RESUMEN
The Fc receptor I for IgA (FcαRI) down-regulates humoral immune responses and modulates the risk of autoimmunity. This study aimed to investigate whether FcαRI targeting can affect progression of pristine-induced lupus nephritis. In the first experiment (early intervention), four groups of animals were evaluated: untreated FcαRI/FcRγ transgenic (Tg) mice and Tg mice administered control antibody (Ctr Fab), saline and anti-FcαRI Fab [macrophage inflammatory protein (MIP)-8a], respectively, three times a week for 29 weeks, after being injected once intraperitoneally with 0·5 ml pristane. In the second experiment, antibody injection started after the onset of nephritis and was carried out for 2 months, with similar groups as described above. MIP-8a improved proteinuria, decreased the amounts of glomerular injury markers, serum interleukin (IL)-6, IL-1 and monocyte chemoattractant protein (MCP)-1, and F4/80 macrophages in the interstitium and glomeruli, in both experiments. When MIP-8a was used as early intervention, a decrease in mouse serum anti-nuclear antibody (ANA) titres and reduced deposition of immunoglobulins in glomeruli were observed. This effect was associated with reduced serum titres of immunoglobulin (Ig)G2a but not IgG1, IgG2b and IgG3. Furthermore, pathological analysis showed lower glomerular activity index and less fibronectin in MIP-8a treated mice. This study suggests that FcαRI targeting could halt disease progression and lupus activation by selective inhibition of cytokine production, leucocyte recruitment and renal inflammation. Our findings provide a basis for the use of FcαRI as a molecular target for the treatment of lupus.
Asunto(s)
Anticuerpos Monoclonales/farmacología , Nefritis Lúpica/prevención & control , Terapia Molecular Dirigida/métodos , Receptores Fc/antagonistas & inhibidores , Animales , Anticuerpos Monoclonales/inmunología , Antígenos CD/genética , Antígenos CD/inmunología , Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Citocinas/sangre , Citocinas/inmunología , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Riñón/efectos de los fármacos , Riñón/patología , Riñón/ultraestructura , Nefritis Lúpica/inducido químicamente , Nefritis Lúpica/inmunología , Ratones Endogámicos C57BL , Ratones Transgénicos , Microscopía Electrónica , Receptores Fc/genética , Receptores Fc/inmunología , Terpenos , Factores de TiempoRESUMEN
BACKGROUND: The perioperative immune status of colorectal robotic surgery (RS), laparoscopic surgery (LS), and open surgery (OS) patients has not been compared. Our aim was to evaluate perioperative stress and immune response after RS, LS and OS. METHODS: This prospective study included 46 colorectal surgery patients from the Department of Surgical Oncology of the University of Tokyo Hospital. Peripheral venous blood samples were obtained preoperatively and on postoperative days 1, 3, and 6. We evaluated expression of HLA-DR (marker of immune competence), C-reactive protein (CRP) levels, and lymphocyte subset counts (natural killers, cytotoxic T cells and helper T cells). RESULTS: Fifteen, 23, and 8 patients underwent RS, LS and OS, respectively. HLA-DR expression was the lowest on day 1 and gradually increased on days 3 and 6 in all the groups. There was no significant difference in postoperative HLA-DR expression between the RS and LS group. However, on day 3, HLA-DR expression in the RS group was significantly higher than in the OS group (p = 0.04). On day 1, CRP levels in the LS group were significantly lower than in the RS group (p = 0.038). There were no significant perioperative changes in the lymphocyte subset cell count between the three groups. CONCLUSIONS: Perioperative surgical stress, as evaluated by immunological parameters, was comparable between robotic and laparoscopic surgery and higher with open surgery. Robotic surgery may be an alternative to laparoscopic surgery, as a minimally invasive surgery option for colorectal cancer.
