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Biomedical subjects

Y Takakura

Publications and source records attributed to Y Takakura.

At least 91 records · Page 5Linked to original sources
PubMed1999

Disposition of radioactivity after injection of liver-targeted proteins labeled with 111In or 125I. Effect of labeling on distribution and excretion of radioactivity in rats.

The effect of radiolabeling liver-specific proteins on the in vivo disposition of radioactivity was investigated. The suitability of 111In and 125I as radiolabels for protein disposition studies in vivo was examined. Galactosylated and cationized bovine serum albumin were labeled with either 125I by the chloramine-T method or 111In, using 1-(4-isothiocyanatobenzyl)ethylenediaminetetraacetic acid (SCN-BZ-EDTA) or diethylenetriaminepentaacetic acid (DTPA) as bifunctional chelating agents (BCAs) and administered intravenously to rats. 125I radioactivity disappeared rapidly from the liver with subsequent excretion in the urine and bile, mainly in the TCA soluble fraction. 111In-associated radioactivity, on the other hand, remained in the hepatic tissue in considerably higher amounts during the experiment and was excreted in the bile and urine to a lower extent when compared with 125I. When the effect of BCA on excretion of 111In radioactivity was compared, no significant differences were observed in the urinary clearances. However, biliary excretion was significantly higher for 111In-SCN-BZ-EDTA-bound radioactivity. In conclusion, when compared with 125I, 111In labeling seems to more accurately characterize the in vivo distribution of liver-targeted proteins after their iv administration in rats and allows a more accurate pharmacokinetic evaluation to be performed.

Albumins↗
PubMed1999

Characterization of plasmid DNA binding and uptake by peritoneal macrophages from class A scavenger receptor knockout mice.

PURPOSE: Plasmid DNA (pDNA) has become an important class of macromolecular agent suitable for non-viral gene therapy as well as DNA vaccination. Our recent study has suggested that pDNA is taken up by mouse peritoneal macrophages via a specific mechanism mediated by a receptor similar to the scavenger receptor (SR). This study was designed to further characterize the pDNA uptake by macrophages in order to elucidate the mechanism. METHODS: The binding and uptake of pDNA labeled with 32P or a fluorescent marker were studied in vitro using cultured Chinese hamster ovary (CHO) cells expressing the class A scavenger receptor (SRA) and peritoneal macrophages from SRA-knockout mice. RESULTS: pDNA binding and uptake by CHO(SRA) cells were minimal and almost identical to that by wild-type CHO cells. Macrophages from the knockout mice showed pronounced pDNA binding and uptake as did the control macrophages. In both types of macrophage, pDNA binding was significantly inhibited by cold pDNA, polyinosinic acid and dextran sulfate but not by polycytidylic acid or Ac-LDL. These results provide direct evidence that SRA is not responsible for the significant binding and subsequent uptake of pDNA by mouse peritoneal macrophages. Further binding experiments revealed that, in addition to polyinosinic acid and dextran sulfate, heparin was a potent inhibitor among a variety of polyanionic compounds such as polynucleotides, anionic polysaccharides and modified proteins including Ox-LDL. CONCLUSIONS: The present study suggest that pDNA binding and uptake by mouse peritoneal macrophages are mediated by a specific mechanism to some defined polyanions not by scavenger receptors. The finding would be an important basis for further studies to elucidate the mechanism(s) of pDNA uptake by macrophages.

Animals↗
PubMed1999

Gene expression and antitumor effects following direct interferon (IFN)-gamma gene transfer with naked plasmid DNA and DC-chol liposome complexes in mice.

