ABSTRACT
It is generally recognized that the accumulation of lipids and immuno-inflammatory cells are early signs of atherosclerosis. In the present study, we have investigated the relationship between the deposition of lipids, of immuno-inflammatory cells and the expression of HLA-DR molecules (a marker of immune activation), the molecules of the class II of major histocompatibility complex (MHC) in diffuse thickening of the intima (DIT). Lipids, including triglycerides, cholesterol esters, free cholesterol and phospholipids were studied by chromatography, Oil Red O histochemisty, as well as by electron microscopic analysis. Immuno-inflammatory cells and the expression of HLA-DR were investigated by immunohistochemistry in consecutive section of the same tissue samples. It has been shown that the lipids were unevenly distributed in DIT. In juxtaluminal sublayer, lipids were detected both in the cytoplasm of intimal cells and extracellularly. In the juxtamedial musculoelastic sublayer of the intima, lipids were present predominantly along elastic fibers. The positive correlation between the presence of lipids and the expression of HLA-DR was revealed (r = 0.79; P < 0.001). Also, a positive correlation was found between the deposition of lipids and the number of immune-inflammatory cells, although correlations was different for different sublayers of the intima. In particular, the correlation between the deposition of lipids and immune-inflammatory cells in the juxtaluminal sublayer of the intima was higher (r = 0.99; P < 0.001) than in the juxtamedial musculoelastic layer (r = 0.28; P < 0.001). These data support the hypothesis that postulates that the accumulation of lipids in the intima is a key factor in the initiation of inflammatory reactions. At the pre-atherosclerotic stage of the development of this disease, earlier pathological processes associated with lipid-dependent activation of immune cells occur mainly in the juxtaluminal portion of the intima.
Subject(s)
Aorta/metabolism , Atherosclerosis/metabolism , Lymphocytes/metabolism , Adult , Aorta/immunology , Aorta/ultrastructure , Atherosclerosis/immunology , Atherosclerosis/pathology , Carotid Intima-Media Thickness , Cholesterol Esters/metabolism , Gene Expression , HLA-DR Antigens/genetics , HLA-DR Antigens/immunology , Humans , Inflammation/immunology , Inflammation/metabolism , Inflammation/pathology , Lymphocytes/immunology , Lymphocytes/pathology , Male , Middle Aged , Phospholipids/metabolism , Triglycerides/metabolismABSTRACT
Dendritic cells were discovered and recognized as antigen-presenting cells in 1973. Since then, large volume of information has accumulated showing role of dendritic cells as a key element connecting the innate and adaptive immunity. Nowadays, dendritic cells are considered to be professional immune sensors capable of recognizing both antigen amounts and antigen persistence via complex mechanisms that involve decoding and integration of various signals received in a receptor-dependant manner. Tissue microenvironment plays an important role in the modulation of effector functions of dendritic cells leading either to activation or to suppression of immune reactions. Dendritic cells maintain the homeostasis and are involved in a number of diseases including infection diseases and cancer. The presence of dendritic cells in arteries has been reported in 1995. And since then the importance of dendritic cells in atherogenesis is intensively studied. This review briefly described current knowledge on dendritic cells and their role in atherogenesis.
Subject(s)
Antigen-Presenting Cells/immunology , Antigens , Atherosclerosis , Dendritic Cells/immunology , Antigen-Presenting Cells/ultrastructure , Antigens/immunology , Antigens/metabolism , Atherosclerosis/immunology , Atherosclerosis/therapy , Cell Communication , Dendritic Cells/ultrastructure , Humans , Immunity, Innate , Plaque, Atherosclerotic/immunology , Plaque, Atherosclerotic/ultrastructureABSTRACT
Increased cell proliferation in early atherosclerotic lesions is recognized as an essential event of atherogenesis but the levels of cell proliferation in different stages of atherosclerotic plague formation in different types of human large arteries are still insufficiently studied. In the present work, we studied intima thickness and proliferation of newly "infiltrates" hematogenous and resident cells in atherosclerotic lesions of the carotid and coronary arteries and compared these parameters with those in the aorta, reported by us in earlier publication. Analysis of intima thickness and proliferation in grossly unaffected intima and in different types pf atherosclerotic lesions (initial lesions, fatty streaks, lipofibrous, plaques, and fibrous plaque) revealed that although there were similar tendencies in the change of the infiltration levels of hematogenous cells and proliferation in different types of arteries, there were significant quantitative differences between different types of arteries. Hematogenous cells in lipofibrous plaques of the coronary and carotid arteries were found to account for a third and almost for a half of the total cell population, respectively, while atherosclerotic lesions in the aorta, as it has been shown by us earlier, to contain no more than 15% ofhematogenous cells. This suggests that the contribution of hematogenous cells to the development of atherosclerosis in the carotid and the coronary artery appears to be more significant than that in the aorta. Despite the differences in numbers of accumulating hematogenous cells in the intima, a similar "bell-shaped" dependence of cell numbers on the lesion type, involved in the following sequence: unaffected intima-initial lesions-fatty streaks-lipofibrous plaques-fibrous plaques, was detected in the coronary and carotid arteries. The visualization of proliferating cells (PCNA-positive) in atherosclerotic and unaffected zones of the coronary and carotid arteries revealed similar patterns. The maximum numbers of PCNA-positive resident cells were identified in lipofibrous plaques. The changes in the total cell numbers were accompanied by the changes in the numbers of both proliferating resident cells and proliferating hematogenous cells.