Asunto(s)
Neoplasias del Colon/cirugía , Laparoscopía , Neoplasias del Recto/cirugía , Procedimientos Quirúrgicos Robotizados , Estrés Fisiológico/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores/metabolismo , Proteína C-Reactiva/metabolismo , Neoplasias del Colon/inmunología , Femenino , Antígenos HLA-DR/sangre , Humanos , Recuento de Linfocitos , Linfocitos/citología , Masculino , Persona de Mediana Edad , Periodo Perioperatorio , Periodo Posoperatorio , Estudios Prospectivos , Neoplasias del Recto/inmunología , Neoplasias del Recto/patologíaRESUMEN
The cell-surface molecule Cd9, a member of the transmembrane-4 superfamily, interacts with the integrin family and other membrane proteins. and is postulated to participate in cell migration and adhesion. Expression of Cd9 enhances membrane fusion between muscle cells and promotes viral infection in some cells. Fertilization also involves membrane fusion, between gametes. In mammals, the sperm binds to microvilli on the egg surface, and sperm-egg membrane fusion first occurs around the equatorial region of the sperm head12. The fused membrane is then disrupted, and the sperm nucleus as well as the cytoplasm is incorporated into the egg. Cd9 is expressed on the plasma membrane of the mouse egg, and an anti-Cd9 monoclonal antibody inhibits sperm-egg surface interactions. We generated Cd9 mice and found that homozygous mutant females were infertile. Sperm-egg binding was normal, but sperm-egg fusion was almost entirely inhibited in eggs from Cd9 females. Intracellular Ca2 oscillations, which signal fertilization, were absent in almost all mutant eggs; in rare cases, a response occurred after a long time period. In normal animals, Cd9 molecules were expressed on the egg microvilli and became densely concentrated at the sperm attachment site. Thus, our results show that Cd9 is important in the gamete fusion process at fertilization.
Asunto(s)
Antígenos CD/fisiología , Fusión Celular , Glicoproteínas de Membrana , Óvulo/metabolismo , Interacciones Espermatozoide-Óvulo , Animales , Antígenos CD/genética , Señalización del Calcio , Adhesión Celular , Femenino , Infertilidad Femenina/genética , Masculino , Ratones , Ratones Noqueados , Microvellosidades/metabolismo , Óvulo/ultraestructura , Tetraspanina 29RESUMEN
Myeloid FcαRI, a receptor for immunoglobulin (Ig)A, mediates cell activation or inhibition depending on the type of ligand interaction, which can be either multivalent or monovalent. Anti-inflammatory signalling is triggered by monomeric targeting using anti-FcαRI Fab or IgA ligand binding, which inhibits immune and non-immune-mediated renal inflammation. The participation of Toll-like receptors (TLRs) in kidney pathology in experimental models and various forms of human glomerular nephritis has been discussed. However, little is known about negative regulation of innate-immune activation. In the present study, we generated new transgenic mice that express FcαRI(R209L) /FcRγ chimeric protein and showed that the monovalent targeting of FcαRI exhibited inhibitory effects in an in vivo model of TLR-9 signalling-accelerated nephritis. Mouse monoclonal anti-FcαRI MIP8a Fab improved urinary protein levels and reduced the number of macrophages and immunoglobulin deposition in the glomeruli. Monovalent targeting using MIP8a Fab attenuates the TLR-9 signalling pathway and is associated with phosphorylation of extracellular signal-related protein kinases [extracellular signal-regulated kinase (ERK), P38, c-Jun N-terminal kinase (JNK)] and the activation of nuclear factor (NF)-κB. The inhibitory mechanism involves recruitment of tyrosine phosphatase Src homology 2 domain-containing phosphatase-1 (SHP-1) to FcαRI. Furthermore, cell transfer studies with macrophages pretreated with MIP8a Fab showed that blockade of FcαRI signalling in macrophages prevents the development of TLR-9 signalling-accelerated nephritis. These results suggest a role of anti-FcαRI Fab as a negative regulator in controlling the magnitude of the innate immune response and a new type of anti-inflammatory drug for treatment of kidney disease.
Asunto(s)
Antígenos CD/inmunología , Glomerulonefritis/inmunología , Inmunoglobulina A/inmunología , Receptores Fc/inmunología , Receptor Toll-Like 9/metabolismo , Animales , Anticuerpos Monoclonales , Antígenos CD/metabolismo , Línea Celular , Ensayo de Inmunoadsorción Enzimática , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Glomerulonefritis/metabolismo , Glomerulonefritis/patología , Humanos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Riñón/inmunología , Riñón/patología , Glomérulos Renales/inmunología , Glomérulos Renales/patología , Macrófagos/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , FN-kappa B/metabolismo , Oligodesoxirribonucleótidos/farmacología , Proteínas Tirosina Fosfatasas/metabolismo , Receptores Fc/metabolismo , Transducción de Señal , Receptor Toll-Like 9/inmunología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
OBJECTIVE: Severe hypoglycemia is an important but uncommon complication of anorexia nervosa. A 35-year-old woman showed severe hypoglycemia after a recovery from severe liver dysfunction due to malnutrition. METHODS: To reveal the cause of severe hypoglycemia, we measured plasma hormones and performed a 75g oral glucose loading test. RESULTS: Fasting serum adrenocorticotropic hormone, cortisol, growth hormone, somatostatin, and active ghrelin were elevated. Serum free triiodothyronine, leptin, and adiponectin were reduced. Plasma glucose fluctuated from 67 to 76 mg/dl after a 75g glucose ingestion, without hyperinsulinemia. Serum growth hormone, somatostatin, and active ghrelin levels were decreased after glucose ingestion. Plasma glucagon levels were increased and remained at high levels at 120 min, and glucose-dependent insulinotropic polypeptide (GIP) levels were continuously and remarkably increased after glucose ingestion. CONCLUSION: We observed a strongly reduced sensitivity in glucagon-induced hepatic glycogenolysis, and significantly elevated fasting and postprandial GIP levels, and a defective GIP-mediated glucagon secretion, in an anorectic patient with severe hypoglycemia.
