The initial immunological research of laryngeal cancer.
Explore the source record for details and available documents.
Adult↗
Biomedical subjects
R Zhong
Publications and source records attributed to R Zhong.
Complement inhibition with an anti-C5 monoclonal antibody prevents hyperacute rejection in a xenograft heart transplantation model.
BACKGROUND: The present study was undertaken to determine whether anti-complement 5 (C5) monoclonal antibodies (mAb) prevent hyperacute rejection (HAR) in a rat-to-presensitized mouse heart transplantation model and whether these mAb, combined with cyclosporine (CsA) and cyclophosphamide (CyP), can achieve long-term graft survival. METHODS: BALB/c mice were presensitized with 2x10(7) splenocytes from Lewis rats 14 days before grafting. Heart grafts from Lewis rats were heterotopically transplanted into BALB/c mice. Presensitized mice were treated with either anti-C5 mAb or a combination of anti-C5 mAb, CsA, and CyP. Controls included: presensitized mice with no treatment, presensitized mice treated with either CsA + CyP or IgG, and nonpresensitized mice with either no treatment or with CsA + CyP treatment. RESULTS: Although typical features of HAR were evident in the presensitized grafts, the mAb completely inhibited complement activation and successfully prevented HAR. Despite complement inactivation, the graft was rejected on postoperative day 6 with acute vascular rejection (AVR) also known as delayed xenograft rejection (DXR). Notably, this type of rejection cannot be effectively overcome by CsA and CyP. CONCLUSIONS: We conclude that (1) anti-C5 mAb prevents HAR, (2) AVR/DXR still occurs when HAR is prevented by complement inactivation, and (3) AVR/DXR cannot be overcome by conventional immunosuppression. These data suggest that anti-C5 mAb may be valuable for preventing HAR in future clinical xenotransplantation and that additional interventions may be required to address AVR/DXR.
Acute Disease↗
A dominant-negative pleiotrophin mutant introduced by homologous recombination leads to germ-cell apoptosis in male mice.
Pleiotrophin (PTN) is an 18-kDa heparin-binding secretory growth/differentiation factor for different cell types. Its gene is differentially expressed in both mesenchyme and central nervous system during development and highly expressed in a number of different human tumors. Recently, a PTN mutant was found to act as a dominant-negative effector of PTN signaling. We have now used homologous recombination to introduce the dominant-negative PTN mutant into embryonic stem cells to generate chimeric mice. All highly chimeric male mice with germinal epithelium exclusively derived from embryonic stem cells with the heterologous PTN mutation were sterile. Their testes were uniformly atrophic, and the spermatocytes were strikingly apoptotic at all stages of development. The results support a central role of PTN signaling in normal spermatogenesis and suggest that interruption of PTN signaling may lead to sterility in males.
Alleles↗
Domain structure of pleiotrophin required for transformation.
The pleiotrophin (PTN) gene (Ptn) is a potent proto-oncogene that is highly expressed in many primary human tumors and constitutively expressed in cell lines derived from these tumors. The product of the Ptn gene is a secreted 136-amino acid heparin binding cytokine with distinct lysine-rich clusters within both the N- and C-terminal domains. To seek domains of PTN functionally important in neoplastic transformation, we constructed a series of mutants and tested their transforming potential by four independent criteria. Our data establish that a domain within PTN residues 41 to 64 and either but not both the N- or C-terminal domains are required for transformation; deletion of both the N and C termini abolishes the transformation potential of PTN. Furthermore, deletion of two internal 5-amino acid residue repeats enhances the transformation potency of PTN 2-fold. Our data indicate that PTN residues 41-64 contain an essential domain for transformation and suggest the hypothesis that this domain requires an additional interaction of the highly basic clusters of the N or C terminus of PTN with a negatively charged "docking" site to enable the transforming domain itself to engage and initiate PTN signaling through its cognate receptor.
3T3 Cells↗
Crystal structures of Nova-1 and Nova-2 K-homology RNA-binding domains.