Gene expression was assessed in three types of mouse solid tumors after direct injection of naked plasmid DNA encoding the luciferase gene (pCMV-Luc) and its DC-chol liposome complexes. Intratumoral injection of 5 or 100 micrograms naked pCMV-Luc into subcutaneously inoculated mouse colon tumor (CT-26), fibrosarcoma (MCA-15) and bladder carcinoma (MBT-2) resulted in significant gene expression. A DC-chol liposome formulation (5 micrograms pCMV-Luc complexed with 25 micrograms DC-chol liposome) showed lower level of gene expression in the tumor models. Based on the results using the reporter gene, we examined the antitumor effect after direct interferon-gamma (IFN-gamma) gene transfer into CT-26 tumors. A significant IFN-gamma production and growth inhibition were obtained following direct intratumoral injection of IFN-gamma gene with naked plasmid DNA (pCMV-Mu gamma). Interestingly, pCMV-Mu gamma/DC-chol liposome complexes exhibited more pronounced growth inhibitory effect despite lower IFN-gamma production. Induction of CT-26 specific antitumor immunity by IFN-gamma gene transfer was confirmed by rejection of a CT-26 tumor challenge in the mice showing complete regression of CT-26 tumors after both treatments. Further analysis demonstrated that a significant cDNA-independent induction of IFN-beta and TNF-alpha occurred following injection with the liposome complexes, suggesting a nonspecific suppressive effect on CT-26 tumor growth by these cytokines. Thus, the present study has demonstrated that tumor tissue might be a promising target for direct IFN-gamma gene transfer with plasmid-based nonviral vectors. It is also suggested that immunomodulatory effects by various cytokines could be involved in antitumor effects after direct intratumoral injection of plasmid DNA formulations.

Animals↗
PubMed1999

Hepatocyte-specific distribution of catalase and its inhibitory effect on hepatic ischemia/reperfusion injury in mice.

To explore the possibility of using catalase for the treatment of reactive oxygen species (ROS)-mediated injuries, the pharmacokinetics of bovine liver catalase (CAT) labeled with 111In was investigated in mice. At a dose of 0.1 mg/kg, more than 70% of 111In-CAT was recovered in the liver within 10 min after intravenous injection. In addition, 111In-CAT was predominantly recovered from the parenchymal cells (PC) in the liver. Increasing the dose retarded the hepatic uptake of 111In-CAT, suggesting saturation of the uptake process. This cell-specific uptake could not be inhibited by coadministration of various compounds which are known to be taken up by liver PC, indicating that the uptake mechanism of CAT by PC is very specific to this compound. The preventive effect of CAT on a hepatic ischemia/reperfusion injury was examined in mice by measuring the GOT and GPT levels in plasma. A bolus injection of CAT at 5 min prior to the reperfusion attenuated the increase in the levels of these indicators in a dose-dependent manner. These results suggest that catalase can be used for various hepatic injuries caused by ROS.

Alanine Transaminase↗
PubMed1999

Long-term results of arthrodesis for osteoarthritis of the ankle.

A bone graft in the shape of a triangular prism was taken from the anterior surface of the tibia and used in the arthrodesis in 43 joints of 42 patients with primary and secondary osteoarthritis of the ankle. The modified anterior sliding inlay graft method was used. The average period of external immobilization was 5.8 weeks (range, 27-84 days), and followup ranged from 2 years 4 months to 14 years 11 months (average, 7 years 2 months). Nonunion was detected in three (7%) patients: one patient returned to work without additional treatment, and the other two patients underwent followup surgery within 7 months, and bony union was achieved. The final rate of nonunion was 2.3%. After the operation, excellent alleviation of pain was obtained. However, dorsiflexion of the foot decreased from the preoperative average value of 10.5 degrees to the postoperative value of 4.2 degrees, and plantar flexion also decreased from the preoperative value of 24.7 degrees to the postoperative value of 14 degrees. In addition, the range of motion of the subtalar and Chopart joints gradually improved with little effect on daily living activities. The most appropriate position of arthrodesis of the ankle appears to be in the neutral position between dorsal and plantar flexion. In addition, the varus and valgus angle of the hindfoot should be in a neutral or slightly valgus position. Degenerative arthritis developed and advanced in the subtalar joint in 32.5% of the patients, and these degenerative changes were exacerbated in many patients if such changes were present before surgery. Tibial stress fracture occurred in two (4.7%) patients as a complication that was specific to the current surgical method. Thus, the surgical procedure was modified to prevent the onset of tibial stress fracture.

Activities of Daily Living↗
PubMed1999

Hindfoot alignment of hallux valgus evaluated by a weightbearing subtalar x-ray view.

A new radiographic view was proposed to evaluate alignment of the hindfoot under weightbearing condition. The ankle joint and the middle and posterior facets of the subtalar joint were clearly visualized in all radiographs. A comparative study was made of 104 feet with hallux valgus in 58 female patients and 67 normal feet in 57 normal female subjects (control group). The mean value of the angle between the axis of the tibia and a line on the surface of the ankle joint on the talus was significantly larger in the group with hallux valgus than in the control group. Likewise, the mean value of the angle between the axis of the tibia and a line on the surface of the posterior facet of the subtalar joint on the calcaneus in the group with hallux valgus was 95.3 degrees, significantly larger than the 87.9 degrees in the control group. These findings showed that the ankle joint and the posterior facet of the subtalar joint in hallux valgus have valgus deviation. The hindfoot in a foot with hallux valgus has a tendency toward pronation. No previous study has measured the inclination of the posterior facet of the subtalar joint directly in weightbearing.

