1-10.PubMed
- Schwartz RS: Immune thrombocytopenic purpura--from agony to agonist. N Engl J Med. 2007, 357: 2299-2301. 10.1056/NEJMe0707126.PubMedView Article
- Toyka KV, Brachman DB, Pestronk A, Kao I: Myasthenia gravis: passive transfer from man to mouse. Science. 1975, 190: 397-399. 10.1126/science.1179220.PubMedView Article
- Anhalt GJ, Labib RS, Voorhees JJ, Beals TF, Diaz LA: Induction of pemphigus in neonatal mice by passive transfer of IgG from patients with the disease. N Engl J Med. 1982, 306: 1189-1196. 10.1056/NEJM198205203062001.PubMedView Article
- Roscoe JT, Diaz L, Sampaio SA, Castro RM, Labib RS, Takahashi Y, et al: Brazilian pemphigus foliaceus autoantibodies are pathogenic to BALB/c mice by passive transfer. J Invest Dermatol. 1985, 85: 538-541. 10.1111/1523-1747.ep12277362.PubMedView Article
- Sams WMJ, Gleich GJ: Failure to transfer bullous pemphigoid with serum from patients. Proc Soc Exp Biol Med. 1971, 136: 1027-1031.PubMedView Article
- Anhalt GJ, Bahn CF, Labib RS, Voorhees JJ, Sugar A, Diaz LA: Pathogenic effects of bullous pemphigoid autoantibodies on rabbit corneal epithelium. J Clin Invest. 1981, 68: 1097-1101. 10.1172/JCI110333.PubMed CentralPubMedView Article
- Anhalt GJ, Diaz LA: Animal models for bullous pemphigoid. Clin Dermatol. 1987, 5: 117-125. 10.1016/0738-081X(87)90056-3.PubMedView Article
- Naito K, Morioka S, Ikeda S, Ogawa H: Experimental bullous pemphigoid in guinea pigs: the role of pemphigoid antibodies, complement, and migrating cells. J Invest Dermatol. 1984, 82: 227-230. 10.1111/1523-1747.ep12260057.PubMedView Article
- Gammon WR, Briggaman RA: Absence of specific histologic changes in guinea pig skin treated with bullous pemphigoid antibodies. J Invest Dermatol. 1988, 90: 495-500. 10.1111/1523-1747.ep12460971.PubMedView Article
- Yamamoto K, Inoue N, Masuda R, Fujimori A, Saito T, Imajoh-Ohmi S, Shinkai H, Sakiyama H: Cloning of hamster type XVII collagen cDNA, and pathogenesis of anti-type XVII collagen antibody and complement in hamster bullous pemphigoid. J Invest Dermatol. 2002, 118: 485-492. 10.1046/j.0022-202x.2001.01683.x.PubMedView Article
- Sesarman A, Oswald E, Chiriac MT, Csorba K, Vuta V, Feldrihan V, Baican A, Bruckner-Tuderman L, Sitaru C: Why human pemphigoid autoantibodies do not trigger disease by the passive transfer into mice. Immunol Lett. 2012, 143: 92-100. 10.1016/j.imlet.2012.01.006.PubMedView Article
- Memar OM, Rajaraman S, Thotakura R, Tyring SK, Fan JL, Seetharamaiah GS, et al: Recombinant desmoglein 3 has the necessary epitopes to adsorb and induce blister-causing antibodies. J Invest Dermatol. 1996, 106: 261-268. 10.1111/1523-1747.ep12340663.PubMedView Article
- Lazarova Z, Yee C, Darling T, Briggaman RA, Yancey KB: Passive transfer of anti-laminin 5 antibodies induces subepidermal blisters in neonatal mice. J Clin Invest. 1996, 98: 1509-1518. 10.1172/JCI118942.PubMed CentralPubMedView Article
- Sitaru C, Mihai S, Otto C, Chiriac MT, Haußer I, Dotterweich B, Saito H, Rose C, Ishiko A, Zillikens D: Induction of dermal-epidermal separation in mice by passive transfer of antibodies to type VII collagen. J Clin Invest. 2005, 115: 870-878.PubMed CentralPubMedView Article
