Xenotransplantation最新文献

{"title":"Characterisation of transgenic pigs expressing a human T cell-depleting anti-CD2 monoclonal antibody.","authors":"Evelyn J Salvaris, Nella Fisicaro, Stephen McIlfatrick, Adwin Thomas, Erin Fuller, Andrew M Lew, Mark B Nottle, Wayne J Hawthorne, Peter J Cowan","doi":"10.1111/xen.12836","DOIUrl":"10.1111/xen.12836","url":null,"abstract":"<p><strong>Background: </strong>Pig islet xenotransplantation is a potential treatment for type 1 diabetes. We have shown that maintenance immunosuppression is required to protect genetically modified (GM) porcine islet xenografts from T cell-mediated rejection in baboons. Local expression of a depleting anti-CD2 monoclonal antibody (mAb) by the xenograft may provide an alternative solution. We have previously reported the generation of GGTA1 knock-in transgenic pigs expressing the chimeric anti-CD2 mAb diliximab under an MHC class I promoter (MHCIP). In this study, we generated GGTA1 knock-in pigs in which MHCIP was replaced by the β-cell-specific porcine insulin promoter (PIP), and compared the pattern of diliximab expression in the two lines.</p><p><strong>Methods: </strong>A PIP-diliximab knock-in construct was prepared and validated by transfection of NIT-1 mouse insulinoma cells. The construct was knocked into GGTA1 in wild type (WT) porcine fetal fibroblasts using CRISPR, and knock-in cells were used to generate pigs by somatic cell nuclear transfer (SCNT). Expression of the transgene in MHCIP-diliximab and PIP-diliximab knock-in pigs was characterised at the mRNA and protein levels using RT-qPCR, flow cytometry, ELISA and immunohistochemistry. Islets from MHCIP-diliximab and control GGTA1 KO neonatal pigs were transplanted under the kidney capsule of streptozotocin-diabetic SCID mice.</p><p><strong>Results: </strong>NIT-1 cells stably transfected with the PIP-diliximab knock-in construct secreted diliximab into the culture supernatant, confirming correct expression and processing of the mAb in β cells. PIP-diliximab knock-in pigs showed a precise integration of the transgene within GGTA1. Diliximab mRNA was detected in all tissues tested (spleen, kidney, heart, liver, lung, pancreas) in MHCIP-diliximab pigs, but was not detectable in PIP-diliximab pigs. Likewise, diliximab was present in the serum of MHCIP-diliximab pigs, at a mean concentration of 1.8 μg/mL, but was not detected in PIP-diliximab pig serum. An immunohistochemical survey revealed staining for diliximab in all organs of MHCIP-diliximab pigs but not of PIP-diliximab pigs. Whole genome sequencing (WGS) of a PIP-diliximab pig identified a missense mutation in the coding region for the dixilimab light chain. This mutation was also found to be present in the fibroblast knock-in clone used to generate the PIP-diliximab pigs. Islet xenografts from neonatal MHCIP-diliximab pigs restored normoglycemia in diabetic immunodeficient mice, indicating no overt effect of the transgene on islet function, and demonstrated expression of diliximab in situ.</p><p><strong>Conclusion: </strong>Diliximab was widely expressed in MHCIP-diliximab pigs, including in islets, consistent with the endogenous expression pattern of MHC class I. Further investigation is required to determine whether the level of expression in islets from the MHCIP-diliximab pigs is sufficient to prevent T cell-mediated islet","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":" ","pages":"e12836"},"PeriodicalIF":3.9,"publicationDate":"2023-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10909556/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"92156865","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Xenoreactive antibodies in α-granules of human platelets bind pig liver endothelial cells.","authors":"Christopher Burlak, Zheng Yu Wang, Greg Martens, Jose Estrada, Luz Reyes, Victor Manuel Novara Gennuso, Rodrigo Vianna, Matt Tector, Alfred Joseph Tector","doi":"10.1111/xen.12834","DOIUrl":"10.1111/xen.12834","url":null,"abstract":"<p><p>Pig liver xenotransplantation is limited by a thrombocytopenic coagulopathy that occurs immediately following graft reperfusion. In vitro and ex vivo studies from our lab suggested that the thrombocytopenia may be the result of a species incompatibility in platelet glycosylation. Realization that platelet α-granules contain antibodies caused us to reevaluate whether the thrombocytopenia in liver