76 results on '"Schuurman HJ"'
Search Results
2. Evaluation of CD38 as target for immunotherapy in multiple myeloma [letter; comment]
- Author
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Vooijs, WC, primary, Schuurman, HJ, additional, Bast, EJ, additional, and de Gast, GC, additional
- Published
- 1995
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3. The immunosuppressant rapamycin blocks in vitro responses to hematopoietic cytokines and inhibits recovering but not steady-state hematopoiesis in vivo
- Author
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Quesniaux, VF, primary, Wehrli, S, additional, Steiner, C, additional, Joergensen, J, additional, Schuurman, HJ, additional, Herrman, P, additional, Schreier, MH, additional, and Schuler, W, additional
- Published
- 1994
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4. Syngeneic leukocytes together with suramin failed to improve immunodeficiency in a case of transfusion-associated AIDS after syngeneic bone marrow transplantation
- Author
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Verdonck, LF, de Gast, GC, Lange, JM, Schuurman, HJ, Dekker, AW, and Bast, BJ
- Abstract
A 22-year-old man who underwent syngeneic bone marrow transplantation (BMT) for acute lymphoblastic leukemia acquired a human immunodeficiency virus (HIV) infection by transfusion of blood products from a donor at risk. The manifestations were acute encephalopathy together with immune thrombocytopenia in the early posttransplant period, and acquired immunodeficiency syndrome (AIDS) developed 20 months after BMT. Because he had a syngeneic donor, the possibility of reconstituting the immune system was investigated by repeated transfer of healthy syngeneic lymphocytes and by combining repeated transfer of syngeneic lymphocytes with the antiviral agent suramin to protect the infused leukocytes from being attacked by HIV. No improvement was observed clinically or in the patient's immune functions by these efforts.
- Published
- 1988
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5. The serological response of adult cattle to vaccination with reduced dose Brucella abortus S19, a trial under Zambian conditions
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Schuurman Hj
- Subjects
Veterinary medicine ,Rose Bengal ,Time Factors ,General Veterinary ,business.industry ,Vaccination ,Age Factors ,Brucella Vaccine ,Brucella abortus ,Brucellosis ,medicine.disease ,Reduced dose ,Virology ,Antibodies, Bacterial ,Serology ,Brucellosis, Bovine ,Agglutination Tests ,medicine ,Animals ,Cattle ,Female ,business ,Immunization Schedule - Abstract
Summary A total of 250 adult cows and 13 (young) heifers, all seronegative for brucellosis, were vaccinated with a reduced dose (3 × 109) Strain 19 (S‐19) vaccine. Twelve months after vaccination all cattle were negative to the serological tests. The results obtained are similar to those found by other authors and support the use of reduced dose S19 under the heat conditions found in Zambia.
- Published
- 1983
6. Modern immunosuppressives.
- Author
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Schuurman, HJ
- Subjects
- *
IMMUNOSUPPRESSIVE agents , *NONFICTION - Abstract
Reviews the book 'Modern Immunosuppressives,' edited by H.J. Schuurman, G. Feutren and J.F. Bach.
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- 2002
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7. Clinically available immunosuppression averts rejection but not systemic inflammation after porcine islet xenotransplant in cynomolgus macaques.
- Author
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Graham ML, Ramachandran S, Singh A, Moore MEG, Flanagan EB, Azimzadeh A, Burlak C, Mueller KR, Martins K, Anazawa T, Appakalai BN, Bansal-Pakala P, Murtaugh MP, O'Brien TD, Papas KK, Spizzo T, Schuurman HJ, Hancock WW, and Hering BJ
- Subjects
- Animals, Graft Rejection etiology, Graft Survival, Heterografts, Humans, Immunosuppression Therapy, Immunosuppressive Agents pharmacology, Immunosuppressive Agents therapeutic use, Inflammation etiology, Macaca fascicularis, Swine, Transplantation, Heterologous methods, Diabetes Mellitus, Islets of Langerhans Transplantation methods
- Abstract
A safe, efficacious, and clinically applicable immunosuppressive regimen is necessary for islet xenotransplantation to become a viable treatment option for diabetes. We performed intraportal transplants of wild-type adult porcine islets in 25 streptozotocin-diabetic cynomolgus monkeys. Islet engraftment was good in 21, partial in 3, and poor in 1 recipient. Median xenograft survival was 25 days with rapamycin and CTLA4Ig immunosuppression. Adding basiliximab induction and maintenance tacrolimus to the base regimen significantly extended median graft survival to 147 days (p < .0001), with three animals maintaining insulin-free xenograft survival for 265, 282, and 288 days. We demonstrate that this regimen suppresses non-Gal anti-pig antibody responses, circulating effector memory T cell expansion, effector function, and infiltration of the graft. However, a chronic systemic inflammatory state manifested in the majority of recipients with long-term graft survival indicated by increased neutrophil to lymphocyte ratio, IL-6, MCP-1, CD40, and CRP expression. This suggests that this immunosuppression regimen fails to regulate innate immunity and resulting inflammation is significantly associated with increased incidence and severity of adverse events making this regimen unacceptable for translation. Additional studies are needed to optimize a maintenance regimen for regulating the innate inflammatory response., (© 2021 The American Society of Transplantation and the American Society of Transplant Surgeons.)
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- 2022
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8. High Prevalence of Recombinant Porcine Endogenous Retroviruses (PERV-A/Cs) in Minipigs: A Review on Origin and Presence.
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Denner J and Schuurman HJ
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- Animals, Cell Line, Humans, Swine virology, Transplantation, Heterologous, Virus Integration, Endogenous Retroviruses genetics, Endogenous Retroviruses isolation & purification, Recombination, Genetic, Swine, Miniature virology
- Abstract
Minipigs play an important role in biomedical research and they have also been used as donor animals for preclinical xenotransplantations. Since zoonotic microorganisms including viruses can be transmitted when pig cells, tissues or organs are transplanted, virus safety is an important feature in xenotransplantation. Whereas most porcine viruses can be eliminated from pig herds by different strategies, this is not possible for porcine endogenous retroviruses (PERVs). PERVs are integrated in the genome of pigs and some of them release infectious particles able to infect human cells. Whereas PERV-A and PERV-B are present in all pigs and can infect cells from humans and other species, PERV-C is present in most, but not all pigs and infects only pig cells. Recombinant viruses between PERV-A and PERV-C have been found in some pigs; these recombinants infect human cells and are characterized by high replication rates. PERV-A/C recombinants have been found mainly in minipigs of different origin. The possible reasons of this high prevalence of PERV-A/C in minipigs, including inbreeding and higher numbers and expression of replication-competent PERV-C in these animals, are discussed in this review. Based on these data, it is highly recommended to use only pig donors in clinical xenotransplantation that are negative for PERV-C.
- Published
- 2021
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9. An Invited Commentary on "Bullying and undermining behaviours in surgery: A qualitative study of surgical trainee experiences in the United Kingdom (UK) & Republic Of Ireland (ROI)" [Int. J. Surg. (2020) Epub ahead of print].
- Author
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Schuurman HJ
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- Humans, Ireland, Qualitative Research, United Kingdom, Bullying prevention & control, Printing, Three-Dimensional
- Published
- 2020
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10. An invited commentary on "Comparative analysis of weight loss and resolution of comorbidities between laparoscopic sleeve gastrectomy and Roux-en-Y gastric bypass: A systematic review and meta-analysis based on 18 studies" (Int J Surg 2020; 76: 101-110).
- Author
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Schuurman HJ
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- Gastrectomy, Humans, Weight Loss, Gastric Bypass, Laparoscopy, Obesity, Morbid surgery
- Abstract
Competing Interests: Declaration of competing interest The author declares no conflict of interest.
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- 2020
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11. Invited Commentary on "Safety and efficacy of Lactobacillus for preventing necrotizing enterocolitis in preterm infants: A systematic review and meta-analysis" (Int J Surg 2020; 76:79-87).
- Author
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Schuurman HJ
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- Humans, Infant, Infant, Newborn, Infant, Premature, Lactobacillus, Enterocolitis, Necrotizing, Probiotics, Sepsis
- Abstract
Competing Interests: Declaration of competing interest The author declares no conflict of interest.
- Published
- 2020
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12. Invited Commentary on: Novel scoring system for recurrence risk classification of surgically resected G1/2 pancreatic neuroendocrine tumors - Retrospective cohort study [Article type: Retrospective cohort Study] (Int J Surg 2020).
- Author
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Schuurman HJ
- Subjects
- Humans, Retrospective Studies, Neuroendocrine Tumors, Pancreatic Neoplasms
- Abstract
Competing Interests: Declaration of competing interest The author declares no conflict of interest.
- Published
- 2020
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13. Invited Commentary on: Efficacy of pulmonary rehabilitation in improving the quality of life for patients with chronic obstructive pulmonary disease: A systematic review and meta-analysis. Review article [Int J Surg 2019].
- Author
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Schuurman HJ
- Subjects
- Humans, Randomized Controlled Trials as Topic, Pulmonary Disease, Chronic Obstructive, Quality of Life
- Abstract
Competing Interests: Declaration of competing interest The author declares no conflict of interest.
- Published
- 2020
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14. Commentary on: Is the renal subcapsular space the preferred site for clinical porcine islet xenotransplantation? Review article (Int J Surg 2019 Jul 30;69:100-107. https://doi.org/10.1016/j.ijsu.2019.07.032. [Epub ahead of print]).
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Schuurman HJ and Graham ML
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- Animals, Heterografts, Swine, Transplantation, Heterologous, Kidney, Printing, Three-Dimensional
- Published
- 2019
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15. Cellular xenotransplantation of animal cells into people: benefits and risk.
