Biomedical subjects

M Otsuka

Publications and source records attributed to M Otsuka.

At least 55 records · Page 3Linked to original sources

PubMed2001

Effects of aging on muscle T2 relaxation time: difference between fast- and slow-twitch muscles.

RATIONALE AND OBJECTIVES: To determine whether the T2 relaxation time of skeletal muscle is affected by aging and to compare the effects of aging between fast- and slow-twitch muscles in a human study. To investigate the mechanisms of age-related changes in T2 relaxation time in an animal (mouse) study. METHODS: T2 relaxation times of the soleus (slow-twitch, rich in type I fiber) and gastrocnemius (fast-twitch, rich in type II fiber) muscles were examined in 59 healthy human subjects, 22 to 76 years of age, by clinical magnetic resonance imaging. In mice, T2 relaxation times, fat ratios, and extracellular space ratios (extracellular space/intracellular plus extracellular space) of the spinalis (fast-twitch, rich in type II fiber) muscles were also examined (group of 7 old mice, 24-26 months; group of 7 young mice, 8-10 weeks). RESULTS: In the human study, the T2 relaxation time of the gastrocnemius muscle increased significantly with aging (r = 0.53, P < 0.01) while that of the soleus muscle did not. In the animal study, the T2 relaxation time of the spinalis muscle was significantly longer (P < 0.05) and the extracellular space ratio of the spinalis muscle significantly wider (P < 0.01) in old than in young mice. No significant difference in fat ratio was observed between old and young mice. A significant, positive correlation was seen between the extracellular space ratio and T2 relaxation time (r = 0.84, P < 0.01). CONCLUSIONS: The T2 relaxation time of fast-twitch muscle increases with aging, due mainly to increased extracellular space, reflecting age-related type II fiber atrophy.

Adult↗

PubMed2001

Long-term results of donor-specific blood transfusion with cyclosporine in living related kidney transplantation.

Donor-specific blood transfusion (DST) was introduced to achieve better graft survival. However, its benefits are controversial considering the immunosuppression of cyclosporine (CYA) or tacrolimus (Tac), and its long-term effects have not been well discussed. Of the 40 patients who received DST with CYA, 3 (7.5%) became cross-match positive. Of the 37 patients with negative cross-match, 34 patients received a one-haplotype-matched kidney and were compared to patients with one-haplotype-matched kidney transplant without preoperative DST (n = 13). Acute rejection within 3 months after transplant was 29.4% in the DST group, and 15.4% in the non-DST group. All rejection episodes were steroid resistant in the non-DST group. If the graft survival rates were calculated excluding non-immunological graft loss, graft survival rate was 91.0 and 72.8% at 5 and 10 years in the DST group, and 83.3% at 5 and 10 years in the non-DST group, respectively. The two graft survival lines converged 7 years and 7 months after transplantation. No beneficial effect of DST was statistically evident under CYA immunosuppression. In terms of the severity of acute rejection or the onset of chronic rejection, DST induced a small benefit, however, which seemed to disappear within 8 years after transplantation.

Acute Disease↗

PubMed2001

Comparative evaluation of tableting compression behaviors by methods of internal and external lubricant addition: inhibition of enzymatic activity of trypsin preparation by using external lubricant addition during the tableting compression process.