Subject(s)
Aorta/pathology , Atherosclerosis/pathology , Carotid Arteries/pathology , Carotid Stenosis/pathology , Coronary Vessels/pathology , Plaque, Atherosclerotic/pathology , Tunica Intima/pathology , Adult , Aged , Aorta/metabolism , Atherosclerosis/metabolism , Autopsy , Carotid Arteries/metabolism , Carotid Stenosis/metabolism , Cell Count , Cell Proliferation , Coronary Vessels/metabolism , Endothelium, Vascular/metabolism , Endothelium, Vascular/pathology , Female , Humans , Immunohistochemistry , Ki-67 Antigen/analysis , Ki-67 Antigen/immunology , Male , Microscopy, Electron , Middle Aged , Organ Specificity , Plaque, Atherosclerotic/metabolism , Proliferating Cell Nuclear Antigen/analysis , Proliferating Cell Nuclear Antigen/immunology , Severity of Illness Index , Tunica Intima/metabolismSubject(s)
Amniotic Fluid/physiology , Anencephaly/embryology , Brain/embryology , Nerve Tissue/embryology , Neurons/cytology , Cell Differentiation/physiology , Cells, Cultured , Culture Media , Embryonic and Fetal Development , Endothelium/cytology , Endothelium/physiology , Female , Humans , Organ Culture Techniques , PregnancyABSTRACT
The kinetics of [3H]-L-glutamate binding to brain synaptic membranes (SM) and to glutamate-binding proteins (GBP) was determined with agonist and monoclonal antibodies (MAbs). It was revealed, that rat and human brain GBP have individual protein components with M(r) from 14 to 92 kDa. Quisqualate inhibited [3H]-L-glutamate binding to solubilized and to purified 68 kDa protein component. MAbs have the most activity, and NMDA was failure. It has been shown that 68 kDa component antigen determinants are similar to those of bovine, frog and rat brain synaptic membranes. Anti-GBP monoclonal antibodies blocked functional non-NMDA receptors in isolated frog spinal cord. Immunocytochemistry was done on rat and human brain sections. Distribution of quisqualate receptors was determined with light and electron microscopy. Some properties of vertebrate CNS non-NMDA receptors are discussed.
Subject(s)
Brain/drug effects , Quisqualic Acid/metabolism , Receptors, Glutamate/drug effects , Adult , Animals , Antibodies, Monoclonal , Brain/metabolism , Brain/ultrastructure , Glutamates/immunology , Glutamates/metabolism , Humans , Immunohistochemistry , Ligands , Membrane Proteins/immunology , Membrane Proteins/metabolism , Microscopy, Immunoelectron , Protein Binding/drug effects , Rana ridibunda , Rats , Receptors, AMPA , Receptors, Glutamate/metabolism , Receptors, Glutamate/ultrastructureABSTRACT
A method has been developed for preparing essentially pure primary cultures of neurons from 10-12 week old fetal human brain. This method is based on the 1) capacity of neurons to form multicellular aggregates, 2) cultivation in chemically defined medium, and 3) treatment with cytosine arabinoside as antimitotic agent. This procedure allowed the preparation of highly purified (95 per cent and more) neuronal aggregate cultures. Ultrastructural analysis of these cultures following one and 8 days has revealed their partial differentiation during cultivation.
