Decline of surface MHC I by adenoviral gene transfer of anti‐MHC I intrabodies in human endothelial cells—new perspectives for the generation of universal donor cells for tissue transplantation

Beyer, Florian; Doebis, Cornelia; Busch, Annette; Ritter, Thomas; Mhashilkar, Abner M.; Marasco, Wayne; Laube, Horst; Volk, Hans‐Dieter; Seifert, Martina

doi:10.1002/jgm.548

Cited by 29 publications

(25 citation statements)

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“…Strategies involving the gene transfer of an anti-HLA I single chain IB (Beyer et al, 2004) or plasmids containing siRNA (Gonzalez et al, 2005) were recently developed to downregulate surface HLA I. Using the IB technology as a second line of capture mechanism for the very few HLA I molecules that still were synthesized despite the use of siRNA caused no retainment of intracellular HLA I molecules.…”

Section: Discussionmentioning

confidence: 99%

Immunobiology of naïve and genetically modified HLA-class-I-knockdown human embryonic stem cells

Deuse

Seifert

Phillips

et al. 2011

Journal of Cell Science

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KD was followed by flow cytometry and showed levels between 1% and 12% of those of naïve hESC between days 7 and 42 (means ± s.e.m.). (b) A total of 10 6 hESCs or hESC KD were transplanted into the gastrocnemius muscle of either immunocompetent Balb/c or severely immunocompromised SCID-beige mice. All ten Balb/c mice rapidly rejected the hESC transplants, whereas the cells survived in ten SCID-beige recipients. Rejection of hESC KD was markedly attenuated and four out of ten hESC KD grafts achieved long-term survival. 4128 Author Correction The first author apologises for these errors.Immunobiology of naïve and genetically modified HLA-class-I-knockdown human embryonic stem cells SummaryHuman embryonic stem cells (hESCs) can serve as a universal cell source for emerging cell or tissue replacement strategies, but immune rejection of hESC derivatives remains an unsolved problem. Here, we sought to describe the mechanisms of rejection for naïve hESCs and upon HLA class I (HLA I) knockdown (hESC KD ). hESCs were HLA I-positive but negative for HLA II and co-stimulatory molecules. Transplantation of naïve hESC into immunocompetent Balb/c mice induced substantial T helper cell 1 and 2 (Th1 and Th2) responses with rapid cell death, but hESCs survived in immunodeficient SCID-beige recipients. Histology revealed mainly macrophages and T cells, but only scattered natural killer (NK) cells. A surge of hESC-specific antibodies against hESC class I, but not class II antigens, was observed. Using HLA I RNA interference and intrabody technology, HLA I surface expression of hESC KD was 88%-99% reduced. T cell activation after hESC KD transplantation into Balb/c was significantly diminished, antibody production was substantially alleviated, the levels of graft-infiltrating immune cells were reduced and the survival of hESC KD was prolonged. Because of their very low expression of stimulatory NK ligands, NK-susceptibility of naïve hESCs and hESC KD was negligible. Thus, HLA I recognition by T cells seems to be the primary mechanism of hESC recognition, and T cells, macrophages and hESC-specific antibodies participate in hESC killing.

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Section: Discussionmentioning

confidence: 99%

Immunobiology of naïve and genetically modified HLA-class-I-knockdown human embryonic stem cells

Deuse

Seifert

Phillips

et al. 2011

Journal of Cell Science

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“…The efficiency of the secreted MB in blocking cell surface transport of the related antigen is comparable to that of intrabodies, which have been used successfully in vitro to down-regulate the expression of transmembrane or secreted target proteins, such as the folate receptor, VEGFR-2 and MHC1 [28][29][30].…”