xenotransplantation could occur because IgM and IgG from inside platelet α-granules bound to pig liver sinusoidal endothelial cells (LSECs). Our in vitro analysis of IgM and IgG from inside α-granules showed that platelets do carry xenoreactive antibodies that can bind to known xenoantigens. This study suggests that thrombocytopenia occurring following liver xenotransplantation could occur because of xenoreactive antibodies tethering human platelets to the pig LSEC enabling the platelet to be phagocytosed. These results suggest genetic engineering strategies aimed at reducing xenoantigens on the surface of pig LSEC will be effective in eliminating the thrombocytopenia that limits survival in liver xenotransplantation.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":" ","pages":"e12834"},"PeriodicalIF":3.3,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136399507","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Open invitation to contribute ideas to a multifaceted approach to ethics in xenotransplantation.","authors":"Kim Solez, Elisa Gordon, Alton Brad Farris, Lynn Cornell","doi":"10.1111/xen.12827","DOIUrl":"10.1111/xen.12827","url":null,"abstract":"","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":" ","pages":"e12827"},"PeriodicalIF":3.3,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10591749","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A message from Mr. David Bennett Jr., the son of the first patient to receive a gene-edited pig heart transplant.","authors":"David Bennett","doi":"10.1111/xen.12839","DOIUrl":"10.1111/xen.12839","url":null,"abstract":"","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"30 6","pages":"e12839"},"PeriodicalIF":3.9,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138804428","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a panel for detection of pathogens in xenotransplantation donor pigs.","authors":"Hikari Otabi, Hiroto Miura, Haruka Uryu, Rana Kobayashi-Harada, Kanako Abe, Kazuaki Nakano, Kazuhiro Umeyama, Koki Hasegawa, Takamitsu Tsukahara, Hiroshi Nagashima, Ryo Inoue","doi":"10.1111/xen.12825","DOIUrl":"10.1111/xen.12825","url":null,"abstract":"<p><p>There have been high expectations in recent years of using xenotransplantation and regenerative medicine to treat humans, and pigs have been utilized as the donor model. Pigs used for these clinical applications must be microbiologically safe, that is, free of infectious pathogens, to prevent infections not only in livestock, but also in humans. Currently, however, the full spectrum of pathogens that can infect to the human host or cause disease in transplanted porcine organs/cells has not been fully defined. In the present study, we thus aimed to develop a larger panel for the detection of pathogens that could potentially infect xenotransplantation donor pigs. Our newly developed panel, which consisted of 76 highly sensitive PCR detection assays, was able to detect 41 viruses, 1 protozoa, and a broad range of bacteria (by use of universal 16S rRNA primers). The applicability of this panel was validated using blood samples from uterectomy-born piglets, and pathogens suspected to be vertically transmitted from sows to piglets were successfully detected. We estimate that, at least for viruses and bacteria, the number of target pathogens detected by the developed screening panel should suffice to meet the microbiological safety levels required worldwide for xenotransplantation and/or regenerative therapy. This panel provides greater diagnosis options to produce donor pigs so that it would render unnecessary to screen for all pathogens listed. Instead, the new panel could be utilized to detect only required pathogens within a given geographic range where the donor pigs for xenotransplantation have been and/or are being developed.