Biomedical subjects

T Sakai

Publications and source records attributed to T Sakai.

At least 217 records · Page 12Linked to original sources

PubMed2000

Proteome analysis of mouse brain: two-dimensional electrophoresis profiles of tissue proteins during the course of aging.

Mouse brain proteins were isolated from five regions (cerebellum, cerebral cortex, hippocampus, striatum, and cervical spinal cord) at five ages from the 10th week to the 24th month, and separated by two-dimensional gel electrophoresis (2-DE). 2-DE was carried out with an immobilized pH gradient bar in the first dimension, and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension. Over one thousand protein spots were visualized by silver staining and quantified by image processing. In the analyses, 58 protein spots were distinguishable among the above five brain regions, and 17 proteins were shown to be varied in quantity in the course of aging. Partial amino-terminal sequences and/or internal sequences for a total of 301 protein spots were analyzed. One hundred and eighty proteins appeared to have blocked N-termini and 122 proteins were identified. Twenty-seven new proteins were identified by sequence homology search. A mouse brain proteome database was constructed, which consists of the 2-DE map images and the respective spot data files with 15 related references.

Acrylic Resins↗

PubMed2000

Comparative analysis of brain proteins from p53-deficient mice by two-dimensional electrophoresis.

p53 is a tumor suppressor protein that regulates many cellular processes including the cell cycle, DNA repair, and apoptosis. It also serves as a critical regulator of neuronal apoptosis in the central nervous system (CNS). To elucidate the role of p53 in the CNS, brain proteins of p53 knock-out mice (p53-/-) were analyzed by two-dimensional gel electrophoresis (2-DE) and compared with those from p53 wild type (p53+/+) mice. Six types of brain tissue (temporal cortex, cerebellum, hippocampus, striatum, olfactory bulb, and cervical spinal cord) and other control tissues (lung and blood) from 18-week-old non-stress-induced mice were analyzed. The morphology of brains from p53-/- mice appeared to be normal and identical to that of p53+/+ mice, although lungs showed diffuse tumors that may have been caused by p53 deficiency. Comparative 2-D gel analysis showed that, on average, 7 of 886 spots from brain tissue were p53-/- specific, whereas 12 of 1008 spots from lung tissue were p53-/- specific. N-terminal amino acid sequence was determined for p53-/- specific proteins. In all brain tissues from p53-/- mice, a newly identified mouse mitochondrial NADH-ubiquinone oxidoreductase 24 kDa subunit showed decreased expression, and apolipoprotein A1 acidic forms showed increased expression. In addition, brain-type creatine kinase B chain and tubulin beta-5 N-terminal fragment were increased in the p53-/- cerebellum, and a new protein in mouse, hydroxyacylglutathione hydrolase (glyoxalase II) was decreased in the temporal cortex of p53-/- mice. The alterations in protein expression identified in this study may imply a p53-related brain function. This is the first proteomic analysis on the p53-/- mouse brain, and further information based on this study will provide new insights into the p53 function in the CNS.

Amino Acid Sequence↗

PubMed2000

Relationship between delta-aminolevulinic acid dehydratase genotypes and heme precursors in lead workers.

BACKGROUND: The objective of this study was to investigate the relationships between genotypes of delta-aminolevulinic acid (ALA) dehydratase (ALAD) and disturbances in the heme biosynthetic pathway by lead exposure. METHODS: The subjects were 192 male lead workers and 125 control subjects. Blood lead concentrations (Pb-B), plasma ALA concentrations (ALA-P), and ALAD genotypes were determined for all subjects. In lead workers, ALAD activity, ALA in urine (ALA-U), and erythrocyte zinc protoporphyrin (ZP) were also determined. RESULTS: The frequency of ALAD2 (minor type of ALAD allele) was calculated to be 0.087 in all subjects. No significant relationship was found between ALAD2 frequency and Pb-B levels in lead workers. ALAD1 homozygotes showed significantly higher levels of ZP and ALA-P in comparison with those of ALAD2 carriers at Pb-B levels more than 20 microg/dL and 40 microg/dL, respectively. CONCLUSIONS: ALAD1 homozygotes might be more susceptible than ALAD2 carriers to disturbances in heme metabolism caused by lead exposure.

Adult↗

PubMed2000

In vivo evaluation of the bond strength of adhesive 4-META/MMA-TBB bone cement under weight-bearing conditions.