BACKGROUND: Nova-1 and Nova-2 are related neuronal proteins that were initially cloned using antisera obtained from patients with the autoimmune neurological disease paraneoplastic opsoclonus-myoclonus ataxia (POMA). Both of these disease gene products contain three RNA-binding motifs known as K-homology or KH domains, and their RNA ligands have been identified via binding-site selection experiments. The KH motif structure has been determined previously using NMR spectroscopy, but not using X-ray crystallography. Many proteins contain more than one KH domain, yet there is no published structural information regarding the behavior of such multimers. RESULTS: We have obtained the first X-ray crystallographic structures of KH-domain-containing proteins. Structures of the third KH domains (KH3) of Nova-1 and Nova-2 were determined by multiple isomorphous replacement and molecular replacement at 2.6 A and 2.0 A, respectively. These highly similar RNA-binding motifs form a compact protease-resistant domain resembling an open-faced sandwich, consisting of a three-stranded antiparallel beta sheet topped by three alpha helices. In both Nova crystals, the lattice is composed of symmetric tetramers of KH3 domains that are created by two dimer interfaces. CONCLUSIONS: The crystal structures of both Nova KH3 domains are similar to the previously determined NMR structures. The most significant differences among the KH domains involve changes in the positioning of one or more of the alpha helices with respect to the betasheet, particularly in the NMR structure of the KH1 domain of the Fragile X disease protein FMR-1. Loop regions in the KH domains are clearly visible in the crystal structure, unlike the NMR structures, revealing the conformation of the invariant Gly-X-X-Gly segment that is thought to participate in RNA-binding and of the variable region. The tetrameric arrangements of the Nova KH3 domains provide insights into how KH domains may interact with each other in proteins containing multiple KH motifs.
Amino Acid Sequence↗
Technical experiences with a microsurgical model of lymphatic reconstruction after intestinal transplantation in rats.
The surgical procedure of intestinal transplantation disrupts the graft's lymphatic drainage. Despite morphological regeneration, the functional impairment of the lymphatic system may contribute to many pathophysiological changes after transplant. We describe the technical details and results of a microsurgical model of lymphatic reconstruction (LR), which immediately reconstitutes lymphatic drainage of the intestinal graft in rats. A total of 42 orthotopic intestinal transplants with LR was performed. The patency of the LR was assessed between postoperative days 2 and 100 using either methylene-blue staining of the lymphatic vessels or mesenteric lymphoradiography. A total patency rate of 88% was achieved. Genetic background, occurrence of rejection, and immunosuppressive treatment with cyclosporine did not affect the patency rate. Technical aspects and potential applications of this model are discussed.
Anastomosis, Surgical↗
Rat and mouse organ transplantation. Introduction.
Explore the source record for details and available documents.
Animals↗
Organ transplantation in mice: current status and future prospects.
With the development of microsurgery and molecular biology in the 1990s, the mouse model for organ transplants has become increasingly popular. In the past 10 years, the number of studies using the mouse model has increased three-fold. All the organ transplants, originally done in the rat model, can now be performed in mice with high success rates. This article reviews the development, advantages, limitations, and unique immunology of the mouse model as well as future prospects.
Animals↗
Surgical technique for vascularized thymus transplantation in mice.
Traditionally, mouse nonvascularized thymus implants have been used to investigate various aspects of thymus function. However, these grafts are easily damaged by ischemia and fail to reproduce the normal anatomy of the thymus. In addition, the function of these grafts has not been fully examined. We have recently developed a vascularized thymus transplant model in mice. The donor operation consists of isolating the right lobe of the thymus and creating a single vascular pathway. In the recipient surgery, end-to-side anastomoses between donor brachycephalic artery and recipient right common carotid artery, and between donor superior caval vein and recipient right external jugular vein, were performed. We performed 10 consecutive isografts in BALB/c mice with a success rate of 90%. The thymus grafts had a normal histology and function. This study illustrates that it is technically possible to transplant a mouse vascular thymus graft. This model has several advantages that make it a useful tool to study many aspects of thymus function. We plan to use this model further to study the potential for induction of tolerance by thymus grafts.
Animals↗
Cyclosporine inhibits long-term survival in cardiac allografts treated with monoclonal antibody against CD45RB.