Subject(s)
Brain/ultrastructure , Neurons/ultrastructure , Brain/metabolism , Cell Aggregation , Cell Differentiation , Cells, Cultured/metabolism , Cells, Cultured/ultrastructure , Culture Media, Serum-Free , Cytarabine/pharmacology , Cytological Techniques , Embryo, Mammalian , Gestational Age , Humans , Immunohistochemistry , Microscopy, Electron , Mitosis/drug effects , Neurons/metabolismABSTRACT
Human thoracic aorta was taken from trauma victims aged from 6 to 40 years. Cell composition of the intimal layer was investigated in the lesion predisposed (LP) areas of the aorta and in the lesion resistant (LR) areas. By double immunofluorescent staining with mono and polyclonal antibodies to monocytes/macrophages (Mn/Mph) and smooth muscle cells (SMC) the presence of Mn/Mph was revealed in all aortas studied. The number of these cells was 2-6 fold higher in LP areas, compared to LR areas in all persons over 21 years of age. Scanning electron microscopy revealed Mn on the luminal surface of the vessel the number of which was frequently higher in LP areas. Mn/Mph in LP areas were heterogeneous in their structure depending on the depth of their localization in the intima. The revealed Mn/Mph infiltration in aortas of young subjects may be the earliest manifestation of the atherosclerotic lesion.
Subject(s)
Aorta, Thoracic/pathology , Arteriosclerosis/pathology , Macrophages/pathology , Monocytes/pathology , Adolescent , Adult , Aorta, Thoracic/ultrastructure , Child , Female , Fluorescent Antibody Technique , Humans , Macrophages/ultrastructure , Male , Microscopy, Electron, Scanning , Monocytes/ultrastructure , Time FactorsSubject(s)
Neuropeptides/therapeutic use , Parkinson Disease/therapy , Animals , Blood Transfusion, Autologous , Cells, Cultured , Chromatography, High Pressure Liquid , Electrochemistry , Humans , Neuropeptides/blood , Neuropeptides/cerebrospinal fluid , Rats , Rats, Inbred Strains , Spinal Cord/cytology , Spinal Cord/drug effects , gamma-Aminobutyric Acid/cerebrospinal fluidABSTRACT
The ultrastructure of human thoracic aortas collected at autopsy from 16 trauma victims aged from 3 to 40 years was investigated. Intercellular contacts were found in all aortas examined. The contacting cells localized predominantly in aortic areas predisposed to atherosclerosis, i. e. in areas situated around the ostia of the intercostal arteries. Most of contacting cells were monocytes--macrophages (Mn/Mph) which interacted with foam cell and intimal smooth muscle cells (SMCs). Only occasional contacts were formed by lymphocytes and endothelial cells. All intimal contacting SMCs were of the synthetic phenotype, i. e. they did not contain myofilaments and had well developed endoplasmic reticulum and Golgi apparatus. In thickened aortic intima the contacting cells were seen with the signs of destruction. The data obtained suggest that the contacts between intimal cells may be the consequence of the immune reaction directed to the transfer of information, cell cytolysis and utilization of cell debris.
Subject(s)
Aorta, Thoracic/ultrastructure , Arteriosclerosis/pathology , Cell Communication/physiology , Adolescent , Adult , Child , Child, Preschool , Foam Cells/pathology , Humans , Macrophages/pathology , Monocytes/pathology , Muscle, Smooth/pathology , Time FactorsSubject(s)
Antibodies, Monoclonal , Neurons, Afferent/physiology , Spinal Cord/physiology , Synaptic Transmission , Animals , Fluorescent Antibody Technique , Glutamates/metabolism , Humans , Membrane Proteins/immunology , Rana ridibunda , Receptors, Glutamate , Receptors, Neurotransmitter/immunology , Spinal Cord/cytologyABSTRACT
The study of the coronary arteries atherogenesis with the use of scanning and transmission electron microscopy was performed on the material of emergency necropsies of 45 patients who suffered the primary cardiovascular pathology. The transformation of macrophages into the foamy cells were studied in vitro with the use of peritoneal mouse macrophages and modified LDL labelled with colloidal gold as well as a method of phase-contrast cinemicrography. Two types of macrophages were distinguished ultrastructurally: scavenger-macrophages participating in the phagocytosis of lipoproteins and transforming into the foamy cells and secreting macrophages determining vascular wall homeostasis and ensuring the T-lymphocyte inclusion into the immune reactions and the contact with target cells. Balanced cell kinetics and cytokine secretion at early stages of atherosclerosis determine whether the lipid spot would regress or would transform into the atherosclerotic plaque.