In order to minimize the problems associated with implant fixation using acrylic bone cement, we studied a new adhesive bone cement that consists of 4-methacryloyloxyethyl trimellitate anhydryde (4-META) and methylmethacrylate (MMA) as monomers, tri-n-butylborane (TBB) as an initiator, and PMMA powder (4-META/MMA-TBB cement). It shows remarkable adhesive properties to metal and bone in vitro. The purpose of this study was to evaluate the strength of the bond of the cement to both metal and bone in vivo under weight-bearing conditions. Metal prostheses were implanted in the right femora of 12 rabbits using either adhesive 4-META/MMA-TBB cement or the conventional PMMA cement, as the control, for fixation. After 4 and 12 weeks, both femora were excised and the same operations were performed in the left femora in vitro. Eighteen femora were sectioned for the mechanical assessment of the bone-cement and cement-implant interfaces. 4-META/MMA-TBB cement had a significantly higher interfacial shear strength than the conventional PMMA cement: 201 N and 90 N, on average, for the implant-cement interface (p<0.01); and 138 N and 89 N, on average, for the bone-cement interface (p<0.01), at 12 weeks. The present results suggest the efficacy of 4-META/MMA-TBB cement in providing greater fixation of implants to bone and promise a firmer intramedullary fixation than the control conventional PMMA cement.

Animals↗

PubMed2000

Prevention of fibrous layer formation between bone and adhesive bone cement: in vivo evaluation of bone impregnation with 4-META/MMA-TBB cement.

We have studied a new adhesive bone cement, that consists of 4-methacryloyloxyethyl trimellitate anhydride (4-META) and methylmethacrylate (MMA) as monomers, tri-n-butyl borane (TBB) as the initiator, and polymethylmethacrylate powder (4-META/MMA-TBB cement). This cement has shown remarkable adhesive properties to bone in vitro. In this study, we assessed the interface in vivo periodically. The femora of rabbits were fenestrated and filled with either the 4-META/MMA-TBB cement or a conventional polymethylmethacrylate cement. The animals were killed after 1, 4, 12, and 24 weeks to analyze the interface by optical microscopy and transmission electron microscopy. Optical microscopic examinations showed that the cured 4-META/MMA-TBB adhesive cement bonded to bone directly for 24 weeks, whereas a fibrous tissue layer was observed between the bone and cured conventional cement at 12 weeks after the operation. The transmission electron microscopy views of 4-META/MMA-TBB cement bonded to bone demonstrated a unique "hybridized bone" with the cement in the subsurface of the substrate in every case. The formation of the hybridized bone indicates the bonding mechanism of the adhesive cement to bone, which prevents the fibrosis intervention between bone and cement. These results suggest that the biomechanical and adhesive properties of 4-META/MMA-TBB cement make it a useful bone-bonding agent in orthopedic surgery.

Animals↗

PubMed2000

Butyrate as a model for "gene-regulating chemoprevention and chemotherapy.".

Recent progress in molecular genetics has facilitated understanding of the mechanisms of carcinogenesis. However, there is not yet any effective therapy or prevention for cancer based on the molecular mechanisms of carcinogenesis. So-called "gene therapy" for cancer is expected to become a new method of treatment, but there are still several serious problems with gene therapy. As a matter of fact, it seems impossible to adopt gene therapy for prevention. We therefore tried to develop a different method of cancer prevention or therapy based on the molecular mechanisms of carcinogenesis. For instance, the tumor-suppressor gene p53 is mutated in about 50% of human malignancies. It is known that p53 stimulates the promoter activities of p21/WAF1, gadd45 and bax genes, resulting in cell cycle arrest, DNA repair and apoptosis, respectively. Therefore, chemical compounds that can stimulate these genes should compensate for the function of p53. As a model of this, we found that histone deacetylase inhibitors such as butyrate or trichostatin A dramatically stimulate the p21/WAF1 gene promoter through the Spl sites, resulting in cell cycle arrest. Interestingly, another group has recently reported that phenylbutyrate, which is also known as a histone deacetylase inhibitor, is very effective for leukemia patients. We therefore consider methods of up-regulating p21/WAF, gadd45 or bax genes should be useful for cancer therapy and termed this method "Gene-regulating chemotherapy". Theoretically, the chemicals up-regulating such genes should be also useful for chemoprevention, and we also termed it as "Gene-regulating chemoprevention". In conclusion, we propose that "Gene-regulating chemotherapy or chemoprevention" may be a promising new method for cancer therapy or prevention and histone deacetylase inhibitor is a good candidate for this method.

