ASP0028 in combination with suboptimal-dose of tacrolimus in Cynomolgus monkey renal transplantation model.

FTY720, a S1P-receptor modulator, has shown to be effective in several transplant and autoimmune disease models, via modulating lymphocyte homing into secondary lymphoid organs (SLOs), and thereby reducing these cells in peripheral blood. ASP0028, a newly developed S1P/S1P-selective agonist, presented comparable efficacy to FTY720 and wider safety margins than FTY720. In this study, we assessed the efficacy and safety of ASP0028 co-administered with suboptimal-dose of tacrolimus in the Cynomolgus monkey renal transplantation model.

Effects of ASKP1240 combined with tacrolimus or mycophenolate mofetil on renal allograft survival in Cynomolgus monkeys.

Background: Blocking the CD40-CD154 signal pathway has previously shown promise as a strategy to prevent allograft rejection. In this study, the efficacy of a novel fully human anti-CD40 monoclonal antibody-ASKP1240, administered as a monotherapy or combination therapy (subtherapeutic dose of tacrolimus or mycophenolate mofetil), on the prevention of renal allograft rejection was evaluated in Cynomolgus monkeys.

Methods: Heterotopic kidney transplants were performed in ABO-compatible, stimulation index 2.

Pharmacokinetics and pharmacodynamics of ASKP1240, a fully human anti-CD40 antibody, in normal and renal transplanted Cynomolgus monkeys.

Background: The purpose of this study was to evaluate the serum concentration of ASKP1240 (pharmacokinetics [PK]) and the CD40 occupancy of ASKP1240 (pharmacodynamics [PD]) in normal and renal transplanted Cynomolgus monkeys to clarify the PK/PD relationship.

Methods: In a 70-day study, two ASKP1240 doses (2 and 5 mg/kg) were evaluated in normal and transplanted monkeys. Full doses were administered during the induction phase, and half doses were administered during the maintenance phase.

Reconstruction of cartilage with clonal mesenchymal stem cell-acellular dermal matrix in cartilage defect model in nonhuman primates.

Objective: Articular cartilage defects are commonly associated with trauma, inflammation and osteoarthritis. Mesenchymal stem cell (MSC)-based therapy is a promising novel approach for repairing articular cartilage. Direct intra-articular injection of uncommitted MSCs does not regenerate high-quality cartilage.

Cartilage regeneration by selected chondrogenic clonal mesenchymal stem cells in the collagenase-induced monkey osteoarthritis model.

Osteoarthritis (OA) is the most common form of arthritis, in which cartilage is irreversibly degraded, causing severe pain and disability. Current therapeutic strategies cannot repair damaged cartilage. We evaluated the repair potential of selected chondrogenic clonal MSCs (sC-MSCs) by delivering them into the injured cartilage site in a collagenase-induced OA model in Cynomolgus monkeys.

Adoptive transfer of CD4+CD25+ regulatory cells combined with low-dose sirolimus and anti-thymocyte globulin delays acute rejection of renal allografts in Cynomolgus monkeys.

Although donor alloantigen specific Treg cells play an important role in transplant tolerance, therapeutic applications are limited by their low frequency. In this study, isolated Tregs from Cynomolgus monkeys were efficiently expanded by a co-culture system, and maintained suppressive function on the proliferation of CD4(+) effector cells in vitro. Adoptive transfer of expanded donor alloantigen specific Tregs without any immunosuppressants could prolong survival of MHC-mismatched allografts in Cynomolgus monkeys.

Surgical complications in kidney transplantation in nonhuman primates.

Surgical complications are important causes of graft loss in the nonhuman primate kidney transplantation model. We reviewed the incidence and intervention methods in 182 kidney transplantations performed in our lab recently 2 years in Cynomolgus monkeys. There were six renal artery thromboses (3.

JAK3 inhibitors in organ transplantation and autoimmune disease.

Janus kinase 3 (JAK3) is a cytoplasmic tyrosine kinase associated with the common gamma chain that is activated by multiple T-cell growth factors including IL-2, -4, -9, -15, and -21. From the recent reports, genetic absence or ablation of JAK3 is associated with defective T-cell immunity that results in severe combined immunodeficiency (SCID) and pharmacological inhibition has prolonged allograft survival in some models of organ transplantation. This review would provide an overview of some patents along with the role of JAK3 in the immune system and efficacy of JAK3 inhibitors in experimental allograft rejection and autoimmune disease.

Combined therapy of CD4(+)CD25(+) regulatory T cells with low-dose sirolimus, but not calcineurin inhibitors, preserves suppressive function of regulatory T cells and prolongs allograft survival in mice.