Subject(s)
Arteriosclerosis/immunology , Macrophages/immunology , Monocytes/immunology , Adult , Arteriosclerosis/pathology , Autoimmune Diseases/pathology , Coronary Artery Disease/immunology , Coronary Artery Disease/pathology , Humans , Macrophages/metabolism , Macrophages/ultrastructure , Middle Aged , Monocytes/metabolism , Monocytes/ultrastructure , Phagocytosis/immunology , PhenotypeABSTRACT
Growth and differentiation of neurons and glia in spinal cord explants of 16 days old rat fetuses with teratogen-induced left-sided micromelia were studied. Progressive destruction of astrocytes that differentiate in interstitial zone of cultures was observed in 37% of explants of the left side, while the development was normal in cultures of the right side. Possible mechanisms leading to destruction of astrocytes in cultures of spinal cord regions that innervate anomalous limbs are discussed.
Subject(s)
Astrocytes/drug effects , Limb Deformities, Congenital , Spinal Cord/drug effects , 2,2'-Dipyridyl/toxicity , Animals , Astrocytes/ultrastructure , Cell Differentiation/drug effects , Cell Division/drug effects , Cell Survival/drug effects , Extremities/innervation , Female , Fetus , Microscopy, Electron , Microscopy, Electron, Scanning , Morphogenesis/drug effects , Organ Culture Techniques , Pregnancy , Prenatal Exposure Delayed Effects , Rats , Spinal Cord/ultrastructureABSTRACT
Dynamics concerning certain intercellular junctions have been followed during the preimplantation period of development in mouse embryos. The morphological analysis of the preimplantational embryos has demonstrated, that at the initial stages of cleavage (2-4 blastomeres) the cells make contacts by means of nonspecific junctions. Specialized intercellular junctions appear at the stage of 8 blastomeres and are presented as dotted tight and gap junctions. When the embryo is developing from the stage of 8 up to the stage of 16 blastomeres, certain connective complexes appear, consisting of dotted or cord-like tight and gap junctions. At the late morula stage, the external blastomeres in the apical part have contacts with each other by means of cingular tight junctions. In this place a connective complex might emerge; it is displayed as a tight junction and one or two gap junctions. At the blastocyst stage desmosomes and adhision zones appear. Between trophectodermal cells a connective complex arises; it is presented in the slice as a tight cingular junction, desmosomes (as a rule two) and an adhision zone. Between cells of the internal cellular mass the intercellular junctions are presented as dotted tight and gap junctions. Cells of the polar trophoectoderm and cells of the internal cellular mass could have contacts by means of gap and dotted tight junctions.
Subject(s)
Blastomeres/ultrastructure , Intercellular Junctions/ultrastructure , Animals , Gestational Age , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Microscopy, ElectronABSTRACT
The authors investigated the expression of cytoskeletal proteins and the ultrastructure of cells in normal intima and atheromatous plaque of human aorta. It has been established, using double immunofluorescent method and a set of antibodies that intimal smooth muscle cells /SMC/ of normal aorta express myosin, vimentin, alpha-actin and actin but not desmin. In seven out of 28 atherosclerotic plaques the cells contained desmin and all other SMC cytoskeletal proteins were found. These cells had the ultrastructural features of SMC, i.e. well-developed endoplasmic reticulum and Golgi apparatus. Besides, some cells in 13 atherosclerotic plaques proved to be myosin-, alpha-actin- and desmin-negative. The cells were stained with monoclonal antibodies specific to SMC but not with macrophage-specific antibody. Ultrastructurally, the cytoplasm of the cells was filled with rough endoplasmic reticulum and a developed Golgi complex, but a certain portion of the cells retained basal lamina and myofilament bundles. The peculiarities of cytoskeletal protein in expression and ultrastructure of cells in human aortic atherosclerotic plaques may be explained by a phenotypic modulation of vascular SMC.
Subject(s)
Arteriosclerosis/pathology , Genetic Variation , Muscle, Smooth, Vascular/ultrastructure , Adolescent , Adult , Aged , Aged, 80 and over , Aorta/metabolism , Aorta/ultrastructure , Arteriosclerosis/metabolism , Cytoskeletal Proteins/metabolism , Fluorescent Antibody Technique , Histocytochemistry , Humans , Microscopy, Electron , Middle Aged , Muscle Proteins/metabolism , Muscle, Smooth, Vascular/metabolism , PhenotypeABSTRACT
Proliferative activity of the aortic intima cells in the course of experimental rabbit hypercholesterolemia is studied. The most intensive DNA synthesis was found in macrophages and smooth cells transforming into the foam cells as shown by electron-microscopic radioautographic analysis. DNA synthesis by damaged and dying foam cells is shown. Cytophotometric analysis showed the small proportion of foam cells in the atherosclerotic lesions to be polyploid.