This study evaluated tableting compression by using internal and external lubricant addition. The effect of lubricant addition on the enzymatic activity of trypsin, which was used as a model drug during the tableting compression process, was also investigated. The powder mixture (2% crystalline trypsin, 58% crystalline lactose, and 40% microcrystalline cellulose) was kneaded with 5% hydroxypropyl cellulose aqueous solution and then granulated using an extruding granulator equipped with a 0.5-mm mesh screen at 20 rpm. After drying, the sample granules were passed through a 10-mesh screen (1680 microm). A 200-mg sample was compressed by using 8-mm punches and dies at 49, 98, 196, or 388 MPa (Mega Pascal) at a speed of 25 mm/min. The external lubricant compression was performed using granules without lubricant in the punches and dies. The granules were already dry coated by the lubricant. In contrast, the internal lubricant compression was performed using sample granules (without dry coating) containing 0.5% lubricant. At 98 MPa, for example, the compression level using the external lubricant addition method was about 13% higher than that for internal addition. The significantly higher compressing energy was also observed at other MPas. By comparison, the friction energy for the external addition method calculated based on upper and lower compression forces was only slightly larger. The hardness of tablets prepared using the internal addition method was 34% to 48% lower than that for the external addition method. The total pore volume of the tablet prepared using the external addition method was significantly higher. The maximum ejection pressure using the no-addition method (ie, the tablet was prepared using neither dry-coated granules nor added lubricant) was significantly higher than that of other addition methods. The order was as follows: no addition, external addition, and then internal addition. The ejection energy (EE) for internal addition was the lowest; for no addition, EE was the highest. In the dissolution test, the tablets obtained using external addition immediately disintegrated and showed faster drug release than those prepared using internal addition. This result occurred because the water penetration rate of the tablet using the external addition was much higher. The trypsin activity in tablets prepared using the external addition method was significantly higher than that produced using the internal addition method at the same pressure. All these results suggest that the external addition method might produce a fast-dissolution tablet. Because the drug will be compressed using low pressure only, an unstable bulk drug may be tableted without losing potency.

Algorithms↗

PubMed2001

Moyamoya disease and coronary artery disease--case report.

A 26-year-old female with idiopathic moyamoya disease developed chest pain with concomitant ST depression on electrocardiography. Coronary angiography detected no stenotic lesions in the epicardial coronary arteries. The clinical diagnosis was vasospastic angina pectoris. She was medicated with calcium antagonists, which reduced the frequency of chest pain episodes. Angina pectoris is a rare occurrence in young patients with moyamoya disease. Coronary artery disease and moyamoya disease may have common etiological factors.

Adult↗

PubMed2001

Ca2+-dependent exocytosis of L-glutamate by alphaTC6, clonal mouse pancreatic alpha-cells.

Pancreatic islet cells express receptors and transporters for L-glutamate and are thus believed to use L-glutamate as an intercellular signaling molecule. However, the mechanism by which L-glutamate appears in the islets is unknown. In the present study, we investigated whether L-glutamate is secreted through exocytosis by alphaTC6 cells (clonal mouse pancreatic alpha-cells). An appreciable amount of L-glutamate was released from cultured cells after the addition of KCl or A23187 in the presence of Ca2+ and 10 mmol/l glucose in the medium. The KCl-induced glutamate release was significantly reduced when assayed in the absence of Ca2+ or when the cells were pretreated with EGTA-AM. The KCl-induced Ca2+-dependent glutamate release was inhibited approximately 40% by voltage-gated Ca2+ channel blockers, such as nifedipine at 20 micromol/l. The degree of KCl-induced Ca2+-dependent glutamate release was correlated with an increase in intracellular [Ca2+], as monitored by fura-2 fluorescence. Botulinum neurotoxin type E inhibited 55% of the KCl-induced Ca2+-dependent glutamate release, followed by specific cleavage of 25 kDa synaptosomal-associated protein. Furthermore, bafilomycin A1, a specific inhibitor of vacuolar H+-ATPase, inhibited 40% of the KCl-induced Ca2+-dependent glutamate release. Immunoelectronmicroscopy with antibodies against synaptophysin, a marker for neuronal synaptic vesicles and endocrine synaptic-like microvesicles, revealed a large number of synaptophysin-positive clear vesicles in cells. Digitonin-permeabilized cells took up L-glutamate only in the presence of MgATP, which is sensitive to bafilomycin A1 or 3,5-di-tert-butyl-4-hydroxybenzylidene-malononitrile (a proton conductor) but insensitive to either oligomycin or vanadate. From these results, it was concluded that alphaTC6 cells accumulate L-glutamate in the synaptophysin-containing vesicles in an ATP-dependent manner and secrete it through a Ca2+-dependent exocytic mechanism. The Ca2+-dependent glutamate release was also triggered when cells were transferred in the medium containing 1 mmol/l glucose, suggesting that low glucose treatment stimulates the release of glutamate. Our results are consistent with the idea that L-glutamate is secreted by alpha-cells through Ca2+-dependent regulated exocytosis.

