Author/Editor		Repnik, Urška
Title		Zorenje dendritičnih celic in njihov vpliv v alogenskem imunskem odzivu in vitro
Translated title		Dendritic cell maturation and their influence in in vitro allogeneic immune response
Type		monografija
Place		Ljubljana
Publisher		Univerza v Ljubljani, Medicinska fakulteta
Publication year		2005
Volume		str. 118
Language		slo
Abstract		Dendritic cells (DCs) are the most potent antigen presenting cells (APCs). They have a major role in the maintenance of tolerance to self antigens and in the initiation and regulation of immune responses against pathogens, tumours and allogeneic grafts. Myeloid DCs reside in peripheral tissues and continuously sample antigens in their microenvironment. Upon the encounter with microbial molecules or inflammatory cytokines, they transform into potent APC, which migrate to draining lymph nodes, where they activate T cells. Development of procedures for ex vivo DC generation prompted their use in immunotherapy. The purpose of our work was to evaluate the effect of different maturation factors and duration of DC maturation on the characteristics of DCs and, consequently, on the nature of the T-cell response they provoke. Our first goal was to develop a protocol for monocyte isolation on Percoll density gradient. We hypothesized that homogenous populations of mature DCs can be prepared by different maturation factors. We wanted to investigate whether differences in concentration of co-stimulatory molecules, observed between DCs matured for different periods of time or with different maturation stimuli, reflect in T-cell proliferation and CTL generation, and whether IL-12 promotes IFN-gamma production by T cells and CTL generation. We suspected that large production of IL-12 by DCs can trigger IL-10 synthesis. Monocytes, isolated from peripheral blood mononuclear cells (PBMC) on Percoll density gradient, were differentiated into immature DCs in a medium supplemented with IL-4 ,and GMCSF. Subsequently, DCs were matured with LPS, a monocyte-conditioned medium (MCM) or TNF-alpha. During DC maturation, morphology of the cells in culture was followed by light microscopy, expression of cell surface markers by flow cytometry and IL-12 production capacity by ELISA. (Abstract truncated at 2000 characters).
Descriptors		DENDRITIC CELLS INTERLEUKIN-12 T-LYMPHOCYTES, CYTOTOXIC INTERFERON TYPE II MONOCYTES LYMPHOCYTE TRANSFORMATION INTERLEUKIN-4 PHENOTYPE ENZYME-LINKED IMMUNOSORBENT ASSAY CULTURE MEDIA FLOW CYTOMETRY