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Scobie L, Galli C, Gianello P, Cozzi E, and Schuurman HJ
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- Animals, Animals, Genetically Modified, Communicable Diseases transmission, Humans, Islets of Langerhans Transplantation adverse effects, Islets of Langerhans Transplantation methods, Tissue and Organ Procurement, Zoonoses, Communicable Diseases veterinary, Islets of Langerhans Transplantation veterinary, Transplantation, Heterologous adverse effects
- Abstract
The main benefit of xenotransplantation is its potential to overcome the worldwide organ shortage experienced in allotransplantation. Allogeneic transplantation is the only successful therapy for several life-threatening diseases, with cell, tissue or organ donation only partially meeting the demand and many patients dying while waiting for treatment. With supply falling short of demand, it is foreseen that the use of porcine material may at some stage overcome the existing gap between organ availability and clinical need. Recently, pig islet cells have been utilised in clinical trials, with safety being demonstrated. Indeed, pig-derived cells present several advantages: i) porcine cells have a stable function and differentiation pattern and are not tumorigenic; ii) pig cells have been shown to meet the physiological needs in large animal models; iii) the source of pig cells can be scaled up to meet demands in a highly standardised manner, and with respect to animal welfare regulations; iv) 'designated-pathogen-free' (DPF) pig lines can be produced, which could result in a higher safety profile than allotransplantation itself; v) the risk of zoonosis, which was raised years ago as the major hurdle, has been recently circumvented and is actually viewed as a controlled risk; and vi) immune risks are being circumvented via the use of genetically modified donor animals and encapsulation of porcine cells, particularly for the treatment of diabetes. Overall, the benefit appears to outweigh potential risks with respect to cellular xenotransplantation and this is discussed further in this review.
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- 2018
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16. Beneficial Effects of Human Mesenchymal Stromal Cells on Porcine Hepatocyte Viability and Albumin Secretion.
- Author
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Montanari E, Pimenta J, Szabó L, Noverraz F, Passemard S, Meier RPH, Meyer J, Sidibe J, Thomas A, Schuurman HJ, Gerber-Lemaire S, Gonelle-Gispert C, and Buhler LH
- Subjects
- Albumins metabolism, Alginates, Animals, Cell Survival, Cells, Cultured, Coculture Techniques, Drug Compounding, Glucuronic Acid, Hepatocytes transplantation, Hexuronic Acids, Humans, Hydrogels, Swine, Transplantation, Heterologous, Hepatocytes physiology, Liver Failure therapy, Mesenchymal Stem Cells physiology
- Abstract
Porcine hepatocytes transplanted during acute liver failure might support metabolic functions until the diseased liver recovers its function. Here, we isolated high numbers of viable pig hepatocytes and evaluated hepatocyte functionality after encapsulation. We further investigated whether coculture and coencapsulation of hepatocytes with human multipotent mesenchymal stromal cells (MSC) are beneficial on hepatocyte function. Livers from 10 kg pigs ( n = 9) were harvested, and hepatocytes were isolated from liver suspensions for microencapsulation using alginate and poly(ethylene-glycol)- (PEG-) grafted alginate hydrogels, either alone or in combination with MSC. Viability, albumin secretion, and diazepam catabolism of hepatocytes were measured for one week. 9.2 ± 3.6 × 10
9 hepatocytes with 95.2 ± 3.1% viability were obtained after isolation. At day 3, free hepatocytes displayed 99% viability, whereas microencapsulation in alginate and PEG-grafted alginate decreased viability to 62% and 48%, respectively. Albumin secretion and diazepam catabolism occurred in free and microencapsulated hepatocytes. Coencapsulation of hepatocytes with MSC significantly improved viability and albumin secretion at days 4 and 8 ( p < 0.05). Coculture with MSC significantly increased and prolonged albumin secretion. In conclusion, we established a protocol for isolation and microencapsulation of high numbers of viable pig hepatocytes and demonstrated that the presence of MSC is beneficial for the viability and function of porcine hepatocytes.- Published
- 2018
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17. Xenotransplantation: Where do we stand in 2016?
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Puga Yung GL, Rieben R, Bühler L, Schuurman HJ, and Seebach J
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- Animals, Humans, Primates, Swine, Transplantation Tolerance, Transplantation, Heterologous mortality, Zoonoses etiology, Zoonoses prevention & control, Disease Models, Animal, Genetic Engineering methods, Tissue and Organ Procurement, Transplantation, Heterologous methods
- Abstract
Worldwide, there is a constant rise in the number of patients with end-stage organ failure in critical need for transplants, but the number of organs/cells available from deceased or living human donors is limited. Xenotransplantation using pig organs/tissues repre-sents a potential solution for this shortage; however, it has been hampered by a number of mainly immuno-logical hurdles. Remarkable progress was presented at the latest biennial (13th) international congress of the International Xenotransplantation Association, November 2015 in Melbourne, Australia, and the American Transplant Congress, May 2016 in Boston, USA. Most importantly, the survival records of pig organ xenografts in nonhuman primate models have strikingly improved with the use of multitransgenic pigs. Moreover, no safety issues were encountered in clinical trials with porcine islets, and the removal of porcine endogenous retroviruses from the genome of a pig cell line by the CRISPR/Cas9 technology offers the perspective to overcome the perceived potential risk of xenozoonosis in the near future. For all these reasons, interest in xenotransplantation has been boosted. This review summarises the current status of xenotransplantation research, including Swiss contri-butions as well as regulatory and safety aspects in the light of upcoming clinical trials.
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- 2017
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18. Current status of hepatocyte xenotransplantation.
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Meier RPH, Navarro-Alvarez N, Morel P, Schuurman HJ, Strom S, and Bühler LH
- Subjects
- Animals, Humans, Immunity, Innate, Immunosuppression Therapy methods, Liver Failure, Acute therapy, Swine, Transplantation, Heterologous methods, Hepatocytes transplantation, Liver Transplantation methods
- Abstract
The treatment of acute liver failure, a condition with high mortality, comprises optimal clinical care, and in severe cases liver transplantation. However, there are limitations in availability of organ donors. Hepatocyte transplantation is a promising alternative that could fill the medical need, in particular as the bridge to liver transplantation. Encapsulated porcine hepatocytes represent an unlimited source that could function as a bioreactor requiring minimal immunosuppression. Besides patients with acute liver failure, patients with alcoholic hepatitis who are unresponsive to a short course of corticosteroids are a target for hepatocyte transplantation. In this review we present an overview of the innate immune barriers in hepatocyte xenotransplantation, including the role of complement and natural antibodies; the role of phagocytic cells and ligands like CD47 in the regulation of phagocytic cells; and the role of Natural Killer cells. We present also some illustrations of physiological species incompatibilities in hepatocyte xenotransplantation, such as incompatibilities in the coagulation system. An overview of the methodology for cell microencapsulation is presented, followed by proof-of-concept studies in rodent and nonhuman primate models of fulminant liver failure: these studies document the efficacy of microencapsulated porcine hepatocytes which warrants progress towards clinical application. Lastly, we present an outline of a provisional clinical trial, that upon completion of preclinical work could start within the upcoming 2-3 years., (Copyright © 2015 IJS Publishing Group Limited. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2015
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19. Regulatory aspects of clinical xenotransplantation.
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Schuurman HJ
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- Animals, Cell Transplantation legislation & jurisprudence, Cell Transplantation standards, Humans, Practice Guidelines as Topic, Risk Management methods, Swine, Transplantation, Heterologous standards, Consumer Product Safety legislation & jurisprudence, Transplantation, Heterologous legislation & jurisprudence
- Abstract
Xenotransplantation attracted interest from regulatory authorities, particularly after the demonstration of pig-to-human transmission of porcine endogenous retrovirus (1996). This added to the risk of a product, resulting in a Guidance of the US Food and Drug Administration (2003). This addresses the full flow chart in product manufacturing, starting with the designated pathogen-free status of the source animal; and special aspects regarding the recipient like informed consent and monitoring for infectious pathogens. Also archiving of records from the donor and recipient, as well as storage of samples is described. The European Medicines Agency issued a Guideline on xenogeneic cell therapy products (2009). Cell-based medicinal products are subject to specific regulations and directives, which apply also to xenogeneic products: the xenotransplant guidances/guidelines are an addition to these regulations. Noteworthy, acellular products like heart valves and decellularized cornea are not considered a cell therapy product, but rather a medical device with its own regulation. WHO issued relevant documents, especially about safety, and the International Xenotransplantation Association published consensus documents, a.o., addressing preclinical efficacy requirements before entering clinical trials. This manuscript presents an overview of the regulatory framework, with special focus on cell therapy products necause these are expected to reach the market first (i.e., pancreatic islets, hepatocytes and cellularized cornea); major illustrations are from the European situation. Albeit being complex, the regulation of xenotransplant products does not form a block in product development, but rather supports the introduction of efficacious and safe products to meet the medical need., (Copyright © 2015 IJS Publishing Group Limited. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2015
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20. Management of adverse side-effects after chemotherapy in macaques as exemplified by streptozotocin: case studies and recommendations.