BACKGROUND: We have previously reported that a monoclonal antibody to CD45RB is a novel immunosuppressive agent; however, the optimal regimen in cardiac allografts remains unknown. The present study was undertaken to determine the optimal protocol of this therapy and its interaction with cyclosporine. METHODS: A heterotopic heart allograft model was used in C57BL/6 to BALB/c mice. The following studies were conducted: 1) dose response study (low, intermediate, and high doses at 1, 3, and 9 mg/kg/day respectively), 2) short course (2 days) therapy vs. long course (9 days) therapy, 3) pretreatment (starting on day -1) vs no pretreatment, 4) daily therapy vs. alternative day therapy, and 5) monoclonal antibody treatment with and without cyclosporine. RESULTS: The efficacy of the CD45RB monoclonal antibody was dose and duration dependent (p<0.01). Pretreatment significantly improved the efficacy of this therapy (74.5+/-13.4 days vs. 30.6+/-1.5 days, p<0.01). Daily therapy was superior to alternate day therapy (74.5+/-13.4 days vs. 30.4+/-1.5 days, p<0.03). Interestingly, we found that administration of cyclosporine prior to, at the same time as, or after administration of the CD45RB monoclonal antibody had a detrimental effect on graft survival compared to mAb treated alone (16.6+/-0.4 days, 25+/-2.3 days, and 35.3+/-0.9 days respectively vs. 74.5 days, p<0.01). CONCLUSIONS: Immunosuppression with CD45RB monoclonal antibody is dose and duration dependent. Pretreatment and daily therapy improves results. Addition of cyclosporine inhibits long-term graft survival achieved by the monoclonal antibody alone.
Animals↗
Rat-to-mouse small bowel xenotransplantation: a novel model for studying acute vascular and hyperacute xenograft rejection and xenogenic cell migration.
The present study was undertaken to establish a rat-to-mouse vascularized small bowel xenotransplantation model to study acute vascular and hyperacute xenograft rejection, and xenogenic cell migration. Lewis rat small bowel grafts were transplanted heterotopically to group 1, Balb/c mice, and group 2, Balb/c mice pre-sensitized with a donor spleen cell injection. The grafts were examined by serial pathology and flow cytometry. In group 1, acute vascular rejection was present by the 5th post-operative day (POD). Immunohistology showed a strong endothelial deposition of IgG, IgM and C3, associated with a minimal lymphocytic infiltrate. There was a vigorous cell migration from the recipient to the graft, in which recipient origin cells comprised 80.1+/-6.9% of the graft mesenteric lymph node by POD 3. However, there was almost no cell migration from the graft to the recipient. The intestinal xenografts in the group 2 showed massive hemorrhage, fibrin deposition, vascular congestion and thrombosis 60 min after transplantation. IgG and C3 were present on the endothelium as early as 1 min after reperfusion. The vigorous humorally-mediated vascular damage and rapid elimination of donor cells seen with intestinal xenograft rejection are distinct from the usual picture of allograft rejection. Hyperacute rejection can be induced by recipient pre-sensitization with donor spleen cells. The potential advantages of studying xenotransplantation in this model include: (1) the wide range of immunologic reagents available for mice; (2) the opportunity to study the progression of vascular damage easily by performing serial biopsies in the same animal; and (3) the opportunity to study, in vivo, two-way cellular response by examining cell trafficking in the mesenteric lymph nodes.
Animals↗
CD40-deficient dendritic cells producing interleukin-10, but not interleukin-12, induce T-cell hyporesponsiveness in vitro and prevent acute allograft rejection.