Adenosine Triphosphate↗

PubMed2001

Reduction of dehydroerythorbic acid in vitamin C-deficient guinea pigs.

A reduction of dehydroerythorbic acid (DERA) to erythorbic acid (ERA) in vitamin C-deficient guinea pigs was evaluated and compared with that of dehydroascorbic acid (DASA). Thirty-six guinea pigs were fed with vitamin C-deficient diets for 18 days. On day 19, the guinea pigs were divided into four groups for the administration of 100 mg of DERA, ERA, ascorbic acid (ASA), or DASA every day. After 12 days of oral administration, the concentration of DERA, ERA, ASA, and DASA in the liver, adrenal, spleen, kidney, and plasma of guinea pigs was determined by HPLC. A recovery from scurvy was measured in terms of weight gain and serum alkaline phosphatase activity. All four groups showed similar recovery, indicating that the oral administration of relatively high concentrations of DERA reversed the effects of scurvy in vitamin C-deficient guinea pigs. In spite of DERA or DASA administration, ERA or ASA was mainly detected in the tissues. The reduction ratios of DEAR and DASA were similar (approximately 80%) in all tissues except spleen. These results suggest that both DASA and DERA are taken up and reduced to ASA or ERA in vivo.

Animals↗

PubMed2001

Effects of ceramic component on cephalexin release from bioactive bone cement consisting of Bis-GMA/TEGDMA resin and bioactive glass ceramics.

The purpose of this study was to elucidate the effect of amount of ceramic cement powder on drug release from bioactive bone cement. The associated bone-bonding strength was also investigated. The bioactive bone cement under investigation consisted of bisphenol-alpha-glycidyl methacrylate (Bis-GMA), triethylene-glycol dimethacrylate (TEGDMA) resin and a combination of apatite- and wollastonite-containing glass-ceramic (A-W GC) powder. A-W GC powder (50%, 70% and 80% w/w) containing 5% cephalexin (CEX) powder hardened within 5 min after mixing with Bis-GMA/TEGDMA resin. The compressive strength of the cement with or without drug increased with increasing the amount of ceramic powder. The compressive strength of the 80% ceramic cement without the incorporation of cephalexin was 194 MPa. This compressive strength was about 3 times higher than that for polymethylmethacrylate cement. After the cement was implanted in the proximal metaphysis of the tibiae of male rabbits, the failure load for the cement was found to increase with increasing of the amount of ceramic powder. This finding suggested that the cement formed a bonding with bone. In vitro CEX release from bioactive bone cement pellets in a simulated body fluid at pH 7.25 and 37 degrees C continued for more than 2 weeks. Drug release profile followed the Higuchi equation initially, but not at later stages. The drug release rate increased with increasing amount of ceramic powder in the mixture. Since the pore volume of the cement increased with increasing of amount of ceramic powder, the drug diffused in the pores between the ceramics particle and polymer matrix. As hydroxyapatite precipitated on the cement surface, the drug release rate decreased, as observed at the later release stage. These results suggest that varying the amount of ceramic powder in the cement system could control the drug release rate from bioactive bone cement.

Absorbable Implants↗

PubMed2001

[New serum markers to monitor treatment of acute exacerbation of interstitial lung disease].

A 58-year-old woman was admitted to our hospital because of recurrent fever, severe cough and sputum. Chest radiological examinations showed diffuse reticulonodular opacities in both lung fields. Interstitial pneumonia with probable polymyositis was diagnosed. Serum surfactant protein (SP)-A, SP-D and KL-6, which are new interstitial lung disease markers, showed values significantly higher than cutoff levels. The markers increased more in parallel with the rapid development of respiratory insufficiency, CPK level, myalgia and proximal muscle weakness. Treatment with a high dose of corticosteroid and the following gradual decrease over 8 months led to clinical and radiological improvement, with normalization of values of the markers. These markers may therefore be reliable indicators of therapeutic success. However, these markers underwent different respective changes during the first 2 months. SP-A reached a maximum at the start of the treatment, while SP-D and KL-6 peaked at 5 and 10 days, respectively, after the treatment was initiated. This discrepancy demonstrates that the markers reach the bloodstream by diverse mechanisms and are useful for analyzing pathophysiological alterations in the lung in the early stages of treatment.