- Author
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Graham ML, Mutch LA, Kittredge JA, Rieke EF, Robinson NA, Zolondek EK, Faig AW, Dufour TA, Munson JW, and Schuurman HJ
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- Acidosis chemically induced, Animals, Cohort Studies, Diabetes Mellitus, Experimental complications, Disease Models, Animal, Female, Insulin therapeutic use, Male, Acidosis therapy, Antibiotics, Antineoplastic adverse effects, Macaca fascicularis metabolism, Macaca mulatta metabolism, Streptozocin adverse effects
- Abstract
The chemotherapeutic streptozotocin is used for induction of diabetes in animal models including non-human primates. Being a cytotoxic nitrosourea compound, it can be associated with adverse events (AEs), mainly nausea and emesis, nephrotoxicity, elevated liver transaminase levels, pulmonary oedema and, most prominently, metabolic acidosis: these can be severe in some cases. The incidence and gravity are to some extent related to the characteristics of the individual animal, diagnostic tools, prompt recognition of symptoms and supportive measures. Careful animal selection, dose adaptation and supportive actions such as renal protective hydration are the main tools in managing AEs, but do not fully eliminate unavoidable and sometimes life-threatening conditions. In our centre we have built experience in a cohort of 78 cynomolgus and rhesus macaques in which six cases manifested severe AEs (8%). This experience has prompted implementation of strategies for early detection and management of adverse effects, together with an animal refinement programme. We present here specific pretreatment regimens, post-infusion laboratory evaluations, and flow charts to assess/treat metabolic acidosis and precipitating factors. Case reports of the six animals with severe AEs are presented to illustrate management of AEs, especially metabolic acidosis, and criteria for early euthanasia where appropriate. We conclude that improved monitoring and validated tools allow for optimal management of adverse effects in an early stage of their manifestation. Reduced morbidity and mortality not only improve individual animal wellbeing but also avoid model-induced confounding that diminishes the translational value of the experimental protocol.
- Published
- 2012
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21. The streptozotocin-induced diabetic nude mouse model: differences between animals from different sources.
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Graham ML, Janecek JL, Kittredge JA, Hering BJ, and Schuurman HJ
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- Analysis of Variance, Animals, Male, Mice, Species Specificity, Diabetes Mellitus, Experimental chemically induced, Diabetes Mellitus, Experimental pathology, Mice, Nude genetics, Mice, Nude metabolism, Streptozocin toxicity
- Abstract
Diabetes is induced in mice by using streptozotocin (STZ), a compound that has a preferential toxicity toward pancreatic β cells. We evaluated nude male mice from various sources for their sensitivity to a single high dose (160 to 240 mg/kg) of STZ. Diabetes was induced in male mice (age: median, 12 wk; interquartile range, 11 to 14 wk; body weight, about 30 g) from Taconic Farms (TAC), Jackson Laboratories (JAX), and Charles River Laboratories (CRL). Mice were monitored for 30 d for adverse side effects, blood glucose, and insulin requirements. In CRL mice given 240 mg/kg STZ, more than 95% developed diabetes within 4 to 5 d, and loss of body weight was relatively low (mean, 0.4 g). In comparison, both TAC and JAX mice were more sensitive to STZ, as evidenced by faster development of diabetes (even at a lower STZ dose), greater need for insulin after STZ, greater body weight loss (mean: TAC, 3.5 g; JAX, 3.7 g), and greater mortality. We recommend conducting exploratory safety assessments when selecting a nude mouse source, with the aim of limiting morbidity and mortality to less than 10%.
- Published
- 2011
22. Refinement of vascular access port placement in nonhuman primates: complication rates and outcomes.
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Graham ML, Mutch LA, Rieke EF, Dunning M, Zolondek EK, Faig AW, Hering BJ, and Schuurman HJ
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- Animals, Catheterization, Peripheral adverse effects, Catheters, Indwelling adverse effects, Catheters, Indwelling microbiology, Retrospective Studies, Catheterization, Peripheral methods, Catheters, Indwelling veterinary, Macaca fascicularis surgery, Macaca mulatta surgery, Papio surgery
- Abstract
Chronic vascular access is often needed in experimental animal studies, and vascular access ports (VAP) have been proposed as an alternative to conventional venipuncture. We previously reported on VAP implantation by using femoral venous cutdown (FVC) and tunneling. In an attempt to decrease the moderate complications associated with the FVC method, we developed the single-incision, peripheral-insertion (SIPI) method. In a retrospective evaluation, 92 FVC procedures were compared with 113 SIPI procedures in cynomolgus and rhesus macaques and baboons with as much as 2.5 y of follow-up. The rate of complications was significantly lower for the SIPI method than for the FVC method (19.4% versus 33.7%), particularly in regard to infectious complications (8.0% versus 27.3%, respectively). In addition, VAP patency for blood sampling and fluid infusion was significantly better for the SIPI method than for the FVC method, with 1-y patency rate of 83% and 46%, respectively, and 2-y patency rate of 74% and 36%, respectively. Additional advantages of the SIPI method include the simplified implantation of the VAP and access in the homecage without any sedation or restraint after appropriate training of animals to cooperate. We conclude that the SIPI method presents an opportunity for refinement and is superior to the FVC method for chronic vascular access.
- Published
- 2010
23. Renal and cardiac endothelial heterogeneity impact acute vascular rejection in pig-to-baboon xenotransplantation.
- Author
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Knosalla C, Yazawa K, Behdad A, Bodyak N, Shang H, Bühler L, Houser S, Gollackner B, Griesemer A, Schmitt-Knosalla I, Schuurman HJ, Awwad M, Sachs DH, Cooper DK, Yamada K, Usheva A, and Robson SC
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- ABO Blood-Group System immunology, Acute Disease, Animals, DNA, Complementary genetics, Graft Rejection mortality, Graft Rejection pathology, Graft Survival immunology, Heart Transplantation mortality, Kidney Transplantation pathology, Oligonucleotide Array Sequence Analysis, Papio, Proteins genetics, Swine genetics, Thymus Gland transplantation, Transplantation Conditioning methods, Graft Rejection immunology, Heart Transplantation immunology, Kidney Transplantation immunology, Transplantation, Heterologous immunology
- Abstract
Xenograft outcomes are dictated by xenoantigen expression, for example, Gal alpha1, 3Gal (Gal), but might also depend on differing vascular responses. We investigated whether differential vascular gene expression in kidney and cardiac xenografts correlate with development of thrombotic microangiopathy (TM) and consumptive coagulation (CC). Immunosuppressed baboons underwent miniswine or hDAF pig kidney (n = 6) or heart (n = 7), or Gal-transferase gene-knockout (GalT-KO) (thymo)kidney transplantation (n = 14). Porcine cDNA miniarrays determined donor proinflammatory, apoptosis-related and vascular coagulant/fibrinolytic gene expression at defined time points; validated by mRNA, protein levels and immunopathology. hDAF-transgenic and GalT-KO xenografts, (particularly thymokidneys) exhibited prolonged survival. CC was seen with Gal-expressing porcine kidneys (3 of 6), only 1 of 7 baboons postcardiac xenotransplantation and was infrequent following GalT-KO grafts (1 of 14). Protective-type genes (heme oxygenase-I, superoxide dismutases and CD39) together with von Willebrand factor and P-selectin were upregulated in all renal grafts. Transcriptional responses in Gal-expressing xenografts were comparable to those seen in the infrequent GalT-KO rejection. In cardiac xenografts, fibrin deposition was associated with increased plasminogen activator inhibitor-1 expression establishing that gene expression profiles in renal and cardiac xenografts differ in a quantitative manner. These findings suggest that therapeutic targets may differ for renal and cardiac xenotransplants.
- Published
- 2009
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24. Rejection of cardiac xenografts transplanted from alpha1,3-galactosyltransferase gene-knockout (GalT-KO) pigs to baboons.
- Author
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Hisashi Y, Yamada K, Kuwaki K, Tseng YL, Dor FJ, Houser SL, Robson SC, Schuurman HJ, Cooper DK, Sachs DH, Colvin RB, and Shimizu A
- Subjects
- Animals, Animals, Genetically Modified, Antibody Formation physiology, CD4-Positive T-Lymphocytes pathology, CD8-Positive T-Lymphocytes pathology, Graft Rejection pathology, Graft Rejection physiopathology, Heart Transplantation pathology, Heart Transplantation physiology, Immunity, Cellular physiology, Immunoglobulin G blood, Immunoglobulin M blood, Killer Cells, Natural pathology, Swine, Swine, Miniature genetics, Thrombosis pathology, Transplantation, Heterologous pathology, Transplantation, Heterologous physiology, Troponin T blood, Galactosyltransferases genetics, Galactosyltransferases physiology, Graft Rejection immunology, Heart Transplantation immunology, Papio hamadryas immunology, Swine, Miniature immunology, Transplantation, Heterologous immunology
- Abstract
The use of alpha1,3-galactosyltransferase gene-knockout (GalT-KO) swine donors in discordant xenotransplantation has extended the survival of cardiac xenografts in baboons following transplantation. Eight baboons received heterotopic cardiac xenografts from GalT-KO swine and were treated with a chronic immunosuppressive regimen. The pathologic features of acute humoral xenograft rejection (AHXR), acute cellular xenograft rejection (ACXR) and chronic rejection were assessed in the grafts. No hyperacute rejection developed and one graft survived up to 6 months after transplantation. However, all GalT-KO heart grafts underwent graft failure with AHXR, ACXR and/or chronic rejection. AHXR was characterized by interstitial hemorrhage and multiple thrombi in vessels of various sizes. ACXR was characterized by TUNEL(+) graft cell injury with the infiltration of T cells (including CD3 and TIA-1(+) cytotoxic T cells), CD4(+) cells, CD8(+) cells, macrophages and a small number of B and NK cells. Chronic xenograft vasculopathy, a manifestation of chronic rejection, was characterized by arterial intimal thickening with TUNEL(+) dead cells, antibody and complement deposition, and/or cytotoxic T-cell infiltration. In conclusion, despite the absence of the Gal epitope, acute and chronic antibody and cell-mediated rejection developed in grafts, maintained by chronic immunosupression, presumably due to de novo responses to non-Gal antigens.
- Published
- 2008
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25. Thrombotic microangiopathy associated with humoral rejection of cardiac xenografts from alpha1,3-galactosyltransferase gene-knockout pigs in baboons.
