Biomedical subjects
Publications and source records attributed to K Kristensson.
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Slices from the brain and liver of rats were prepared and upon incubation exhibited a continuous and high capacity for incorporation of radioactive precursors into proteins and lipids. Using [3H]mevalonate as precursor, the rates of biosynthesis of cholesterol, ubiquinone, dolichol and dolichyl-P in brain slices were determined and found to be 5.5, 0.25, 0.0093 and 0.0091 nmol/h/g, respectively. Dolichol and dolichyl-P accumulate to a limited extent, but almost all of these lipids in the brain originate from de novo synthesis. The calculated half-lives for cholesterol, ubiquinone, dolichol and dolichyl-P were 4076, 90, 1006 and 171 h, respectively. The results indicate that lipids formed via the mevalonate pathway in the brain have an active and independently regulated biosynthesis.
The effect of intrathecally (i.t.) applied substance P (SP) analogue, (D-NicLys1,3-Pal3,D-Cl2Phe5,Asn6,D-Trp7,9,Nle 11)-SP (Spantide II), was examined in rats. Spantide II even at a high dose (10 micrograms) did not evoke any behavioural responses and caused no motor disturbances, but it did have a brief antinociceptive effect on the hot-plate test. Spantide II dose-dependently reduced the caudally directed scratching/biting behaviour, evoked by 1 microgram i.t. SP for over 30 min, but did not block the caudally directed scratching behaviour evoked by i.t. somatostatin. Histological examination revealed no pathological changes in the spinal cord after treatment with Spantide II. The results indicate that Spantide II is an effective tachykinin antagonist in the central nervous system and that it causes no neural damage.
The protein and glycoprotein composition of a sucrose gradient fraction from vaccinia infected cells treated with rifampicin was studied. This particulate fraction contained cytoplasmic membranes and pleomorphic membranous structures. The glycoproteins (89, 42 and 20-23 kDa, respectively) were identified as the same glycoproteins that are found in plasma membranes of infected cells and the envelope of extracellular enveloped vaccinia (EEV). These glycoproteins could be solubilized by 0.1% NP-40. The Golgi membrane associated 41K acylated vaccinia protein was also NP-40 soluble. In contrast, most particulate fraction proteins (125, 100, 86, 65, 41, 39, 31, 27, 25, 14 and 12.5 kDa) with the exception of the 33 and 29 kDa proteins remained essentially insoluble after NP-40 treatment. The 86 and 65 kDa proteins are the rifampicin inhibited precursors to INV core proteins while the 33 and 29 kDa proteins are INV surface proteins. Twelve proteins behaved like their respective comigrating INV proteins when extracted with NP-40 and 2ME. Electron microscopy showed that a centrifuged sediment from NP-40 treated cells contained pleomorphic protein containing membranous structures that we have called rifampicin bodies. We conclude that (1) the major glycoproteins found in the particulate fraction from sucrose gradients are vaccinia glycoproteins residing in cytoplasmic membranes while (2) the major non-glycosylated proteins are components of the rifampicin bodies and that (3) the rifampicin bodies represent an intermediate in the morphogenetic process leading to mature INV.
Dysfunction of subcortical serotoninergic neurons has been implicated in some behaviour disturbances. The serotoninergic neurons in the dorsal and median raphe project widely in the brain. They innervate the olfactory bulbs and can be targets for exogenous agents attacking the olfactory epithelium and bulbs. We report here an injury to the serotoninergic neurons after intranasal infection in 12-day-old rats with a temperature-sensitive mutant of vesicular stomatitis virus. The brain infection was focal and transient. Viral antigens could no longer be detected 13-15 days after infection. In spite of this the animals, as adults, had a severe serotonin depletion in the cerebral cortex and hippocampus, and showed abnormal locomotor and explorative behaviour as well as learning deficits. The neocortex was histologically intact and parameters related to other neurotransmitters such as dopamine, noradrenaline, GABA and acetylcholine showed no marked changes. A relatively selective damage to serotoninergic nuclei as a result of virus neuroinvasion through a natural portal of entry, may constitute a new pathogenetic mechanism for cortical dysfunction and behavioural deficits.
Sprague-Dawley rats infected with Trypanosoma brucei brucei showed a strong and rapid induction of splenocyte IFN-gamma (within 12 h post-infection) as measured by a single cell assay for IFN-gamma secretion. Depletion of CD8+ cells in infected rats abrogated the IFN-gamma production, suppressed parasite growth and increased survival of the animals. Induction of MHC class I antigens in the paraventricular and supra-optic hypothalamic nuclei caused by the trypanosome infection was also inhibited by the CD8+ cell depletion. It is suggested that the CD8+ cells are involved directly or indirectly in growth regulation of the parasite and that IFN-gamma induced by the parasite may be one of the factors that trigger MHC expression and immunosuppression.