Acute Disease↗

PubMed2001

[Recent progress and prospective view of chronic hepatitis C research].

Hepatitis C is a major public health problem because of the high incidence of its related hepatocellular carcinoma. With the progress in molecular biology, the mechanisms of persistent infection, chronic inflammation, and hepatocarcinogenesis have been described in terms of virus, host, and virus-cell interactions. On the other hand, clinically, some recent studies using a large number of subjects with long-term observation after interferon therapy showed that improving hepatic inflammation might be associated with regression or retardation of fibrosis. However, current therapy for hepatitis C, although effective in some patients, is problematic even though the efficacy of combination therapy, interferon plus ribavirin, or PEG-interferon has been reported. Here we review the progress and discuss the prospective view of hepatitis C virus research from a point of view of both basic and clinical aspects.

Animals↗

PubMed2000

Calcium level-responsive in-vitro zinc release from zinc containing tricalcium phosphate (ZnTCP).

The in vitro Zn release from tricalcium phosphate containing Zn (ZnTCP; 0.63, 6.17, and 12.05 Zn w/w%) was investigated. The rates of release from ZnTCP powders were measured in 25 mL of simulated body fluid (SBF) containing 10 mg/100mL Ca (SBF/H), 5 mg/100mL Ca (SBF/L), or no Ca (SBF/-) at pH 7.25, 37.0+/-0.1 degrees C. The release from 6 and 12% ZnTCP was initially very fast. The rate of release from ZnTCP decreased as the concentration of Ca in the dissolution media increased, but increased as concentration of Zn in TCP increased. The dissolution kinetics of ZnTCP followed the Hixon-Crowell equation at the initial stage of dissolution, and the initial dissolution rate constant (IDR) was calculated by the least-squares method. The effect of Ca concentration on percent IDR of ZnTCP suggested that Zn release from 0.6% ZnTCP was significantly high compared to that from 6 or 12% ZnTCP. The relationship between the amount of Ca precipitated and Zn release of various ZnTCP samples suggested that the release from 0.6% ZnTCP was significantly different compared to that from 6 and 12% ZnTCP, consistent with the data for percent Zn IDR. X-ray diffraction data suggested that 0.6% ZnTCP contained 5% hydroxyapatite, a low solubility material, which acted as seed crystal during the dissolution test.

Body Fluids↗

PubMed2000

Large isoform of hepatitis delta antigen activates serum response factor-associated transcription.

Hepatitis delta virus infection sometimes causes severe and fulminant hepatitis as a coinfection or superinfection along with the hepatitis B virus. To elucidate the underlying mechanism of injury caused by hepatitis delta virus, we examined whether two isoforms of the hepatitis delta antigen (HDAg) had any effect on five well defined intracellular signal transduction pathways: serum response factor (SRF)-, serum response element (SRE)-, nuclear factor kappaB-, activator protein 1-, and cyclic AMP response element-dependent pathways. Reporter assays revealed that large HDAg (LHDAg) activated the SRF- and SRE-dependent pathways. In contrast, small HDAg (SHDAg) did not activate any of five pathways. LHDAg enhanced the transcriptional ability of SRF without changing its DNA binding affinity in an electrophoretic mobility shift assay. In addition, LHDAg activated a rat SM22alpha promoter containing SRF binding site and a human c-fos promoter containing SRE. In conclusion, LHDAg, but not SHDAg, enhances SRF-associated transcriptions. Despite structural similarities between the two HDAgs, there are significant differences in their effects on intracellular signal transduction pathways. These results may provide clues that will aid in the clarification of functional differences between LHDAg and SHDAg and the pathogenesis of delta hepatitis.

Animals↗

PubMed2000

Ionotropic glutamate receptors expressed in human retinoblastoma Y79 cells.