- Author
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Shimizu A, Hisashi Y, Kuwaki K, Tseng YL, Dor FJ, Houser SL, Robson SC, Schuurman HJ, Cooper DK, Sachs DH, Yamada K, and Colvin RB
- Subjects
- Animals, Animals, Genetically Modified, Coronary Thrombosis pathology, Endothelial Cells pathology, Graft Rejection prevention & control, Hemorrhage immunology, Hemorrhage pathology, Immunosuppressive Agents therapeutic use, Microcirculation immunology, Microcirculation pathology, Myocardial Ischemia immunology, Myocardial Ischemia pathology, Myocardium pathology, Necrosis, Papio, Swine, Swine, Miniature, Transplantation, Heterologous, Coronary Thrombosis immunology, Galactosyltransferases genetics, Graft Rejection immunology, Heart Transplantation immunology
- Abstract
Heterotopic cardiac xenotransplantation from alpha1,3-galactosyltransferase gene-knockout (GalT-KO) swine to baboons was performed to characterize immunological reaction to the xenograft in the absence of anti-Gal antibody-mediated rejection. Eight baboons received heterotopic cardiac xenografts from GalT-KO porcine donors. All baboons were treated with chronic immunosuppressive therapy. Both histological and immunohistochemical studies were performed on biopsy and graftectomy samples. No hyperacute rejection was observed. Three baboons were euthanized or died 16 to 56 days after transplantation. The other five grafts ceased beating between days 59 and 179 (median, 78 days). All failing grafts exhibited thrombotic microangiopathy (TM) with platelet-rich fibrin thrombi in the microvasculature, myocardial ischemia and necrosis, and focal interstitial hemorrhage. TM developed in parallel with increases in immunoglobulin (IgM and IgG) and complement (C3, C4d, and C5b-9) deposition, as well as with subsequent increases in both TUNEL(+) endothelial cell death and procoagulant activation (increased expression of both tissue factor and von Willebrand factor and decreased expression of CD39). CD3(+) T-cell infiltration occurred in all grafts and weakly correlated with the development of TM. In conclusion, although the use of GalT-KO swine donors prevented hyperacute rejection and prolonged graft survival, slowly progressive humoral rejection--probably associated with non-Gal antibodies to the xenograft--and disordered thromboregulation represent major immunological barriers to long-term xenograft survival.
- Published
- 2008
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26. Life-supporting function of genetically modified swine lungs in baboons.
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Nguyen BN, Azimzadeh AM, Zhang T, Wu G, Schuurman HJ, Sachs DH, Ayares D, Allan JS, and Pierson RN 3rd
- Subjects
- Animals, Blood Cell Count, Complement Activation, Complement C3a analysis, Galactosyltransferases genetics, Immunologic Factors, Lung pathology, Membrane Cofactor Protein metabolism, Organisms, Genetically Modified, Papio anubis, Peptide Fragments blood, Platelet Activation, Prothrombin, Pulmonary Circulation, Swine, Vascular Resistance, Graft Survival, Lung Transplantation, Transplantation, Heterologous, beta-Thromboglobulin analysis
- Abstract
Objective: During ex vivo perfusion with human blood, homozygous galactosyl transferase knockout swine lungs exhibit prolonged survival (approximately 2 hours) relative to wild-type (<15 minutes) and swine lungs expressing human decay accelerating factor (<1 hour). In this study, the in vivo behavior of galactosyl transferase knockout lungs was evaluated., Methods: Three galactosyl transferase knockout swine left lungs were transplanted into baboons in a life-supporting model. One baboon lung allograft and two swine lung xenografts transgenic for human membrane cofactor protein (CD46) served as controls., Results: Whereas two membrane cofactor protein lungs exhibited high pulmonary vascular resistance (>500 mm Hg x min/L) and failed to support life within 21 minutes, two of three galactosyl transferase knockout lungs supported life, for 90 and 215 minutes, and displayed low peripheral vascular resistance (48 +/- 12 mm Hg x min/L at 60 minutes), similar to the allogeneic control. Complement activation (delta C3a < 250 ng/mL through 60 minutes) and C5b-9 deposition were minimal in both galactosyl transferase knockout and membrane cofactor protein lungs. Neutrophils, monocytes, and platelets were rapidly sequestered in galactosyl transferase knockout and human membrane cofactor protein lung recipients, unlike the allogeneic control (<20%); and thrombin formation (delta plasma fraction 1+2 > 0.5 nmol/L) was seen in the galactosyl transferase knockout recipients. Platelet activation (beta-thromboglobulin rise > 200) and appearance of capillary congestion and vessel thrombosis confirmed coagulation activation associated with galactosyl transferase knockout lung failure., Conclusions: Galactosyl transferase knockout swine lungs are significantly protected in vivo from the physiologic consequences (increased pulmonary vascular resistance, capillary leak) associated with hyperacute lung rejection. As during ex vivo perfusion, dysregulated coagulation-thrombin elaboration, platelet activation, and intravascular thrombosis-mediates galactosyl transferase knockout lung xenograft injury.
- Published
- 2007
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27. Prolonged survival of porcine hepatocytes in cynomolgus monkeys.
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Nagata H, Nishitai R, Shirota C, Zhang JL, Koch CA, Cai J, Awwad M, Schuurman HJ, Christians U, Abe M, Baranowska-Kortylewicz J, Platt JL, and Fox IJ
- Subjects
- Animals, Animals, Outbred Strains, Antibodies, Heterophile blood, Asialoglycoprotein Receptor metabolism, Cell Transplantation methods, Feasibility Studies, Hepatocytes immunology, Hepatocytes physiology, Immunosuppressive Agents pharmacology, Macaca fascicularis, Male, Radionuclide Imaging, Serum Albumin, Spleen diagnostic imaging, Swine, Technetium, Graft Survival, Hepatocytes transplantation, Transplantation, Heterologous methods
- Abstract
Background & Aims: Management of patients with liver failure can be a significant medical challenge, and transplantation of the liver is the only definitive therapy. Whole liver allotransplantation is limited by a shortage of human donors and the risks of the surgery in those most ill. Transplants consisting of xenogeneic hepatocytes might overcome these problems, and work in rodents indicates that such transplants can correct some metabolic deficiencies and can prevent the complications and mortality associated with hepatic failure. As a prelude to clinical application, we tested the feasibility of hepatocyte xenotransplantation in nonhuman primates., Methods: One to 2 billion hepatocytes from outbred swine were transplanted into the spleens of cynomolgus monkeys using conventional immunosuppression to control rejection. Duration of graft function was determined based on assay for porcine albumin., Results: Following a single infusion, xenogeneic hepatocytes functioned for more than 80 days and, following re-transplantation, for more than 253 days. Engraftment in the spleen was confirmed 40 days after transplantation by asialoglycoprotein receptor-directed nuclear scanning. The humoral immune response to the transplanted porcine cells had no discernible impact on the survival of the grafts., Conclusions: Xenotransplantation of hepatocytes should be explored as a readily available, minimally invasive form of therapy for hepatic failure.
- Published
- 2007
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28. Acute and chronic vascular rejection in nonhuman primate kidney transplantation.
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Wieczorek G, Bigaud M, Menninger K, Riesen S, Quesniaux V, Schuurman HJ, Audet M, Blancher A, Mihatsch MJ, and Nickeleit V
- Subjects
- Acute Disease, Animals, Biomarkers blood, Chronic Disease, Disease Models, Animal, Endarteritis immunology, Endarteritis pathology, Female, Kidney Transplantation mortality, Kidney Transplantation pathology, Macaca fascicularis, Male, Postoperative Period, Primates, Survival Analysis, Vascular Diseases etiology, Graft Rejection immunology, Kidney Transplantation immunology, Vascular Diseases immunology
- Abstract
A nonhuman primate (NHP) study was designed to evaluate in nonlife-supporting kidney allografts the progression from acute rejection with transplant endarteritis (TXA) to chronic rejection (CR) with sclerosing vasculopathy. Group G1 (n = 6) received high cyclosporine A (CsA) immunosuppression and showed neither TXA nor CR during 90 days post-transplantation. Group G2 (n = 6) received suboptimal CsA immunosuppression and showed severe TXA with graft loss within 46 days (median). Arterial intimal changes included infiltration of macrophages and T lymphocytes (CD3, CD4, CD8) with few myofibroblasts, abundant fibronectin/collagen IV, scant collagens I/III, high rate of cellular proliferation and no C4d accumulation along peritubular capillaries. Group G3 (n = 12) received suboptimal CsA and anti-rejection therapy (rabbit ATG + methylprednisolone + CsA) of TXA. Animals developed CR and lost grafts within 65 days (median). As compared to G2, the arterial intimal changes showed less macrophages and T lymphocytes, an increased number of myofibroblasts, abundant fibronectin/collagen IV and scar collagens I/III, C4d deposition along capillaries in 60% of animals and transplant glomerulopathy in 80% of animals. In conclusion, CR is an immune stimulated process initiated during TXA with the accumulation and proliferation of myofibroblasts, and progressive deposition of collagens in the intima. Our experimental design appears well suited to study events leading to CR.
- Published
- 2006
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29. Prolonged diabetes reversal after intraportal xenotransplantation of wild-type porcine islets in immunosuppressed nonhuman primates.