Asunto(s)
Anorexia Nerviosa/complicaciones , Polipéptido Inhibidor Gástrico/sangre , Hipoglucemia/etiología , Adulto , Anorexia Nerviosa/sangre , Glucemia/metabolismo , Femenino , Glucagón/metabolismo , Hormonas/sangre , Humanos , Hepatopatías/etiología , Pruebas de Función HepáticaRESUMEN
BACKGROUND: Signals from the B-cell antigen receptor (BCR) help to determine B-cell fate, directing either proliferation, differentiation, or growth arrest/apoptosis. The protein tyrosine phosphatase SHP-1 is known to regulate the strength of BCR signaling. Although the B-cell co-receptor CD22 binds SHP-1, B cells in CD22-deficient mice are much less severely affected than those in SHP-1-deficient mice, suggesting that SHP-1 may also regulate B-cell signaling by affecting other signaling molecules. Moreover, direct substrates of SHP-1 have not been identified in any B-cell signaling pathway. RESULTS: We identified the B-cell transmembrane protein CD72 as a new SHP-1 binding protein and as an in vivo substrate of SHP-1 in B cells. We also defined the binding sites for SHP-1 and the adaptor protein Grb2 on CD72. Tyrosine phosphorylation of CD72 correlated strongly with BCR-induced growth arrest/apoptosis in B-cell lines and in primary B cells. Preligation of CD72 attenuated BCR-induced growth arrest/death signals in immature and mature B cells or B-cell lines, whereas preligation of CD22 enhanced BCR-induced growth arrest/apoptosis. CONCLUSIONS: We have identified CD72 as the first clear in vivo substrate of SHP-1 in B cells. Our results suggest that tyrosine-phosphorylated CD72 may transmit signals for BCR-induced apoptosis. By dephosphorylation CD72. SHP-1 may have a positive role in B-cell signaling. These results have potentially important implications for the involvement of CD72 and SHP-1 in B-cell development and autoimmunity.
Asunto(s)
Antígenos CD/metabolismo , Antígenos de Diferenciación de Linfocitos B/metabolismo , Linfocitos B/fisiología , Proteínas Tirosina Fosfatasas/metabolismo , Receptores de Antígenos de Linfocitos B/fisiología , Animales , Linfocitos B/inmunología , División Celular , Línea Celular , Células Cultivadas , Péptidos y Proteínas de Señalización Intracelular , Ratones , Ratones Endogámicos BALB C , Fosforilación , Fosfotirosina , Proteína Tirosina Fosfatasa no Receptora Tipo 11 , Proteína Tirosina Fosfatasa no Receptora Tipo 6 , Proteínas Recombinantes de Fusión/metabolismo , Proteínas Tirosina Fosfatasas con Dominio SH2 , Transducción de Señal , Bazo/inmunología , Especificidad por Sustrato , Transfección , Dominios Homologos srcRESUMEN
To study the effects of IL-1 alpha in arthritis, we generated human IL-1 alpha (hIL-1 alpha). Transgenic mice expressed hIL-1 alpha mRNA in various organs, had high serum levels of hIL-1 alpha, and developed a severe polyarthritic phenotype at 4 weeks of age. Not only bone marrow cells but also synoviocytes from knee joints produced biologically active hIL-1 alpha. Synovitis started 2 weeks after birth, and 8-week-old mice showed hyperplasia of the synovial lining layer, the formation of hyperplastic synovium (pannus) and, ultimately, destruction of cartilage. Hyperplasia of the synovial lining was due to the accumulation of macrophage-like cells expressing F4/80 molecules. hIL-1 alpha was widely distributed in macrophage- and fibroblast-like cells of the synovial lining cells, as well as synovial fluid monocytes. T and B cells were rare in the synovial fluid, and analysis of marker expression suggests that synoviocytes were directly histolytic and did not act as antigen-presenting cells. In the joints of these mice, we found elevated levels of cells of the monocyte/macrophage and granulocyte lineages and of polymorphonuclear neutrophils (PMNs), most of which expressed Gr-1, indicating that they were mature, tissue-degrading PMNS: Cultured synoviocytes and PMNs from these animals overexpress GM-CSF, suggesting that the hematopoietic changes induced by IL-1 and the consequent PMN activation and joint destruction are mediated by this cytokine.