Adolescent↗
PubMed1999

Pharmacokinetic evaluation of biodistribution data obtained with radiolabeled proteins in mice.

Radiolabeling of proteins is a widely used approach to study their in vivo disposition patterns. However, the obtained results may largely depend on the radiolabeling method used. The purpose of the present study was to investigate the effect of the radiolabeling method on the pharmacokinetic analysis of liver targeted protein in mice. Galactosylated bovine serum albumin (Gal-BSA) was labeled with 125I or 111In, using diethylenetriamine-pentaacetic dianhydride (cDTPA) or 1-(4-isothiocyanobenzyl)ethylenediaminetetraacetic acid (SCN-Bz-EDTA) as bifunctional chelating agents. The Gal-BSA was then injected in mice by a bolus intravenous injection. Samples of plasma, urine, liver, kidney, intestine and feces were collected at various time intervals and their radioactivity was measured. In none of the samples examined was there any significant difference in radioactivity distribution originating from the radiolabeling methods within 5 min after administration. After this period, 125I radioactivity in the liver started to decrease significantly faster than that of 111In, which would indicate the intracellular degradation of the protein. Consequently, the reappearance of trichloracetic acid (TCA) soluble 125I radioactivity in the plasma occurred. But whereas the hepatic uptake clearance (CLliver) of [111In]DTPA-Gal-BSA remained constant during 8 h postinjection, the CLliver of [125I]Gal-BSA at 30 min represented only one eighth of its initial values. The CLliver of [111In]SCN-Bz-EDTA-Gal-BSA resembled that of [111In]DTPA-Gal-BSA within 1 h of the experiment but it started to decline after this interval. The observed discrepancies most probably resulted from the formation of different radiolabeled metabolites in the hepatocytes and their different capability of crossing biological membranes. Our findings indicate that among the three methods employed, [111In]DTPA radiolabeling of Gal-BSA is the most appropriate method to study its tissue disposition.

Animals↗
PubMed1999

Development of the ball-and-socket ankle as assessed by radiography and arthrography. A long-term follow-up report.

We studied the development of ball-and-socket deformity of the ankle by arthrography and radiography in 14 ankles of ten patients with congenital longitudinal deficiency of the fibula accompanied by various anomalies. The mean follow-up was for 18 years 10 months. In three ankles in infants less than one year old the lateral and medial sides of the ankle were already slightly round. In another seven ankles the ball-and-socket appearance developed before the age of five years. This was thought to be due to osseous coalition which limits eversion and inversion. In another four ankles in children who were over the age of one year at the initial examination, the deformity was demonstrated by arthrography and radiography at their first examination. Ball-and-socket deformity accompanied by tarsal coalition is an acquired deformity secondary to limitation of movement of the subtalar and midtarsal joints. It has completely developed by about five years of age.

Adolescent↗
PubMed1999

Arthroscopic drilling for the treatment of osteochondral lesions of the talus.

BACKGROUND: An osteochondral lesion of the talus is a relatively rare disorder of the ankle. While a number of treatment options have been reported, it appears to be difficult to manage all lesions with a single approach. We evaluated the indications for and the results of arthroscopic drilling for the treatment of an osteochondral lesion of the talus. METHODS: Eighteen ankles (seventeen patients) with a symptomatic osteochondral lesion of the talus were examined. The ages of the patients ranged from ten to seventy-eight years (mean, 28.0 years) at the time of the operation, and the patients were followed postoperatively for two to 9.5 years (mean, 4.6 years). After the continuity of the cartilage overlying the lesion and the stability of the lesion had been confirmed, arthroscopic drilling was performed with use of a Kirschner wire that was 1.0 to 1.2 millimeters in diameter. A cast was not applied postoperatively, and full weight-bearing was allowed six weeks after the procedure. RESULTS: The clinical result was good for thirteen ankles and fair for five; all ankles had improvement. Twelve of the thirteen ankles that were in patients who were less than thirty years old had a good result. In contrast, only one of the five ankles in patients who were fifty years old or more had a good result. Thus, the clinical results tended to be better for younger patients. Improvement was seen radiographically in fifteen ankles. However, the three ankles in patients who were more than sixty years old were found to have no improvement on radiographic examination. Analysis of the group of patients who had a history of trauma revealed that the mean interval between the injury and the operation was 6.3 months for the three ankles that had a good radiographic result and 11.3 months for the six that had a fair result. Thus, the radiographic results tended to be better when the interval between the injury and the operation was shorter. CONCLUSIONS: Arthroscopic drilling for the treatment of medial osteochondral lesions of the talus does not require osteotomy of the medial malleolus or postoperative immobilization; thus, the procedure is less invasive than other types of operative treatment for the condition and it allows early resumption of daily activities and sports. On the basis of the results in this study, we believe that the procedure is effective and useful in young patients, especially those who have not yet had closure of the epiphyseal plate. A specific indication for the procedure is an early lesion with only mild osteosclerosis of the surrounding talar bone, continuity of the cartilaginous surface, and stability of the osteochondral fragment.

