Resistance of neonatal porcine Sertoli cells to human xenoantibody and complement-mediated lysis is associated with low expression of α-Gal and high production of clusterin and CD59

Yin, Zhongyuan; Wang, Lu; Xiang, Ying; Ruan, Yongle; Li, Junhua; Wang, Ximo; Ichim, Thomas E.; Shi, Chen; Chen, Gang

doi:10.1111/j.1399-3089.2010.00581.x

Cited by 11 publications

(10 citation statements)

References 33 publications

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“…MCP and DAF accelerate the degradation of C3 and C5 convertases as well as limit the production of proinflammatory molecules, while clusterin and CD59 inhibit the assembly of the MAC 21 . The mRNA for these inhibitors has been detected in mouse and porcine SCs 15,22,23 and was confirmed by this study. Here we report for the first time that NPSCs express significantly higher levels of clusterin, MCP, and DAF mRNA as well as significantly higher levels of MCP and DAF proteins compared to NPIs.…”

Section: Discussionsupporting

confidence: 82%

Xenotransplanted Pig Sertoli Cells Inhibit Both the Alternative and Classical Pathways of Complement-Mediated Cell Lysis While Pig Islets Are Killed

et al. 2016

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Xenotransplantation has vast clinical potential but is limited by the potent immune responses generated against xenogeneic tissue. Immune-privileged Sertoli cells (SCs) survive xenotransplantation long term (>90 days) without immunosuppression, making SCs an ideal model to identify xenograft survival mechanisms. Xenograft rejection includes the binding of natural and induced antibodies and the activation of the complement cascade. Using an in vitro cytotoxicity assay, wherein cells were cultured with human serum and complement, we demonstrated that neonatal pig SCs (NPSCs) are resistant to complement mediated cell lysis and express complement inhibitory factors, membrane cofactor protein (MCP; CD46), and decay- accelerating factor (DAF; CD55) at significantly higher levels than neonatal pig islets (NPIs), which served as non-immune-privileged controls. After xenotransplantation into naive Lewis rats, NPSCs survived throughout the study, while NPIs were rejected within 9 days. Serum antibodies, and antibody and complement deposition within the grafts were analyzed. Compared to preformed circulating anti-pig IgM anti bodies, no significant increase in IgM production against NPSCs or NPIs was observed, while IgM deposition was detected from day 6 onward in both sets of grafts. A late serum IgG response was detected in NPSC (days 13 and 20) and NPI (day 20) recipients. Consistently, IgG deposition was first detected at days 9 and 13 in NPSC and NPI grafts, respectively. Interestingly, C3 was deposited at days 1 and 3 in NPI grafts and only at day 1 in NPSC grafts, while membrane attack complex (MAC) deposition was only detected in NPI grafts (at days 1–4). Collectively, these data suggest NPSCs actively inhibit both the alternative and classical pathways of complement-mediated cell lysis, while the alternative pathway plays a role in rejecting NPIs. Ultimately, inhibiting the alternative pathway along with transplanting xenogeneic tissue from transgenic pigs (expressing human complement inhibitory factors) could prolong the survival of xenogeneic cells without immunosuppression.

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Section: Discussionsupporting

confidence: 82%

Xenotransplanted Pig Sertoli Cells Inhibit Both the Alternative and Classical Pathways of Complement-Mediated Cell Lysis While Pig Islets Are Killed

et al. 2016

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“…988,1000,1001 Sertoli cells produce several complement inhibitors, as well as inhibitors of granzyme B, which is a lytic molecule produced by cytotoxic lymphocytes. [1002][1003][1004] Furthermore, IL6, which is secreted by the Sertoli cell under hormonal control, has a number of immunoregulatory properties, stimulating the production of anti-inflammatory cytokines by T cells, 125 and inhibiting the maturation of dendritic cells from circulating monocytes. 234 Cultured murine Sertoli cells progressively produce indoleamine 2,3-dioxygenase (IDO), an enzyme which catalyzes the metabolism of tryptophan, and which has been shown to stimulate dendritic cellinitiated tolerance, 1005 and to stimulate the development of Treg cells in tumors and in pregnancy.…”

Section: The Role Of the Sertoli Cellmentioning

confidence: 99%

The Immunophysiology of Male Reproduction

Hedger

2015

Knobil and Neill's Physiology of Reproduction

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“…Therefore, this in vitro xenoreactive CDC assay may be used to test the inhibitory effects on complement activation after pretreatment of human or NHP serum with compstatin or its derivatives in vitro or for ex vivo monitoring of residual complement activity in inhibitor-treated animals during preclinical studies. Because a large number of homogeneous PECs can be easily obtained from the SV40-immortalized porcine aorta endothelial cell line (iPEC) [20], these cells were used as target cells in the present study.…”

Section: Discussionmentioning

confidence: 99%

“…Staining for α-Gal was performed as described previously [20]. In brief, iPECs (2 × 10 5 ) were incubated with fluorescein isothiocyanate (FITC)-conjugated Griffonia simplicifolia isolectin B4 (GS-IB4) (5 μg/ml, Sigma-Aldrich Corp., USA) for 30 min at 4 °C.…”

Section: Methodsmentioning

confidence: 99%

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Using an in vitro xenoantibody-mediated complement-dependent cytotoxicity model to evaluate the complement inhibitory activity of the peptidic C3 inhibitor Cp40

Wang

Xiang³

et al. 2016

Clinical Immunology

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Simple and reliable methods for evaluating the inhibitory effects of drug candidates on complement activation are essential for preclinical development. Here, using an immortalized porcine aortic endothelial cell line (iPEC) as target, we evaluated the feasibility and effectiveness of an in vitro xenoantibody-mediated complement-dependent cytotoxicity (CDC) model for evaluating the complement inhibitory activity of Cp40, a potent analog of the peptidic C3 inhibitor compstatin. The binding of human xenoantibodies to iPECs led to serum dilution-dependent cell death. Pretreatment of the human serum with Cp40 almost completely inhibited the deposition of C3 fragments and C5b-9 on the cells, resulting in a dose-dependent inhibition of CDC against the iPECs. Using the same method to compare the effects of Cp40 on complement activation in humans, rhesus and cynomolgus monkeys, we found that the inhibitory patterns were similar overall. Thus, the in vitro xenoantibody-mediated CDC assay may have considerable potential for future clinical use.

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Resistance of neonatal porcine Sertoli cells to human xenoantibody and complement-mediated lysis is associated with low expression of α-Gal and high production of clusterin and CD59

Abstract: These data demonstrate that the resistance of NPSCs to human xenoantibody and complement-mediated lysis is associated with low expression of xenoantigen alpha-Gal and high production of the complement inhibitors clusterin and CD59.

Cited by 11 publications

References 33 publications

Xenotransplanted Pig Sertoli Cells Inhibit Both the Alternative and Classical Pathways of Complement-Mediated Cell Lysis While Pig Islets Are Killed

Xenotransplanted Pig Sertoli Cells Inhibit Both the Alternative and Classical Pathways of Complement-Mediated Cell Lysis While Pig Islets Are Killed

The Immunophysiology of Male Reproduction

Using an in vitro xenoantibody-mediated complement-dependent cytotoxicity model to evaluate the complement inhibitory activity of the peptidic C3 inhibitor Cp40

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