Asunto(s)
Artritis/etiología , Interleucina-1/biosíntesis , Macrófagos , Neutrófilos , Membrana Sinovial/patología , Animales , Artritis/genética , Artritis/inmunología , Artritis/patología , Linaje de la Célula , Factor Estimulante de Colonias de Granulocitos y Macrófagos , Células Madre Hematopoyéticas , Humanos , Interleucina-1/genética , Ratones , Ratones Transgénicos , Monocitos , Fenotipo , Proteínas Recombinantes/biosíntesisRESUMEN
Six murine monoclonal antibodies were found to react with ganglioside GD2 lactone as well as purified ganglioside GD2. However, the reactivities of these antibodies to various ganglioside lactones were found to differ from each other. Four antibodies only reacted with GD2 lactones, while the other two cross-reacted with lactones of other gangliosides such as GD1b and GT1b.
Asunto(s)
Anticuerpos Monoclonales , Gangliósidos/inmunología , Animales , Química Encefálica , Bovinos , Cromatografía en Capa Delgada , Gangliósidos/aislamiento & purificación , Lactonas , Relación Estructura-ActividadRESUMEN
Alcoholic hepatitis leads to major derangements in lipoprotein metabolism. This study defines the characteristics of the abnormal high density lipoprotein and very low density lipoprotein in relation to the severity of the disease. In severely affected subjects very low density lipoprotein apolipoproteins were deficient in apolipoprotein E and apolipoprotein C. The concentration of high density lipoprotein was markedly reduced, although the proportion of high density lipoprotein 1 was substantially elevated when compared to normal subjects. High density lipoproteins were deficient in apolipoprotein AI and apolipoprotein AII but enriched in apolipoprotein E, apolipoprotein E complexes and apolipoprotein C, and contained a mixture of particles. The high density lipoprotein of subjects with alcoholic hepatitis contained a high proportion of material which bound to heparin affinity columns. This bound fraction contained a group of particles rich in apolipoprotein E, apolipoprotein E complexes and apolipoprotein C and was deficient in apolipoprotein AI and apolipoprotein AII. Examination by electron microscopy showed the presence of both discoidal and spherical particles, which varied in concentration according to the severity of the disease. Another fraction of high density lipoprotein, not bound to heparin, contained reduced amounts of apolipoprotein AI and apolipoprotein AII, consisted of disc-shaped particles and showed a higher esterified: free cholesterol ratio than the other high density lipoprotein fraction.
Asunto(s)
Apolipoproteínas A , Apolipoproteínas/sangre , Hepatitis Alcohólica/sangre , Apolipoproteína A-I , Apolipoproteína A-II , Apolipoproteínas B , Apolipoproteínas C , Apolipoproteínas E , Humanos , Lipoproteínas HDL/sangre , Lipoproteínas VLDL/sangre , Microscopía ElectrónicaRESUMEN
Four hybridoma clones (ACV-1, -3, -4, and -5) were established for Chinemys reevesii (Reeves' turtle) vitellogenin (VTG) as a precursor protein of egg yolk and a biomarker of environmental pollution. Binding-inhibition experiments indicated that the epitopes of four mAbs were distinct. No binding of ACV-4 to C. reevesii VTG in the Western blot suggests that the epitope of ACV-4 would be dependent on the three-dimensional structure. ACV-1, -3, and -5 bound to C. reevesii VTG in the Western blot. The signal for ACV-1 and -5 disappeared by reduction of the VTG, suggesting that the construction of the epitopes for ACV-1 and -5 were dependent on the disulfide bridge in the VTG molecule. All four mAbs recognized Trachemys scripta and Mauremys japonica VTGs in the ELISA. The yolk proteins were tested for the binding of the mAbs in the Western blot. ACV-1 being capable to bind to the VTG in the reduced condition did not bind to any protein bands of the yolk. This indicates that ACV-1 recognizes a part of the VTG molecule that is not incorporated in the oocytes. Both ACV-3 and -5 bound to the 32- and 70-kDa yolk proteins. Since a mAb recognizes only one site (epitope) on a protein molecule, the 32-kDa protein originated from the 70-kDa one. An ELISA system using ACV-5 as the capture antibody and ACV-3 as the detecting antibody showed the lower detectable concentration (2 ng/mL) and a wide detectable range to 1000 ng/mL (R2=0.999). The system was used to determine serum VTG levels of juvenile turtles treated with estradiol-17beta or vehicle (corn oil). By the use of the mAbs described in this paper, basic and applied studies for turtle VTGs would be improved.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos/inmunología , Tortugas/inmunología , Vitelogeninas/inmunología , Animales , Anticuerpos Monoclonales/biosíntesis , Anticuerpos Monoclonales/química , Western Blotting/métodos , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Estrógenos/farmacología , Masculino , Vitelogeninas/químicaRESUMEN
Binding data on inbred mouse strains and immunochemical isolation of Ia antigens with subsequent separation on non-reduced/reduced two-dimensional gels provide evidence for the cross-reactivity of monoclonal antibodies with I-A and I-E products. Thus two monoclonal antibodies were found to react with A alpha A beta as well as E alpha E beta dimers. One of these mAbs, K22 -42, reacts with the precursor form of E beta chain of B10.GD mice which is associated with the invariant chain (Ii). This indicates that the respective determinant on E beta is formed prior to association of E beta with E alpha.