Our previous study proved that sirolimus is a potent immunosuppressant which induces long-term allograft survival depends on persistence of alloantigens. CD4(+)CD25(+) regulatory T (Treg) cells are potent suppressors in transplantation. Our objectives focus on whether combined-therapy of Tregs with immunosuppressants could prolong allograft survival in mice.

Dysfunction of IL-10-producing type 1 regulatory T cells and CD4(+)CD25(+) regulatory T cells in a mimic model of human multiple sclerosis in Cynomolgus monkeys.

CD4(+)CD25(+) Treg and IL-10(+) Tr1 cells play a major role in controlling autoimmunity by suppressing self-reactive T cells. Dysfunction of Tregs appears to be a critical factor in the pathogenesis of autoimmune diseases. Multiple sclerosis (MS) is an inflammatory demyelinating disorder of CNS, where CD4(+) T cells result in nervous tissue damage.

New look at therapeutic strategies for blocking costimulatory signal in experimental and pre-clinical transplantation.

The activation of T cells depends upon two signals, antigen-specific signal through the T cell receptor and non-antigen-specific costimulatory signal through antigen present cell surface molecules. In clinical transplantation, activated T cells orchestrate the immune response and result in allograft acute rejection. Allograft chronic rejection is also a serious complication and a major cause of late allograft loss after the transplantation.

Active Chinese mistletoe lectin-55 enhances colon cancer surveillance through regulating innate and adaptive immune responses.

Aim: To investigate the potential role of active Chinese mistletoe lectin-55 (ACML-55) in tumor immune surveillance.

Methods: In this study, an experimental model was established by hypodermic inoculating the colon cancer cell line CT26 (5 x 10(5) cells) into BALB/c mice. The experimental treatment was orally administered with ACML-55 or PBS, followed by the inoculation of colon cancer cell line CT26.

Cryopreservation of vascularized ovary: an evaluation of histology and function in rats.

Cryopreservation of organs has been investigated to sustain the reproductive function of patients undergoing sterilizing chemotherapy and radiotherapy or reproductive surgery. A modified protocol for whole organ cryopreservation was described and the outcome of cryopreservative ovaries was evaluated, and apoptosis of cryopreservative cells stored for different time period and the viability of cryopreserved cells stored at different temperature was examined in rats. Lewis rat ovarian grafts were perfused for 30 min at 0.

Antioxidant therapy for prevention of inflammation, ischemic reperfusion injuries and allograft rejection.

Oxidative stress results from an imbalance, an excess of oxidants, depletion of antioxidants or failure to repair oxidative damage induced by reactive oxygen species. A vast amount of evidence implicates oxygen-derived free radicals and high-energy oxidants as mediators in many pathological conditions of inflammation, shock, and organ responses to ischemia/reperfusion, which arise during a number of clinical surgical interventions, including transplant graft rejection and coronary bypass surgery, and in such diseases as, diabetes, atherosclerosis, hypertension, organ ischemia/reperfusion, cardiovascular inflammation, cardiac/brain infarction, cancer, pulmonary emphysema and autoimmune diseases. To eliminate or attenuate oxidative stress, antioxidant therapies have been developed and may be of great help to these patients.

A lectin from Chinese mistletoe increases gammadelta T cell-mediated cytotoxicity through induction of caspase-dependent apoptosis.

In this study, a mistletoe lectin (ML) was purified from Chinese mistletoe and the effect of this 60 kDa Chinese ML on human gammadelta T cell cytotoxicity, apoptosis and modulation of the cytokine network was studied. The cytotoxic properties of delta T cells was evaluated by using a (51)Cr release test and employed fluorescence-activated cell sorting and enzyme-linked immunosorbent assay analysis to quantify translocation of the cell membrane phospholipid, phosphatidylserine and nuclear DNA fragmentation during apoptosis. It was found that: (i) ML effectively stimulated gammadelta T cell proliferation in a dose- and time-dependent manner; (ii) ML increased gammadelta T cell cytotoxicity; (iii) ML could modulate lipopolysaccharide-induced cytokine release in a pro-inflammatory manner by increasing tumor necrosis factor (TNF)-alpha release and inhibiting the release of anti-inflammatory interleukin (IL)-10; (iv) ML induced apoptosis in caspase-dependent and CD95-independent manner.

Effect of Baohuoside-1 aglycone and tacrolimus monotherapy and combination therapy on prevention of acute heart allograft rejection in the rat.

