The main goal of this thesis was to study cancer immunotherapy based on combination of ligands of TLR and phagocytic receptor agonists. This study is focused on looking for proper and save TLR stimulation and specific anchoring of phagocytic ligands.

This thesis is focused on analysis of immune processes during cancer immunotherapy which is based on synergistic combination of TLR agonists and phagocytosis stimulating ligands anchored into the tumour cell membrane. This immunotherapy was tested in murine melanoma B16-F10 and murine pancreatic adenocarcinoma Panc02 models. The aims of this thesis were to analyse the tumor infiltration, elucidate the role of innate immunity in this immunotherapy, and study the possibility of strengthening immunotherapeutic effect using anti-CD40 and anti-CTLA-4.

Lung cancer is the leading cause of cancer mortality worldwide. Understanding biological processes of specific antitumor immune response remains of an eminent interest and represents necessity for designing successful antitumor immunotherapeutic strategies. The theoretical part of the thesis describes components of the immune system that are involved in antitumor response and discusses their role in the hitherto known and used lung cancer immunotherapy. In the practical part of the thesis, two studies studying different aspects of anticancer immune response are described. Both studies were conducted in cooperation with 3rd Surgical Department 1st Faculty of Medicine, Charles University and University Hospital Motol and with the biotechnology company Sotio a.s. The first study is focused on the humoral component of the specific antitumor response and prospectively analyses serum frequency of antitumor antibodies against NY-ESO-1, Her2/neu and MAGE-A4 antigens in 121 patients with NSCLC. Here it was shown for the first time that tobacco smoking significantly increases the frequency of NY- ESO-1 antibodies in sera of smokers in comparison to ex-smokers and non-smokers. The second study is focused on the cellular component of the specific antitumor response investigating the activity of the dendritic...

In the first part of the theses, I studied therapeutical effect of intratumoral application of various types of bacteria on melanoma B16-F10 bearing mice alone or in combination with another immunostimulatory compounds. Tumour size, metastasis and survival were monitored. The second part was focused on study of cytotoxic effect of neutrophils on melanoma cells and the role of neutrophil extracellular traps formation.

The aim of this thesis was to investigate the possible role of neutrophil granulocytes in antitumor reactions. Most of our experiments were focused on in vitro studies assessing the cytotoxic effect of mouse neutrophils on B16-F10 melanoma cells labelled with PAMPs. We put an emphasis on activation and generating a prime state of neutrophils. Moreover, a release rate of enzyme myeloperoxidase from azurophil granules was detected as a marker of neutrophil degranulation. We also attempted to attract neutrophils into tumor microenvironment using thioglycolate medium and its main compound casein.

The aim of thesis was to study murine melanoma B16- F10 therapy based on the use of TLR agonists combinedwith activation of phagocytosis. Mechanisms of this therapy were studied on the bases of analysis of tumor infiltrating immune cells and evaluationof thein effect on tumor cells.

Transduction of melanoma cells with the aim to induce avidine expression on tumor cell surface was studied. Subsequently the method enabling quantification of binding of ligands to the cell surface was developed.

The main goal of the thesis was to optimize the use of agonists of the FPR receptors for cancer therapy. The aim was: (1) to find the best anchor of ligands to the cancer cells and the optimal timing of the treatment, (2) to verify the universality of agonists of FPR receptors for different types of cancer, (3) to explain the synergy between ligands of FPR and ligands of TLR by flow cytometry and (4) to verify the applicability of cytotoxic test CD45+/ PI for f-MLF-DOPE ligand.

The aim of this thesis was to study cancer immunotherapy based on combination of TLR7 signalling and activation of phagocytosis of cancer cells. This thesis was focused on the possibility of enhancing its effect. The possibilities of removing immunosuppressive barriers (esp. MDSCs and adenosine) were studied, using a combination with chemotherapy or enzyme - adenosine deaminase and purine nucleoside phosphorylase. The effect of enzymes was studied in both in vivo and in vitro experiments. As another way of optimizing the therapy used so far, the modulation of the therapeutic mixture using lower molecular weight substances was tried.

The immunotherapeutic drug DCVAC/OvCa is being tested in the treatment of ovarian cancer patients within the SOV02 clinical trial (Eudra CT number: 2013-001323-38). Ovarian cancer belongs to gynaecological malignancies with the highest mortality rate. Around 60% of patients are diagnosed at advanced stages. Despite the initial successful treatment, relapses occur in most cases, and the disease often becomes resistant to chemotherapy. Effective therapy for relapsed or metastatic patients is still missing. The solution could be immunotherapeutic treatment. DCVAC is an active cellular immunotherapy based on autologous dendritic cells. The aim of this diploma thesis was monitoring of immune parameters in samples from clinical trial SOV02 patients during the time period defined in the study protocol. We have monitored the presence of antigen specific T lymphocytes, tumor specific antibodies, immunosuppressive populations of regulatory T cells and MDSC cells, and also the expression of inhibitory molecules on the surface of T lymphocytes. We observed higher levels of Her-2, Muc-1 and MAGE-A1 antibodies in the DCVAC/OvCa treated group of patients versus the control group. Significant differences in the other monitored parameters were not observed. However, a large amount of data have been obtained that...