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":" ","pages":"e12825"},"PeriodicalIF":3.3,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41138219","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lessons learned from the first cardiac xenotransplant in a consciously consenting human: Psychiatric considerations and the impact of media exposure.","authors":"Nithya Cherukuru, Argyro Athanasiadi, Rachel LeMalefant, David Mancini, Anique Forrester, David Glovinsky, Pinar Miski, Catherine Harrison-Restelli, Charles Robinson","doi":"10.1111/xen.12830","DOIUrl":"10.1111/xen.12830","url":null,"abstract":"","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":" ","pages":"e12830"},"PeriodicalIF":3.3,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49682899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modeling human anti-pig xenoimmune responses in a pig artery tissue grafted humanized mouse model.","authors":"Minghui Fang, Jun Zou, Fei Xu, Xue Wang, Shucheng Hua, Qi Zhou, Yong-Guang Yang, Zheng Hu","doi":"10.1111/xen.12824","DOIUrl":"10.1111/xen.12824","url":null,"abstract":"<p><strong>Background: </strong>Blood vessels that contain endothelial cells (ECs) on the surface are in direct contact with host blood and are the first target of xenograft rejection. Currently, our understanding of human anti-pig vessel immune responses is primarily based on in vitro assays using pig ECs. Therefore, it is necessary to develop an animal model that permits in vivo study of human immunological rejection of pig vessels.</p><p><strong>Methods: </strong>Pig artery tissues (PAT) were transplanted into human immune system (HIS) mice or immunodeficient NSG mice (as controls). Intragraft human immune cell infiltration and antibody deposition were quantified using histology and immunohistochemistry. Donor antigen-specific immune responses were quantified using a mixed lymphocyte reaction and a complement-dependent killing assay.</p><p><strong>Results: </strong>Pig CD31⁺ ECs were detected and increased 2-fold from weeks 3 to 5 in PAT xenografts from immunodeficient NSG mice. However, compared with NSG mice, PAT xenografts in HIS mice had significantly lower numbers of porcine CD31⁺ ECs and showed a marked reduction from week 3 to week 5. PAT xenograft rejection in HIS mice is associated with intensive infiltration of human immune cells, deposition of human IgM and IgG antibodies, and the formation of a tertiary lymphoid structure. Robust donor pig antigen-specific human T cells and antibody responses were detected in PAT-transplanted HIS mice.</p><p><strong>Conclusion: </strong>We have developed a humanized mouse model to evaluate human anti-pig xenoimmune responses by PAT transplantation in vivo. This model is expected to facilitate the refinement of pig gene-editing strategies (the expression on EC surface) and the testing of local immunosuppressive strategies for clinical pig organ xenotransplantation.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":" ","pages":"e12824"},"PeriodicalIF":3.3,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10200810","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acceptance of xenotransplantation by patients waiting for organ donation: A qualitative study.","authors":"Özlem Şahin Akboğa, Akarsu Rukiye Hobek","doi":"10.1111/xen.12813","DOIUrl":"10.1111/xen.12813","url":null,"abstract":"<p><strong>Background: </strong>The aim of this study was to determine the acceptance and barriers to xenotransplantation in patients waiting for organ transplantation.</p><p><strong>Methods: </strong>It is qualitative and descriptive research. It was completed with 18 patients receiving treatment and waiting for organ transplantation in a dialysis center located in the inner region of Turkey between January 26, 2023 and February 3, 2023. Data were collected with an introductory information form and a semi-structured interview form. The research data were collected through face-to-face in-depth interviews. The content analysis method was used to analyze the data. In line with the goal of reaching data saturation, in-depth interviews were conducted with 18 participants who were open to communication.</p><p><strong>Results: </strong>Two main themes, \"Values\" and \"Thoughts\", and five sub-themes, \"social and religious values, positive, negative and future thoughts\", were identified. Thirteen codes were created including \"not being understood by the society, xenotransplantation prejudice, fear of ridicule and exclusion, religious pressure, desire for unconditional acceptance/rejection\" and \"thought of survival, hope, thought of sinning, submission to doctors, the attitude of religious men and excessive demand\".</p><p><strong>Conclusions: </strong>Patients awaiting organ transplantation need religious, social, and community support for xenotransplantation.