Volume 45 (1999) No. 4
Granulocyte-Macrophage Colony-Stimulating Factor Gene-Modified Vaccines
for Immunotherapy of Cancer
JAN BUBENÍK.................................................115
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague,
Czech Republic
Corresponding author: Jan Bubeník, Institute of Molecular Genetics, Academy of Sciences
of the Czech Republic, Flemingovo nám. 2, 166 37 Prague 6, Czech Republic.
Tel.: (420 2) 20 18 32 34. Fax: (420 2) 24 31 09 55.
Articles
Therapeutic Effect of Heat Shock on T-Cell Lymphoma in Inbred Sprague-Dawley Rat
B. OTOVÁ1, M. MRÁZ2, V. MANDYS3,4, A. PANCZAK1, J. SCHRAMLOVÁ5,
I. VOTRUBA6, A. HOLÝ6.......................................................121
1Institute of Biology, 2Department of Pharmacology, 3Hlava 1st Department of Pathology, 1st Medical Faculty, Charles University, Prague, Czech Republic
4Department of Teratology, Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague, Czech Republic
5National Institute of Public Health, Prague, Czech Republic
6Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Berta Otová, Institute of Biology, 1st Medical Faculty, Charles University, Albertov 4, 128 00 Praha 2, Czech Republic. E-mail: boto(zavináč)lf1.cuni.cz.
Abstract.
Essential Fatty Acids and Related Molecular and Cellular Mechanisms in Multiple Sclerosis:
New Looks at Old Concepts
M. MAYER........................................133
Neurorehabilitation Unit, Department of Rehabilitation, Faculty Hospital, Olomouc, Czech Republic
Corresponding author: Michal Mayer, Department of Rehabilitation, Faculty Hospital,
779 00 Olomouc, Czech Republic. E-mail: michal@tunw.upol.cz.
Abstract.
EGF Receptor-like Determinants on Human Spermatozoa and Their Possible Cytoskeletal
Association
MARKOVA, M. D., MARINOVA, T. T................................143
Cell Biology Laboratory, Department of Biology, Medical Faculty, Sofia Bulgaria
Corresponding author: Maya D. Markova, Department of Biology, Medical Faculty,
2 Zdrave Str., 1431 Sofia, Bulgaria. Tel. (+359 2) 51 66 507. Fax (+359 2) 51 71 76. E-mail: mmarkova@ns.medfac.acad.bg.
Abstract.
Detection of đ(Đ)-N-Acetyl-D-Galactosamine-Binding Sites in Kidney - Relation to
Henoch-Schönlein Associated IgA Nephropathy
A. ŠEDIVÁ1*, J. STEJSKAL, J. BARTŮŇKOVÁ1, K. SMETANA, Jr.3,
H.-J. GABIUS4..........................................147
1Institute of Immunology, 2Institute of Pathology, 2nd Faculty of Medicine, Charles University,
Prague, Czech Republic
3Institute of Anatomy, 1st Faculty of Medicine, Charles University, Prague, Czech Republic
4Institute of Physiological Chemistry, Faculty of Veterinary Medicine,
Ludwig-Maximilians University, Munich, Germany
Corresponding author: Anna Šedivá, Institute of Immunology, 2nd Faculty of Medicine, Charles University, V Úvalu 84, 150 18 Prague 5, Czech Republic. Tel. (420 2) 24 43 59 60.
Fax (420 2) 24 43 59 62. E-mail: anna.sediva@lfmotol.cuni.cz.
Abstract.
Diversity Among Wild Type and Vaccination Strains of Trichophyton verrucosum
Investigated Using Random Amplified Polymorphic DNA Analysis
M. HAJDÚCH1, J. DRÁBEK2, V. RACLAVSKÝ3, V. KOTALA1, T. MICHÁLEK1,
I. ZELENKOVÁ4.......................................151
1Laboratory of Experimental Medicine, Department of Paediatrics, Faculty of Medicine, Palacký University and Faculty Hospital in Olomouc, Olomouc, Czech Republic
2Institute of Immunology, 3Centre of Molecular Biology and Medicine, 4Institute of Biology,
Faculty of Medicine, Palacký University, Olomouc, Czech Republic
Corresponding author: Marián Hajdúch, Laboratory of Experimental Medicine, Department of Paediatrics, Faculty of Medicine, Palacký University and Faculty Hospital in Olomouc, Puškinova 6, 775 20 Olomouc, Czech Republic. Tel.: +420-68-585 4441, Fax: +420-68-585 2527, e-mail: marian@risc.upol.cz.
Abstract.