Mammalian retinal photoreceptors and pinealocytes have common characteristic that they secrete melatonin and L-glutamate as chemical transmitters. Although pinealocytes express glutamate receptors and receive glutamate signals, whether or not photoreceptors express glutamate receptors is unknown. Here, we investigated the expression of the glutamate receptors in cultured Y79 clonal human retinoblastoma cells, as model systems of photoreceptors. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that GluR1, GluR5, GluR7, EAA2, NR1, NR2A and NR2D mRNAs were present in the cultured cells. Northern analysis confirmed the presence of GluR7, EAA2, NR1, NR2A and NR2D mRNAs, while other mRNAs were under the detection limit. Addition of (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid or kainate increases intracellular (Ca(2+)) in Fura-2 loaded cells, which is blocked by 6-cyano-7-nitroquinoxaline-2,3-dione. N-methyl-D-aspartate also increases intracellular (Ca(2+)). These results demonstrated the presence of functional ionotropic receptors in Y79 cells.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed2000

Hepatitis C virus core protein enhances p53 function through augmentation of DNA binding affinity and transcriptional ability.

Hepatitis C virus (HCV) causes a persistent infection, chronic hepatitis, and hepatocellular carcinoma. Since there are several reports indicating that some viruses influence the tumor suppressor p53 function, we determined the effects of HCV proteins on p53 function and its mechanism determined by use of a reporter assay. Among seven HCV proteins investigated (core, NS2, NS3, NS4A, NS4B, NS5A, and NS5B), only core protein augmented the transcriptional activity of p53 and increased the expression of p21(waf1) protein, which is a major target of p53. Core protein increased both DNA-binding affinity of p53 in electrophoretic morbidity shift assay and transcriptional ability of p53 itself in a reporter assay. The direct interaction between core protein and C terminus of p53 was also shown by glutathione S-transferase fusion protein binding assay. In addition, core protein interacted with hTAF(II)28, a component of the transcriptional factor complex in vivo and in vitro. These results suggest that HCV core protein interacts with p53 and modulates p53-dependent promoter activities during HCV infection.

3T3 Cells↗

PubMed2000

Pharmacologic graft protection without donor pretreatment in liver transplantation from non-heart-beating donors.

BACKGROUND: Non-heart-beating donors (NHBDs) are considered potential sources of transplant organs in an effort to alleviate the problem of donor shortage in clinical liver transplantation. We investigated the possibility of pharmacologic protection of hepatic allograft function from NHBDs without donor pretreatment. METHODS: Orthotopic liver transplantation was performed using pigs. In donors, cardiac arrest was induced by stopping the respirator. Forty-five minutes after cessation of the respirator, the liver was flushed with cold lactated Ringer's solution including heparin and with the University of Wisconsin (UW) solution, and then preserved for 8 hr at 4 degrees C in the UW solution. The pigs were divided into two groups: a control group and a treated group. In the treated group, an endothelin antagonist TAK-044 was added to the UW solutions (10 mg/L), and TAK-044 (10 mg/kg body weight) and a platelet activating factor antagonist E5880 (0.3 mg/kg body weight) were also administered to the recipients. RESULTS: TAK-044 and E5880 treatment significantly increased the 7-day survival rate of the recipients (100% vs. 17%, P<0.05). In the treated group, portal venous pressure immediately after reperfusion of the graft was significantly lower than in the control group, and postoperative increase in serum concentrations of glutamic oxaloacetic transaminase and total bilirubin was attenuated. Moreover, the energy charge and adenosine triphosphate concentration of the liver were rapidly restored after reperfusion. CONCLUSIONS: Pharmacologic modulation with TAK-044 and E5880 avoiding donor pretreatment can improve the viability of hepatic allografts procured from NHBDs.

Animals↗

PubMed2000

Calcium-level responsive controlled drug delivery from implant dosage forms to treat osteoporosis in an animal model.

