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Effect of various forms of the C1 esterase inhibitor (C1‐INH) and DAF on complement mediated xenogeneic cell lysis

Effect of various forms of the C1 esterase inhibitor (C1‐INH) and DAF on complement mediated... Fukuta D, Miyagawa S, Yamada M, Matsunami K, Kurihara T, Shirasu A, Hattori H, Shirakura R. Effect of various forms of the C1 esterase inhibitor (C1‐INH) and DAF on complement mediated xenogeneic cell lysis. Xenotransplantation 2003; 10: 132–141. © Blackwell Munksgaard, 2003 The purpose of the present study was to assess the effect of various forms of the surface‐bound form of the C1 esterase inhibitor (C1‐INH‐PI) and decay accelerating factor (DAF) on xenogenic cells. cDNAs of various deletion mutants of the C1‐INH‐PI, such as delta‐1‐99 amino acid (AA), delta‐108‐183AA loop, delta‐whole loop, delta‐exon5, delta‐exon6 + 7, and delta‐exon5 + 6 + 7, and that of DAF, the delta‐short consensus repeat (SCR) 1‐DAF were established. While all deletion mutants of C1‐INH‐PI except the delta‐1‐99AA were expressed in the cytoplasm but not on the cell surface, the delta‐1‐99AA was clearly expressed on the xenogeneic cell surface. Amelioration of complement‐mediated xenogeneic cell lysis by delta‐1‐99AA was next tested, and compared with delta‐SCR1 DAF. Both molecules blocked human complement‐mediated cell lysis by approximately 57 to 90 and 93 to 98%, respectively, in Chinese hamster ovarian tumor (CHO) cells and pig endothelial cells (PECs). The CHO cell transfectants were incubated with 20% normal human serum, and the amounts of C4 and C3 deposition on the cell surface were analysed by flow cytometry. The DAF transfectant showed a large amount of C4‐deposition and much less C3‐deposition than the controls (approximately 85% suppression), whereas the delta‐1‐99AA showed approximately a 40% suppression in both C4‐ and C3‐deposition. Consequently, both the delta‐1‐99AA C1‐INH‐PI and delta‐SCR1 DAF molecules are quite effective in down‐regulating the xenogeneic cell lysis, but accomplished this in different manners. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Xenotransplantation Wiley

Effect of various forms of the C1 esterase inhibitor (C1‐INH) and DAF on complement mediated xenogeneic cell lysis

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References (53)

Publisher
Wiley
Copyright
Copyright © 2003 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0908-665X
eISSN
1399-3089
DOI
10.1034/j.1399-3089.2003.01120.x
Publisher site
See Article on Publisher Site

Abstract

Fukuta D, Miyagawa S, Yamada M, Matsunami K, Kurihara T, Shirasu A, Hattori H, Shirakura R. Effect of various forms of the C1 esterase inhibitor (C1‐INH) and DAF on complement mediated xenogeneic cell lysis. Xenotransplantation 2003; 10: 132–141. © Blackwell Munksgaard, 2003 The purpose of the present study was to assess the effect of various forms of the surface‐bound form of the C1 esterase inhibitor (C1‐INH‐PI) and decay accelerating factor (DAF) on xenogenic cells. cDNAs of various deletion mutants of the C1‐INH‐PI, such as delta‐1‐99 amino acid (AA), delta‐108‐183AA loop, delta‐whole loop, delta‐exon5, delta‐exon6 + 7, and delta‐exon5 + 6 + 7, and that of DAF, the delta‐short consensus repeat (SCR) 1‐DAF were established. While all deletion mutants of C1‐INH‐PI except the delta‐1‐99AA were expressed in the cytoplasm but not on the cell surface, the delta‐1‐99AA was clearly expressed on the xenogeneic cell surface. Amelioration of complement‐mediated xenogeneic cell lysis by delta‐1‐99AA was next tested, and compared with delta‐SCR1 DAF. Both molecules blocked human complement‐mediated cell lysis by approximately 57 to 90 and 93 to 98%, respectively, in Chinese hamster ovarian tumor (CHO) cells and pig endothelial cells (PECs). The CHO cell transfectants were incubated with 20% normal human serum, and the amounts of C4 and C3 deposition on the cell surface were analysed by flow cytometry. The DAF transfectant showed a large amount of C4‐deposition and much less C3‐deposition than the controls (approximately 85% suppression), whereas the delta‐1‐99AA showed approximately a 40% suppression in both C4‐ and C3‐deposition. Consequently, both the delta‐1‐99AA C1‐INH‐PI and delta‐SCR1 DAF molecules are quite effective in down‐regulating the xenogeneic cell lysis, but accomplished this in different manners.

Journal

XenotransplantationWiley

Published: Mar 1, 2003

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