- Chiriac MT, Roesler J, Sindrilaru A, Scharffetter-Kochanek K, Zillikens D, Sitaru C: NADPH oxidase is required for neutrophil-dependent autoantibody-induced tissue damage. J Pathol. 2007, 212: 56-65. 10.1002/path.2157.PubMedView Article
- Sitaru C, Chiriac MT, Mihai S, Büning J, Gebert A, Ishiko A, Zillikens D: Induction of complement-fixing autoantibodies against type VII collagen results in subepidermal blistering in mice. J Immunol. 2006, 177: 3461-3468.PubMedView Article
- Kobayashi M, Amagai M, Kuroda-Kinoshita K, Hashimoto T, Shirakata Y, Hashimoto K, Nishikawa T: BP180 ELISA using bacterial recombinant NC16a protein as a diagnostic and monitoring tool for bullous pemphigoid. J Dermatol Sci. 2002, 30: 224-232. 10.1016/S0923-1811(02)00109-3.PubMedView Article
- Liu Z, Giudice GJ, Swartz SJ, Fairley JA, Till GO, Troy J, et al: The role of complement in experimental bullous pemphigoid. J Clin Invest. 1995, 95: 1539-1544. 10.1172/JCI117826.PubMed CentralPubMedView Article
- Nelson KC, Zhao M, Schroder PR, Wetsel RA LN, Diaz LA, Liu Z: Role of different pathways of the complement cascade in experimental bullous pemphigoid. J Clin Invest. 2006, 10.1172/JCI17891.
- Mihai S, Chiriac MT, Takahashi K, Thurman JM, Holers VM, Zillikens D, Botto M, Sitaru C: The alternative pathway of complement activation is critical for blister induction in experimental epidermolysis bullosa acquisita. J Immunol. 2007, 178: 6514-6521.PubMedView Article
- Di Zenzo G, Thoma-Uszynski S, Calabresi V, Fontao L, Hofmann SC, Lacour JP, Sera F, Bruckner-Tuderman L, et al: Demonstration of epitope-spreading phenomena in bullous pemphigoid: results of a prospective multicenter study. J Invest Dermatol. 2011, 131: 2271-2280. 10.1038/jid.2011.180.PubMedView Article
- Oswald E, Sesarman A, Franzke CW, Wolfle U, Bruckner-Tuderman L, Jakob T, Martin SF, Sitaru C: The flavonoid luteolin inhibits Fcgamma-dependent respiratory burst in granulocytes, but not skin blistering in a new model of pemphigoid in adult mice. PLoS One. 2012, 7: e31066-10.1371/journal.pone.0031066.PubMed CentralPubMedView Article
- Haase C, Budinger L, Borradori L, Yee C, Merk HF, Yancey K, Hertl M: Detection of IgG autoantibodies in the sera of patients with bullous and gestational pemphigoid: ELISA studies utilizing a baculovirus- encoded form of bullous pemphigoid antigen 2. J Invest Dermatol. 1998, 110: 282-286.PubMed
- Dopp R, Schmidt E, Chimanovitch I, Leverkus M, Bröcker EB, Zillikens D: IgG4 and IgE are the major immunoglobulins targeting the NC16A domain of BP180 in Bullous pemphigoid: serum levels of these immunoglobulins reflect disease activity. J Am Acad Dermatol. 2000, 42: 577-583.PubMed
- Schmidt E, Obe K, Bröcker EB, Zillikens D: Serum levels of autoantibodies to BP180 correlate with disease activity in patients with bullous pemphigoid. Arch Dermatol. 2000, 136: 174-178. 10.1001/archderm.136.2.174.PubMed
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