The effects of plasma calcium levels on estradiol release from a self-setting apatite bone cement containing 0.5% estradiol and on the bone mineral density (BMD) of ovariectomized rats were investigated. Apatite cement consisting of an equimolar mixture of tetracalcium phosphate, dicalcium phosphate dihydrate and 0.5% beta-estradiol was prepared. The in vitro release profiles from the cements in simulated body fluid containing 0, 5 and 10 mg/100 ml calcium indicated that estradiol release rate decreased with increasing calcium concentration in the dissolution medium. After subcutaneous implantation of the cement, in vivo estradiol release in diseased rats (ovariectomized rats on a low calcium diet) was significantly higher than that in normal rats. The diseased rats maintained a low calcium level during drug release. The bone mass of the recovery model rat was greater after the experiment than before. The results suggested that the severity of osteoporosis in this animals can be reduced by the implantation of this estradiol-loaded apatite cement.

Animals↗

PubMed2000

Solid dosage form preparations from oily medicines and their drug release. Effect Of degree of surface-modification of silica gel on the drug release from phytonadione-loaded silica gels.

A surface-modified silica gel was produced to improve the surface affinity to an oily medicine, phytonadione (VK1). The effect of the degree of surface modification of the silica gel on the drug release behavior from the silica porous matrix was investigated. The silica gels were surface-modified using the silan coupling agent, 3-methacryloxypropyltrimethoxysilane (C7), octadecyltriethoxysilane (C18), or 3,3,3-trifluoropropyltrimethoxysilane (F3). A mixture of VK1 solution and surface-modified silica gel was evaporated under reduced pressure at room temperature, then the resulting powder was dried in vacuo. The degree of surface modification was evaluated based upon elementary analysis. The dissolution profiles of the samples were investigated in Japanese Pharmacopoeia XII, 1st fluid buffer (pH 1.2, 37+/-0.5 degrees C) containing 1.5% sodium lauryl sulfate. The FT-IR spectra of VK1-loaded surface-modified silica gels suggested that the amount of hydrogen-bonded VK1 with the silanol group on the gel surface decreased with increasing hydrophobicity of the silica gel. Since the modified group was rotating on the silica gel surface, and inhibited the adsorption of VK1 to the surface, the attractive molecular interaction between VK1 and the silica gel surface might decrease with increasing length of the modified functional group. However, the characteristics of the affinity of VK1 to the functional groups significantly differed among the groups. The VK1 release from the modified silica gels was initially rapid, slowed markedly after 1 h, and continued for more than 24 h. The amount of VK1 released from the modified surface silica gels by C7, C18 or F3 increased with increasing density of the surface modification group. The mean drug release moment (MDT) decreased with an increase in surface-modified group density.

Adsorption↗

PubMed2000

Effects on human and nonhuman primate immune response of a new rat anti-CD2 monoclonal antibody.

BACKGROUND: Nonhuman primate models are highly clinically relevant in transplantation. The development of immunosuppressive tools or a tolerogenic regimen for primate models therefore represents an important goal of transplantation immunological research. Hence, we have developed a rat monoclonal antibody (mAb) that recognizes the CD2 molecule (LO-CD2b) on both human and nonhuman primate cells. METHODS: The LO-CD2b mAb has been characterized by flow cytometry, E-rosetting inhibition, and Western blotting. In vitro inhibition of immune responses by LO-CD2b was assessed after both mitogenic and allogeneic stimulation in mixed lymphocyte reactions (MLR). Several LO-CD2b dose and time responses were tested. In vivo, peripheral and lymph node T-cell depletion was examined both by flow cytometry and immunohistology in 10 baboons that received intravenous injection of LO-CD2b at different doses and time courses. Xenosensitization (anti-rat) was assessed by ELISA. Renal allograft survival was followed in two baboons treated with iterative LO-CD2b injections. RESULTS: In vitro, LO-CD2b binds a lymphocyte antigenic determinant of 52 kDa that is recognized by other well-characterized anti-CD2 mAbs (T11, Leu5b). LO-CD2b recognized natural killer CD2+ cells. Administration of 200 ng/ml LO-CD2b almost completely inhibited human and baboon mitogenic stimulation. Allogeneic baboon and human MLR were completely inhibited by the addition of LO-CD2b (at 312 ng/ml) on the day of the initiation of culture; when added after 1 or 2 days, LO-CD2b still provided a significant MLR inhibition (>50%). Incubation of LO-CD2b with baboon peripheral blood mononuclear cells produced very low cytokine levels (interferon-y, tumor necrosis factor-alpha, interleukin 2). In secondary MLR, baboon peripheral blood mononuclear cells previously incubated with LO-CD2b were unable to respond to a second allogeneic stimulation but were able to react to mitogens. In vivo, within the first hour after LO-CD2b injection (at 0.15, 0.5, and 2 mg/kg), an 85-90% peripheral depletion of CD2+ cells was observed. A partial T-cell depletion in inguinal lymph nodes was seen after 1 week. The mechanism of peripheral T-cell depletion could have been antibody-dependent cell cytotoxicity or opsonization but was complement independent. Iterative LO-CD2b injections (12 days at 0.35 mg/kg) slightly prolonged the renal allograft survival in two baboons. CONCLUSION: LO-CD2b is a nonactivating rat anti-CD2 mAb able to strongly inhibit both mitogenic and allogeneic responses in human and nonhuman primates. In vivo, LO-CD2b provides a rapid peripheral T-cell depletion, which is reversible within days after the cessation of injections. This rat mAb represents a very important tool for in vivo experimental investigation in nonhuman primates because it similarly reacts against human T cells in vitro.