Animals↗

PubMed2000

Midazolam premedication reduces propofol requirements for sedation during regional anesthesia.

PURPOSE: Propofol is often used for sedation during spinal anesthesia. We investigated the effects of midazolam premedication on the propofol requirements and incidence of complications during sedation. METHODS: In a prospective randomized, controlled, and single-blinded study, 50 patients undergoing elective gynecological surgery were randomly divided into control and midazolam groups. Patients in the midazolam group received 2 mg midazolam im 30 min before arrival at the operation room. After spinal anesthesia was instituted with intrathecal injection of hyperbaric tetracaine, we provided sedation using continuous infusion of propofol. The level of sedation was controlled at a level between "eyes closed but rousable to command" and "eyes closed but rousable to mild physical stimulation" by adjusting the infusion rate. During sedation, the propofol requirements and complications were recorded and patients were asked, two hours after the end of operation, whether they remembered intraoperative events. RESULTS: In the midazolam group, the loading dose, steady state infusion rate, and overall infusion rate of propofol were 0.74 mg x kg(-1), 2.86 mg x kg(-1) x hr(-1), and 3.32 mg x kg(-1) x hr(-1), respectively, which were about 17% lower than those in the control group (P<0.05). Moreover, midazolam premedication reduced the incidence of intraoperative memory (P < 0.05), but had no effects on other complications. CONCLUSION: Midazolam premedication reduced propofol requirements and the incidence of intraoperative memory during sedation. These effects on sedation using propofol during spinal anesthesia are considered beneficial for patients.

Adult↗

PubMed2000

Ketamine suppresses endotoxin-induced NF-kappaB expression.

PURPOSE: Ketamine reduces endotoxin-induced production of proinflammatory cytokines, including tumour necrosis factor-alpha (TNF), in several types of inflammatory cells, including monocytes and macrophages. Transcription of the genes that encode production of these proinflammatory cytokines is regulated by nuclear factor-kappa B (NF-kappaB). Cytoplasmic B protein is activated by endotoxin (LPS) as well as by TNF, allowing B protein to migrate into the cell nucleus to activate gene transcription for these inflammatory mediators. Because NF-kappaB is likely involved in brain injury and inflammatory neurodegenerative disease, such as multiple sclerosis, we examined whether ketamine inhibits LPS-induced activation of NF-kappaB in human glioma cells in vitro and intact mouse brain cells in vivo. METHODS: Endotoxin-induced NF-kappaB expression in both the human glioma cells in vitro and the intact mouse brain cells in vivo was determined by electrophoretic mobility shift assays (EMSA) of nuclear extracts and measurement of NF-kappaB expression by densitometry. Endotoxin was injected intracerebroventricularly in vivo and intact brain was harvested. Klenow fragment labeling was used to identify NF-kappaB protein for both the in vivo and vitro experiments. RESULTS: Endotoxin treatment increased NF-kappaB expression (P < 0.05) both in vivo and vitro compared with control (untreated) cells. Ketamine suppressed endotoxin-induced neuronal NF-kappaB activation in a dose-dependent manner (P < 0.05, except for the 10(-5) M concentration in vitro) both in vivo and vitro. CONCLUSION: Ketamine inhibits endotoxin-induced NF-kappaB expression in brain cells in vivo and vitro and it is suggested that this may have implications in the neuroprotective effects of ketamine reported by other investigators.

Animals↗

PubMed2000

MR findings of necrotic lesions and the extralesional area of osteonecrosis of the femoral head.

OBJECTIVE: To investigate the MR findings of necrotic lesions and the extralesional area of osteonecrosis of the femoral head (ONFH) for each of the radiological stages. DESIGN AND PATIENTS: Forty-nine hips in 29 patients (15 female, 14 male; mean age 38 years, range 17-59 years) were imaged using a 1.0-T superconducting magnet. T2-weighted spin echo pulse sequences (T2WI), spoiled gradient recalled echo pulse sequences (SPGR) and fat suppression SPGR (FS-SPGR), followed by Gd-DTPA enhanced fat suppression SPGR (Gd-FS-SPGR), were all obtained with the aid of a TORSO surface coil. RESULTS AND CONCLUSIONS: While a normal fat intensity area with a low-intensity band on SPGR (band pattern) was seen in 16 of 16 stage 1 (100%), nine of 11 stage 2 (82%), four of 17 stage 3 (24%), and none of five stage 4 hips, all hips showed peripheral rim enhancement on Gd-FS-SPGR (100%). This enhancement band on Gd-FS-SPGR corresponded to histological findings of necrotic trabecular bone, repaired marrow, and fibrous reparative tissue. Bone marrow edema was also clearly demonstrated as a diffuse, high-intensity area outside this enhancement band on Gd-FS-SPGR in two stage 2 (18%), 12 stage 3 (71%), and one stage 4 hip (20%). In cases at stage 2 or more advanced stages with homogeneous or inhomogeneous low intensity on nonenhanced MRI, the reparative process both inside and outside the necrotic lesion, including bone marrow edema, was detected clearly on contrast-enhanced MRI.