The induction of an immune response or tolerance is mediated by corresponding subsets of dendritic cells (DC). However, the property of tolerogenic DC is not clear. Recently, we have characterized a population of CD11c+ splenic DC derived from long-term mixed leucocyte culture (LT-MLC), which are able to proliferate upon stimulation and have a strong primary mixed leucocyte reaction (MLR)-stimulating activity in conventional MLR. In this study, we show that, in contrast to the irradiated ones, non-irradiated LT-MLC-derived DC induce polyclonal antigen-specific T-cell hyporesponsiveness when cocultured with allogeneic splenocytes for 3-11 days. The degree of the hyporesponsiveness increased with the length of coculture. Although these DC expressed major histocompatibility complex class II and B7 costimulatory molecules, which are down-regulated during coculture, they expressed very low or undetectable CD40 before and after coculture, respectively. The CD40-deficient DC spontaneously produce interleukin-10 (IL-10), but not IL-12. The skewed balance between IL-10 and IL-12 is associated with their capability to induce T-cell hyporesponsiveness, because a neutralizing antibody to IL-10, exogenous recombinant IL-12 or lipopolysaccharide (LPS) significantly blocked the hyporesponsiveness. Accordingly, infusion of a small number of non-irradiated LT-MLC-derived DC (5x105) significantly prolonged the survival of a vascularized heterotopic murine heart transplant, whereas irradiated DC accelerated graft rejection. These data suggest that CD40-deficient DC producing IL-10, but not IL-12 can induce T-cell hyporesponsiveness in vitro and in vivo.
Adoptive Transfer↗
Activation of heat shock transcription factor 1 to a DNA binding form during the G(1)phase of the cell cycle.
The heat shock transcription factor (HSF) genes encode proteins that bind to the heat shock elements (HSE) of stress-inducible genes. We have observed the induction of HSF1, the ubiquitous member of the HSF family from a latent cytoplasmic state to a form competent to bind HSE during early G(1)in HeLa cells in the absence of stress. The induction of DNA-binding HSF1 coincided with a burst in cellular protein synthesis in early G(1)and inhibition of this translational peak prevented the formation of DNA binding-activated HSF1. A potential role for HSF1 in cell cycle regulation was suggested by the finding that cell lines stably overexpressing HSF1 showed an increased proportion of G(1)cells relative to other cell cycle phases. However, in contrast to the effects of heat shock, entry into G(1)did not lead to HSF1 hyperphosphorylation or increased activity of a heat shock promoter-reporter gene and did not cause the induction of heat shock protein 70 expression. Thus HSF1, previously implicated in the heat shock response is activated to a DNA binding from in G(1)under non-stress conditions and may play a role in G(1)regulation that does not involve the transcription of heat shock genes.
Cycloheximide↗
Transformation of the collateral vascular bundles into amphivasal vascular bundles in an Arabidopsis mutant.
Arabidopsis inflorescence stems develop a vascular pattern similar to that found in most dicots. The arrangement of vascular tissues within the bundle is collateral, and vascular bundles in the stele are arranged in a ring. Although auxin has been shown to be an inducer of vascular differentiation, little is known about the molecular mechanisms controlling vascular pattern formation. By screening ethyl methanesufonate-mutagenized populations of Arabidopsis, we have isolated an avb1 (amphivasal vascular bundle) mutant with a novel vascular pattern. Unlike the collateral vascular bundles seen in the wild-type stems, the vascular bundles in the avb1 stems were similar to amphivasal bundles, i.e. the xylem completely surrounded the phloem. Furthermore, branching vascular bundles in the avb1 stems abnormally penetrated into the pith, which resulted in a disruption in the ring-like arrangement of vascular bundles in the stele. The avb1 mutation did not affect leaf venation pattern and root vascular organization. Auxin polar transport assay indicated that the avb1 mutation did not disrupt the auxin polar transport activity in inflorescence stems. The avb1 mutation also exhibited pleiotropic phenotypes, including curled stems and extra cauline branches. Genetic analysis indicated that the avb1 mutation was monogenic and partially dominant. The avb1 locus was mapped to a region between markers mi69 and ASB2, which is covered by a yeast artificial chromosome clone, CIC9E2, on chromosome 5. Isolation of the avb1 mutant provides a novel means to study the evolutionary mechanisms controlling the arrangement of vascular tissues within the bundle, as well as the mechanisms controlling the arrangement of vascular bundles in the stele.
Arabidopsis↗
IFL1, a gene regulating interfascicular fiber differentiation in Arabidopsis, encodes a homeodomain-leucine zipper protein.
