Volume 45 (1999) No. 6
Volume 45 (1999) No. 6
Interleukin 2 Gene Therapy of Surgical Minimal Residual Tumour Disease:
Characterization of Cytolytic Effector Cells from Tumour Progressors and Regressors
R. HÁJKOVÁ, M. INDROVÁ, T. JANDLOVÁ, J. BUBENÍK, M. REINIŠ.............................227
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague,
Czech Republic
Corresponding author: Romana Hajkova, Institute of Molecular Genetics, Academy of Sciences
of the Czech Republic, Flemingovo nám. 2, 166 37 Prague 6, Czech Republic.
Abstract.
A Treshold Level of gag Expression is Required for Particle Formation in Rat Cells
Transformed by Avian Retroviruses
M. POLIANOVA1, M. ALEXANDROV2, V. SOVOVÁ3, K. SIMEONOV2, R. ROUSSEV2,
D. BESHKOV1, T. G. TODOROV2...................................233
1National Reference AIDS Laboratory, National Center of Infections and Parasitic Diseases, Sofia
2Institute of Experimental Pathology and Parasitology, Bulgarian Academy of Sciences, Sofia
3Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague
Corresponding author: Maria Polianova, National Reference AIDS Laboratory, 44A Stoletov Str.,
1233 Sofia, Bulgaria.
Abstract.
The Asymmetric Distribution of Interphasic Silver-Stained Nucleolus Organizer Regions
in Human and Rat Proerythroblasts
K. SMETANA1, Z. LIKOVSKÝ2, I. JIRÁSKOVÁ,1, J. ČERMÁK1..................243
1Institute of Hematology and Blood Transfusion, Prague, Czech Republic
2Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague,
Czech Republic
Corresponding author: Karel Smetana, Institute of Hematology and Blood Transfusion, U Nemocnice 1,
128 20 Prague 2, Czech Republic. Fax: 420 2 299821; e-mail: karel.smetana@uhkt.c
Abstract.
Hematocrit and Hemoglobin Values Are Negatively Correlated with Insulin Resistance
in Spontaneous Hypertension
V. ZÍDEK1, A. FUČÍKOVÁ2, A. MUSILOVÁ1, V. BÍLÁ3, V. KŘEN1,3
M. PRAVENEC1,3.......................................247
1Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic
2Agronomical Faculty, Czech Agricultural University, Prague, Czech Republic
3Institute of Medical Genetics and Biology, 1st Medical Faculty, Charles University, Prague,
Czech Republic
Corresponding author: Michal Pravenec, Institute of Physiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20 Prague 4, Czech Republic. Phone/Fax: (4202) 475 2297; e-mail: pravenec@biomed.cas.cz.
Abstract.
Sexing of Chicken Feather Follicle, Blastodermal and Blood Cells
P. TREFIL1, M. M. BRUNO1, T. MIKUS1, P. THORAVAL2........................253
1Biopharm, Research Institute of Biopharmacy and Veterinary Drugs, Inc., Jílové u Prahy, Czech Republic
2Institut National de la Recherche Agronomique, Nouzilly, France
Corresponding author: Pavel Trefil, Biopharm, Research Institute of Biopharmacy and Veterinary Drugs, Inc., Pohoří-Chotouň, 254 49 Jílové u Prahy, Czech Republic.
Abstract.
Short Communication
CD10 Expression in Cultured Human Osteoblast-like Cells
C. REYES-BOTELLA1, M. J. MONTES2, A. C. ABADÍA-MOLINA2,
M. F. VALLECILLO-CAPILLA1, C. RUIZ3.....................................257
1Departamento Estomatología, 2Departamento Bioquímica y Biología Molecular, Sección de Inmunología, 3Departamento de Enfermería, Sección de Fisiología, Instituto de Neurociencias, Universidad de Granada, Spain.
Corresponding author: Concepcion Ruiz, Institute of Neuroscience. F. Medicine. U. Granada, 18012 Granada, Spain. Fax +34-58-242894. E-mail: crr@goliat.ugr.es.
Abstract.
Articles
Interleukin 2 Gene Therapy of Surgical Minimal Residual Tumour Disease:
Characterization of Cytolytic Effector Cells from Tumour Progressors and RegressorsR. HÁJKOVÁ, M. INDROVÁ, T. JANDLOVÁ, J. BUBENÍK, M. REINIŠ Experiments were designed to characterize cytolyticeffector cells from mice
with SMRTD treated with IL-2 gene therapy. Mice were inoculated with syngeneic
murine MK16 carcinoma cells. When the tumours reached 8--12 mm in diameter, the were excised and the operated mice were randomized into two groups. The first group without any further treatment was designated as operated-only; the second group, vaccinated 3 days after the operation with IL-2-producing tumour vaccine, is referred to as operated-vaccinated. Tumour recurrence rate in the operated-only mice was 90 percent; in the operated-vaccinated group the recurrence rate was 38.5 percent (progressors). The remaining 61.5 percent of mice were permanently protected (regressors).