Animals↗

PubMed2000

A 12-day course of FK506 allows long-term acceptance of semi-identical liver allograft in inbred miniature swine.

BACKGROUND: Spontaneous tolerance to liver allograft has been reported previously in outbred pig models, but the lack of genetic background did not allow to analyze the impact of the major histocompatibility complex (MHC) on tolerance induction. A model of semi-identical liver allograft was therefore developed in inbred miniature swine in order to mimic the clinical situation of living related liver transplant (parent into infant) and to study a protocol for inducing tolerance to liver allograft. METHODS: SLAdd (class Id/d, class IId/d) pigs received orthotopic liver allograft from heterozygous SLAcd (class Ic/d, class IIc/d) miniature swine. Eight animals did not receive immunosuppression. Fourteen SLAdd animals had a 12-day course of FK506 and were divided in two subgroups. In subgroup FK-1, six pigs received a daily intramuscular injection of FK506 at 0.1-0.4 mg/kg, in order to reach daily trough levels between 7 and 20 ng/ml; in subgroup FK-2, eight additional animals received two daily injections of FK506 at 0.05 mg/kg regardless of the daily trough levels. Graft survival, liver biological tests, histology, cellular and humoral immune responses, as well as detection of microchimerism were assessed in all groups. RESULTS: All untreated animals rejected their allograft and died within 28.1 +/- 9.5 days. These rejector animals developed a significant anti-donor cellular and humoral immune response. No peripheral or lymphoid tissue microchimerism was detected in this group. In contrast, long-term survival was obtained in five FK-treated animals (112, 154, 406, 413, and 440 days), whereas several pigs died with a normal allograft function from either overimmunosuppression or intercurrent causes. All FK-treated pigs developed a specific anti-donor unresponsiveness in both cell mediated lymphocytotoxicity and mixed lymphocyte reaction and did not develop anti-donor alloantibodies. The study of the anti-donor immune response by mixed lymphocyte reaction, during the first postoperative week, demonstrated a specific anti-donor unresponsiveness in the peripheral blood from the first posttransplant day. Although microchimerism was detectable in the peripheral blood for several postoperative weeks (maximum 10 weeks) in FK-treated animals, donor cells or DNA were not detected during the long-term follow-up in peripheral blood or lymphoid tissues. CONCLUSIONS: Spontaneous tolerance to semi-identical orthotopic liver allograft did not occur, whereas a 12-day course of FK506 allowed long-term graft acceptance. All FK-treated animals developed in vitro signs of specific immune unresponsiveness and transient peripheral microchimerism. The specific anti-donor cellular unresponsiveness occurred on the first postoperative day after surgery and was of long-term duration. The study of the early immunological events in this model could be of major importance regarding clinical living related liver transplantation.

Animals↗