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Establishment and characterization of novel IL-6-dependent myeloma cell line ÚHKT-893
Vančurová, Irena ; Stöckbauer, Petr (advisor) ; Škvor, Jiří (referee)
Multiple myeloma is an incurable fatal neoplasm of plasma cells affecting mainly elderly people. There are many research laboratories in the world where multiple myeloma is studied. Permanent cell lines are indispensable tools for both basic and applied research. However, the establishment of new cell lines is difficult with poor success. We established 96 primary cultures of bone marrow samples from myeloma patients. Only one culture succeeded in permanet myeloma cell line. The novel plasmacytic cell line ÚHKT-893 was established from bone marrow sample of 57 years old female relapsed with multiple myeloma of IgGκ isotype. The cell line is however dependent on the continuous presence of interleukin-6 in culture media. ÚHKT-893 cells grow continuously already more than 1 year. The growth of cells was regularly monitored according to growth curves and by measurement of cell viability. Surface antigenic profile was repeatedly determined by flow cytometry. The simultaneous expression of both CD138 and CD38 surface molecules confirmed the plasma cell origin of cells. The establishment of the ÚHKT-893 myeloma cell line will extend the panel of existing myeloma cell lines as a new ex vivo model for the study of the etiology and pathogenesis of multiple myeloma. It will also provide the source of new...
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Application of molecular and cellular biology methods in research of protozoa Eimeria
Vrba, Vladimír ; Škvor, Jiří (advisor) ; Čepička, Ivan (referee) ; Hampl, Vladimír (referee)
Eimeria is an apicomplexan parasite causing disease coccidiosis that is most prominent in poultry farming industry. This thesis is aimed to develop new molecular tools and resolve issues that would be a valuable contribution in the field from both research and industry perspective. Because immunity to Eimeria is strictly species- specific, it is important to know and recognize correctly all species that parasitize the host. Traditional diagnostic approaches rely on classical methods such as oocyst morphology determination under the microscope, measurement of prepatent period or in-vivo assessment of lesions caused by this parasite. However, diagnostics of individual species using these methods is very time-consuming and it is often unreliable, especially when mixture of multiple species whose parameters overlap is analyzed. Methods utilizing conventional PCR to distinguish species already exist, however, they lack advantages offered by quantitative real-time PCR (qPCR). The first aim of this thesis was to develop qPCR assays for detection and quantification of seven Eimeria species which infect chicken utilizing single-copy non-polymorphic targets in order to ensure maximal specifity and coverage of all strains of each species. Usefulness of this method was demonstrated by analysis of field...
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Immunologic and epigenic modulation of gene expression of human leukemia cell lines
Elknerová, Klára ; Stöckbauer, Petr (advisor) ; Špíšek, Radek (referee) ; Škvor, Jiří (referee)
Therapy by monoclonal antibody to CD34 molecule Growth-inhibitory and proapoptotic effect of the monoclonal antibody to CD34 molecule, clone 4H11, were tested on CD34+ leukemic cell lines (MOLM-9, JURL-MK1) and CD34- cell line (PS-1). We have found that the monoclonal antibody to CD34 inhibited the proliferation and induced apoptosis of all CD34+ cell lines tested, however it has similar effect on CD34- leukemic cell line (PS-1) in concentration higher than 32 µg/ml. We did not observe induction of differentiation by anti-CD34 antibody, but a growth arrest of cells in the G0/G1 phase of the cell cycle was detected in CD34+ cell lines. Combinations of anti-CD34 antibody with both type I (α, β) or type II ( ) interferons did not enhance the effects on the cell growth or inhibition of cellular proliferation of the antibody alone. Combinations of anti-CD34 antibody with hematopoietic cytokines or INF-γ did not induced diferentiation. Our data suggest that the monoclonal antibody to CD34 molecule prepared from clone 4H11, after sufficient experimental and preclinical testing on laboratory animals, may provide a new basis for possible targeted antibody therapy of acute or chronic myeloid leukemia. Therapy by histone deacetylase inhibitors Histone deacetylase inhibitors (HDACi) are emerging new class of...
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