Section: Discussionmentioning

confidence: 99%

Controlling complement resistance in cancer by using human monoclonal antibodies that neutralize complement-regulatory proteins CD55 and CD59

et al. 2005

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Expression of the complement-regulatory proteins (CRP) CD46, CD55 and CD59 represents a strategy used by tumor cells to evade complement-dependent cell cytoxicity stimulated by monoclonal antibodies. We have isolated two single-chain variable fragments (scFv) to CD55 and CD59 from a human phage-display library and from these scFv we have produced two miniantibodies (MB), MB-55 (against CD55) and MB-59 (against CD59), containing the human hinge-CH2-CH3 domains of IgG1. The specificity of the two MB for the corresponding CRP was assessed by ELISA using purified CD46, CD55 and CD59. MB-55 and MB-59 neutralized the inhibitory activity of CD55 and CD59, respectively, restoring the complement-mediated lysis of sheep and guinea pig erythrocytes that was otherwise inhibited by the two CRP. FACS analysis revealed binding of MB-55 and MB-59 to the lymphoma cell line Karpas 422. The two MB induced a two-fold increase in the complement-dependent killing of these cells stimulated by Rituximab, a chimeric anti-CD20 monoclonal antibody. Transfection of HEK293T cells with vectors encoding MB-55 or MB-59 markedly reduced the expression of CD55 and CD59. We conclude that the human antibodies MB-55 and MB-59 may represent a therapeutic tool to increase the complement-dependent killing activity of Rituximab in the treatment of non-Hodgkin's lymphoma.

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“…Strategies involving the gene transfer of an anti-HLA I single-chain IB 9 or plasmids containing siRNA 22 recently were introduced to downregulate surface HLA I. Although we could demonstrate in the present study that either technique worked with hESC, only their combination provided the needed level of HLA I knockdown to generate hypoantigeneic hESC siRNAϩIB .…”

Section: Discussionmentioning

confidence: 53%

Human Leukocyte Antigen I Knockdown Human Embryonic Stem Cells Induce Host Ignorance and Achieve Prolonged Xenogeneic Survival

Deuse

Seifert²,

Phillips³

et al. 2011

Circulation

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Background-Although human embryonic stem cells (hESC) have enormous potential for cell replacement therapy of heart failure, immune rejection of hESC derivatives inevitably would occur after transplantation. We therefore aimed to generate a hypoantigeneic hESC line with improved survival characteristics. Methods and Results-Using various in vivo, nonischemic, hindlimb xenotransplant models (immunocompetent and defined immunodefective mouse strains) as well as human in vitro T-cell and natural killer (NK)-cell assays, we revealed a central role for T cells in mediating hESC rejection. The NK-cell susceptibility of hESC in vivo was found to be low, and the NK response to hESC challenge in vitro was negligible. To reduce the antigenicity of hESC, we successfully generated human leukocyte antigen (HLA) I knockdown cells (hESC siRNAϩIB ) using both HLA I RNA interference (siRNA) and intrabody (IB) technology. HLA I expression was Ϸ99% reduced after 7 days and remained low for weeks. Cellular immune recognition of these hESC siRNAϩIB was strongly reduced in both xenogeneic and allogeneic settings. Immune rejection was profoundly mitigated after hESC siRNAϩIB transplantation into immunocompetent mice, and even long-term graft survival was achieved in one third of the animals without any immunosuppression. The survival benefit of hESC siRNAϩIB was further confirmed under ischemic conditions in a left anterior descending coronary artery ligation model. Conclusions-HLA I knockdown hESC siRNAϩIB provoke T-cell ignorance and experience largely mitigated xenogeneic rejection. By generating hypoantigeneic hESC lines, the generation of acceptable hESC derivatives may become a practical concept and push cell replacement strategies forward.

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Decline of surface MHC I by adenoviral gene transfer of anti‐MHC I intrabodies in human endothelial cells—new perspectives for the generation of universal donor cells for tissue transplantation

Abstract: Our data indicate that HUVEC with reduced levels of MHC I might be used as universal donor cells for the seeding of vascular grafts.

Cited by 29 publications

References 46 publications

Immunobiology of naïve and genetically modified HLA-class-I-knockdown human embryonic stem cells

Immunobiology of naïve and genetically modified HLA-class-I-knockdown human embryonic stem cells

Controlling complement resistance in cancer by using human monoclonal antibodies that neutralize complement-regulatory proteins CD55 and CD59

Human Leukocyte Antigen I Knockdown Human Embryonic Stem Cells Induce Host Ignorance and Achieve Prolonged Xenogeneic Survival

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