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":" ","pages":"e12813"},"PeriodicalIF":3.3,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9867688","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expert opinion on the identification, risk assessment, and mitigation of microorganisms and parasites relevant to xenotransplantation products from pigs.","authors":"Huybert Groenendaal, Solenne Costard, Reid Ballard, Stephen Bienhoff, Diana C Challen, Brandon J Dominguez, Douglas R Kern, Dan Miller, Jeske Noordergraaf, Larisa Rudenko, Henk-Jan Schuurman, Tom Spizzo, Matthew Sturos, Bill Zollers, Jay A Fishman","doi":"10.1111/xen.12815","DOIUrl":"10.1111/xen.12815","url":null,"abstract":"<p><p>Xenotransplantation has the potential to address shortages of organs available for clinical transplantation, but concerns exist regarding potential risks posed by porcine microorganisms and parasites (MP) to the health of human recipients. In this study, a risk-based framework was developed, and expert opinion was elicited to evaluate porcine MP based on swine exposure and risk to human health. Experts identified 255 MP to include in the risk assessment. These were rated by experts for five criteria regarding potential swine exposure in the USA and human health risks. MP were subsequently categorized into three risk mitigation groups according to pre-defined rules: disqualifying porcine MP (due to their pathogenic potential, n = 130); non-disqualifying porcine MP (still relevant to consider for biosecurity or monitoring efforts, n = 40); and alert/watch list (not reported in the USA or MP not in swine, n = 85). Most disqualifying (n = 126) and non-disqualifying (n = 36) porcine MP can effectively be eliminated with high biosecurity programs. This approach supports surveillance and risk mitigation strategies for porcine MP in swine produced for xenotransplantation, such as documentation of freedom from porcine MP, or use of porcine MP screening, monitoring, or elimination options. To the authors' knowledge, this is the first effort to comprehensively identify all relevant porcine MP systematically and transparently evaluate the risk of infection of both donor animals and immunosuppressed human recipients, and the potential health impacts for immunosuppressed human recipients from infected xenotransplantation products from pigs.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":" ","pages":"e12815"},"PeriodicalIF":3.3,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10067984","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Auxiliary liver xenotransplantation technique in a transgenic pig-to-non-human primate model: A surgical approach to prolong survival.","authors":"Kyo Won Lee, Sean S W Park, Dong Suk Kim, Kimyung Choi, Joohyun Shim, Jihun Kim, Sung Joo Kim, Jae Berm Park","doi":"10.1111/xen.12814","DOIUrl":"10.1111/xen.12814","url":null,"abstract":"<p><p>Xenotransplantation using pigs' liver offers a potentially alternative method to overcome worldwide donor shortage, or more importantly as a bridge to allotransplantation. However, it has been challenged by profound thrombocytopenia and fatal coagulopathy in non-human primate models. Here we suggest that a left auxiliary technique can be a useful method to achieve extended survival of the xenograft. Fifteen consecutive liver xenotransplants were carried out in a pig-to-cynomolgus model. Right auxiliary technique was implemented in two cases, orthotopic in eight cases, and left auxiliary in five cases. None of the right auxiliary recipients survived after surgery due to hemorrhage during complex dissection between the primate's right lobe and inferior vena cava. Orthotopic recipients survived less than 7 days secondary to profound thrombocytopenia and coagulopathy. Two out of five left auxiliary xenotransplants survived more than 3 weeks without uncontrolled thrombocytopenia or anemia, with one of them surviving 34 days, the longest graft survival reported to date. Left auxiliary xenotransplant is a feasible approach in non-human primate experiments, and the feared risk of thrombocytopenia and coagulopathy can be minimized. This may allow for longer evaluation of the xenograft and help better understand histopathological and immunological changes that occur following liver xenotransplantation.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":" ","pages":"e12814"},"PeriodicalIF":3.3,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9870619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}