Volume 45 (1999) No. 3

Volume 45 (1999) No. 3

Editorial
Dendritic Cell-Based Cancer Vaccines
J. BUBENÍK..............71
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.

Articles

Baculovirus Expression System Cells Expressing v-myb Oncogene: The Distribution of RNA
and DNA in Specific Nuclear Compartments with Respect to Structures Interacting with anti-v-Myb Antibody.
J. ŠTOKROVÁ, J. KORB, M. DVOŘÁKOVÁ...................................75
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Jitka Štokrová, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo n. 2, 166 37 Prague 6, Czech Republic.
Abstract.

Genetic Analysis of the Rat Hypodactylous Mutation
D. KŘENOVÁ¹, Z. JIRSOVÁ², D. HOUSA¹, F. LIŠKA¹, L. ŠOLTYSOVÁ³, R. KAŠPÁREK¹,
V. BÍLÁ¹, M. PRAVENEC^4,1, V. KŘEN^1,4........................................81
¹Institute of Biology and Medical Genetics, 1st Medical Faculty, Charles University, Prague
²Institute of Histology and Embryology, 1st Medical Faculty, Charles University, Prague
³Institute of Pharmacology, 1st Medical Faculty, Charles University, Prague
⁴Institute of Physiology, Academy of Sciences of the Czech Republic, Prague
Corresponding author: Vladimír Křen, Institute of Biology and Medical Genetics, 1st Medical Faculty, Charles University, Albertov 4, Prague 2, 128 00, Czech Republic. E-mail vkren(zavináč)lf1.cuni.cz.
Abstract.

Experimental Lesion of Medial Frontal Cortex Mediates Inhibition of Expression
of Carbohydrate-Binding Sites in the Spleen Macrophages in Rat
K. SMETANA, Jr.^1, P. ZACH¹, V. NĚMCOVÁ¹, P. PETROVICKY¹,
H.-J. GABIUS².......................................87
¹Institute of Anatomy, 1^st Faculty of Medicine, Charles University, Prague, Czech Republic
²Institute of Physiological Chemistry, Faculty of Veterinary Medicine, Ludwig-Maximilians-University, Munich, Germany
Corresponding author: Karel Smetana, Jr., Institute of Anatomy, 1^st Faculty of Medicine, Charles University, 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.

Review
Mechanisms of Horizontal Gene Transfer
D. ŽGUR-BERTOK..........................................91
Department of Biology, Biotechnical Faculty, Ljubljana, Slovenia
Corresponding author: Darja Žgur-Bertok, Department of Biology, Biotechnical Faculty, Večna pot 111, 1000 Ljubljana, Slovenia. Fax: 386 61 372 390; e-mail: Darja.Zgur@uni-Lj.Si.
Abstract.

Short Communication

FreeTree - Freeware Program for Construction of Phylogenetic Trees on the Basis
of Distance Data and Bootstrap/Jackknife Analysis of the Tree Robustness. Application
in the RAPD Analysis of Genus Frenkelia.
A. PAVLÍČEK¹, Š. HRDÁ^2, J. FLEGR² ...............................97
¹Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
²Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic
Corresponding author: Jaroslav Flegr, Department of Parasitology, Faculty of Science, Charles University, Viničná 7, 128 44 Prague 2, Czech Republic. Fax (4202) 299 713; Tel. (4202) 2195 3289; e-mail flegr@beba.cesnet.cz.
Abstract.

Monoclonal Antibody Register

A Signalling Accessory Molecule Revealed by a New
Anti-Fibroblastoid L Cell Monoclonal Antibody
A. PANCZAK¹, F. HIRSCH^2, D. HAGERTY³, R. BLUMBERG, P. PONCET⁵...............101
¹Institute of Biology and Medical Genetics, 1^st Medical Faculty, Charles University Prague, Czech Republic
²UPR 420 CNRS, Equipe d'Immunologie Cellulaire et de Transplantation, Villejuif, France
³Washington University School of Medicine, St Louis, USA
⁴Brigham and Women's Hospital, Gastroenterology Division, Boston, USA
⁵Institut Pasteur, Paris, France
Corresponding author: Pascal Poncet, Institut Pasteur, Unité d'Immuno-Allergie, 28 rue du Dr. Roux, 75724 Paris Cedex 15, France. Tel.: 33 01 45 68 82 42; Fax: 33 01 40 61 31 60; e-mail: pponcet@pasteur.fr.
Abstract.

