In vitro generation and evaluation of atypical B cells
The B cell response is strongly affected during P. falciparum malaria. This results in the generation of an atypical B cell compartment that has been associated with poor formation of long-lived B cell-mediated immunity. The role the atypical B cells play in immunity remains unknown. Recently, these cells have also been identified in many more infectious diseases, such as HIV, Tuberculosis, Influenza etc, indicating a general mechanism for their generation and suggest that they could haven an important function. Mouse studies have shown similar cells to be excellent antigen-presenting cells, initiating T cell responses, but also to have a regulatory B cell phenotype, that is important for controlling the strength of the immune response. The aim of this study is to investigate: 1. the capacity of in vitro derived atypical B cells to present antigen to T cells, 2. production of the regulatory cytokine IL-10 in response to stimulation with TLR-ligands. The main methods include to derive atypical B cell in vitro using interferon-g and BCR stimulation from purified B cells obtained from human buffy coats. The cells will then be fed whole CMV antigen or purified peptides, which are then presented to T cells added to the culture. The regulatory effect of atypical B cells will be investigated by stimulating the cells with the toll-like receptor 9-ligand, CpG, after which interleukin 10 will be measured in the culture supernatant. |