On day 53, the tumour progressors, regressors and healthy controls were sacrificed, and their spleen cells were used for 51Cr microcytotoxicity assay. Splenocytes from any group of mice were not cytolytic when allowed to react with MK16, YAC-1 (NK sensitive) and C1498 (NK resistant) targets. However, when grown for 3 days in IL-2-containing medium, the splenocytes from all groups of mice could develop cytolytic activity. The cytolytic activity of splenocytes from tumour progressors and regressors was substantially lower then that of splenocytes from healthy controls. In addition, significantly lower cytolytic activity was observed with IL-2-activated splenocytes from tumour progressors as compared to that of tumour regressors. Depletion of NK1.1+ cells or CD4+ plus CD8+ cells prevented the induction of significant IL-2-stimulated cytotoxicity directed against MK16 and C1498 targets in spleen cell cultures from tumour progressors, regressors, and healthy control mice, indicating that both, NK 1.1+ and CD4+ plus CD8+, cells participate in the antitumour effect of IL-2 gene therapy. This was further supported by the finding that after depletion of CD4+ plus CD8+ cells, a residual cytolytic activity directed exclusively against NK-sensitive YAC-1 cells was observed.
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A Treshold Level of gag Expression is Required for Particle Formation in Rat Cells
Transformed by Avian Retroviruses
To test the hypothesis that the block of polyprotein precursor processing and particle formation in RSV-transformed mammalian cells is due to a low level of pr76gag expression, rat tumor cell lines with different amounts of precursor molecules were used. The wild type forms of pr76gag have been expressed at a high level by use of SV40-based vector and thirty-two stable transfected cell clones were isolated. The gag protein expression was detected in the cell lysate by immunoblotting. Untransfected cells released no proteins that could be detected by immunoprecipitation with anti-RSV serum. Membrane-enclosed gag precursor-polyprotein molecules and infectious virus particles from different stably transfected clones have been found in the medium. Both immature and mature virions of type C morphology were directly detected by transmission electron microscopy. Surprisingly, virus-like particles of morphology similar to mature type C retroviruses were found enclosed within intracellular membranes in a stably transfected nonproducing clone.
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The Asymmetric Distribution of Interphasic Silver-Stained Nucleolus Organizer Regions
in Human and Rat Proerythroblasts
K. SMETANA, Z. LIKOVSKÝ, I. JIRÁSKOVÁ, J. ČERMÁK
The distribution of SSPs representing AgNORs was studied in human as well as rat proerythroblasts to provide an information on the distribution of these nucleolar components in highly immature and proliferating non-neoplastic cells. The distribution of SSPs was asymmetric and most of the cells contained one nucleolus which possessed a larger number of these nucleolar components than the remaining nucleoli. Such nucleolus might be functionally dominant, since the number of nucleolar SSPs is apparently related to the nucleolar biosynthetic activity. On the other hand, when a proerythroblast possessed only one nucleolus, the number of SSPs in such a cell was very similar to the sum of SSPs in a polynucleolar cell. The asymmetric distribution of SSPs characteristic for most proerythroblasts disappeared in the terminal stages of the erythroblastic development. Cells in such stages, as described previously, were characterized by the presence of a limited number of single SSPs.
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Hematocrit and Hemoglobin Values Are Negatively Correlated with Insulin Resistance
in Spontaneous Hypertension
V. ZÍDEK, A. FUČÍKOVÁ, A. MUSILOVÁ, V. BÍLÁ, V. KŘEN, M. PRAVENEC
It has been recently reported that increased hematocrit and hemoglobin values often accompany insulin resistance and compensatory hyperinsulinemia in humans. In the current study, we analysed the relationship between hematocrit/hemoglobin on the one hand and insulin resistance, dyslipidemia, and hypertension on the other hand in HXB/BXH recombinant inbred (RI) strains derived from the spontaneously hypertensive rat (SHR) and the Brown Norway (BN) rat. The SHR progenitor strain had a significantly increased hematocrit values and it was also hypertensive and insulin-resistant when compared with the BN progenitor. The distribution of hematocrit and hemoglobin values among RI strains was continuous, suggesting a polygenic mode of inheritance. Analysis of RI strains revealed that hemoglobin was negatively correlated with insulin and insulin/glucose ratio, and that hematocrit was negatively correlated with insulin-stimulated glucose uptake in isolated adipocytes. There was no relationship between hematological parameters and blood pressure or lipid phenotypes in RI strains. The findings of the current study suggest that hematocrit and hemoglobin values might be added to the clustering variables related to insulin resistance syndrome in the SHR strain.
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P. TREFIL, M. M. BRUNO, T. MIKUS, P. THORAVAL
The production of chicken chimeras using donor and acceptor cells which can be of opposite sex has necessitated the utilization of methods developed to distinguish the sex of chickens. We demonstrate one of these methods, based on the polymerase chain reaction which amplifies the EcoRI repeat unit of the fowl W chromosome, and how this technique may be used to sex various cell types in chickens as well as small numbers of blastodermal cells. Our results demonstrate the ability to sex chickens using EcoRI primers, specific for the W chromosome, from as little as 2 ng of female genomic DNA isolated from blood and feathers - the latter being the result of DNA extraction from only one feather. Also evident in this study is the detection of the W chromosome by PCR from approximately 50 blastodermal cells originating from the developing blastodisc at stage X.
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CD10 Expression in Cultured Human Osteoblast-like Cells
C. REYES-BOTELLA, M. J. MONTES, A. C. ABADÍA-MOLINA,
M. F. VALLECILLO-CAPILLA, C. RUIZ
Morphological features, bone nodule formation and alkaline phosphatase activity are currently used to identify osteoblasts. CD10 (cALLa antigen) is a glycoprotein with endopeptidase activity and it is present on the surface of many cell types. We have studied the expression of CD10 in osteoblast-like cells by immunocytochemistry and flow cytometry in order to identify other markers of the osteoblast lineage. We isolated osteoblast-like cells from specimens obtained in the course of oral surgery. Expression of the cALLa antigen (CD10) may also be an indicator of the osteoblast phenotype.
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