Simultaneous Detection of Edogenous Lectins and Their Binding Capacity at
the Single-Cell Level - a Technical Note
V. FROŇKOVÁ1*, Z. HOLÍKOVÁ1, FU-TONG LIU2, J. HOMOLKA3, D. C. RIJKEN4,
S. ANDRÉ5, N. V. BOVIN6, K. SMETANA, Jr.1, H.-J. GABIUS5
1Institute of Anatomy, 1st Faculty of Medicine, Charles University, Prague, Czech Republic
2La Jolla Institute for Allergy and Immunology, San Diego, CA, USA
31st Department of Tuberculosis and Respiratory Diseases, 1st Faculty of Medicine,
Charles University, Prague, Czech Republic
4Gaubius Laboratory, TNO Prevention and Health, Leiden, Netherlands
5Institute of Physiological Chemistry, Faculty of Veterinary Medicine,
Ludwig-Maximilians-University, Munich, Germany
6Shemyakin Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia
Corresponding auhor: Karel Smetana Jr., Charles University, 1st Faculty of Medicine, Institute of Anatomy, U nemocnice 3, 128 00 Prague 2, Czech Republic. Tel: + 420 2 24 91 50 03. Fax: + 420 2 29 76 92. E-mail: ksmet(zavináč)lf1.cuni.cz.
Abstract.
Monoclonal Antibody Register
Monoclonal Antibody NF-09 Specific for Neurofilament Protein NF-M
E. DRÁBEROVÁ, V. SULIMENKO, V. KUKHARSKYY, P. DRÁBER
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague,
Czech Republic
Corresponding author: Eduarda Dráberová, Institute of Molecular Genetics,
Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20 Praha 4,
Czech Republic. Fax: 420-2 475 2632. E-mail: drabere@biomed.cas.cz.
Background.
Therapeutic Effect of Heat Shock on T-Cell Lymphoma in Inbred Sprague-Dawley Rat
B. OTOVÁ, M. MRÁZ, V. MANDYS, A. PANCZAK, J. SCHRAMLOVÁ, I. VOTRUBA, A. HOLÝ
Anticancer effect of heat shock, either alone or in combination with the drug PMEDAP, and cold water immersion stress were studied in an in vivo model of s.c. transplanted rat T-cell lymphomas in an inbred Sprague-Dawley rat line (SD/cub). Significant anticancer effect was induced by repeated sessions of heat shock; decrease of s.c. lymphoma weight and prolongation of survival time of treated rats was found to be dependent on the number of HS sessions. Much stronger therapeutic effect was observed after repeated heat shock in combination with PMEDAP administration. Light and electron microscopy studies were performed to characterize the alterations within the lymphomas. Morphologically, cellular alterations corresponding with apoptosis were observed in lymphoma cells after repeated heat shock. Indirect immunoperoxidase technique was used to detect HSP 72--73 protein(s), p53 and Bcl2 proteins in lymphomas heated directly or undirectly. The induction of HSP 72--73 protein(s) was found in the lymphoma tissues from autopsied animals exposed to heat shock; the intensity of its expression was dependent on the experimental design. The expression of p53 and Bcl2 proteins was not changed in lymphoma cells of HS treated animals as compared to that of untreated lymphoma bearing controls; the Bcl2 protein was present in both treated and untreated lymphomas, and the p53 protein remained undetectable in all samples. Contrary to the heat shock, the cold stress did not suppress growth of lymphomas and, furthermore, accelerated the infiltration of parenchymatous organs with lymphoma cells.
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Essential Fatty Acids and Related Molecular and Cellular Mechanisms in Multiple Sclerosis: New Looks at Old Concepts
M. MAYER
Essential polyunsaturated fatty acids, their metabolism and their effects represent a ubiquitous system important for cellular membrane stability, for immunological and inflammatory regulations, for inter- and intracellular signalling. There are complex links to other regulatory and effector mechanisms - to the reactive oxygen species, nitric oxide and peroxynitrite anion, to eicosanoids and related compounds. The relation of polyunsaturated acids to multiple sclerosis was extensively discussed some time ago. Recently, a number of data placing these concepts in a new light have emerged, which are reviewed in the present article. The recent knowledge indicates that this topic deserves further attention in both the basic and the clinical research of multiple sclerosis.
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EGF Receptor-like Determinants on Human Spermatozoa and Their Possible Cytoskeletal Association
MARKOVA, M. D., MARINOVA, T. T.
The study was focused on the localization of human sperm epidermal growth factor receptor and its resistance to Triton X-100 extraction, indicating possible cytoskeletal association. Human spermatozoa were subjected to immunofluorescence and pre-embedding immunoelectron microscopy using anti-EGF receptor monoclonal antibody as a probe. In detergent-untreated cells, the entire head was stained with intensity declining towards the acrosomal tip. Triton-extracted spermatozoa were stained predominantly in the equatorial segment region. Electron microscopy performed after extraction revealed that the label was also present in the equatorial segment and localized to electron-dense material overlying the acrosomal remnants. This Triton-resistant staining could imply cytoskeletal association of the EGF receptor in the equatorial segment, with possible importance in fertilization.
