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Experimental and clinically used vaccines based on vaccinia virus
Pilná, Hana ; Mělková, Zora (advisor) ; Šroller, Vojtěch (referee)
Vaccinia virus (VACV) is an enveloped DNA virus belonging in the Orthopoxviridae genus. It is a laboratory virus in which the natural host and exact origin remain unclear. However it is of great significance for human kind. First of all, different VACV strains were used for preparation of vaccines used in the smallpox eradication campaign. Even today a significant effort is made to prepare more efficient and safer vaccines against smallpox, namely because of still remaining concerns that variola virus - causative agent of smallpox - could be misused as a biological weapon. Advances in genetic engineering allowed use of VACV for additional purposes, namely as a vaccination and expression vector. VACV enables insertion of large pieces of foreign DNA into its genome and expression of this DNA in a host. Furthermore VACV replicates exclusively in a cytoplasm, decreasing a risk of incorporation of the viral DNA into the host genome. These and other features make VACV an ideal candidate as a vector for preparation of recombinant vaccines against various infectious and oncological diseases. This thesis provides a summary of both clinically used and experimental vaccines derived from VACV. Powered by TCPDF (www.tcpdf.org)
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In vitro evaluation of novel Toll-like receptor ligands I.
Hudáková, Kristína ; Trejtnar, František (advisor) ; Soukup, Ondřej (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Kristína Hudáková Supervisor: doc. PharmDr. František Trejtnar, CSc. Title of diploma thesis: In vitro evaluation of novel Toll-like receptor ligands I Vaccination against preventable infections prevents millions of deaths each year. Their immunity enhancing activity is strengthened by the presence of vaccine adjuvants. Development of vaccine adjuvants leads to improved safety profile and also can play a vital role in the research of new vaccines against pathogens against which the vaccines currently do not exist. The main aim of this diploma thesis was to verify the ability of rationally developed small molecule ligands to influence Toll-like receptors and thus their potential to be utilized as vaccine adjuvants. The assay was carried out using modified cell lines continually expressing the human TLR4 or TLR8 whose activation leads to production of secreted embryonic alkaline phosphatase. Ten analyzed substances labelled as DM 001 - DM 010 were examined for their agonistic and also antagonistic properties while interacting with the TLRs. Immunomodulatory activity of these tested samples was then determined by quantification of secreted alkaline phosphatase with the help of a colorimetric...
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In vitro evaluation of novel Toll-like receptor ligands III
Tamášiová, Linda ; Trejtnar, František (advisor) ; Pejchal, Jaroslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Linda Tamášiová Supervisor: doc. PharmDr. František Trejtnar, CSc. Title of diploma thesis: In vitro evaluation of novel Toll-like receptor ligands III Research on new ligands with potential immunomodulatory activity is a tool for the development of new immunological adjuvants for use in vaccines, or as separate immunomodulators in the treatment of various diseases. The aim of this thesis was to analyze the ability of new ligands to stimulate toll-like receptors 4 (TLR4) and to assess the potential for further use of these substances based on established immunomodulatory activity. The analysis was performed on modified TLR4-expressing cell lines whose activation was subsequently detected using a colorimetric-enzymatic reaction. Sixteen new substances were tested for TLR4 receptor activation in comparison with a standard agonist. The results showed a significant TLR4 agonistic activity in several of the test substances, suggesting that they are activating ligands of the receptor tested. However, due to their low solubility, some of these substances are not suitable candidates for further use and testing. Taking all parameters into consideration, among of all of the evaluated substances that...
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In vitro evaluation of novel Toll-like receptor ligands II.
Machalová, Vanda ; Trejtnar, František (advisor) ; Janočková, Jana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Vanda Machalová Supervisor: doc. PharmDr. František Trejtnar, CSc. Title of diploma thesis: In vitro evaluation of novel Toll-like receptor ligands II. The aim of this diploma thesis was to test the new substances at Toll-like receptors (TLR), namely TLR4 and TLR8 subtypes, as potential vaccine adjuvants. Adjuvants are required for new subunit vaccines to promote stimulation of a sufficient immune response. Stimulation of TLR receptors is safe and leads to the activation of both innate and adaptive immunity and to subsequent specific immune response. Testing was carried out on cell lines stable co-expressing the respective receptors, with colorimetric activity detection. In this work, both agonist and antagonist activity at TLR4 and TLR8 receptors was investigated and a half maximal effective concentration (EC50) of resiquimod was determined. Based on the results, the respective substances cannot be considered as potential adjuvants to the TLR8 receptor and DM014 exhibits the potential for agonist activity on the TLR4 receptor, whereas the substance DM015 can be used on further investigation of immunosuppression in autoimmune diseases and diseases associated with excessive activation of the...
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In vitro evaluation of novel Toll-like receptor ligands I.
Hudáková, Kristína ; Trejtnar, František (advisor) ; Soukup, Ondřej (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Kristína Hudáková Supervisor: doc. PharmDr. František Trejtnar, CSc. Title of diploma thesis: In vitro evaluation of novel Toll-like receptor ligands I Vaccination against preventable infections prevents millions of deaths each year. Their immunity enhancing activity is strengthened by the presence of vaccine adjuvants. Development of vaccine adjuvants leads to improved safety profile and also can play a vital role in the research of new vaccines against pathogens against which the vaccines currently do not exist. The main aim of this diploma thesis was to verify the ability of rationally developed small molecule ligands to influence Toll-like receptors and thus their potential to be utilized as vaccine adjuvants. The assay was carried out using modified cell lines continually expressing the human TLR4 or TLR8 whose activation leads to production of secreted embryonic alkaline phosphatase. Ten analyzed substances labelled as DM 001 - DM 010 were examined for their agonistic and also antagonistic properties while interacting with the TLRs. Immunomodulatory activity of these tested samples was then determined by quantification of secreted alkaline phosphatase with the help of a colorimetric...