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Hering BJ, Wijkstrom M, Graham ML, Hårdstedt M, Aasheim TC, Jie T, Ansite JD, Nakano M, Cheng J, Li W, Moran K, Christians U, Finnegan C, Mills CD, Sutherland DE, Bansal-Pakala P, Murtaugh MP, Kirchhof N, and Schuurman HJ
- Subjects
- Animals, Cell- and Tissue-Based Therapy, Diabetes Mellitus, Experimental chemically induced, Diabetes Mellitus, Experimental immunology, Graft Rejection immunology, Graft Survival immunology, Islets of Langerhans cytology, Portal System immunology, Diabetes Mellitus, Experimental surgery, Immunosuppression Therapy, Islets of Langerhans Transplantation, Macaca immunology, Macaca surgery, Swine, Transplantation, Heterologous
- Abstract
Cell-based diabetes therapy requires an abundant cell source. Here, we report reversal of diabetes for more than 100 d in cynomolgus macaques after intraportal transplantation of cultured islets from genetically unmodified pigs without Gal-specific antibody manipulation. Immunotherapy with CD25-specific and CD154-specific monoclonal antibodies, FTY720 (or tacrolimus), everolimus and leflunomide suppressed indirect activation of T cells, elicitation of non-Gal pig-specific IgG antibody, intragraft expression of proinflammatory cytokines and invasion of infiltrating mononuclear cells into islets.
- Published
- 2006
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30. Marked prolongation of porcine renal xenograft survival in baboons through the use of alpha1,3-galactosyltransferase gene-knockout donors and the cotransplantation of vascularized thymic tissue.
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Yamada K, Yazawa K, Shimizu A, Iwanaga T, Hisashi Y, Nuhn M, O'Malley P, Nobori S, Vagefi PA, Patience C, Fishman J, Cooper DK, Hawley RJ, Greenstein J, Schuurman HJ, Awwad M, Sykes M, and Sachs DH
- Subjects
- Animals, Animals, Genetically Modified, Creatinine metabolism, Disaccharides immunology, Disaccharides metabolism, Galactosyltransferases metabolism, Papio, Swine, Time Factors, Galactosyltransferases genetics, Kidney Transplantation, Thymus Gland transplantation, Transplantation, Heterologous immunology
- Abstract
The use of animal organs could potentially alleviate the critical worldwide shortage of donor organs for clinical transplantation. Because of the strong immune response to xenografts, success will probably depend upon new strategies of immune suppression and induction of tolerance. Here we report our initial results using alpha-1,3-galactosyltransferase knockout (GalT-KO) donors and a tolerance induction approach. We have achieved life-supporting pig-to-baboon renal xenograft survivals of up to 83 d with normal creatinine levels.
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- 2005
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31. Heart transplantation in baboons using alpha1,3-galactosyltransferase gene-knockout pigs as donors: initial experience.
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Kuwaki K, Tseng YL, Dor FJ, Shimizu A, Houser SL, Sanderson TM, Lancos CJ, Prabharasuth DD, Cheng J, Moran K, Hisashi Y, Mueller N, Yamada K, Greenstein JL, Hawley RJ, Patience C, Awwad M, Fishman JA, Robson SC, Schuurman HJ, Sachs DH, and Cooper DK
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- Animals, Animals, Genetically Modified, Disaccharides metabolism, Fluorescent Antibody Technique, Galactosyltransferases metabolism, Myocardium pathology, Papio, Swine, Disaccharides immunology, Galactosyltransferases genetics, Heart Transplantation, Transplantation, Heterologous immunology
- Abstract
Hearts from alpha1,3-galactosyltransferase knockout pigs (GalT-KO, n = 8) were transplanted heterotopically into baboons using an anti-CD154 monoclonal antibody-based regimen. The elimination of the galactose-alpha1,3-galactose epitope prevented hyperacute rejection and extended survival of pig hearts in baboons for 2-6 months (median, 78 d); the predominant lesion associated with graft failure was a thrombotic microangiopathy, with resulting ischemic injury. There were no infectious complications directly related to the immunosuppressive regimen. The transplantation of hearts from GalT-KO pigs increased graft survival over previous studies.
- Published
- 2005
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32. The anti-CD2 monoclonal antibody BTI-322 generates unresponsiveness by activation-associated T cell depletion.
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Xu Y, Kolber-Simonds D, Hope JA, Bazin H, Latinne D, Monroy R, White-Scharf ME, and Schuurman HJ
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- Animals, Antibodies, Monoclonal, Humanized, Antibody Specificity immunology, CD3 Complex immunology, Cell Division immunology, Cells, Cultured, Epitopes immunology, Humans, Immune Tolerance immunology, Leukocytes, Mononuclear immunology, Lymphocyte Activation immunology, Lymphocyte Culture Test, Mixed methods, Lymphocyte Depletion, Monocytes immunology, Receptors, Antigen, T-Cell, alpha-beta immunology, Swine, Antibodies, Monoclonal immunology, T-Lymphocytes immunology
- Abstract
The antihuman CD2 MoAb BTI-322 (Lo-CD2a) effectively inhibits T cell responses in vitro to allogeneic cells, which is followed by unresponsiveness to the original stimulator in secondary stimulation. We studied the xenogeneic human antiporcine mixed lymphocyte reaction (MLR), and utilized anti-T cell receptor (TCR) Vbeta family antibody-induced cell proliferation to determine the specificity and mechanism. BTI-322 and its humanized version, MEDI-507, effectively inhibited the primary xenogeneic MLR. After suboptimal primary stimulation using lower numbers of xenogeneic stimulator cells, the unresponsiveness in secondary culture was apparent only for xenogeneic stimulator cells of the original SLA haplotype, and not for third-party stimulators or allogeneic cells. The inhibition of primary MLR was not observed for nylon-wool-purified T cells, but was seen after reconstitution of purified T cells with monocytes. Similarly, anti-Vbeta family-specific stimulation showed family-specific unresponsiveness in secondary culture. This required the presence of the whole BTI-322 molecule: a F(ab')2 fragment was not effective. T cells of a distinct Vbeta family were depleted after stimulation with an anti-Vbeta family-specific antibody and BTI-322. We conclude that the inhibition by BTI-322 of a primary xenogeneic MLR or the response to an anti-TCR Vbeta antibody is associated with unresponsiveness upon restimulation, due to activation-associated cell depletion. In this process, the interaction between monocytes and the Fc part of the antibody is involved. This unique characteristic of BTI-322 suggests the potential of the antibody for tolerance induction in vivo, besides the potential use as a T cell depleting agent.
- Published
- 2004
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33. Porcine endogenous retrovirus transmission characteristics of galactose alpha1-3 galactose-deficient pig cells.
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Quinn G, Wood JC, Ryan DJ, Suling KM, Moran KM, Kolber-Simonds DL, Greenstein JL, Schuurman HJ, Hawley RJ, and Patience C
- Subjects
- Animals, Cell Line, Disaccharides antagonists & inhibitors, Humans, Disaccharides physiology, Endogenous Retroviruses physiology, Swine virology
- Abstract
Galactose alpha1-3 galactose (Gal) trisaccharides are present on the surface of wild-type pig cells, as well as on viruses particles produced from such cells. The recognition of Gal sugars by natural anti-Gal antibodies (NAb) in human and Old World primate serum can cause the lysis of the particles via complement-dependent mechanisms and has therefore been proposed as an important antiviral mechanism. Recently, pigs have been generated that possess disrupted galactosyl-transferase (GGTA1) genes. The cells of these pigs do not express Gal sugars on their surface, i.e., are Gal null. Concerns have been raised that the risk of virus transmission from such pigs may be increased due to the absence of the Gal sugars. We investigated the sensitivity of porcine endogenous retrovirus (PERV) produced from Gal-null and Gal-positive pig cells to inactivation by purified NAb and human serum. PERV produced in Gal-null pig cells was resistant to inactivation by either NAb or human serum. In contrast, although Gal-positive PERV particles were sensitive to inactivation by NAb and human serum, they required markedly higher concentrations of NAb for inactivation compared to the Gal-positive cells from which they were produced. Complete inactivation of Gal-positive PERV particles was not achievable despite the use of high levels of NAb, indicating that NAb-mediated inactivation of cell-free PERV particles is an inefficient process.
- Published
- 2004
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34. Suppression of natural and elicited antibodies in pig-to-baboon heart transplantation using a human anti-human CD154 mAb-based regimen.
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Kuwaki K, Knosalla C, Dor FJ, Gollackner B, Tseng YL, Houser S, Mueller N, Prabharasuth D, Alt A, Moran K, Cheng J, Behdad A, Sachs DH, Fishman JA, Schuurman HJ, Awwad M, and Cooper DK
- Subjects
- Animals, Antibodies, Monoclonal immunology, Graft Rejection immunology, Graft Rejection metabolism, Graft Survival, Immunosuppressive Agents immunology, Myocardium immunology, Myocardium pathology, Papio, Swine, Time Factors, Antibodies, Monoclonal pharmacology, CD40 Ligand immunology, Heart Transplantation immunology, Immunosuppressive Agents pharmacology, Transplantation, Heterologous immunology
- Abstract
Natural and elicited antipig antibodies (Abs) lead to acute humoral xenograft rejection (AHXR). Ten baboons underwent heterotopic heart transplantation (Tx) from human decay-accelerating factor (hDAF) pigs. Depletion of anti-Galalpha1, 3Gal (Gal) Abs was achieved by the infusion of a Gal glycoconjugate from day-1. Immunosuppression included induction of antithymocyte globulin, thymic irradiation, and cobra venom factor, and maintenance with a human antihuman CD154 mAb, mycophenolate mofetil, and methylprednisolone; heparin and prophylactic ganciclovir were also administered. Pig heart survival ranged from 4 to 139 (mean 37, median 27) days, with three functioning for >50 days. Graft failure (n = 8) was from classical AHXR [4], thrombotic microangiopathy [3], or intragraft thrombosis [1], with death (n = 2) from pneumonia [1], or possible drug toxicity (with features of thrombotic microangiopathy) [1]. Anti-Gal Abs (in microg/mL) were depleted by Gal glycoconjugate before graft implantation from means of 41.3 to 6.3 (IgM) and 12.4-4.6 (IgG), respectively, and at graft excision were 6.3 and 1.7 microg/mL, respectively. No elicited Abs developed, and no cellular infiltration was seen. The treatment regimen was effective in maintaining low anti-Gal Ab levels and in delaying or preventing AHXR. The combination of costimulatory blockade and heparin with Tx of a Gal-negative pig organ may prolong graft survival further.