Adolescent↗

PubMed2000

Melatonin stimulates thyroid-stimulating hormone accumulation in the thyrotropes of the rat pars tuberalis.

We have reported that the unique thyroid-stimulating hormone-immunoreactive cells (TSH cells) in the intact adult and fetal rat pars tuberalis (PT) show an intense spot-like TSH immunoreaction in the perinuclear region. The present study was designed to investigate the relationship between melatonin and these unique TSH cells. We classified TSH cells in the PT (PT-TSH cells), on the basis of immunoreactivity, into spot-like stained cells (SC) and whole cytoplasm stained cells (WC), and estimated the proportion of each TSH cell type to total cells in the experimental rats by morphometry. Chronic administration of melatonin to control rats leads to an increase of WC in number but a decrease of SC. On the other hand, the intensity of TSH immunoreactivity and the number of rat PT-TSH cells significantly decreased after pinealectomy and recovered by melatonin administration. Radioimmunoassay showed that melatonin treatment increased the TSH content in the PT. Moreover, electron microscopy showed that the number of TSH secretory granules in the PT-TSH cells increased in the melatonin-replaced rats. These results demonstrated that melatonin stimulates the accumulation of TSH in the rat PT-TSH cells via secretory granule formation and suggest that melatonin regulates TSH release from PT-TSH cells.

Animals↗

PubMed2000

Japanese patients with sporadic Hirschsprung: mutation analysis of the receptor tyrosine kinase proto-oncogene, endothelin-B receptor, endothelin-3, glial cell line-derived neurotrophic factor and neurturin genes: a comparison with similar studies.

UNLABELLED: We report on mutation analysis of five genes involved in the receptor tyrosine kinase (RET) or the endothelin-signalling pathways in 28 sporadic Japanese patients with Hirschsprung disease. Analysis of DNA obtained from peripheral blood cells revealed six mutations in the RET proto-oncogene, four of which were disease-causing mutations in exon 9 (D584G), the splice donor site of intron 10 (+2T to A), exon 11 (A654T), and exon 12 (T706A). A heterozygous A to G transition was found in 47 bases upstream from the 5' end of exon 2 in two HD patients but was also seen in one control subject (2/28; 1/24). A silent 2307T to G transversion was observed in exon 13. Two disease-causing mutations were detected in the endothelin receptor (EDNRB) gene, in the non-coding region of exon 1 (-26 G to A) and in exon 4 (A301T); the latter mutation was a novel one. One silent mutation was observed in exon 4 (codon 277). One heterozygous T to C mutation was found in the glial cell line-derived neurotrophic factor gene in 25 bases upstream of the coding region in exon 1. No nucleotide changes were detected in either the endothelin-3 or neurturin genes. Disease-causing mutation rates in the RET proto-oncogene and the EDNRB gene were estimated at 14.3% (4/28) and 10.7% (3/28), respectively. In addition to mutations in the RET and EDNRB genes, embryonic environmental factors and/or other genetic factors appear to be involved in the development of Hirschsprung disease. Further systematic studies of genetic variation in a large series of patients and controls are necessary for elucidating the pathogenesis of this disorder. CONCLUSION: This study provides further gene alterations as disease-causing mutations in Japanese cases of sporadic Hirschsprung disease. However, the low mutation rate of the susceptibility genes may indicate that Hirschsprung disease arises from a combination of genetic and environmental factors.

Asian People↗

PubMed2000

Optimizing femoral anteversion and offset after total hip arthroplasty, using a modular femoral neck system: an experimental study.

To investigate the effectiveness of a modular femoral neck system, consisting of two neutral and four types of retroverted necks for the correction of femoral anteversion and offset in total hip arthroplasty, an experimental study was carried out, using sawbones with four different angles of femoral anteversion (16 degrees, 34 degrees, 47 degrees, and 59 degrees ). With the neutral neck, reconstruction of the preoperative anteversion and offset in the normal femur was achieved. While the 15 degrees retroverted long neck was effective for the mildly or moderately anteverted femur, this retroverted neck showed insufficient correction for the severely anteverted femur. This modular neck system proved to be useful for correction of the medial component of femoral offset in femora with anteversion of less than 47 degrees. For patients with greater anteversion, a feature which is rarely seen in the clinical situation, femoral necks with a greater degree of retroversion may be useful.