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Therapeutic approaches to Ebola virus infections
Tomanová, Tereza ; Drda Morávková, Alena (advisor) ; Matyska Lišková, Petra (referee)
Ebola virus is a very dangerous virus because of its high mortality rate and unavailability of an approved vaccine or medicament. There was more than 31 000 infected people since the first epidemic in 1976 and almost 13 000 of them died (cdc.gov). Ebola virus usually occures in the west Africa, its occure on the other continents was rare in the past and it was always just one case. There is one exception though, strain Reston of Ebola virus. Strain Reston was spread from Philippines to the USA while deporting macaques. Strain Reston is able to infect people, but there is no case of death after being infected with this strain (who.int). This occurence in the USA was a warning sign of how easily could Ebola virus travel to other continents. If it was one of the deadly strains, the results would be fatal. Ebola virus is a threat and it is necessary to develop effective vaccine and drug. This bachelor's thesis is about the Ebola virus, its genome, life cycle and mainly about the options of therapy of the ebola virus disease. It is focused on presently tested vaccines and drugs against the Ebola virus.
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Experimental and clinically used vaccines based on vaccinia virus
Pilná, Hana ; Mělková, Zora (advisor) ; Šroller, Vojtěch (referee)
Vaccinia virus (VACV) is an enveloped DNA virus belonging in the Orthopoxviridae genus. It is a laboratory virus in which the natural host and exact origin remain unclear. However it is of great significance for human kind. First of all, different VACV strains were used for preparation of vaccines used in the smallpox eradication campaign. Even today a significant effort is made to prepare more efficient and safer vaccines against smallpox, namely because of still remaining concerns that variola virus - causative agent of smallpox - could be misused as a biological weapon. Advances in genetic engineering allowed use of VACV for additional purposes, namely as a vaccination and expression vector. VACV enables insertion of large pieces of foreign DNA into its genome and expression of this DNA in a host. Furthermore VACV replicates exclusively in a cytoplasm, decreasing a risk of incorporation of the viral DNA into the host genome. These and other features make VACV an ideal candidate as a vector for preparation of recombinant vaccines against various infectious and oncological diseases. This thesis provides a summary of both clinically used and experimental vaccines derived from VACV. Powered by TCPDF (www.tcpdf.org)
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Development of vaccines against porcine circovirus type 2
Janovec, Václav ; Drda Morávková, Alena (advisor) ; Pokorná, Dana (referee)
Porcine circovirus type 2 is a single stranded DNA virus that belongs to the genus Circovirus in the family Circoviridae. This virus is associated with many kinds of diseases in pigs and causes significant economic losses in swine-breeding. In this study, two approaches of vaccination were tested in order to develop an effective vaccine against PCV2. The first approach was to test DNA vaccines. For this purpose, eukaryotic expression plasmids encoding two form of PCV2 Cap protein were constructed. The expression plasmids encoding murine TNF-α and IFN-α1 were also prepared for co-immunization with antigen encoding plasmid to enhance the immune response. The second approach is based on the previous finding that chimeric pentamers of VP1 mouse polyomavirus capsid protein fused with PCV2 can induce protective immunity against PCV2. These chimeric pentamers were further modified by AA substitutions in PCV2 Cap immunodominant epitope in order to enhance protective antibody response directed against PCV2. The chimeric pentamers and DNA vaccines were tested for ability to induce antibody immune response against PCV2 in mice. The results showed that chimeric pentamers are more potent inducers of protective antibody immune response against PCV2 compared to DNA vaccines. However, the protective antibody...
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Inovation in influenza vaccines
Satoriová, Martina ; Drda Morávková, Alena (advisor) ; Motlová, Lucia (referee)
Influenza viruses annually infect 3 to 5 millions of people worldwide, cause annually many hospitalizations, 250 000-500 000 deaths and significant economical losses. The vaccines still remain the most efficient way of prevention of this infectious disease. Conventional egg- based vaccines are used for more than 60 years. Although they are safe, they have many disadvantages. Their main disadvantage is the relatively low effectiveness and time-limited induced immunity. The need for annually updates of their composition due to an antigenic variability of viral surface proteins can be considered a disadvantage as well. What is more, the dependence on an egg suply and a way too long time of preparation might be limiting in the case of pandemic. The development of an universal vaccine that would induce a broad imunne response against different strains of Influenza and longlasting protection is a worldwide priority.
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Fish Rhabdoviruses and its prevention
Satoriová, Martina ; Drda Morávková, Alena (advisor) ; Novák, Josef (referee)
The rise of aquaculture has been one of the most significant changes in global food production over the last 100 years. Driven by rising demand for seafood, the growth of population, the farming of aquatic animals has expanded rapidly to become a major global industry. A number of aquatic animal species is kept in high densities in freshwater, brackish and marine systems where they are exposed to a new environment and potentially new diseases. On-farm stresses may negatively affect their ability to defeat infection. Impefect farming practices often facilitate fast transmission of disease. Viral pathogens, whether they have been known for decades or whether they are newly emerging, are particularly challenging since there are few, if any, efficacious treatments. The development of effective viral vaccines for delivery in aquatic systems still remains almost elusive. This thesis reviews a few of the more significant Rhabdoviral pathogens of finfish, their impacts, their prevention and the protective immune mechanisms that fish mount in response to rhabdovirus infections.
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