ARTICLES

The distribution of RNA, total DNA and newly synthesized DNA within nucleoli-like structures in insect cells overexpressing v-myb oncogene was investigated. Three types of these structures which revealed interaction with anti-v-Myb oncoprotein antibody were found on the ultrastructural level. Specific staining by toluidine blue at pH 5.2 showed the presence of RNA in these nucleoli-like structures. To detect total DNA, the in situ terminal deoxynucleotidyl transferase-immunogold technique was used. In addition to an expected labeling of host condensed chromatin, the labeling of the three types of nucleoli-like structures differed from each other. While the compact (type I) and ring-shaped (type II) nucleoli-like structures were labeled only on their periphery and in the proximity of baculovirus particles that interacted with them, the structures with an appearance of nucleolonemas (type III) were labeled strongly. Incorporation of 5-bromo-2-deoxyuridine, in spite of a poor labeling of newly synthesized DNA, confirmed these results. We suggest that the nucleoli-like structures of type I and II are of nucleolar origin. The type III more likely represents virogenic stroma or viral DNA storage site.
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Genetic Analysis of the Rat Hypodactylous Mutation
D. KŘENOVÁ¹, Z. JIRSOVÁ², D. HOUSA¹, F. LIŠKA¹, L. ŠOLTYSOVÁ³, R. KAŠPÁREK¹,
V. BÍLÁ¹, M. PRAVENEC^4,1, V. KŘEN^1,4

Autosomal recessive rat hypodactylous mutation Hd leads in homozygous condition to reductive changes of the digital arch of all feet. There is a variable preaxial reduction of the number of fingers in both sexes. Moreover, homozygous males are sterile. Testes of homozygous Hd/Hd and +/Hd adult rats were examined in the light and electron microscopes. Spatial organization of stages of the spermatogenetic cycle was not confirmed in Hd/Hd testes comparing with +/Hd males. Significant decrease in the number of germ cells in seminiferous tubule of Hd/Hd testes was accompanied with loosening and vacuolization of the seminiferous epithelium. The assignment of the Hd locus to RNO10 excluded the suspected homology between rat and mouse Hd mutations. More precise mapping using microsatellite markers revealed close linkage of the Hd locus with the D10Mit8 marker defining Syb2 gene coding for synaptobrevin 2. A chromosomal segment of RNO10 carrying Hd and Syb genes is being incrossed onto BN and SHR inbred strains in order to examine the modifying influences of different genetic backgrounds.
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The fundamental concept of a neuroimmunological network is well appreciated although detailed description of the individual mechanisms has not yet been attained. In an effort to close this gap, the effect of electrolesion of the frontal medial cortex, a structure with the known "immunoreactivity", on selected phenotypic features of spleen macrophages was studied. Since sugar receptors (lectins) are pivotal for recognition, custom-made tools termed neoglycoproteins were employed to delineate any injury-induced changes of their expression. The total number of macrophages in the spleen red pulp was assessed using the ED-1 monoclonal antibody. The results showed that after lesion of the medial frontal cortex, the extent of expression of carbohydrate-binding sites in red pulp spleen macrophages significantly decreased without affecting the total number of these cells. These data intimate that distinct brain regions are involved in the control of the phenotype of macrophages in the central lymphoid organs by currently elusive biochemical mechanisms.
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Short Communication

The Win95 program for computation of distance matrixes and construction of phylogenetic or phenetic trees on the basis of RAPD, RFLP and allozyme data was presented. In contrast with other presently available software, the program FreeTree can also assess the robustness of the tree topology by bootstrap, jackknife or OTU-Jackknife analysis. Moreover, the program can be used also for an analysis of data obtained in several independent experiments performed with nonidentical subsets of taxa. The function of the program was demonstrated by an analysis of RAPD data from 22 strains of Frenkelia. The program is available as an autoextractive archive containing the installation files of FreeTree and TreeView, manual in MS Word format and a sample of the input file at http://www.natur.cuni.cz/$flegr/programs/freetree.
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Monoclonal Antibody Register

A Signalling Accessory Molecule Revealed by a New
Anti-Fibroblastoid L Cell Monoclonal Antibody
A. PANCZAK¹, F. HIRSCH^2, D. HAGERTY³, R. BLUMBERG, P. PONCET⁵

Fibroblastoid mouse L-cells are widely used in immunological models because when transfected with class II-coding genes they become efficient antigen presenting cells. Little is known, however, about the cell surface markers borne by L-cells and their putative involvement/interference with the experimental models studied. Rats were immunized against DAP.3 cells (subclone of L-cells) and monoclonal antibodies (mAbs) were prepared. One of them, 4D4, was studied in detail. It recognizes an epitope which is neither cell lineage- nor strain- nor species-restricted since, in addition to DAP.3 cells, it binds, as determined by flow cytometry and immunohistochemistry, to various cells such as CD8⁺ T cells from thymus, spleen, lymph node or intestinal epithelium, mouse peritoneal B cells and various tissues such as renal, pulmonary or intestinal epithelia. 4D4 mAb immunoprecipitates an undescribed 68 kDa protein. Functionally, this mAb inhibits the IL-2 secretion of a T cell clone in response to its peptide presented by appropriate class II-transfected L-cells and induces a negative selection of double positive CD4⁺CD8⁺ thymocytes. Since the 4D4 ligand is found on cells which are submitted to selection (T cells) and on cells which mediate selection (epithelial and antigen presenting cells), we conclude that 4D4 mAb defines a cell surface antigen involved, as an accessory molecule, in a cell selection process.
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