Subject(s)
Arteriosclerosis/metabolism , Blood Vessels/metabolism , DNA/biosynthesis , Foam Cells/metabolism , Macrophages/metabolism , Animals , Arteriosclerosis/pathology , Autoradiography , Blood Vessels/ultrastructure , Cell Division/physiology , DNA/analysis , Diet, Atherogenic , Foam Cells/ultrastructure , Male , Microscopy, Electron , Photometry , Polyploidy , RabbitsABSTRACT
Data are provided on cytodifferentiation of the cerebral cortex cultured cells taken from 10-12 week old embryos of man. It is shown that low differentiated neuroblasts well survive in culture for 21 days. Mature granular cells and middle pyramidal neurons are revealed in cultures. The number of morphological criteria may testify to the maturity of neurons: the presence of the Nissl substance, differentiation of dendrites and axons; the presence of various types of synapses. The absence of myelinized fibres testifies to the insufficient maturity of the cultures, that is probably associated with employing the low differentiated nervous tissue for cultivation and with insufficient cultivation period.
Subject(s)
Cerebral Cortex/ultrastructure , Neurons/ultrastructure , Cell Differentiation , Cerebral Cortex/embryology , Humans , Microscopy, Electron , Organ Culture Techniques/methods , Time FactorsABSTRACT
In the tissue culture of the newborn rat cerebrum structural peculiarities of the glial growth have been investigated, using scanning and transmissive microscopy. Glioblasts are the first to migrate into the growth zone from the explant. Formation of the glial network is accompanied with formation of multilayered structures. In the cerebral tissue culture 3 zones are distinguished differing from each other by their morphofunctional peculiarities and behavior of the glial cells: growth zone, intermediate and central ones. The growth zone is characterized with permanent moving and modification of glial cells, participating in formation of cellular cords, networks and layers. Glioblasts are predominate cells in this zone. The intermediate zone consists of multilayered glial elements, possessing a high proliferative activity. The central zone, where neuropil and neurons consisting of numerous processes are situated, is characterized with a high degree of differentiation of glial elements. The central part of the explants is covered with epitheliomorphic layer of glial cells, predominantly consisting of cytoplasmic and fibrous astrocytes. The data presented demonstrate that differentiation of glial elements into various types occurs in the cerebral cultures exclusively in the area of the neural cells localization and, possibly, under their immediate influence.
Subject(s)
Animals, Newborn , Brain/cytology , Neuroglia/cytology , Animals , Cell Differentiation , Culture Techniques , Mitosis , RatsABSTRACT
The complex virological, biochemical and morphological study permitted to obtain various characteristics of mice herpes encephalitis. The reaction of astrocyte glia at different stages of herpes encephalitis was revealed and analyzed in detail. New data on the dynamics of desoxyribonuclease activity changes in neuroglia and the glial complex formation were obtained. It was shown that the increased DNA-ase activity in neuroglia and the astrocyte activation which morphological manifestation was the formation of glial complexes, may be referred to as processes dealing with barrier and elimination neuroglia functions in herpes encephalitis. The results presented allows to suppose that the severity of the development and outcome of herpes encephalitis mainly depends on the astrocyte glia condition, its potential abilities in appearing of barrier and eliminative functions.
Subject(s)
Encephalitis/enzymology , Herpes Simplex/enzymology , Neuroglia/enzymology , Animals , Astrocytes/enzymology , Astrocytes/ultrastructure , Deoxyribonucleases/metabolism , Encephalitis/pathology , Herpes Simplex/pathology , Histocytochemistry , Lysosomes/enzymology , Lysosomes/ultrastructure , Mice , Mice, Inbred CBA , Neuroglia/ultrastructure , Neurons/enzymology , Neurons/ultrastructure , Ribonucleases/metabolism , Time FactorsABSTRACT
The aim of the study was to analyse the distribution and localization of glutamate receptors in the cultured cells of the neonatal rat motor cortex, using immunoelectron microscopic technique, and monoclonal antibodies preliminary labeled with colloidal gold. Monoclonal antibodies against glutamate-binding proteins of the adult rat cerebral cortex were produced by means of hybridization of immune splenocytes with plasmocytoma cells. It was found that monoclonal antibodies labeled with colloidal gold could reveal selectively the localization of glutamate receptors on the membranes of neurons. Glutamate receptors were detected on differentiating neuron membranes only, being absent from the glia cell surface.