Recovery of a human natural antibody against the noncollagenous-1 domain of type IV collagen using humanized models

Inge M Worni-Schudel, Amy G Clark, Tiffany Chien, Kwan-Ki Hwang, Benny J Chen, Mary H Foster
2015 Journal of Translational Medicine  
Anti-glomerular basement membrane nephritis and Goodpasture syndrome result from autoantibody (Ab)-mediated destruction of kidney and lung. Ab target the noncollagenous 1 (NC1) domain of alpha3(IV) collagen, but little is known about Ab origins or structure. This ignorance is due in part to the inability to recover monoclonal Ab by transformation of patients' blood cells. The aim of this study was to assess the suitability of two humanized models for this purpose. Methods: NOD-scid-gamma
more » ... eficient mice were engrafted either with human CD34+ hematopoietic stem cells (HSC) (Hu-HSC mice) and immunized with alpha3(IV)NC1 collagen containing the Goodpasture epitopes or with nephritis patients' peripheral blood leukocytes (PBL) (Hu-PBL mice). After in vivo immune cell development and/ or expansion, recovered human B cells were Epstein Barr virus (EBV)-transformed, screened for antigen (Ag) binding, electrofused with a mouse-human heterohybridoma, subcloned, and human Ab RNA sequenced by PCR after reverse transcription to cDNA. Flow cytometry was used to assess human B cell markers and differentiation in Hu-PBL mice.
doi:10.1186/s12967-015-0539-4 pmid:26048777 pmcid:PMC4467618 fatcat:p2wyhhw3r5gdro4tbyhjqbbv6y