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Epítopos/genética , Antígenos de Histocompatibilidad Clase II/inmunología , Antígenos de Histocompatibilidad/inmunología , Polimorfismo Genético , Animales , Anticuerpos Monoclonales/inmunología , Reacciones Cruzadas , Electroforesis en Gel de Poliacrilamida , Epítopos/análisis , Ratones , Ratones Endogámicos BALB CRESUMEN
Some forms of familial Alzheimer's disease are caused by mutations in the amyloid beta protein precursor (beta APP), and there is excellent evidence that these mutations foster amyloid deposition by increasing secretion of total amyloid beta protein (A beta) or the highly amyloidogenic A beta 1-42 form. These observations provide a powerful rationale for developing an animal model of AD by generating transgenic mice in which cerebral amyloid deposition is induced by A beta overproduction. To produce substantial A beta in vivo, we generated mice expressing the transgene of signal peptide and 99 residues of carboxyl-terminal fragment (CTF) of beta APP under control of the cytomegalovirus enhancer/chicken beta-actin promoter. The transgenic mRNA was detected in many tissues of these mice, but the levels of transgenic mRNA, CTF, and A beta did not correlate well indicating that tissue-specific posttranslational processing may play an important role in determining the amount of A beta that accumulates in various tissues. A beta was detected biochemically in brain, kidney, and pancreas with the largest amount present in pancreas. In transgenic plasma, there was a marked accumulation of human A beta 1-40 and A beta 1-42(43) to levels over 30-times those observed in normal human plasma. Thus, the transgenic mice produce and secrete considerable A beta. Despite this increase in A beta secretion and the elevated A beta in brain, immunohistochemistry revealed no consistent cerebral A beta deposition. In pancreas, however, intracellular A beta deposits were detected immunohistochemically in acinar cells and interstitial macrophages, some of which showed severe degeneration. In addition, examination of these cells by immunoelectron microscopy revealed many putative amyloid fibrils (7-12 nm) that were stained by anti-A beta antibodies. Overall, our findings indicate that tissue-specific posttranslational processing may play a pivotal role in A beta production and amyloid fibril formation in vivo. By carefully analyzing the changes that occur in the transgenic mice described here as compared to the transgenic line that has recently been shown to form extracellular amyloid plaques in brain, it may be possible to gain considerable insight into the factors that determine the location and amount of A beta that accumulates as amyloid.
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Péptidos beta-Amiloides/metabolismo , Neurofibrillas/metabolismo , Páncreas/metabolismo , Péptidos beta-Amiloides/genética , Animales , Secuencia de Bases , Northern Blotting , Western Blotting , Citomegalovirus/genética , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunohistoquímica , Ratones , Ratones Transgénicos , Microscopía Electrónica , Datos de Secuencia Molecular , Neurofibrillas/ultraestructura , Páncreas/ultraestructura , Reacción en Cadena de la Polimerasa , TransgenesRESUMEN
We have identified and analyzed a 27-nucleotide sequence (U5 repressive element, designated as U5RE) at the U5 region of the human T-cell leukemia virus type I (HTLV-I) long terminal repeat (LTR) which is required for HTLV-I basal transcriptional repression. The basal promoter strength of constructs that contained deletions in the U5 region of the LTR was analyzed by chloramphenicol acetyltransferase (CAT) assays following transfection of HeLa cells or Jurkat T-cells in the presence or absence of viral transactivator tax protein. We consistently observed a 2- to 5-fold increase in basal promoter activity when sequences between +277 to +306 were deleted. In vivo competition experiments suggested that the U5 DNA fragment from +269 to +295 contains a functional repressive element (U5RE). Using gel mobility shift assays, we have purified a highly enriched fraction that could specifically bind U5RE. This DNA affinity column fraction contained three major detectable proteins on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with silver staining: 110-, 80- and 70-kDa proteins. The 110-kDa protein appeared to be a novel DNA-binding protein whose characteristics are still obscure, while the 70- and 80-kDa proteins were shown to be related to the human autoantigen Ku, the Ku (p70/p80) complex, as demonstrated by amino acid sequencing and immunological analyses. As Ku is known to be involved in transcriptional regulation, the specific interaction of Ku with U5RE raises intriguing possibilities for its function in HTLV-I basal transcriptional repression.
