Generation and phenotyping of mCd59a and mCd59b double‐knockout mice

Abstract

CD59 is a membrane protein inhibitor of the membrane attack complex (MAC) of complement. Humans express only one, whereas mice express two CD59 genes. We previously reported the targeted deletion of the mCd59b gene in which absence of mCd59b together with an unintended down regulation of mCd59a caused hemolytic anemia with spontaneous platelet activation. To confirm the complement role in the hemolytic anemia caused by abrogation of mCd59 function, we have developed a mCd59a and mCd59b double knock out mice and analyzed its phenotype in complement sufficient and deficient (C3−/−). We report here that total abrogation of mCd59 function in mCd59ab−/− mice results in complement-mediated hemolytic anemia that is rescued by the deficiency of C3 in compound mCd59ab−/−/C3−/− mice.

The Protective Role of CD59 and Pathogenic Role of Complement in Hepatic Ischemia and Reperfusion Injury

“…Although the protective role of CD59 in hepatic IRI has not been defined previously, the protective role of CD59 in renal IRI 26 and in cerebral ischemia 48 has been elucidated by utilization of deficiency of the mCd59a, a primary murine. 27,49,50 These results, together with the fact that CD59 is universally expressed in almost of all cells in a variety of organs, indicate that CD59 may protect against the IRI in organs other than liver, kidney, and brain. Furthermore, CR2-CD59, a novel targeting complement inhibitors for complement activation sites, recently developed by Tomlinson's research group 51 may provide a new avenue to protect organs and donor grafts, including liver, from IRI.…”

“…All mice on a C57BL/6 (B6) genetic background were 8 to 12 weeks old, with 20 to 30 g body weight. mCd59a and mCd59b double-knockout mice (mCd59ab Ϫ/Ϫ ) and mCd59a, mCd59b, and mC3 triple-knockout mice (mCd59ab Ϫ/Ϫ /mC3 Ϫ/Ϫ ) in a B6 background were generated at the Laboratory for Translational Research at Harvard Medical School as previously described by Qin et al 27 In the rat experiment, all adult male Wistar rats (200 to 250 g body weight) were purchased from the Animal Facility of Shanghai Jiao Tong University. The mouse experiments were conducted at the Laboratory for Translational Research at Harvard Medical School and the rat experiments at the Department of General Surgery of Shanghai First People's Hospital at Shanghai Jiao Tong University.…”

“…To assess the role of CD59 and MAC in warm IRI, we used mCd59a and mCd59b double-knockout mouse model 27 and our own mCd59a, mCd59b, and C3 tripleknockout mouse model, together with a complement depletion method [ie, preadministration of mCd59ab Ϫ/Ϫ mice with cobra venom factor (CVF)]. Furthermore, to extend the present study to a clinically relevant context of liver transplantation, rats with complement deficiency induced by CVF were used to investigate the role of complement activation in IRI after syngeneic OLT.…”

The Role of Complement in the Pathogenesis of Artery Aneurysms

“…This subject expressed a severe PNH phenotype from the unusually young age of thirteen and also had a stroke that left him with permanent neurological damage (Yamashina et al, 1990). Not only does the Cd59 knockout out mouse exhibit a full PNH-like anemia as well as platelet activation, but it also exhibits progressive loss of fertility (Holt et al, 2001;Qin et al, 2009;Qin et al, 2003). Also, although the role of S-protein as an inhibitor of MAC formation in tissues has been suggested, the Sprotein knockout mouse did not show any detectable phenotype and instead developed normally and was fully fertile (Zheng et al, 1995).…”

“…Moreover, human studies have suggested that MAC may play a role in the formation and rupture of aorta aneurysms as we discussed above. Taken together, this evidence prompted us to explore the role of MAC in the pathogenesis of AAA with our recent generation of both anti-MAC inhibitor CD59 deficient and overexpressing mice (Qin et al, 2009;Wu et al, 2010). In an angiotensin (Ang) II-induced AAA model, deficiency of both mouse Cd59a and Cd59b (mCd59ab -/-) in ApoE-null mice accelerated the disease development, while transgenic over-expression of human CD59 in the endothelial and circulating cells(macrophages and platelets) (hCD59 ICAM-2+/-) attenuated AAA progression (Wu et al, 2010), these results suggest a protective role of CD59 in AAA development.…”