Aberrant expression of α‐Gal on primary human endothelium does not confer susceptibility to NK cell cytotoxicity or increased NK cell adhesion

He, Zhong; Ehrnfelt, Cecilia; Kumagai‐Braesch, Makiko; Islam, Khalid B.; Holgersson, Jan

doi:10.1002/eji.200324683

Cited by 26 publications

(30 citation statements)

References 46 publications

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“…Therefore, we conclude that there is no direct correlation between the absence of ␣Gal and VCAM-1 and that ␣Gal does not play a crucial role in the adhesion process. In line with our findings, naive human NK cells adhered, activated, and lysed pEC independently of ␣Gal (62), and no differences regarding NK adhesion and lysis were observed upon introduction of ␣Gal in human aortic endothelial cells (63). In contrast, using ␣1,3GT-transfected COS-7 cells, soluble carbohydrates, and F(abЈ) 2 of xenoreactive NAb, Inverardi et al (37) showed a role for ␣Gal in Ab-independent NK adhesion to and destruction of pEC.…”

Section: Discussionsupporting

confidence: 90%

Lack of Galactose-α-1,3-Galactose Expression on Porcine Endothelial Cells Prevents Complement-Induced Lysis but Not Direct Xenogeneic NK Cytotoxicity

Baumann

Forte

Hawley³

et al. 2004

The Journal of Immunology

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The galactose-α-1,3-galactose (αGal) carbohydrate epitope is expressed on porcine, but not human cells, and therefore represents a major target for preformed human anti-pig natural Abs (NAb). Based on results from pig-to-primate animal models, NAb binding to porcine endothelial cells will likely induce complement activation, lysis, and hyperacute rejection in pig-to-human xenotransplantation. Human NK cells may also contribute to innate immune responses against xenografts, either by direct recognition of activating molecules on target cells or by FcγRIII-mediated xenogeneic Ab-dependent cellular cytotoxicity (ADCC). The present study addressed the question as to whether the lack of αGal protects porcine endothelial cells from NAb/complement-induced lysis, direct xenogeneic NK lysis, NAb-dependent ADCC, and adhesion of human NK cells under shear stress. Homologous recombination, panning, and limiting dilution cloning were used to generate an αGal-negative porcine endothelial cell line, PED2*3.51. NAb/complement-induced xenogeneic lysis of PED2*3.51 was reduced by an average of 86% compared with the αGal-positive phenotype. PED2*3.51 resisted NK cell-mediated ADCC with a reduction of lysis ranging from 30 to 70%. However, direct xenogeneic lysis of PED2*3.51, mediated either by freshly isolated or IL-2-activated human NK cells or the NK cell line NK92, was not reduced. Furthermore, adhesion of IL-2-activated human NK cells did not rely on αGal expression. In conclusion, removal of αGal leads to a clear reduction in complement-induced lysis and ADCC, but does not resolve adhesion of NK cells and direct anti-porcine NK cytotoxicity, indicating that αGal is not a dominant target for direct human NK cytotoxicity against porcine cells.

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Section: Discussionsupporting

confidence: 90%

Lack of Galactose-α-1,3-Galactose Expression on Porcine Endothelial Cells Prevents Complement-Induced Lysis but Not Direct Xenogeneic NK Cytotoxicity

Baumann

Forte

Hawley³

et al. 2004

The Journal of Immunology

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“…17 For transduction, FACS-sorted cells were transferred to 10-mL tubes with 1-mL culture medium and were incubated with concentrated pHRЈEF1␣ GFPSIN virions at a multiplicity of infection (MOI) of 0.1 at 37°C in a 5% CO 2 atmosphere overnight. After virus incubation, cells were washed once in PBS and were directly injected in the denuded femoral artery of nude mice (see "Mice").…”

Section: Lentiviral Transduction Of Facs-sorted Cd14 ؉ /Vegfr-2 ؉ Andmentioning

confidence: 99%

Only a specific subset of human peripheral-blood monocytes has endothelial-like functional capacity

Elsheikh

Uzunel

et al. 2005

Blood

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106

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The monocyte population in blood is considered a possible source of endothelial precursors. Because endothelial-specific receptor tyrosine kinases act as regulators of endothelial cell function, we investigated whether expression of the vascular endothelial growth factor receptor-2 (VEGFR-2) on monocytes is important for their endothelial-like functional capacity. Peripheral-blood monocytes expressing vascular endothelial growth factor receptor-2 (VEGFR-2), or CD14 ؉ /VEGFR-2 ؉ , were isolated, and their phenotypic, morphologic, and functional capacities were compared with those of monocytes nega-

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“…Gal on porcine EC constitutes potential binding sites for human PMN, NK cells, and monocytes. Human NK cells may induce porcine EC activation by direct interaction in the absence of Gal (24,25) and no significant reduction of direct NK cytotoxicity or NK cell and PMN adhesion on porcine ECs was found in the absence of, or upon blocking, Gal. In contrast, monocyte adhesion to porcine ECs depends on Gal-mediated activation (26).…”

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confidence: 90%

Cytokine Secretion Depends on Galα(1,3)Gal Expression in a Pig-to-Human Whole Blood Model

Sæthre

Schneider

Lambris

et al. 2008

The Journal of Immunology

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Transplants from α1,3-galactosyltransferase (Gal) gene-knockout pigs to nonhuman primates are largely protected from hyperacute but not acute humoral xenograft rejection. The present study investigates the role of Gal in cytokine responses using a novel pig-to-human whole blood in vitro model, developed for species-specific analysis of porcine and human cytokines. Porcine (n = 7) and human (n = 27) cytokines were measured using ELISA or multiplex technology, respectively. Porcine aortic endothelial cells from control (Gal+/+) and Gal-deficient (Gal−/−) pigs were incubated with human lepirudin anticoagulated whole blood from healthy donors. E-selectin expression was measured by flow cytometry. The C3 inhibitor compstatin and a C5aR antagonist were used to study the role of complement. Cytokine species specificity was documented, enabling detection of 2 of 7 porcine cytokines and 13 of 27 human cytokines in one single sample. Gal+/+ porcine aortic endothelial cells incubated with human whole blood showed a marked complement C5b-9 dependent up-regulation of E-selectin and secretion of porcine IL-6 and IL-8. In contrast, Gal−/− cells responded with E-selectin and cytokine expression which was so weak that the role of complement could not be determined. Human IL-6, IL-8, IFN-γ, MIP-1α, MIP-1β, eotaxin, and RANTES were detected in the Gal+/+ system, but virtually no responses were seen in the Gal−/− system (p = 0.03). The increase in human cytokine release was largely complement dependent and, in contrast to the porcine response, mediated through C5a. Species-specific analysis of cytokine release revealed a marked, complement-dependent response when Gal+/+ pig cells were incubated with human whole blood, compared with Gal−/− cells which induced virtually no cytokine release.

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Aberrant expression of α‐Gal on primary human endothelium does not confer susceptibility to NK cell cytotoxicity or increased NK cell adhesion

Cited by 26 publications

References 46 publications

Lack of Galactose-α-1,3-Galactose Expression on Porcine Endothelial Cells Prevents Complement-Induced Lysis but Not Direct Xenogeneic NK Cytotoxicity

Lack of Galactose-α-1,3-Galactose Expression on Porcine Endothelial Cells Prevents Complement-Induced Lysis but Not Direct Xenogeneic NK Cytotoxicity

Only a specific subset of human peripheral-blood monocytes has endothelial-like functional capacity

Cytokine Secretion Depends on Galα(1,3)Gal Expression in a Pig-to-Human Whole Blood Model

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