Adolescent↗
PubMed1999

Pharmacokinetic evaluation of mannosylated bovine serum albumin as a liver cell-specific carrier: quantitative comparison with other hepatotropic ligands.

To assess the feasibility of mannosylated macromolecules as a liver-specific carrier system, hepatic uptake characteristics of mannosylated bovine serum albumin (Man-BSA) were pharmacokinetically investigated. After intravenous injection, 111In-Man18-BSA accumulated in the liver up to 70% of dose at 2h; the endothelial cells and Kupffer cells contributed about 66% and 21% of the uptake, respectively. In single-pass perfusion experiments using rat liver at varying inflow concentrations (0.1-2.0 microg/ml), 111In-Man18-BSA and 111In-Man33-BSA were continuously extracted by the liver and their extraction ratios decreased with the increasing inflow concentrations. The outflow curves of each 111In-Man-BSA at three concentrations were simultaneously fitted to a pharmacokinetic model including a binding to the cell surface and an internalization, by using a nonlinear regression program MULTI(RUNGE). The binding constant augmented with the increase in the number of mannose per BSA, whereas the internalization rate constant was quite comparable for both derivatives. The pharmacokinetic analysis has demonstrated that the uptake process of 111In-Man-BSA is characterized to possess fewer binding sites and a greater internalization rate in comparison with other liver-specific carriers such as galactosylated, succinylated and cationized BSAs. These results will provide useful information in designing drug targeting systems to the liver nonparenchymal cells via mannose receptors.

Animals↗
PubMed1999

Uptake by hepatocytes and biliary excretion of intravenously administered polystyrene microspheres in rats.

The in vivo uptake by hepatocytes and biliary excretion of fluorescein isothiocyanate-labeled polystyrene microsphere with a particle size of 50 nm (MS-50) after intravenous administration was studied in rats. It was confirmed by using confocal laser scanning microscopy that MS-50 was partially phagocytosed by the hepatocytes and that MS-50 taken up by the hepatocytes existed exclusively inside the cells 1 h after intravenous administration. Studies on the mechanism of the uptake of MS-50 by the hepatocytes using the liver perfusion technique revealed that a process mediated by apo-E was involved. After intravenous administration of MS-50, about 4% of dose was excreted into bile in 24 h. Pharmacokinetic evaluation of the excretion rate of MS-50 into bile showed that the process followed first-order kinetics. Qualitative evaluation of the fluorescence detected in the bile after intravenous administration of MS-50 revealed that the particles were certainly excreted into bile in an intact form. From these results, it was suggested that intravenously administered MS-50 would be partially phagocytosed by hepatocytes through a process mediated by apo-E and that MS-50 ingested by hepatocytes would be partially excreted into the bile.

Animals↗
PubMed1999

Targeted delivery and improved therapeutic potential of catalase by chemical modification: combination with superoxide dismutase derivatives.

