Porcine Endothelium Supports Transendothelial Migration of Human Leukocyte Subpopulations: Anti-Porcine Vascular Cell Adhesion Molecule Antibodies as Species-Specific Blockers of Transendothelial Monocyte and Natural Killer Cell Migration1

Hauzenberger, Elenor; Hauzenberger, Dan; Hultenby, Kjell; Holgersson, Jan

doi:10.1097/00007890-200005150-00018

Cited by 35 publications

(31 citation statements)

References 39 publications

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“…2). This is in sharp contrast to the appearance of the endothelium following migration of IL-2 activated NK cells, after which it is completely destroyed revealing the underlying filter with its pores [12].…”

Section: Discussionmentioning

confidence: 78%

“…The endothelial surface, however, was intact without any detectable membrane pores. In cases where the endothelium is lysed, the underlying membrane with its pores becomes visible [12]. The increased adhesion and cell aggregation seen in the presence of anti- § -Gal IgG was dependent on Ab binding to the porcine endothelium, because the same concentration of anti- § -Galdepleted human IgG had no dramatic effect on the adhesion of PBMC to porcine endothelium (Fig.…”

Section: Migration Of Pbmc Across Porcine Endothelium In the Presencementioning

confidence: 93%

“…The mean percentage ± S.D of NK cells in each fraction (nonadhered, adhered and migrated) relative to the total number of NK cells added to the upper well is shown. HAEC) [12]. No apparent difference in the ability of porcine and human endothelium to support migration of human leukocytes was observed [12].…”

Section: Introductionmentioning

confidence: 95%

“…EC were isolated from the abdominal aorta of an outbred pig (Swedish landrace) using 0.4% (w/v) collagenase A (Boehringer Mannheim, Mannheim, Germany) as described [12]. Primary PAEC were cultured in plastic tissue culture flasks coated with porcine fibronectin (10 ?…”

Section: Primary Porcine Aortic Endothelial Cellsmentioning

confidence: 99%

“…HAEC) [12]. No apparent difference in the ability of porcine and human endothelium to support migration of human leukocytes was observed [12]. Thus, there appears to be an overall compatibility between porcine and human adhesion receptors in the context of leukocyte extravasation [12].…”

Section: Introductionmentioning

confidence: 97%

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Anti‐Gal IgG potentiates natural killer cell migration across porcine endothelium via endothelial cell activation and increased natural killer cell motility triggered by CD16 cross‐linking

2004

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Xenoreactive antibodies (Ab) are important for the development of acute vascular rejection (AVR) of xenografts characterized by monocytes, natural killer (NK) cells and neutrophils infiltrating the graft. The mechanisms by which anti-galactose § 1,3galactose ( § -Gal) IgG influence NK cell migration across porcine aortic endothelium (PAEC) were investigated. NK cell migration across PAEC increased in the presence of anti- § -Gal IgG. Anti- § -Gal IgG exposure activated PAEC as shown by an increased expression of CD62E and CD106. NK cells adhered, spread and showed motile forms on plastic surfaces coated with human IgG, IgG Fc and on mAb against CD16, but not on mouse IgG or BSA, suggesting that CD16 cross-linking can mediate increased adhesiveness. Increased NK cell motility was observed on Boyden filters coated with human IgG, IgG Fc, and mAb against CD16 and the § 4, § 5, § L, g 1 and g 2 integrin chains. No motile response was seen on mouse IgG or CD7, CD56 and § 6 integrin mAb. NK cell migration on human IgG and anti-CD16 Ab was blocked by anti-CD16 or anti-g 2, but not anti-g 1 Ab, implying that the motile response triggered by CD16 cross-linking is mediated via g 2 integrins. Preformed or induced anti- § -Gal IgG may therefore contribute to AVR by stimulating innate immune cell infiltration of the graft.

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Section: Discussionmentioning

confidence: 78%

Section: Migration Of Pbmc Across Porcine Endothelium In the Presencementioning

confidence: 93%

Section: Introductionmentioning

confidence: 95%

Section: Primary Porcine Aortic Endothelial Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

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Anti‐Gal IgG potentiates natural killer cell migration across porcine endothelium via endothelial cell activation and increased natural killer cell motility triggered by CD16 cross‐linking

2004

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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

He¹,

Ehrnfelt²,

Kumagai‐Braesch³

et al. 2004

Eur J Immunol

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The contribution of Gal § 1,3Gal ( § -Gal) to cell-mediated organ xenograft rejection is controversial. We have used recombinant lentiviruses encoding a porcine § 1,3 galactosyltransferase ( § 1,3GalT) to obtain § -Gal-expressing primary human aortic endothelial cells (HAEC) at a frequency of 70-90%. These cells were compared to non-transduced and mock-transduced HAEC with regard to their susceptibility to human NK cell-mediated lysis, ability to stimulate IFN-+ production by NK cells, and support of NK cell adhesion under static and dynamic conditions. Using green fluorescent protein (GFP) as a reporter gene, it was shown that the frequency of green fluorescent HAEC increased until day 5 posttransduction, and at a multiplicity of infection of 2.5, it reached 98%. Lentiviral transduction did not result in activation of HAEC, and transduced HAEC responded as expected to TNF- § and IFN-+ stimulation. No differences were detected between non- § -Gal-and § -Galexpressing HAEC in terms of their susceptibility to NK cell-mediated lysis, ability to stimulate IFN-+ production by NK cells, or ability to support NK cell adhesion under static and dynamic conditions. In conclusion, these data argue against an important role for the § -Gal epitope in the direct interaction between endothelium and NK cells and prove that recombinant lentiviruses are efficient gene carriers for primary human endothelial cells.

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Cellular Studies for In Vitro Modeling of Xenogeneic Immune Responses

Sommaggio

Pérez-Cruz

Costa

2012

Xenotransplantation

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Cellular studies are essential in the xenotransplantation field in order to investigate the cellular immune responses triggered by xenogeneic cells and identify the key molecules involved. A series of functional studies can be conducted with this purpose that include treatment with proinflammatory cytokines and xenogeneic cell-based assays that put together pig cells and human leukocytes such as monocytes, NK cells, and T cells. The choice of the pig cell type is critical to appropriately model the transplant setting of interest. Thus, pig endothelial cells are commonly used for studying the rejection process of vascularized organs. Treatment with cytokines allows studying the regulation of adhesion, costimulatory molecules, and receptors involved in triggering the immune response in an attempt to reproduce the more complex in vivo situation. The adhesion assays are used to determine the capacity of human leukocytes to adhere to porcine cells under various conditions. Furthermore, we describe coculture, costimulatory, and cytotoxicity assays for investigating the cellular and molecular mechanisms that take place during the xenogeneic immune response.

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Porcine Endothelium Supports Transendothelial Migration of Human Leukocyte Subpopulations: Anti-Porcine Vascular Cell Adhesion Molecule Antibodies as Species-Specific Blockers of Transendothelial Monocyte and Natural Killer Cell Migration1

Cited by 35 publications

References 39 publications

Anti‐Gal IgG potentiates natural killer cell migration across porcine endothelium via endothelial cell activation and increased natural killer cell motility triggered by CD16 cross‐linking

Anti‐Gal IgG potentiates natural killer cell migration across porcine endothelium via endothelial cell activation and increased natural killer cell motility triggered by CD16 cross‐linking

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

Cellular Studies for In Vitro Modeling of Xenogeneic Immune Responses

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