Volume 46 (2000) No. 3

Articles
Peritumoral Administration of Antigen-Unstimulated Bone Marrow-Derived Dendritic Cells
Inhibits Tumour Growth
L. MENDOZA¹, M. INDROVÁ¹, R. HÁJKOVÁ¹, M. REINIŠ¹, M. ŠMAHEL², V. VONKA²,
J. BUBENÍK¹, T. JANDLOVÁ¹......................................................91
¹Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
²Institute of Haematology and Blood Transfusion, Prague, Czech Republic
Corresponding author: Luis Mendoza, Institute of Molecular Genetics, Academy of Sciences of
the Czech Republic, Flemingovo nám. 2, 166 37 Prague 6, Czech Republic. Tel: 420 (2) 20183461; Fax: 420 (2) 24310955.
Abstract.
Full text. 91-92  93-94 95-97

Rat Inbred PD/Cub Strain as a Model of Dyslipidemia and Insulin Resistance
L. ŠEDOVÁ¹, L. KAZDOVÁ², O. ŠEDA³, D. KŘENOVÁ³, V. KŘEN^3,4......................99
¹Institute of Pharmacology, 1^st Medical Faculty, Charles University, Prague
²Institute for Clinical and Experimental Medicine, Prague
³Institute of Biology and Medical Genetics, 1^st 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, Prague, Albertov 4, 12800 Prague 2, Czech Republic. Tel: + 420 (2) 24914958; email: vkren(zavináč)lf1.cuni.cz.
Abstract.
Full text.99-101 102-104 105-106

Perichondrial Chondrogenesis and Periosteal Osteogenesis by Localized Graft-vs-Host
Reaction in Mice
M. LUCZAK¹, K. WLODARSKI², K. GALUS².....................................107
¹Department of Medical Microbiology, and ²Department of Histology and Embryology,
Biostructure Center of Medical Academy, Warsaw, Poland
Corresponding author: Krysztof Wlodarski, Department of Histology and Embryology,
Biostructure Center of Medical, Academy, 02-004 Warszawa, Chalubinskiego 5, Poland.
Abstract.
Full text. 107-112

Synthetic Hydrogel Capacity to Induce Formation of Foreign-Body Giant Multinucleate
Cells Differs in Vivo and in Vitro
K. SMETANA, Jr.^1,2, Z. HOLÍKOVÁ¹, U. SEITZER³, H. HAAS⁴, J. VACÍK²..............113
¹Charles University, 1^st Faculty of Medicine, Institute of Anatomy, Prague, Czech Republic
²Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Prague,
Czech Republic
³Division of Immunology and Cell Biology and ⁴Division of Clinical Medicine, Research Center Borstel, Germany
Corresponding author: 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 24 91 95 26; e-mail: ksmet(zavináč)lf1.cuni.cz.
Abstract.
Full text. 113-114  115-116  117-118

Phenotype, Immunological Reactivity and Cytokine Production Profile of Peyer's Patch Cells
from Mice Immunized Orally with Allogeneic Cells
J. FRIČ^1,2, M. KRULOVÁ^1,2, A. ZAJÍCOVÁ¹, V. HOLÁŇ^1,2.................................119
¹Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
²Faculty of Sciences, Charles University, Prague, Czech Republic
Corresponding author: Vladimír Holáň, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 37 Prague 6, Czech Republic. Tel.: 420 (2) 20183226;
Fax: 420 (2) 24310955; e-mail: holan@img.cas.cz.
Abstract.
Full text. 119-121  122-123 124-125