Asunto(s)
Antígenos Nucleares , Autoantígenos/metabolismo , ADN Helicasas , ADN Viral/metabolismo , Proteínas de Unión al ADN/metabolismo , Regulación Viral de la Expresión Génica , Virus Linfotrópico T Tipo 1 Humano/genética , Proteínas Nucleares/metabolismo , Secuencias Repetitivas de Ácidos Nucleicos , Secuencia de Aminoácidos , Secuencia de Bases , Línea Celular , Cloranfenicol O-Acetiltransferasa/genética , Proteínas de Unión al ADN/aislamiento & purificación , Productos del Gen tax/metabolismo , Células HeLa , Virus Linfotrópico T Tipo 1 Humano/metabolismo , Humanos , Autoantígeno Ku , Datos de Secuencia Molecular , Sondas de Oligonucleótidos , Unión Proteica , Secuencias Reguladoras de Ácidos Nucleicos , Linfocitos T , Transcripción GenéticaRESUMEN
Nineteen hypercholesterolemic patients (10 without and 9 with hypertriglyceridemia) were given evening primrose oil rich in gammalinolenic acid (GLA, 18: 3n - 6), in a placebo controlled cross-over design, over 16 weeks (8 + 8 weeks), with safflower oil as the placebo. During supplementation with evening primrose oil, dihomogammalinolenic acid (20: 3n - 6) increased in plasma lipids and red blood cells, and in subjects without hypertriglyceridemia there was a significant decrease in low density lipoprotein-cholesterol and plasma apolipoprotein B compared with the levels observed during safflower oil administration. Our results confirmed that evening primrose oil is effective in lowering low density lipoprotein in hypercholesterolemic patients.
Asunto(s)
Apolipoproteínas/sangre , Grasas Insaturadas en la Dieta/farmacología , Hipercolesterolemia/sangre , Ácidos Linolénicos/farmacología , Lipoproteínas/sangre , Adulto , Colesterol/sangre , Método Doble Ciego , Ácidos Grasos Esenciales/farmacología , Femenino , Humanos , Hipercolesterolemia/dietoterapia , Ácidos Linoleicos , Masculino , Persona de Mediana Edad , Oenothera biennis , Fosfolípidos/sangre , Aceites de Plantas , Distribución Aleatoria , Aceite de Cártamo/farmacología , Triglicéridos/sangre , Ácido gammalinolénicoRESUMEN
Several antihypertensive agents such as thiazide diuretics and some beta-blockers have recently been shown to adversely affect lipid metabolism. Moreover, there is a growing suspicion that the adverse effect on plasma lipids might outweigh the favourable effect of lowering blood pressure. The effect of ketanserin tartrate (20 to 60 mg daily), a new antihypertensive drug, on blood lipids was evaluated in a 12-week non-comparative clinical trial in 34 patients with mild or moderate hypertension. Ketanserin reduced systolic and diastolic blood pressure by 12.2 and 9.8%, respectively, without altering heart rates. Total cholesterol and low density lipoprotein (LDL)-cholesterol levels in the fasting plasma were observed to decrease significantly by 6.3 and 8.8% respectively, whereas mean triglyceride and high density lipoprotein (HDL)-cholesterol remained almost unchanged. These changes were consistent irrespective of their initial values. Significant decrease in apolipoprotein B and E was also observed. Apolipoprotein A-I, A-II, C-II and C-III were not altered significantly. It is speculated that ketanserin affects mainly LDL-cholesterol. Based on these findings, ketanserin is considered to have a potentially beneficial effect on coronary risk profile and should be given full consideration when drug therapy is selected for patients with mild to moderate hypertension.