- Published
- 2004
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35. Absence of replication-competent human-tropic porcine endogenous retroviruses in the germ line DNA of inbred miniature Swine.
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Scobie L, Taylor S, Wood JC, Suling KM, Quinn G, Meikle S, Patience C, Schuurman HJ, and Onions DE
- Subjects
- Amino Acid Sequence, Animals, Cloning, Molecular, Endogenous Retroviruses chemistry, Endogenous Retroviruses genetics, Genomic Library, Humans, Molecular Sequence Data, Proviruses genetics, Proviruses physiology, Endogenous Retroviruses physiology, Germ Cells virology, Swine, Miniature genetics, Swine, Miniature virology, Virus Replication
- Abstract
The potential transmission of porcine endogenous retroviruses (PERVs) has raised concern in the development of porcine xenotransplantation products. Our previous studies have resulted in the identification of animals within a research herd of inbred miniature swine that lack the capacity to transmit PERV to human cells in vitro. In contrast, other animals were capable of PERV transmission. The PERVs that were transmitted to human cells are recombinants between PERV-A and PERV-C in the post-VRA region of the envelope (B. A. Oldmixon, J. C. Wood, T. A. Ericsson, C. A. Wilson, M. E. White-Scharf, G. Andersson, J. L. Greenstein, H. J. Schuurman, and C. Patience, J. Virol. 76:3045-3048, 2002); these viruses we term PERV-A/C. This observation prompted us to determine whether these human-tropic replication-competent (HTRC) PERV-A/C recombinants were present in the genomic DNA of these miniature swine. Genomic DNA libraries were generated from one miniature swine that transmitted HTRC PERV as well as from one miniature swine that did not transmit HTRC PERV. HTRC PERV-A/C proviruses were not identified in the germ line DNAs of these pigs by using genomic mapping. Similarly, although PERV-A loci were identified in both libraries that possessed long env open reading frames, the Env proteins encoded by these loci were nonfunctional according to pseudotype assays. In the absence of a germ line source for HTRC PERV, further studies are warranted to assess the mechanisms by which HTRC PERV can be generated. Once identified, it may prove possible to generate animals with further reduced potential to produce HTRC PERV.
- Published
- 2004
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36. Identification of exogenous forms of human-tropic porcine endogenous retrovirus in miniature Swine.
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Wood JC, Quinn G, Suling KM, Oldmixon BA, Van Tine BA, Cina R, Arn S, Huang CA, Scobie L, Onions DE, Sachs DH, Schuurman HJ, Fishman JA, and Patience C
- Subjects
- Animals, Cells, Cultured, Endogenous Retroviruses genetics, Endogenous Retroviruses physiology, Female, Humans, In Situ Hybridization, Fluorescence, Male, Proviruses genetics, Proviruses physiology, RNA, Viral analysis, RNA, Viral genetics, Recombination, Genetic genetics, Swine, Miniature genetics, Transplantation, Heterologous adverse effects, Virus Replication, Endogenous Retroviruses isolation & purification, Swine, Miniature virology
- Abstract
The replication of porcine endogenous retrovirus subgroup A (PERV-A) and PERV-B in certain human cell lines indicates that PERV may pose an infectious risk in clinical xenotransplantation. We have previously reported that human-tropic PERVs isolated from infected human cells following cocultivation with miniature swine peripheral blood mononuclear cells (PBMC) are recombinants of PERV-A with PERV-C. Here, we report that these recombinants are exogenous viruses in miniature swine; i.e., they are not present in the germ line DNA. These viruses were invariably present in miniature swine that transmitted PERV to human cells and were also identified in some miniature swine that lacked this ability. These data, together with the demonstration of the absence of both replication-competent PERV-A and recombinant PERV-A/C loci in the genome of miniature swine (L. Scobie, S. Taylor, J. C. Wood, K. M. Suling, G. Quinn, C. Patience, H.-J. Schuurman, and D. E. Onions, J. Virol. 78:2502-2509, 2004), indicate that exogenous PERV is the principal source of human-tropic virus in these animals. Interestingly, strong expression of PERV-C in PBMC correlated with an ability of the PBMC to transmit PERV-A/C recombinants in vitro, indicating that PERV-C may be an important factor affecting the production of human-tropic PERV. In light of these observations, the safety of clinical xenotransplantation from miniature swine will be most enhanced by the utilization of source animals that do not transmit PERV to either human or porcine cells. Such animals were identified within the miniature swine herd and may further enhance the safety of clinical xenotransplantation.
- Published
- 2004
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37. Genotyping of porcine endogenous retroviruses from a family of miniature swine.
- Author
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Quinn G, Wood J, Suling K, Arn S, Sachs DH, Schuurman HJ, and Patience C
- Subjects
- Animals, Animals, Inbred Strains, Base Sequence, Cell Line, Deoxyribonuclease HpaII metabolism, Deoxyribonucleases, Type II Site-Specific metabolism, Endogenous Retroviruses physiology, Genotype, Humans, Leukocytes, Mononuclear virology, Lymphocyte Activation, Molecular Sequence Data, Polymorphism, Restriction Fragment Length, Swine virology, Endogenous Retroviruses classification, Endogenous Retroviruses genetics, Swine, Miniature virology
- Abstract
The identification of animals in an inbred miniature swine herd that consistently fail to produce replication- competent humantropic porcine endogenous retrovirus (PERV) has prompted studies on the biology of PERV in transmitter and nontransmitter animals. We analyzed PERV RNA transcript profiles in a family of inbred miniature swine (SLA(d/d) haplotype) in which individual members differed in their capacity to generate humantropic and ecotropic (i.e., pigtropic) virus. We identified unique HaeIII and HpaII gag restriction fragment length polymorphism (RFLP) profiles resulting from single nucleotide polymorphisms in blood cells; these were found only in animals that produced humantropic PERV. These HaeIII and HpaII gag RFLP profiles proved to be components of humantropic PERV as they were transmitted to 293 human target cells in vitro. The humantropic HaeIII and HpaII gag RFLP genotypes in the family of study were not present in other miniature swine in the herd that produced humantropic PERV, indicating that these RFLP profiles relate specifically to this family's lineage.
- Published
- 2004
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38. Correlation of biochemical and hematological changes with graft failure following pig heart and kidney transplantation in baboons.
- Author
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Knosalla C, Gollackner B, Bühler L, Mueller NJ, Houser S, Mauiyyedi S, Sachs DH, Robson SC, Fishman J, Schuurman HJ, Awwad M, and Cooper DK
- Subjects
- Animals, Disseminated Intravascular Coagulation immunology, Graft Rejection blood, Hematocrit, Hemoglobins metabolism, L-Lactate Dehydrogenase metabolism, Papio immunology, Swine immunology, Time Factors, Graft Rejection immunology, Heart Transplantation immunology, Kidney Transplantation immunology, Transplantation, Heterologous immunology
- Abstract
We have explored biochemical and hematologic parameters that might indicate acute humoral xenograft rejection (AHXR) following pig organ transplantation in baboons. Baboons (n = 15) received an immunosuppressive regimen, and underwent a miniature swine or hDAF kidney (Group 1, n = 6) or heart (Group 2, n = 7) transplantation. Control baboons (Group 3, n = 2) received the immunosuppressive regimen without organ transplantation. Blood chemistry and hematologic parameters were measured daily. Baboon and porcine cytomegalovirus were monitored. In Groups 1 and 2, organ grafts survived for up to 29 days. A plasma fibrinogen of <80 mg/dL on 2 consecutive days, and a serum lactate dehydrogenase of >600 U/L and aspartate transaminase of >300 U/L, were associated with the development of AHXR in both heart and kidney grafts. In Group 1, a decrease in platelet count of >150,000/microL within 3 days, or a count of <50,000/microL, were associated with AHXR. In Group 2, a creatine phosphokinase of >500 U/L was associated with graft failure. In Group 3, no abnormalities were observed. The possibility that porcine CMV may play a role in graft injury could not be excluded. Noninvasive parameters were identified that have predictive potential for AHXR. Monitoring of these might enable therapeutic intervention to reverse rejection.
- Published
- 2003
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39. Porcine endogenous retrovirus transmission characteristics of an inbred herd of miniature swine.
- Author
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Oldmixon BA, Wood JC, Ericsson TA, Wilson CA, White-Scharf ME, Andersson G, Greenstein JL, Schuurman HJ, and Patience C
- Subjects
- Animals, Animals, Inbred Strains, Base Sequence, Cell Line, Cells, Cultured, Coculture Techniques, Genes, env, Humans, Leukocytes, Mononuclear, Molecular Sequence Data, Recombination, Genetic, Sequence Analysis, DNA, Virus Replication, Endogenous Retroviruses genetics, Endogenous Retroviruses physiology, Swine, Miniature virology
- Abstract
Here we report the identification of inbred miniature swine that failed to produce human-tropic replication-competent porcine endogenous retroviruses (HTRC PERVs), using in vitro coculture assays. When HTRC PERVs were isolated from transmitting animals, all were recombinant viruses, with the receptor-binding domain of PERV-A combining with PERV-C-related sequences.
- Published
- 2002
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40. Efficacy and toxicity of plasma-cell-reactive monoclonal antibodies B-B2 and B-B4 and their immunotoxins.