Murine BM cells from C57BL/6 (B6) mice were grown in vitro in medium supplemented with GM-CSF and IL-4 to differentiate DC from DC precursors. After 10 days of culture, approximately 20% of the cell population exhibited a characteristic morphology of BMDC. In cytofluorometric analysis the morphological changes of cells were accompanied by upregulation of the expression of the MHC class II, CD11c, CD80, and CD86 molecules. The BMDC were pulsed with a lysate of syngeneic MK16 carcinoma cells and used for in vitro activation of SC. Co-cultivation of the carcinoma lysate-pulsed BMDC with SC induced a proliferative response of the syngeneic SC. Priming of the proliferative responses was more efficient when the BMDC were grown in the presence of GM-CSF and IL-4 for 10 days than for 7 days. The in vivo effect of mature, tumour lysate-unstimulated BMDC was examined in mice carrying syngeneic MK16 carcinoma transplants. It has been found that local pretreatment with BMDC inhibits growth of a subsequent challenge inoculum of the MK16 cells. Similarly, treatment of mice carrying small MK16 tumours and of those after MK16 surgical minimal residual disease with BMDC significantly inhibited tumour growth. It can be concluded from these results that local concentration of mature BMDC at the tumour site can control the development and growth of the transplanted tumour inoculum.
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Genome scan and metabolic profile of the PD/Cub rat inbred strain in comparison with SHR and BN strains is presented. The PD/Cub strain has been bred by brother sister mating for more than 60 generations since 1969. Its highly inbred status has been confirmed by PCR genotyping with more than 170 microsatellite markers. No case of residual heterozygosity has been recorded. Accordingly, the values obtained by the analysis of metabolic phenotypes are homogeneous with low variance. The PD/Cub strain has significantly higher triglyceride levels and epididymal fat weight as compared to both SHR and BN strains. The PD strain also displays the lowest incorporation of ¹⁴C-U glucose into the epididymal adipose tissue. The data on glucose metabolism clearly indicate that the PD/Cub strain is insulin resistant. Genetic homogeneity and reproducibility of experimental results qualify the PD/Cub rats as an animal model for analysis of the syndrome X.
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Localized GvH reaction produced by the subcutaneous injection of parental lymphoid cells into the ear lobe of Fl recipient mice results in local activation of perichondrial chondrogenesis. Similarly, when the GvH reaction was elicited in shanks, the periosteal membranes at the site of immune reaction were stimulated to proliferate and to produce new bone. However, syngeneic lymphocytes activated in vivo by Con A and intact spleen and bone marrow cells administered locally produced similar response of bone and cartilage. Thus the lymphocytes activated either in the course of GvH reaction or syngeneic lymphocytes grafted into heterotopic sites presumably release mediators capable to stimulate periosteal/perichondrial membranes.
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The granulomatous reaction accompanied with MGC formation represents the most striking feature of the non-favourable biological tolerance of implanted devices. We compared MGC formation in the course of the granulomatous reaction in vitro and in vivo employing three types of hydrogels whose biocompatibility had been well studied earlier. The efficiency of the in vitro assay for the granulomatous reaction, including MGC formation, was verified employing the nematode Nippostrongylus brasiliensis, a well-known inductor of
MGC formation in vitro. The in vitro results demonstrated a very low level of MGC formation in reaction against all three types of hydrogels without polymer-specific differences in comparison with the nematode experiment characterized by a high extent of MGC formation. On the other hand, the extent of MGC formation was implant type-specific in vivo: pHEMA-coDMAEMA > pHEMA > pHEMA-co-NaMA. These results indicate that in the in vitro assay it was not possible to discriminate among the types of polymers used in the experiment in comparison with the animal experiment. They also indicate potential differences between granuloma formation induced by parasites and by foreign bodies.
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Mice of inbred strain BALB/c were immunized orally for 10 consecutive days with fresh spleen
cells from allogeneic C57BL/10 (B10) donors. The immunized mice displayed significant allotransplantation immunity in vivo as demonstrated by resistance to the growth of allogeneic tumours induced by high doses of tumour cells. No significant changes in the proportion of the major T cell
subsets in PP of immunized mice were found 1 or 7 days after the last immunization dose. However, PP cells from orally immunized mice displayed stronger proliferative response after stimulation with cells used for oral immunization than the cells from control animals. Similarly, after stimulation in vitro with specific alloantigens, PP cells from orally immunized mice produced more IFN-gamma than the cells from control recipients. On the contrary, the production of IL-4 was significantly decreased in the immunized mice. The production of IL-2 by PP cells after oral immunization was not significantly changed and IL-10 was only slightly increased. The results thus show that oral immunization with allogeneic cells induces systemic transplantation immunity which can be demonstrated already in Peyer's patches by increased cell proliferation after immunization with specific alloantigens and by changes in cytokine production.
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