Baohuoside-1 (B-1), a recently introduced novel immunosuppressant that was proved to be potent in inhibition of T and B cell proliferation and B-1, also prevents cardiac allograft rejection in rodents. The present study further proved that monotherapy of B-1's analogue B-1 aglycone effectively prolongs cardiac allograft survival and combination therapy of B-1 aglycone with tacrolimus (FK506) produces synergistic effect in prevention acute cardiac allograft rejection in the rat. .

Effect of a novel inducible nitric oxide synthase inhibitor, FR260330, in prevention of renal ischemia/reperfusion injury in vervet monkeys.

Cytotoxic nitric oxide (NO) is produced during ischemia/reperfusion (I/R) injury by the expression of inducible NO synthase (iNOS). Therefore, continuous iNOS inhibition might prevent early graft dysfunction. FR260330, a potent and selective inhibitor of iNOS activity, impedes the dimmerization of iNOS monomer.

Baohuoside-1 inhibits activated T cell proliferation at G(1)-S phase transition.

Background: The effect of baohuoside-1 (B1), a novel flavonoid, on cell proliferation and the cell cycle was evaluated in this study.

Methods: The antiproliferative properties of B1 were evaluated by proliferation assay. Western blotting and flow cytometric analysis were employed to investigate the expression of cyclins and cyclin-dependent kinase proteins.

Effect of a novel inducible nitric oxide synthase inhibitor in prevention of rat chronic aortic rejections.

Background: Cytotoxic nitric oxide (NO) is produced during ischemia-reperfusion injury and acute and chronic rejection in allografts by expression of inducible (i) NO synthase (NOS). Therefore, continuous inhibition of iNOS may prevent early graft dysfunction and immune injury (rejection) and consequently improve graft survival. FR260330 is a potent and selective inhibitor of iNOS activity that works by preventing iNOS monomers from dimerization.

Role of soluble Fas ligand in autoimmune diseases.

Aim: To investigate the role of soluble Fas ligand in autoimmune diseases.

Methods: RT-PCR was performed to amplify sFasL cDNA from the total RNA extracted from activated human peripheral blood lymphocytes. DNA fragments were cloned into PCR vector.

Baohuoside-1, a novel immunosuppressive molecule, inhibits lymphocyte activation in vitro and in vivo.

Background: We evaluated the in vitro and in vivo immunosuppressive effects of baohuoside-1 (B1), a novel flavonoid isolated from Epimedium davidii.

Methods: Proliferation assay was used to determine the antiproliferative properties on T-cell and B-cell proliferation. Flow cytometry analysis was applied to detect changes of phenotypes on activated cells.

[Study of HLA molecules and its associated genes expression in human ovarian cancer cells].

Aim: To explore the correlation between the expressions of HLA gene and its related gene, and expression of class II transactivator (CIITA) gene induced by IFN-gamma in human ovarian cells.

Methods: The expression of HLA molecule in the tumor cells was detected by Western blot, immunohistochemical staining and flow cytometry. The expressions of transporters associated with antigen processing (TAP), low molecular weight peptides (LMP), and CIITA genes were analyzed by RT-PCR.

[The expression and application of human Fas ligand in E.coli].

Aim: To express recombinant human FasL molecule in E.coli.

Methods: RT-PCR was applied to amplify FasL cDNA from the total RNA extracted from activated human peripheral blood lymphocytes.

[Purification and identification of human recombinant IFN-beta expressed in yeast Pichia pastoris].

To find an effective and quick way of purifying and identifying recombinant human IFN-beta (rhIFN-beta) expressed in yeast Pichia pastoris, Blue Sepharose 6 fast flow (Blue S6FF) and immunological affinity chromatography (IAC) were compared in this report. rhIFN-beta was produced in 15 liter bioreactor and purified using the two methods mentioned above. The protein concentrations of rhIFN-beta and residual mouse IgG in purified rhIFN-beta were determined with ELISA.

Combination therapy of malononitrilamide FK778 with tacrolimus on cell proliferation assays and in rats receiving renal allografts.

Background: Malononitrilamide FK778, an analogue of leflunomide's active metabolite, is a promising novel small molecule with immunosuppressive and immunomodulatory properties. In this study, we evaluated the ability of combination therapy of FK778 with tacrolimus to inhibit lymphocyte proliferation and to prevent acute allograft rejection.

Methods: Proliferation assay was used to evaluate the effect of FK778 plus tacrolimus on murine splenocytes, monkey lymphocytes, and human peripheral blood mononuclear cells, after activation with T or B cell-specific mitogens.