- Author
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Vooijs WC, Post J, Wijdenes J, Schuurman HJ, Bolognesi A, Polito L, Stirpe F, Bast EJ, and de Gast GC
- Subjects
- Antibodies, Monoclonal metabolism, Antibodies, Monoclonal toxicity, Bone Marrow drug effects, Bone Marrow Cells, Cell Division drug effects, Cell Line, Endothelium cytology, Endothelium drug effects, Epithelial Cells, Epithelium drug effects, Hematopoietic Stem Cells cytology, Hematopoietic Stem Cells drug effects, Humans, Immunohistochemistry, Immunotoxins metabolism, Immunotoxins toxicity, Liver cytology, Liver drug effects, Multiple Myeloma drug therapy, Plasma Cells metabolism, Sensitivity and Specificity, Antibodies, Monoclonal pharmacology, Immunotoxins pharmacology, Plasma Cells drug effects
- Abstract
Immunotherapy based on the delivery of toxic agents to the tumor site using monoclonal antibodies (mAb) may be a promising modality in the treatment of hematological malignancies. In the selection of mAb, both for ex vivo but even more for in vivo therapy, not only their reactivity to the neoplastic cells should be considered, but also reactivity to other body constituents. Here we describe the screening of two human plasma-cell-reactive mAb B-B2 and B-B4, which may be used for immunotherapy of multiple myeloma. Cross-reactivity of B-B2 and B-B4 was determined by immunohistochemistry on a series of tissues. This revealed for both B-B2 and B-B4 a strong staining of epithelial cells in various organs, e.g. lung, liver, skin, kidney and gut, while only a weak and diffuse staining was seen with endothelial cells. In bone marrow reactivity was only found with plasma cells and not with hemopoietic precursors (CD34+ cells). Immunotoxins from B-B2 and B-B4 were constructed by coupling them to the plant-derived ribosome-inactivating protein saporin. Both B-B2 and B-B4 immunotoxins appeared to be efficient in specific inhibition of protein synthesis in plasma cell lines (IC50 respectively 1 nM and 0.1 nm). The immunotoxins were also tested on epithelial cell line A431, on liver cell line HepG2 and on human umbilical vein endothelial cells. The epithelial cell line A431 was reactive with both B-B2 and B-B4, but was only inhibited by B-B4 immunotoxin. Cell line HepG2 was reactive with both mAb, but was not inhibited by either immunotoxin. The endothelial cells showed no reactivity with B-B2 and B-B4 and were not inhibited by either immunotoxin. Bone marrow treated with B-B2 and B-B4 immunotoxin did not show a decrease in colonies of hemopoietic precursor cells. Incubation of multiple-myeloma-derived bone marrow with these immunotoxin resulted in a clear decrease of the number of plasma cells. From these data we conclude that B-B2 and B-B4 immunotoxin can be used for ex vivo bone marrow purging. Discrepancies were found between immunohistochemistry, binding assays and cytotoxicity assays with the mAb and the immunotoxin, which underlines the necessity for these various assays as a preclinical screening.
- Published
- 1996
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41. Juvenile chronic arthritis: T cell reactivity to human HSP60 in patients with a favorable course of arthritis.
- Author
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de Graeff-Meeder ER, van Eden W, Rijkers GT, Prakken BJ, Kuis W, Voorhorst-Ogink MM, van der Zee R, Schuurman HJ, Helders PJ, and Zegers BJ
- Subjects
- Adolescent, Arthritis, Juvenile classification, Arthritis, Juvenile etiology, Biomarkers, Child, Child, Preschool, Chronic Disease, Female, Humans, Immunohistochemistry, Lymphocyte Activation, Male, Synovial Fluid cytology, Synovial Membrane blood supply, Synovial Membrane immunology, Time Factors, Arthritis, Juvenile immunology, Chaperonin 60 immunology, Synovial Fluid immunology, T-Lymphocytes immunology
- Abstract
Synovial fluid and peripheral blood mononuclear cell proliferative responses to the 60-kD human heat shock protein (HSP60) were studied in 23 patients with juvenile chronic arthritis (JCA) and 7 non-JCA control patients. All patients showed active arthritis at the time of study. The patients were divided into two groups according to the presence (group A) or absence (group B) of T lymphocyte reactivity to human HSP60. We show that reactivity to human HSP60 is primarily, though not exclusively, occurring in patients with a remitting course of disease, i.e., the subgroup of HLA-B27 negative JCA patients with an oligoarticular onset. Immunohistochemical analysis of HSP expression in synovial membranes showed a significantly higher intensity of staining in JCA patients than in non-JCA controls. The results suggest that, in accordance with the earlier observation made in experimental models, T lymphocyte reactivity to human HSP60 in this subgroup of JCA patients may be part of T cell regulatory mechanisms that control the development of arthritis.
- Published
- 1995
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42. Lymphodepletion of the thymus cortex in rats after single oral intubation of 2,3,7,8-tetrachlorodibenzo-p-dioxin.
- Author
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De Heer C, Schuurman HJ, Vos JG, and Van Loveren H
- Subjects
- Administration, Oral, Animals, Dose-Response Relationship, Drug, Lymphopenia chemically induced, Male, Organ Size drug effects, Polychlorinated Dibenzodioxins administration & dosage, Rats, Rats, Wistar, Thymus Gland pathology, Polychlorinated Dibenzodioxins toxicity, Thymus Gland drug effects
- Abstract
Atrophy of the thymus and thymic lymphodepletion occurs after exposure to sublethal doses of TCDD. Previous studies using high doses of TCDD (150 micrograms/kg) have indicated a preferential lymphodepletion of the thymus cortex on day 4 after a single oral intubation. Here we describe the effects of a single oral intubation of lower dose levels of TCDD (1, 5, and 25 micrograms/kg) on thymic weights and thymocyte subpopulations in Wistar rats on day 4 after exposure. A single administration of 1 micrograms/kg TCDD induced a significant reduction in the number of immature CD4+CD8+ double-positive (DP) thymocytes. At this time point the numbers of mature CD3high medullary thymocytes were not affected at any dose level tested. We conclude that also lower dose levels of TCDD induce thymic atrophy via a preferential lymphodepletion of the thymus cortex.
- Published
- 1994
- Full Text
- View/download PDF
43. Assessment of immunotoxicity of buprenorphine.
- Author
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Van Loveren H, Gianotten N, Hendriksen CF, Schuurman HJ, and Van der Laan JW
- Subjects
- Analysis of Variance, Animals, Blood Cell Count drug effects, Body Weight drug effects, Bone Marrow Cells, Immunoglobulin Isotypes blood, Killer Cells, Natural drug effects, Lymphocyte Activation drug effects, Lymphoid Tissue cytology, Male, Organ Size drug effects, Rats, Rats, Wistar, Bone Marrow drug effects, Buprenorphine toxicity, Immunoglobulin Isotypes drug effects, Lymphoid Tissue drug effects
- Abstract
In order to use buprenorphine as an analgesic in immunological experiments, we have studied the potential immunotoxicity of buprenorphine. Three-week-old male Wistar Riv:TOX rats were subcutaneously treated with buprenorphine by injection of 0.1, 0.4, or 1.6 mg/kg body weight per day over a period of 4 weeks. Concentrations used were within the range for analgesia in rats. A slight decrease of body weight gain was observed at the highest dose in one but not in a duplicate study. Decreased liver weights were observed in all dose groups. Histopathologically glycogen storage was decreased and fatty vacuolation was found to be increased starting from the lowest dose group. The relative but not absolute weight of the lungs was slightly increased at the lowest dose, this phenomenon was therefore not dose-dependent. Histopathologically, a dose-dependent increase in interstitial pneumonia in the lung was found. At the 2 higher dose levels the weight of the adrenal glands was increased. No haematological changes were found, nor were there effects on bone marrow. In one of 2 studies indications of potential immunotoxicity noted were: an increased weight of the thymus, as well as an increased weight of popliteal and mesenteric lymph nodes. No effects on the weight of the spleen were found. Histologically, there were no changes in the lymphoid organs tested. Total immunoglobulin A concentrations in serum were significantly decreased in the highest dose group, whereas IgG concentrations were increased, albeit not statistically significantly. IgM and IgE concentrations showed no alterations. Two types of immune function assays were carried out: determination of natural killer cell activity and of mitogen responsiveness of spleen cells.(ABSTRACT TRUNCATED AT 250 WORDS)
- Published
- 1994
- Full Text
- View/download PDF
44. Soluble CD8 and CD25 in serum of patients after heart transplantation.
- Author
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Wijngaard PL, Van der Meulen A, Gmelig Meyling FH, De Jonge N, and Schuurman HJ
- Subjects
- Graft Rejection prevention & control, Humans, Immunosuppressive Agents therapeutic use, Lymphocyte Activation, Myocardium immunology, Solubility, CD8 Antigens blood, Heart Transplantation immunology, Receptors, Interleukin-2 immunology
- Abstract
To evaluate the diagnostic value of serum cytokine levels and cytokine receptor levels in the diagnosis of acute rejection after heart transplantation, we measured soluble CD8 and soluble CD25 in the serum of heart transplant recipients. The results were compared with endomyocardial biopsy (EMB) histopathology, lymphocyte activation by morphologic inspection of peripheral blood cells (cytoimmunologic monitoring), clinically manifested infections, and the maintenance immunosuppressive therapy. Significantly increased levels were observed in cases of lymphocyte activation in cytoimmunologic monitoring indicative of either rejection or infection. In clinically documented cytomegalovirus (CMV), bacterial, and Pneumocystis carinii infections, increased levels of soluble CD25 were observed. Soluble CD8 was only increased in a single case of P. carinii infection. A statistically significant correlation was calculated between the levels of soluble CD8 and whole blood cyclosporin A level. Considering chemotherapy, the levels of soluble CD8 showed an inverse correlation with the daily dosage of azathioprine. In conclusion, the levels of soluble CD8 and CD25 are associated with lymphocyte activation in peripheral blood, but do not differentiate between lymphocyte activation indicative of rejection or infection. No relationship was observed between levels of soluble CD8 and CD25, and EMB histopathology. Therefore, the assessment of these two cell products has no diagnostic potential for monitoring acute rejection after heart transplantation.