Four types of bovine liver catalase (CAT) derivatives, succinylated (Suc-CAT), galactosylated (Gal-CAT), mannosylated (Man-CAT), and polyethylene glycol conjugate (PEG-CAT), were synthesized and their pharmacokinetics and therapeutic potential in a hepatic ischemia/reperfusion injury model were studied in mice. About 90% of the CAT enzymatic activity was retained after chemical modification. Biodistribution studies showed that 111indium (111In)-Gal-CAT accumulated selectively in the liver parenchymal cells as 111In-CAT, whereas an increased amount of 111In-Suc-CAT and 111In-Man-CAT was delivered to liver nonparenchymal cells. 111In-PEG-CAT exhibited prolonged retention in plasma. Pharmacokinetic analysis revealed that the hepatic uptake clearances of 111In-Suc-CAT, 111In-Gal-CAT, and 111In-Man-CAT were much greater than that of 111In-CAT, whereas that of 111In-PEG-CAT was very small. In the ischemia/reperfusion injury model, in which hepatic injury was induced by occlusion of the portal vein for 30 min followed by 1 h reperfusion, the elevation of plasma glutamic pyruvic transaminase and glutamic oxaloacetic transaminase levels was slightly inhibited by treatment with native CAT or Gal-CAT. PEG-CAT was less potent. In contrast, Suc-CAT and Man-CAT effectively suppressed the increase in plasma glutamic pyruvic transaminase and glutamic oxaloacetic transaminase. Coinjection of mannosylated superoxide dismutase marginally improved the inhibitory effects of CAT derivatives. These results demonstrate that targeted CAT delivery to liver nonparenchymal cells via chemical modification is a promising approach to prevent hepatic injuries caused by reactive oxygen species. The potential usefulness of combining of CAT and superoxide dismutase derivatives is also demonstrated.

Alanine Transaminase↗
PubMed1999

Development of a hepatocyte-specific prostaglandin E(1) polymeric prodrug and its potential for preventing carbon tetrachloride-induced fulminant hepatitis in mice.

A polymeric prodrug of prostaglandin E(1) (PGE(1)) was synthesized using galactosylated poly(L-glutamic acid hydrazide) (Gal-HZ-PLGA) as a biodegradable and targetable carrier to hepatocytes. Poly(L-glutamic acid hydrazide) was prepared by reacting poly(gamma-benzyl-L-glutamate) with hydrazine monohydrate, followed by reaction with 2-imino-2-methoxyethyl-1-thiogalactosides to obtain Gal-HZ-PLGA after i.v. injection. (111)In-labeled galactosylated poly(L-glutamic acid hydrazide) ((111)In-Gal-HZ-PLGA) rapidly accumulated in the liver in a dose-dependent manner, whereas (111)In-poly(L-glutamic acid hydrazide) did not, indicating the involvement of a galactose-specific mechanism in the uptake of (111)In-Gal-HZ-PLGA. Fractionation of liver cells revealed that (111)In-Gal-HZ-PLGA was preferentially taken up by liver parenchymal cells. After being taken up by the liver, (111)In-Gal-HZ-PLGA was gradually degraded, and radioactive metabolites with low molecular weight were detected within 10 min after injection. Then, PGE(1) or [(3)H]PGE(1) was coupled to Gal-HZ-PLGA via a hydrazone bond under mild conditions to obtain PGE(1) conjugate. After i.v. injection, [(3)H]PGE(1) conjugate was rapidly taken up by the liver (more than 80% of the dose). After injection of the conjugate, (3)H radioactivity remained in the liver, representing about 70% of the dose, even at 24 h, whereas little radioactivity remained in the organ at 1 h after the injection of free [(3)H]PGE(1). Finally, its pharmacological activity was examined in mice with fulminant hepatitis induced by peritoneal injection of carbon tetrachloride. The i.v. injection of PGE(1) conjugate at a dose of 1 mg (0.074 mg PGE(1))/kg effectively inhibited the increase of plasma glutamic pyruvic transaminase activity, whereas twice this dose (0.15 mg/kg) of free PGE(1) had little effect. These results suggest that the PGE(1) conjugate is an excellent polymeric prodrug of PGE(1) for hepatitis therapy.

Acute Disease↗
PubMed1998

Asialoglycoprotein receptor-mediated gene transfer using novel galactosylated cationic liposomes.