Cultured spinal ganglion and spinal cord neurons were used to examine the intraneuronal distribution of five structural proteins of Sendai virus by immunohistochemistry. In spinal ganglion cells the internal, cytosolic viral proteins (the nucleocapsid, polymerase and matrix proteins) were confined to the perikarya, while the envelope glycoproteins (the haemagglutinin-neuraminidase and fusion proteins) also appeared in the axon-like processes. All five proteins occurred in the dendrite-like processes of spinal cord neurons. In both types of neuron the cytosolic viral proteins showed a pattern of distribution similar to that observed for the microtubule-associated protein MAP2. The segregated occurrence of the viral envelope and cytosolic proteins in axons may prevent virus assembly in axons and limit long-distance spread of paramyxoviruses in the nervous system.
Human CD4+ T cells differ in their expression of the leucocyte common antigen. Antibodies detecting certain forms (CD45RA and CD45RO) of this antigen have been used to identify and isolate subpopulations of the CD4+ T cells. These isolated subsets have been shown to have different abilities concerning lymphokine production and provision of help to B cells for Ig production. When these T-cell subsets were activated in vitro with polyclonal activators, the production. When these T-cell subsets were activated in vitro with polyclonal activators, the CD45RA+ cells lost this marker and gained the expression of CD45RO. This was true for all mitogens used in this report, i.e. accessory cell-dependent stimulation with SEA and accessory cell-independent activation with PMA or PHA. A correlation between proliferation and differentiation was observed, but this was probably not causative as stimulation with PMA in the absence of DNA synthesis resulted in the acquisition of CD45RO and loss of the CD45RA antigen. Moreover, cells proliferating vigorously for long periods of time expressed both markers at significant levels, which suggests that proliferation did not automatically result in complete loss of the CD45RA marker. The phenotypical differentiation was associated with a functional differentiation which induced the stimulated cells' ability to act as helper cells for Ig production and to produce gamma interferon (IFN-gamma). The results obtained in this study support the contention that the CD45RA+ cells are precursors of the CD45RO+ cells and that the two subsets represent different maturational stages of the same lineage.
The neutral and phospholipid compositions of various regions of the human brain were analyzed using autopsy material covering the life period between 33 and 92 years of age. The protein content was also measured and, on a weight basis, this content is unchanged in the cerebellum, pons, and medulla oblongata, whereas in the 90-year-old group it decreases in the hippocampus, gray matter, and nucleus caudatus. In white matter, the protein content decreases continuously with age. The phospholipid composition is characteristic of the region investigated, but remains unchanged during aging. The total phospholipid content exhibits only a 5-10% decrease in the oldest age group. The content of dolichol and its polyisoprenoid pattern are also characteristic of the region analyzed. Between 33 and 92 years of age, the amount of dolichol in all portions of the brain increases three- to fourfold, but the isoprenoid pattern remains constant. The level of dolichyl-P varies between different regions, but only a moderate increase is seen with age. Ubiquinone content is highest in the nucleus caudatus, gray matter, and hippocampus, and in all areas this content is decreased to a great extent in the oldest age groups. All regions of the human brain are rich in cholesterol, but alterations in the amount of this lipid are highly variable during aging, ranging from no change to a 40% decrease.
Interferon-gamma (IFN-gamma)-like-immunoreactivity was observed in subsets of rat skeletal muscle fibres with 3 different mouse monoclonal antibodies directed to rat IFN-gamma and a rabbit polyclonal anti-rat IFN-gamma-antibody. These strongly IFN-gamma-immunoreactive fibres also expressed antigen defining T-cells of the cytotoxic/suppressor phenotype (CD8) and major histocompatibility complex (MHC) class I antigen.
After a peripheral nerve lesion (rat facial and sciatic) an induction of major histocompatibility complex (MHC) antigens class I was detected immunohistochemically in skeletal muscle fibers and motor neurons. This MHC expression was transient after a nerve crush, when regeneration occurred, but persisted after a nerve cut, when regeneration was prevented. Since the time course of MHC class I expression correlates to that of regeneration a role for this cell surface molecule in regeneration may be considered.
Sections from the nasal cavity of 12-day-old Swiss albino mice (NMRI strain) were subjected to lectin histochemistry. A panel of biotinylated lectins (Con A, WGA, s-WGA, PNA, SBA, DBA and UEA I) and a horseradish peroxidase-conjugated lectin (GSA II) showed marked differences in binding to the respiratory and the neuroepithelial cells. SBA (affinity for galactose and N-acetylgalactosamine), PNA (galactose) and WGA (sialic acids and N-acetylglucosamine) labelled the receptor neurons in the olfactory and vomeronasal epithelium. DBA (N-acetylgalactosamine) labelled a subgroup of about 5% of the olfactory receptor neurons, but most neurons in the vomeronasal organ. UEA I (fucose) and s-WGA (N-acetylglucosamine) intensely labelled the entire nerve cell population in the vomeronasal organ, but in the olfactory epithelium the labelling with these lectins was stratified. In the respiratory epithelium the ciliated cells were labelled with WGA and s-WGA, while the secretory cells bound most of the lectins. Thus different sugars are exposed on the surface of the different types of epithelia in the nasal cavity, providing a basis for selectivity in microbial attacks on these areas.