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Hipertensión/tratamiento farmacológico , Ketanserina/uso terapéutico , Lípidos/sangre , Adulto , Anciano , Ensayos Clínicos como Asunto , Femenino , Humanos , Hipertensión/sangre , Ketanserina/efectos adversos , Ketanserina/farmacología , Masculino , Persona de Mediana EdadRESUMEN
A new kind of myeloproliferative disorder (L-8313) has been discovered. It was transplantable into syngeneic mice with spleen cells. The mice showed hepato-splenomegaly with a marked leukocytosis and anemia 3 weeks after transplantation of L-8313 cells. The number of GM-CFU and CFU-S per spleen increased to more than 40 times normal. The results of chromosomal and PGK analysis demonstrated that these increased stem cells were of host origin. Both the culture medium of the spleen cells and the serum from L-8313 bearing mice showed high levels of IL-3, BPA and CSF. Consequently, hematopoietic cells of the host mice underwent remarkable proliferation in response to these stimulating factors when L-8313 cells were transplanted. We also have been successful in establishing an in-vitro cell line and have maintained it for over one year. The phenotype of L-8313 cells was Thy 1.2 positive. Some L-8313 cells showed a positive acid phosphatase reaction but the cytochemical character of myeloid lineage was not observed. Therefore, L-8313 is considered to be a T-cell derived hematopoietic regulatory cell neoplasm with the ability to produce several hematopoietic stimulating factors.
Asunto(s)
Factores Estimulantes de Colonias/biosíntesis , Sustancias de Crecimiento/biosíntesis , Interleucina-3/metabolismo , Trastornos Mieloproliferativos/etiología , Anemia/etiología , Animales , Antígenos de Superficie/análisis , Línea Celular , Femenino , Factores de Crecimiento de Célula Hematopoyética , Células Madre Hematopoyéticas/patología , Masculino , Ratones , Ratones Endogámicos C3H , Trastornos Mieloproliferativos/sangre , Trastornos Mieloproliferativos/patología , Proteínas de Neoplasias/metabolismo , Trasplante de Neoplasias , Fenotipo , Fosfoglicerato Quinasa/análisis , Bazo/patología , Esplenomegalia/etiología , Linfocitos T/metabolismo , Linfocitos T/patología , Antígenos Thy-1RESUMEN
We have established a novel monoclonal antibody that recognises mouse and rat CD157, and uncovered striking differences in both the level and stage of expression of this antigen in the primary lymphoid organs between these two species. Unlike mouse, the majority of rat thymocytes express CD 157. SHR and WKY rats were the exception, having unusually low levels (similar to those of the mouse) of these cells. However, in both species, a subset of CD3- CD4- CD8- thymocytes exhibited high levels of CD157. Surprisingly, these CD157high cells temporarily upregulated MHC class I molecules in both species. Furthermore, a third of CD157high rat thymocytes were CD45RC+, a marker found on immature thymocytes with regenerative capacity. Examination of the bone marrow lymphoid population shows that the expression of rat CD157 is largely observed at the CD45R+ IgM- pre-B-II cell stage, and unlike mouse, extension of expression into the IgM+ immature B cell stage was marginal. Similar to CD157high immature thymocytes, these immature B cells also expressed high levels of MHC class I. With the exception of the LEC, SHR and WKY rat strains, which have three- to four-fold less CD157+ bone marrow myeloid cells, percentages of these cells are similar between these two species. Thus, marked differences in the level and stage(s) of CD157 expression on lymphoid cells in mouse and rat indicate that CD157 may not, as previously thought, have a direct role in T or B cell differentiation.