We synthesized three novel galactosylated cholesterol derivatives, cholesten-5-yloxy-N-(4-((1-imino-c-beta-D-thiogalactosyl+ ++-ethyl)amino) butyl)formamide (Gal-C4-Chol) and its ethyl formamide and hexyl formamide analogues (Gal-C2-Chol, Gal-C6-Chol), to prepare liposomal gene carriers possessing the cationic charge necessary for plasmid DNA binding and galactose residues as a targetable ligand for liver parenchymal cells. Liposome/DNA complexes prepared with these lipids showed low cytotoxicity in human hepatoma HepG2 cells. Gal-C4-Chol/DC-Chol/DOPE(3:3:4) liposomes, consisting of 3:3:4 mixtures of Gal-C4-Chol, 3beta[N',N', N'-dimethylaminoethane)-carbamoyl] cholesterol (DC-Chol), and dioleoylphosphatidylethanolamine (DOPE), showed higher transfection activity and [32P]DNA uptake than DC-Chol/DOPE(6:4) liposomes. The presence of 20 mM galactose significantly inhibited both transfection efficiency and uptake of DNA of Gal-C4-Chol/DC-Chol/DOPE(3:3:4) and Gal-C4-Chol/DOPE(6:4) liposomes, but not those of DC-Chol/DOPE(6:4) liposomes. These results indicate that the liposome/DNA complexes prepared using novel galactosylated cholesterol derivatives are efficiently recognized by asialoglycoprotein receptors and internalized and lead to gene expression. In addition, we found that the galactosylated cholesterol derivative with a longer spacer showed higher transfection activity.

3T3 Cells↗
PubMed1998

Extravasation of macromolecules.

Macromolecules can extravasate across the normal endothelium by transcapillary pinocytosis as well as by passage through interendothelial cell junctions, gaps or fenestrae. The main biological factors that control extravasation of a solute include regional differences in the capillary structures, the disease state of the organ or tissue, and the rate of blood and lymph supply. Physicochemical properties that are of profound significance in the extravasation of macromolecules are molecular size, shape, charge and hydrophilic/lipophilic balance (HLB) characteristics. Extravasation of small drugs, proteins, oligonucleotides and genes can be controlled by conjugating or forming complexes with macromolecular carriers. This requires a thorough understanding of the relationship between the chemical structures, physicochemical properties and the pharmacokinetics of both carrier and active molecules. This review article discusses the extravasation of macromolecules from the view points of pharmacokinetics and drug delivery systems, with the main emphasis on the extravasation across the liver, kidney and tumor capillaries.

Journal Article↗
PubMed1998

Targeted delivery of plasmid DNA complexed with galactosylated poly(L-lysine).

Galactose was introduced to poly(L-lysine) (PLL) with an average molecular weight of 13,000 to develop a hepatocyte-specific carrier for gene drugs. The pharmacokinetic characteristics of a model plasmid, pCAT (plasmid DNA encoding chloramphenicol acetyltransferase reporter gene), complexed with galactosylated PLL (Gal-PLL) was studied in mice in relation to its physicochemical properties. pCAT/Gal-PLL complex at a ratio of 1:0.6 (w/w) has a zeta potential of -20 mV and a mean particle size of about 180 nm. After intravenous injection, [32P]pCAT/Gal-PLL was rapidly eliminated from the circulation and preferentially taken up by the liver's parenchymal cells. The hepatic uptake of [32P]pCAT/Gal-PLL was significantly inhibited by prior administration of Gal-bovine serum albumin, suggesting that the uptake was mediated by the asialoglycoprotein receptors on hepatocytes. In vitro transfection experiments using a hepatoma cell line expressing the asialoglycoprotein receptor revealed that pCAT/Gal-PLL gave a high CAT gene expression whereas pCAT complexed with unmodified PLL failed to transfect the cells.

Animals↗
PubMed1998

Involvement of specific mechanism in plasmid DNA uptake by mouse peritoneal macrophages.

The binding and uptake of plasmid DNA encoding luciferase reporter gene (pCMV-Luc) were studied in vitro using cultured mouse peritoneal macrophages. A significant and time-dependent cellular association of [32P]pCMV-Luc with resident macrophages was observed at 37 degrees C and this decreased at 4 degrees C. The binding at 4 degrees C was saturable and a Scatchard plot gave a maximum binding capacity of 0.81 microgram/mg-protein and a dissociation constant of 0.30 microgram/ml. The binding of [32P]-pCMV-Luc was inhibited by polyinosinic acid, dextran sulfate and salmon sperm DNA, but not by polycytidylic acid, dextran and EDTA. A confocal microscopic study demonstrated that fluorescein-labeled pCMV-Luc was internalized at 37 degrees C while only cell surface binding occurred at 4 degrees C. No significant luciferase gene expression was obtained after incubation with a high concentration (100 micrograms/ml) of pCMV-Luc. These data suggest that plasmid DNA is taken up by macrophages via a mechanism mediated by a receptor like the macrophage scavenger receptor.

Animals↗