- Published
- 1994
- Full Text
- View/download PDF
45. Breaking of transplantation tolerance after reduction of immunosuppression.
- Author
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Wijngaard PL, Schuurman HJ, Meyling FH, Jambroes G, and Borleffs JC
- Subjects
- Adult, Female, Humans, Heart Transplantation immunology, Immune Tolerance immunology, Immunity, Cellular immunology, Immunosuppression Therapy methods
- Published
- 1993
46. Host cell membrane proteins on human immunodeficiency virus type 1 after in vitro infection of H9 cells and blood mononuclear cells. An immuno-electron microscopic study.
- Author
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Meerloo T, Sheikh MA, Bloem AC, de Ronde A, Schutten M, van Els CA, Roholl PJ, Joling P, Goudsmit J, and Schuurman HJ
- Subjects
- Antigens, CD analysis, CD11 Antigens, Cell Adhesion Molecules analysis, Cell Line, HIV Core Protein p24 analysis, HIV Envelope Protein gp120 analysis, HIV Envelope Protein gp41 analysis, HLA Antigens analysis, Humans, Intercellular Adhesion Molecule-1, Microscopy, Immunoelectron, Antigens, Surface analysis, HIV-1 chemistry, Leukocytes, Mononuclear microbiology, Membrane Proteins analysis
- Abstract
Human immunodeficiency virus type 1 (HIV-1)-infected H9 and blood mononuclear cells (MNCs) were studied by immunogold electron microscopy for the presence of HIV-1 gag p24 protein, env gp41 and gp120 proteins, and host cell molecules CD4, CD11a, CD25, CD54, CD63, HLA class I and HLA-DR. Uninfected H9 cells and MNC membranes labelled for CD4, HLA class I and class II, and, at low density, CD11a and CD54; lysosomal structures in the cytoplasm labelled for CD63. The infected cell surface showed immunolabelling for HIV-1 proteins, as did budding particle-like structures. Immunogold labelling of the cell membrane for CD4 was almost non-existent. The level of immunolabelling for CD11a and CD54 on infected cells was greater than that on uninfected cells; this is presumably related to a state of activation during virus synthesis. Budding particle-like structures and free virions in the intercellular space were immunogold-labelled for all host cell markers investigated. This was confirmed by double immunogold labelling using combinations of HIV-1 gag p24 labelling and labelling for the respective host cell molecule. We conclude that virions generated in HIV-1-infected cells concentrate host-derived molecules on their envelope. Also molecules with a prime function in cellular adhesion concentrate on the virion.
- Published
- 1993
- Full Text
- View/download PDF
47. Cytoimmunologic monitoring for rejection and infection after lung transplantation.
- Author
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Schuurman HJ, Wijngaard PL, Jansen EW, van der Meulen A, Slootweg PJ, and Meyling FH
- Subjects
- Adult, Humans, Male, Middle Aged, Graft Rejection, Immunophenotyping, Infections blood, Leukocyte Count, Lung Transplantation immunology, Postoperative Complications blood
- Published
- 1993
48. Epithelium-free area in the thymic cortex of rats.
- Author
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Bruijntjes JP, Kuper CF, Robinson JE, and Schuurman HJ
- Subjects
- Animals, CD4 Antigens analysis, CD8 Antigens analysis, Cell Division, Epithelium anatomy & histology, Histocompatibility Antigens Class II analysis, Histocytochemistry, Histocytological Preparation Techniques, Macrophages cytology, Rats, Rats, Wistar, T-Lymphocytes cytology, Thymus Gland growth & development, T-Lymphocytes physiology, Thymus Gland anatomy & histology
- Abstract
The histology of epithelium-free areas in the subcapsular region of the thymus was studied in Wistar rats. Lymphocytes in these areas were CD4/CD8 double-positive, TCR alpha/beta positive in low intensity, and in CD5 labeling either negative or positive in low intensity. There was a high proliferative activity as assessed by bromodeoxyuridine incorporation in vivo and detected by immunohistochemistry. Various macrophage types were observed. They were either large and round to slightly dendritic, or small and dendritic. Most large cells were positive for MHC Class II, and labeled by the antimacrophage antibodies ED1 and ED2. A few cells were strongly positive for Sudan black, Oil red O, nonspecific esterase, and acid phosphatase; they resembled the large rounded macrophages in the corticomedullary zone, although their MHC Class II and ED2 staining was more intense. A few cells showed features of tangible body macrophages, as they contained cellular debris. Serial sections showed that epithelium-free areas run from the subcapsular area to deep in the cortex, and often border the medulla. This opens the opportunity for immature lymphocytes to move into the medulla and corticomedullary zone without contacting and potential selection with cortical stromal elements other than macrophages in the epithelium-free areas. In this case, the epithelium-free areas may offer a separate intrathymic pathway for T lymphocytes.
- Published
- 1993
- Full Text
- View/download PDF
49. Epitopes of human immunodeficiency virus regulatory proteins tat, nef, and rev are expressed in normal human tissue.
- Author
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Parmentier HK, van Wichen DF, Meyling FH, Goudsmit J, and Schuurman HJ
- Subjects
- Antibodies, Monoclonal, Child, Dendritic Cells chemistry, Gene Products, nef genetics, Gene Products, rev genetics, Gene Products, tat genetics, Humans, Immunohistochemistry, Intestines chemistry, Pancreas chemistry, RNA, Messenger analysis, RNA, Messenger genetics, Skin chemistry, Spleen chemistry, Thymus Gland chemistry, Trachea chemistry, Epitopes analysis, Gene Products, nef immunology, Gene Products, rev immunology, Gene Products, tat immunology, Lymph Nodes chemistry
- Abstract
The expression of regulatory proteins tat, rev, and nef of human immunodeficiency virus type-1 (HIV-1) and tat of HIV-2 was studied in frozen sections of lymph nodes from HIV-1-infected individuals, and various tissues from uninfected persons. In HIV-1-positive lymph nodes, monoclonal antibodies to HIV-1-tat stained solitary cells in the germinal centers and interfollicular zones, and vascular endothelium. Staining by an anti-nef monoclonal antibody was restricted to follicular dendritic cells, whereas anti-rev antibody bound to fibriohistiocytes and high endothelial venules. The antibodies used labeled several cell types in tissues from uninfected individuals. Anti-HIV-1-tat antibodies labeled blood vessels and Hassall's corpuscles in skin and thymus; goblet cells in intestinal tissue and trachea; neural cells in brain and spinal cord; and zymogen-producing cells in pancreas. Anti-rev antibody stained high endothelial venules, Hassall's corpuscles and histiocytes. One anti-nef antibody solely stained follicular dendritic cells in spleen, tonsil, lymph node and Peyer's patches, whereas two other anti-nef antibodies bound to astrocytes, solitary cells in the interfollicular zones of lymph nodes, and skin cells. The current results hamper the immunohistochemical study for pathogenetic and diagnostic use of HIV regulatory protein expression in infected tissue specimens or cells.
- Published
- 1992
50. T-cell receptor V beta-family usage in primary cutaneous and primary nodal T-cell non-Hodgkin's lymphomas.
- Author
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Preesman AH, Hu HZ, Tilanus MG, de Geus B, Schuurman HJ, Reitsma R, van Wichen DF, van Vloten WA, and de Weger RA
- Subjects
- Base Sequence, Gene Expression, Gene Rearrangement, beta-Chain T-Cell Antigen Receptor, Humans, Immunohistochemistry, Immunophenotyping, Lymphoma, T-Cell ultrastructure, Lymphoma, T-Cell, Cutaneous ultrastructure, Molecular Sequence Data, Polymerase Chain Reaction, RNA, Messenger analysis, Tumor Cells, Cultured, Lymphoma, T-Cell genetics, Lymphoma, T-Cell, Cutaneous genetics, Receptors, Antigen, T-Cell, alpha-beta genetics
- Abstract
To evaluate whether the expression of T-cell receptor (TCR) V beta families in eight cases of malignant T-cell lymphomas took place in a preferential manner, we analyzed four cases of mycosis fungoides (MF), the most common form of primary cutaneous T-cell non-Hodgkin's lymphomas (NHL), and four cases of primary nodal T-cell NHL. The usage of V beta families in T-cell populations was investigated on mRNA that was transcribed to cDNA using a C beta primer and reverse transcriptase. Subsequently, the specific usage of the families was analyzed by polymerase chain reaction (PCR) using combinations of the selected C beta-oligonucleotide primer and one of the family-specific V beta primers. Peripheral blood lymphocytes from four healthy volunteers and 1 "reactive" lymph node served as a control and expressed all 20 V beta families tested for. In T-cell lines, with restricted V beta expression, and in three patients with advanced MF, only one or two V beta families were expressed at the mRNA level. In an early MF lesion this monoclonal expression was absent: several V beta families were expressed with a weak intensity. This may indicate either a polyclonal origin of MF, or that too few monoclonal neoplastic cells were present in the tissue specimen. In the four nodal T-cell NHL, only one family could be clearly distinguished, whereas some of the other V beta families showed only a weak expression. These latter families represent the reactive T-cell component in the nodal T-cell NHL. Both in nodal T-cell NHL and in MF there was no preferential expression of a particular V beta family. There was a good correlation between PCR data and the expression of V beta-family protein products observed by immunohistochemistry on tissue sections of the T-cell lymphomas. All T-cell lines, three cases of MF, and three cases of nodal T-cell NHL showed a rearrangement of the TCR beta chain on DNA level.
- Published
- 1992
- Full Text
- View/download PDF
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