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Anti-collagen XVII single-chain Fv antibody blocks the autoimmune reaction mediated by pathogenic autoantibodies in bullous pemphigoid.
J Dermatol Sci. 2013 Oct; 72(1):25-31.JD

Abstract

BACKGROUND

Pathogenic autoantibodies in bullous pemphigoid (BP) recognize the non-collagenous 16A domain (NC16A) of collagen XVII (COL17), a hemidesmosomal component at the skin membrane. This immune inflammation involves activation of the complement cascade via the classical pathway. With similar antigen binding activity, Fab and single-chain variable fragments (scFv) of pathogenic anti-COL17 antibodies can interfere with COL17 binding of autoantibodies, blocking subsequent complement activation and granulocyte activation.

OBJECTIVE

To characterize the biological functions of human anti-COL17 scFv antibody.

METHODS

We constructed scFv antibodies against the corresponding antigen from parental Fab by expression in Escherichia coli. IgG autoantibodies against COL17 were purified by affinity chromatography from serum of BP patients. The inhibitory effects of anti-COL17 scFv on binding of BP autoantibodies to the NC16A domain of human COL17 antigen were observed by inhibition ELISA, immunofluorescence, and inhibition of complement activation. Reactive oxygen production assay and BP cryosection model were performed to assess the inhibitory effect of scFv on granulocyte activation and then the dermal-epidermal separation.

RESULTS

ELISA and Western blot showed specific binding of scFv to COL17. We found that anti-COL17 scFv can inhibit the binding of intact IgG purified from BP parents to the corresponding COL17 antigen and then subsequent C1q and C3 activation and granulocyte activation in vitro. Most importantly, we confirmed that recombinant scFv can inhibit BP-IgG induced dermal-epidermal separation by BP cryosection model.

CONCLUSION

The anti-COL17 scFv antibody can inhibit the binding of BP-IgG autoantibodies to COL17, thereby affecting subsequent complement activation and granulocyte activation in vitro. Our results suggest that blocking pathogenic epitopes using engineered scFv is an efficient BP therapy.

Authors+Show Affiliations

Department of Dermatology, Xijing Hospital, Fourth Military Medical University, 127 Changlexi Road, Xi'an 710032, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

23827201

Citation

Wu, Yan, et al. "Anti-collagen XVII Single-chain Fv Antibody Blocks the Autoimmune Reaction Mediated By Pathogenic Autoantibodies in Bullous Pemphigoid." Journal of Dermatological Science, vol. 72, no. 1, 2013, pp. 25-31.
Wu Y, Sun NN, Dang EL, et al. Anti-collagen XVII single-chain Fv antibody blocks the autoimmune reaction mediated by pathogenic autoantibodies in bullous pemphigoid. J Dermatol Sci. 2013;72(1):25-31.
Wu, Y., Sun, N. N., Dang, E. L., Jin, L., Liu, Z. F., Zhang, W., Yang, L. T., & Wang, G. (2013). Anti-collagen XVII single-chain Fv antibody blocks the autoimmune reaction mediated by pathogenic autoantibodies in bullous pemphigoid. Journal of Dermatological Science, 72(1), 25-31. https://doi.org/10.1016/j.jdermsci.2013.05.010
Wu Y, et al. Anti-collagen XVII Single-chain Fv Antibody Blocks the Autoimmune Reaction Mediated By Pathogenic Autoantibodies in Bullous Pemphigoid. J Dermatol Sci. 2013;72(1):25-31. PubMed PMID: 23827201.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Anti-collagen XVII single-chain Fv antibody blocks the autoimmune reaction mediated by pathogenic autoantibodies in bullous pemphigoid. AU - Wu,Yan, AU - Sun,Na-Na, AU - Dang,Er-Le, AU - Jin,Liang, AU - Liu,Zhen-Feng, AU - Zhang,Wei, AU - Yang,Lu-Ting, AU - Wang,Gang, Y1 - 2013/06/13/ PY - 2012/12/17/received PY - 2013/04/22/revised PY - 2013/05/01/accepted PY - 2013/7/6/entrez PY - 2013/7/6/pubmed PY - 2014/4/26/medline KW - Autoantibodies KW - Bullous pemphigoid KW - Complement KW - Single-chain Fv KW - Therapy SP - 25 EP - 31 JF - Journal of dermatological science JO - J Dermatol Sci VL - 72 IS - 1 N2 - BACKGROUND: Pathogenic autoantibodies in bullous pemphigoid (BP) recognize the non-collagenous 16A domain (NC16A) of collagen XVII (COL17), a hemidesmosomal component at the skin membrane. This immune inflammation involves activation of the complement cascade via the classical pathway. With similar antigen binding activity, Fab and single-chain variable fragments (scFv) of pathogenic anti-COL17 antibodies can interfere with COL17 binding of autoantibodies, blocking subsequent complement activation and granulocyte activation. OBJECTIVE: To characterize the biological functions of human anti-COL17 scFv antibody. METHODS: We constructed scFv antibodies against the corresponding antigen from parental Fab by expression in Escherichia coli. IgG autoantibodies against COL17 were purified by affinity chromatography from serum of BP patients. The inhibitory effects of anti-COL17 scFv on binding of BP autoantibodies to the NC16A domain of human COL17 antigen were observed by inhibition ELISA, immunofluorescence, and inhibition of complement activation. Reactive oxygen production assay and BP cryosection model were performed to assess the inhibitory effect of scFv on granulocyte activation and then the dermal-epidermal separation. RESULTS: ELISA and Western blot showed specific binding of scFv to COL17. We found that anti-COL17 scFv can inhibit the binding of intact IgG purified from BP parents to the corresponding COL17 antigen and then subsequent C1q and C3 activation and granulocyte activation in vitro. Most importantly, we confirmed that recombinant scFv can inhibit BP-IgG induced dermal-epidermal separation by BP cryosection model. CONCLUSION: The anti-COL17 scFv antibody can inhibit the binding of BP-IgG autoantibodies to COL17, thereby affecting subsequent complement activation and granulocyte activation in vitro. Our results suggest that blocking pathogenic epitopes using engineered scFv is an efficient BP therapy. SN - 1873-569X UR - https://www.unboundmedicine.com/medline/citation/23827201/Anti_collagen_XVII_single_chain_Fv_antibody_blocks_the_autoimmune_reaction_mediated_by_pathogenic_autoantibodies_in_bullous_pemphigoid_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0923-1811(13)00211-9 DB - PRIME DP - Unbound Medicine ER -