| Abstract [eng] |
One of the most widely used tools for T lymphocyte activation is magnetic beads with immobilized IgG antibodies against CD3 and CD28 receptors. These antibodies provide two essential signals for T lymphocyte activation and stimulate their proliferation. Currently, there is an effort to use not only recombinant single-domain antibodies instead of conventional IgG antibodies, but also alternative carriers instead of magnetic beads. This allows to avoid certain drawbacks of existing activators, reduce their cost, simplify the production process and create an alternative tool, more suitable for other applications. The aim of this master thesis is to compare different T lymphocyte activators, comparing not only their function, but their activation mechanism as well. During the study, at first, activation of CD3 and CD28 receptors was determined by stimulating modified T lymphocytes. Afterwards, using T lymphocytes isolated from peripheral blood mononuclear cells, the ability of activators to induce phenotype changes, stimulate cytokine synthesis and cell division was analyzed. Subsequently, the activation mechanism at the genetic level was determined using microarray technology. The results obtained from these studies were used to determine the relative effectiveness of the investigated T lymphocyte activators. |