Arthroplasty, Replacement, Hip↗

PubMed2000

Atherosclerosis and omega-3 fatty acids in the populations of a fishing village and a farming village in Japan.

The effect of different dietary habits on atherosclerosis was investigated by examining the content of ordinary diets and relevant risk factors through a mass health survey on two village populations in Japan. In total, 261 inhabitants in the fishing village and 209 in the farming village were examined for body build, blood pressure, and blood chemistry. Information on smoking habits and food consumption was obtained using a semi-quantitative item-frequency questionnaire. Pulse wave velocity of the aorta, intima-media thickness of the carotid artery, and atherosclerotic plaques as obtained by ultrasonography were used as measures of atherosclerosis. All measures of atherosclerosis are lower in the fishing village than in the farming village in both men and women. There is a striking 5-8-fold difference in the number of atherosclerotic plaques (P < 0.0001) between the populations. The observed differences in atherosclerosis parallels differences in dietary habits and differences in the serum essential fatty acids. Evaluation of the omega-3 fatty acids over the combined populations reveals a negative association with the number of plaques in the common carotid while the omega-6 fatty acids shows a weak positive association with plaques.

Adult↗

PubMed2000

Albumin inhibits apolipoprotein AI and AII production in human hepatoblastoma cell line (Hep G2): additive effects of oleate-albumin complex.

Although the role of multiple humoral agents (such as plasma albumin, glucose, hormones etc.) are implicated in lipoprotein metabolism, the mechanism of action of these agents on various steps of the synthesis and secretion of lipoproteins and apolipoproteins (protein moieties of lipoproteins) are not completely understood. Specifically, the hepatocellular mechanisms of the effect of albumin and fatty acids on apolipoprotein (apo) AI and AII [major proteins of high density lipoproteins (HDL)] synthesis and secretion are not known. Using human hepatoblastoma cells (Hep G2) as an in vitro model system, this study examined the effect of albumin and fatty acids on the synthesis, secretion, and the steady-state mRNA expression of apo AI and AII. The data indicated that the incubation of Hep G2 cells with albumin, dose-dependently, inhibited apo AI and AII accumulation (secretion) in the media, de novo synthesis, and the steady-state mRNA expression. Albumin did not alter total protein synthesis; thus the effect of albumin appeared to be specific for the synthesis and secretion of apo AI and apo AII. Free fatty acids (FFA) are transported by albumin and diseases characterized by enhanced FFA mobilization (e.g. diabetes mellitus) are associated with low HDL levels. Studies were therefore performed to examine the effect of albumin-bound-oleic acid on apo AI and apo AII production. The results showed that the albumin-oleate complex further increased the inhibitory effects of albumin on apo AI and apo AII production. These data suggest how HDL metabolism may be affected at the hepatocellular level by alterations in plasma albumin concentrations and/or fatty acid mobilization in clinical situations characterized by altered HDL levels.

Albumins↗

PubMed2000

Construction of a bioengineered cardiac graft.

OBJECTIVES: Currently available graft materials for repair of congenital heart defects cause significant morbidity and mortality because of their lack of growth potential. An autologous cell-seeded graft may improve patient outcomes. We report our initial experience with the construction of a biodegradable graft seeded with cultured rat or human cells and identify their 3-dimensional growth characteristics. METHODS: Fetal rat ventricular cardiomyocytes, stomach smooth muscle cells, skin fibroblasts, and adult human atrial and ventricular cardiomyocytes were isolated and cultured in vitro. These cells were injected into or laid onto biodegradable gelatin meshes, and their rate of proliferation and spatial location within the mesh was evaluated by using a cell counter and histologic analysis. RESULTS: Rat cardiomyocytes, smooth muscle cells, and fibroblasts demonstrated steady proliferation over 3 to 4 weeks. The gelatin mesh was slowly degraded, but this process was most rapid after seeding with fibroblasts. Human atrial cardiomyocytes proliferated within the gelatin meshes but at a slower rate than that of fetal rat cardiomyocytes. Human ventricular cardiomyocytes survived within the gelatin mesh matrix but did not increase in number during the 2-week duration of evaluation. Grafts seeded with rat ventricular cells exhibited spontaneous rhythmic contractility. All cell types preferentially migrated to the uppermost surface of each graft and formed a 300- to 500-microm thick layer. CONCLUSIONS: Fetal rat ventricular cardiomyocytes, gastric smooth muscle cells, skin fibroblasts, and adult human atrial cardiomyocytes can grow in a 3-dimensional pattern within a biodegradable gelatin mesh. Similar autologous cell-seeded constructs may eventually be applied to repair congenital heart defects.