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A. ŠEDIVÁ, J. STEJSKAL, J. BARTŮŇKOVÁ, K. SMETANA, Jr., H.-J. GABIUS
Henoch-Schönlein purpura is the most common vasculitis of childhood, accompanied by the deposition of IgA1 immunoglobulins into the glomerular mesangium. The actual molecular mechanism of IgA deposition is not clear, but the altered glycosylation of O-linked oligosaccharides of the hinge region of IgA1 is generally considered as the crucial etiopathogenic factor. The oligosaccharides of this glycoprotein from healthy persons are principally of mucin-type GalĐ1,3GalNAcđ-O-glycan core structure, frequently sialylated. The patient's IgA hinge region saccharide is an incomplete GalNAcđ-O-glycan only. This study investigates the presence of binding sites for đ-GalNAc and Đ-GalNAc in frozen sections of kidney with and without nephropathy prompted by the possibility for a lectin mechanism of IgA deposition to mesangium. Neoglycoproteins prepared as conjugates with derivatized đ- or Đ-GalNAc moieties as histochemically crucial ligands and biotinylated bovine serum albumin as a carrier were employed for this purpose. The result of the experiments demonstrated expression of specific and accessible binding sites for both đ- and Đ-GalNAc in tubules but not in glomerules of kidney samples both with and without nephropathy. These findings imply no involvement of a lectin mechanism of IgA1 binding to mesangium, unless a temporary alteration of accessibility of binding sites for probes in glomerules occurs or the linkage region beyond the monosaccharide is pivotal for a receptor whose binding site may accommodate a peptide epitope in addition to the O-linked đ-GalNAc residue.
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Diversity Among Wild Type and Vaccination Strains of Trichophyton verrucosum Investigated Using Random Amplified Polymorphic DNA Analysis
M. HAJDÚCH, J. DRÁBEK, V. RACLAVSKÝ, V. KOTALA, T. MICHÁLEK, I. ZELENKOVÁ
We initially tested 20 primers for their ability to amplify genomic DNA of Trichophyton verrucosum using RAPD. Six of these were selected for further study aimed at discrimination of wild type and vaccination strains of T. verrucosum. The results indicate that RAPD successfully distinguished all strains included in the study. In addition, results of corrected cluster analysis were consistent with the fact that the avirulent vaccination strains (T. verrucosum TV-M9 and T. verrucosum TV-M-130) were prepared by ultraviolet (UV) light induced mutagenesis of the standard wild type strain T. verrucosum Strážnice. No marker for a/virulence was detected. These outcomes suggest new possibilities for epidemiological analyses, for discrimination among different vaccination strains and studies of fungal population in vaccinated/infected hosts.
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Simultaneous Detection of Edogenous Lectins and Their Binding Capacity at the Single-Cell Level - a Technical Note
V. FROŇKOVÁ, Z. HOLÍKOVÁ, FU-TONG LIU, J. HOMOLKA, D. C. RIJKEN, S. ANDRÉ, N. V. BOVIN, K. SMETANA, Jr., H.-J. GABIUS
Endogenous lectins are proteins/glycoproteins which selectively recognize distinct saccharide ligands and are different from immunoglobulins and carbohydrate-utilizing enzymes. Expression of these molecules can be detected immunohistochemically using nonblocking monoclonal antibodies (A1D6: anti-galectin-3, MR-15-2-2: anti-175 kD mannose receptor). Alternatively, biotinylated (neo)glycoconjugates which are recognized by a studied lectin can be employed as convenient probes to demonstrate specific binding of sugar epitopes by carbohydrate recognition domains (CRD) of endogenous lectins. In this study, we describe a new procedure for immunocytochemical visualization of the expression of endogenous lectins and glycochemical visualization of the reactivity of carbohydrate recognition domain(s), performed simultaneously at the single-cell level.
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Monoclonal Antibody Register
Monoclonal Antibody NF-09 Specific for Neurofilament Protein NF-M
E. DRÁBEROVÁ, V. SULIMENKO, V. KUKHARSKYY, P. DRÁBER
Background
The cytoskeleton of mammalian cells is a complex filamentous network of microtubules, microfilaments and intermediate filaments. Intermediate filament proteins can be divided into six types according to their sequence homologies, immunological cross-reactivities, compatibilities of self-assembly, and with respect to their occurrence in various cell types (Franke, 1993). Neurofilaments are the intermediate filaments specific to nervous tissue. They are composed of neurofilament (NF) triplet proteins - NF-L (light), NF-M (middle) and NF-H (heavy) - differing in their apparent molecular masses. Mammalian neurofilament proteins, namely NF-M and NF-H, are highly phosphorylated in axons. Neurofilament functions depend on the state of phosphorylation of these proteins (Julien and Mushynski, 1982). Monoclonal antibodies against various phosphorylated forms of neurofilament epitopes are markers of cells of neuronal origin and are useful in studying the role of neurofilament protein phosphorylation during neuronal development (Lukáš et al., 1993).
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