Arabidopsis inflorescence stems develop extraxylary fibers at specific sites in interfascicular regions. The spatial specification of interfascicular fiber differentiation is regulated by the INTERFASCICULAR FIBERLESS1 (IFL1) gene because mutation of that gene abolishes the formation of normal interfascicular fibers in Arabidopsis stems. To understand further the role of IFL1 in the specification of fiber differentiation, we cloned the IFL1 gene by using a positional cloning strategy. Sequence analysis showed that the IFL1 gene encodes a transcription factor that has the same features as a family of homeodomain-leucine zipper (HD-ZIP) proteins found only in plants. The predicted IFL1 protein is composed of three distinct domains, including a 60-amino acid HD at the N terminus followed by a 28-amino acid ZIP motif and a 724-amino acid C-terminal region. A nuclear targeting assay showed that IFL1 is able to direct a beta-glucuronidase fusion protein into the nucleus, which is consistent with IFL1's presumed function as a transcription factor. Gene expression analysis demonstrated that the IFL1 gene is expressed in the interfascicular regions in which fibers differentiate, which is consistent with its role in the control of interfascicular fiber differentiation. Furthermore, the IFL1 gene was shown to be expressed in the vascular regions, indicating its possible role in the regulation of vascular tissue formation. This possibility is supported by the observation that differentiation of both xylary fibers and vessel elements is altered in the vascular bundles of ifl1 mutants. Our results provide direct evidence that an HD-ZIP protein plays a role in the spatial control of fiber differentiation.
Amino Acid Sequence↗
Adoptively transferable tolerance induced by CD45RB monoclonal antibody.
The phenomenon of rejection remains the most serious problem in transplantation. The ultimate goal in transplant immunology is to develop therapeutic strategies that lead to tolerance. It has been shown that two injections of a monoclonal antibody to CD45RB leads to indefinite acceptance of renal allografts in mice. Moreover, the CD45RB monoclonal antibody reverses acute rejection and still induces tolerance. The purpose of this study was to assess mechanisms that could underlie this therapeutic benefit. It was shown that splenic lymphocytes from tolerant animals augmented proliferation in allogeneic mixed lymphocyte reactions against donor alloantigens, and the serum of tolerant mice contained donor-specific antibodies, mainly of the IgG1 isotype, suggesting the presence of TH2 cytokines. Tolerance could not be broken by interleukin-2 infusion, but tolerance could be adoptively transferred by transfusion of tolerant mouse CD4+ splenic lymphocytes into naive allografted animals. These data suggest that an active immunoregulatory mechanism is partly responsible for the therapeutic effect. CD45RB-directed therapy may find clinical application in organ transplantation in human patients.
Adoptive Transfer↗
Xenotransplantation.
As transplantation waiting lists lengthen because of the shortage of donor organs, the death rates of patients continue to rise. Xenotransplantation offers the potential to solve the problem of organ shortage br providing an unlimited supply of healthy donor organs. However, there are several barriers to xenotransplantation, including graft rejection, potential xenozoonosis, physiologic incompatibilities and ethical concerns. Experimental xenotransplantation studies continue in several areas, ranging from tissue to whole- organ grafting. Clinical studies continue in the area of tissue xenotransplantation. Trials with extracorporeal xenografts in an acute setting to support fulminant organ failure are likely to begin in the near future. The reintroduction of whole-organ xenotransplantation must be based on sound scientific analysis with broad societal input so as to offer the maximal benefit to transplant recipients and their families.
Animals↗
[Elimination of CD28+ T cell in patients with chronic active hepatitis].
OBJECTIVE: To study the role of toxicyte T cell in active hepatitis. METHODS: We examined CD28+ T cell in patients with chronic active/remission hepatitis by flow cytometry. RESULTS: It shows that CD28+, CD8+CD28+ T cell in patients with chronic active hepatitis decreased significantly, compared with normal control or patients with liver cirrhosis and heptoma. CONCLUSION: It implies that B7-CD28 costimulation pathway contribute to the tissue injury in hepatitis.
CD28 Antigens↗