Adult↗

PubMed2000

Autologous porcine heart cell transplantation improved heart function after a myocardial infarction.

OBJECTIVE: Fetal cardiomyocyte transplantation improved heart function after cardiac injury. However, cellular allografts were rejected despite cyclosporine (INN: ciclosporin) therapy. We therefore evaluated autologous heart cell transplantation in an adult swine model of a myocardial infarction. METHODS: In 16 adult swine a myocardial infarction was created by occlusion of the distal left anterior descending coronary artery by an intraluminal coil. Four weeks after infarction, technetium 99m-sestamibi single photon emission tomography showed minimal perfusion and viability in the infarcted region. Porcine heart cells were isolated and cultured from the interventricular septum at the time of infarction and grown in vitro for 4 weeks. Through a left thoracotomy, either cells (N = 8) or culture medium (N = 8) was injected into the infarct zone. RESULTS: Four weeks after cell transplantation, technetium 99m-sestamibi single photon emission tomography demonstrated greater wall motion scores in the pigs receiving transplantation than in control animals (P =.01). Pigs receiving transplantation were more likely to have an improvement in perfusion scores (P =.03). Preload recruitable stroke work (P =.009) and end-systolic elastance (P =. 02) were greater in the pigs receiving transplantation than in control animals. Scar areas were not different, but scar thickness was greater (P =.02) in pigs receiving transplantation. Cells labeled with bromodeoxyuridine in vitro could be identified in the infarct zone 4 weeks after transplantation. Swine receiving transplantation gained more weight than control animals (P =.02). CONCLUSION: Autologous porcine heart cell transplantation improved regional perfusion and global ventricular function after a myocardial infarction.

Analysis of Variance↗

PubMed2000

A single plate ultrasonic gyroscope in plane motion

We have proposed a vibration gyroscope made of a single piezoelectric plate in plane motion. The gyroscope makes use of the degenerated modes associated with a square plate. In the present paper, the experimental verification using an enlarged piezoelectric ceramic plate is presented, whose results are compared with the numerical prediction based on two-dimensional and three-dimensional finite element modelling.

Journal Article↗

PubMed2000

Purification and characterization of cytokine-inducing protein of seed extract from Aeginetia indica L., a parasitic plant.

We have isolated 55 kDa protein from the seed extract of Aeginetia indica L. (AIL), a parasitic plant, by an affinity chromatography on N-hydroxysuccinimide (NHS)-activated Sepharose High Performance column bound F3 monoclonal antibody which neutralizes cytokine-inducing and antitumor effect of AIL. In in vitro model using human peripheral blood mononuclear cells (PBMC), the 55 kDa protein (AILb-A) induced multiple cytokines, such as IFN-gamma, tumor necrosis factor (TNF)-alpha, granulocyte macrophage-colony stimulating factor (GM-CSF), IL-2, IL-6, IL-10, IL-12 and IL-18, and also accelerated killer cell activities of PBMC. When compared with a commonly used immunotherapeutic agent OK-432, AILb-A induced Th1 cytokines are greater than OK-432. Of the Th2 cytokines, the amounts of IL-6 and IL-10 induced by AILb-A were lower than those by OK-432. No significant induction of IL-4 and IL-13 was observed in AILb-A-stimulated PBMC. TNF family including TNF-alpha, TNF-beta, Fas ligand (FasL) and TNF-related apoptosis-inducing ligand (TRAIL) were suggested to be important for AILb-A-induced killing activity of PBMC by reverse transcription-polymerase chain reaction (RT-PCR) analysis. Furthermore, the neutralizing test using cytokine-specific antibodies demonstrated that IL-18 plays a most significant role for IFN-gamma- and killer cell-inducing ability of AILb-A among the cytokines tested. These findings clearly indicated that AILb-A, a 55 kDa protein of AIL, is a potent Th1 cytokine inducer and may be a useful immunotherapeutic agent for the patients with malignancies.

Adjuvants, Immunologic↗