Asunto(s)
ADP-Ribosil Ciclasa , Antígenos CD , Biomarcadores/análisis , Linfocitos/inmunología , Glicoproteínas de Membrana/farmacología , Ratones Endogámicos/inmunología , Ratas Endogámicas/inmunología , Animales , Anticuerpos Monoclonales , Especificidad de Anticuerpos , Células de la Médula Ósea/inmunología , Células CHO , Diferenciación Celular , Células Cultivadas , Cricetinae , Proteínas Ligadas a GPI , Subgrupos Linfocitarios/inmunología , Ratones , Ratas , Especificidad de la Especie , Timo/inmunología , Distribución TisularRESUMEN
The fine structural specificities of six monoclonal antibodies (MAbs) to ganglioside GD2, GalNAc beta 1----4(NeuAc alpha 2----8NeuAc alpha 2----3)Gal beta 1----4Glc-Cer, were studied. The binding specificities of these MAbs were found to differ from each other by virtue of their binding to structurally related authentic standard glycolipids as revealed by three different assay systems, including enzyme immunostaining on thin-layer chromatography, enzyme-linked immunosorbent assay, and immune adherence inhibition assay. The MAbs examined could be divided into three binding types. MAbs A1-201, A1-410, and A1-425 bound specifically to ganglioside GD2 and none of the other gangliosides tested. Two other MAbs (A1-245 and A1-267) reacted not only with GD2, but also with several other gangliosides having the sequence NeuAc alpha 2----8NeuAc alpha 2----3Gal (GD3, GD1b, GT1a, GT1b, and GQ1b). The reactivities with these gangliosides varied to some degree. In addition, these MAbs were found to react with both GD3(NeuAc-NeuAc) and GD3(NeuGc-NeuAc), but not with GD3(NeuAc-NeuGc) or GD3(NeuGc-NeuGc). The last MAb (A1-287) also reacted with several other gangliosides but with lower avidity than A1-245 and A1-267. These findings suggest that each MAb to ganglioside GD2 may have an individual binding specificity and avidity. These MAbs represent potentially useful reagents for analyzing the function of GD2 on cell surface membranes, and provide a system for precisely studying the interactions between an anti-ganglioside antibody and the binding epitope of the antigenic determinant.
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Anticuerpos Monoclonales/inmunología , Gangliósidos/inmunología , Animales , Línea Celular , Cromatografía en Capa Delgada , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Glucolípidos/inmunología , Glucolípidos/aislamiento & purificación , Humanos , Reacción de Inmunoadherencia , Inmunoquímica , RatonesRESUMEN
We previously reported the binding specificities of two anti-ganglioside GD2 murine monoclonal antibodies (MAbs), A1-425 and A1-267, both of which are of IgG3 isotype. A1-425 reacts specifically with ganglioside GD2, whereas A1-267 binds preferentially to GD2 but also reacts with GD3 [Tai, T., Kawashima, I., Tada, N., & Dairiki, K. (1988) J. Biochem. 103, 682-687]. In this paper, they were used for comparative analyses of antibody-mediated cytotoxicity, i.e., antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) against human melanoma and neuroblastoma cell lines. Melanoma cells were found to contain GD2 and/or GD3, whereas neuroblastoma cells expressed only GD2. Both antibodies induced high levels of ADCC and CDC to GD2/GD3-positive cells with human peripheral large granular lymphocytes (LGL) as effector cells and in the presence of human serum, respectively. A good correlation was obtained between the contents of disialogangliosides and the binding level of the antibodies; both melanoma and neuroblastoma cells with larger amounts of GD2/GD3 showed a higher level of antibody binding than did the cells with a smaller amount of GD2/GD3. Surprisingly, ADCC did not correlate well with the binding level of the antibodies. Thus, A1-425 showed stronger lytic activity than A1-267 in spite of the binding level of A1-425 being similar to or lower than that of A1-267 on the cell surfaces. Antigen-antibody complexes composed of GD2 and A1-425 showed higher binding levels to LGL than complexes of GD2 and A1-267. In contrast, free MAb molecules gave minimum binding to LGL.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Anticuerpos Monoclonales , Citotoxicidad Celular Dependiente de Anticuerpos , Gangliósidos/inmunología , Melanoma/inmunología , Neuroblastoma/inmunología , Animales , Química Encefálica , Bovinos , Cromatografía en Capa Delgada , Pruebas Inmunológicas de Citotoxicidad , Glicoesfingolípidos/análisis , Humanos , Técnicas para Inmunoenzimas , Ratones , Receptores Fc/inmunología , Receptores Fc/metabolismo , Células Tumorales Cultivadas/inmunologíaRESUMEN
Protein C (PC) is a vitamin K-dependent serine protease, a deficiency of which results in thrombus. There is no spontaneously occurring mouse model of the disease. Attempts to create such a model in mice by using anti-sense gene technology requires isolation of a normal mouse PC cDNA. When a mouse liver (BALB/c) cDNA library was screened using a human PC cDNA as a probe, nine overlapping cDNA clones were isolated and sequenced. The cloned mouse PC cDNA comprised 1,512 nucleotides and the open reading frame of the cDNA encoded a polypeptide of 461 amino acids residues including a leader peptide composed of 41 amino acids. Mouse PC exhibited high homology to both human and bovine PCs. Mouse PC also had several structural features common in other PCs; locations of 23 Cys residues, location of putative beta-hydroxy Asp71, possible carbohydrate attachment sites involving Asp residues at amino acid positions 249, 314, and 330, and location of active sites such as His212, Asp258, and Ser361. Northern blot hybridization analysis identified a single species of mouse PC mRNA (2.0 kb in length) in mouse liver.