The monoclonal antibody W3/13, which is directed against leukosialin (a molecule present on certain types of leukocytes), was found to react immunohistochemically with distinct regions and structures in the rat CNS. The leukosialin (W3/13)-like immunoreactivity occurred on neurite-like fibres and on bouton-like structures on nerve cell bodies and their dendrites. This labelling was most pronounced in certain subcortical areas and brain stem nuclei including different thalamic nuclei, olfactory tubercle, superior olive, cochlear and vestibular nuclei, motor and somatosensory nuclei. In the cerebellum, immunoreactive fibres were prominent in the molecular layer. In the spinal cord there was a dense labelling of fibres in the whole grey matter.
Patients with distal myopathy (Welander) were subjected to muscle biopsy from the anterior tibial muscle (n = 4) and to nerve biopsy from the sural nerve at the ankle (n = 5) in order to elucidate a possible neurogenic component of the disease. The type I muscle fibres had a larger mean cross-sectional area as compared to normal controls and an increased variation in fibre size with both hypertrophic and atrophic fibres in one and the same biopsy. A normal muscle fibre type composition was found in the patients. Structural muscle fibre abnormalities such as atrophic fibres, mainly angulated, split fibres, rimmed vacuoles and centrally located nuclei were found in all biopsies. A disorganization and loss of myofibrils as well as autolytic vacuoles were the most prominent findings at the ultrastructural level. Two patients had a moderate loss of myelinated sural nerve fibres. The mean nerve fibre density was decreased as compared to normal controls while the mean nerve fibre area and circular diameter were increased due to selective loss of small diameter (A-delta) nerve fibres. The muscle and nerve fibre abnormalities are discussed with reference to neuropathic and myopathic changes. Some muscle fibre abnormalities are typical for a neurogenic disorder while some others are mostly seen in myopathies but may also appear in neurogenic conditions. A neurogenic etiology in Welander distal myopathy is further supported by the finding of loss of small diameter nerve fibres in the sural nerve.
Standardised skin biopsies followed by immunohistochemical examination for the presence of terminal nerve fibres reacting for neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) were evaluated. Healthy subjects regularly displayed free nerve endings of both fibre types in the papillary and reticular dermis. Both fibre types were present close to blood vessels, while CGRP immunoreactive fibres were more often encountered near sweat gland acini compared to SP fibres. Diabetes mellitus complicated by polyneuropathy was accompanied by marked reduction of SP and CGRP reactive fibres in the dermis layers. Five type I diabetes patients without clinical or neurophysiological evidence of polyneuropathy also had reduced density of both fibre types, being significant for CGRP fibres when compared with controls. Skin biopsy with immunohistochemical staining for neuropeptides may represent a sensitive tool in evaluation of patients with peripheral neuropathies.
The effect of unilateral peripheral nerve lesions on the inflammatory response of experimental allergic encephalomyelitis (EAE) in rat central nervous system (CNS) was studied. Immunostaining for major histocompatibility complex (MHC) antigens and T-cell subsets demonstrated that MHC class I expression was markedly enhanced in as well as around axotomized motor neurons and that MHC class II expression was induced on several cells, probably microglial cells, in close proximity to the axotomized motor neurons. There was also a pronounced increase in interleukin 2 receptor-positive lymphocytes as well as T-cells and the T-cell subsets on the injured as compared to the non-injured contralateral side. These effects were present particularly in the initial phase of EAE and persisted for several weeks. The results suggest that neurons may communicate immunoregulatory signals to their microenvironment and that retrograde axonal signals from the distant periphery may alter the immune response locally within the CNS.
Sprague-Dawley rats were injected intraperitoneally with a suspension of Trypanosoma brucei brucei. An early induction of major histocompatibility complex (MHC) class I antigens as well as an infiltration of macrophage-like cells and cytotoxic T-cells was detected with immunohistochemical techniques in circumventricular organs, such as the median eminence, neurohypophysis, subfornical organ, pineal gland and area postrema. These areas, which lack a blood-brain barrier, correspond to those showing early invasion of trypanosomes. In addition, there was a marked induction of MHC class I in neurons in two hypothalamic nuclei, the paraventricular and supraoptic nuclei. Neurons in these two nuclei are located behind the blood-brain barrier, but project to the neurohypophysis and to the median eminence, thereby exposing their axon terminals to factors circulating in the blood or released locally from invading trypanosomes or from macrophages or cytotoxic T-cells. It is suggested that the alteration in the nerve cell bodies in the hypothalamic nuclei is caused by retrograde axonal signals from these target areas.
Fourteen-day-old Lewis rats were injected intracerebrally with the hamster neurotropic (HNT) strain of measles virus. At the same time, CD8+ T cytotoxic cells were eliminated by a single injection of a mouse monoclonal antibody (Ox8) directed against this lymphocyte phenotype. The lymphocyte depletion, which endured for more than 7 weeks, markedly reduced the elimination of measles virus antigen from the brain, but did not affect the induction of major histocompatibility complex (MHC) molecules in the early phase of infection. These results demonstrate a role for MHC class I-restricted CD8+ T cells in controlling persistence of measles virus infection in neurons.