Background
Vaccination of vertebrates induce the production of protective antibodies and memory cells. The adaptive immune system responsible for this response is more than 500 million years old and conserved from sharks to humans. A handful of genes introduced during the “big bang” of vertebrate immune evolution are required for these responses and are found in virtually all sequenced vertebrates. Surprisingly, Atlantic cod (and a few other fishes) have lost genes that are essential for functional immunity in mammals. These genes are called MHC II and CD4 and are involved in the presentation of antigens to T cells by antigen-presenting cells. In humans and mouse, T cells express either CD4 or CD8 known as helper cells or killer cells. Through single-cell RNA sequencing we have identified T-cell subsets that express neither CD4 (expected as the gene is missing) nor CD8 and other unusual T cells.
The project
To study T cells (and other cells) in non-model organism, we need to make antibody reagents. We have previously made monoclonal antibodies (mAb) against IgM and B cells to investigate the antibody system of cod. Now we are looking for a masters’ student interested in making antibodies against cod T cells.
Methodology
Generation of monoclonal antibodies involve immunization of mice and harvesting of B cells from the spleen. Spleen B cells are then fused with a leukemia cell line to generate hybridomas, which produce antibodies. Antibodies from clones of hybridomas can subsequently be screened for binding to the desired target, which was used for immunization. The student will work with supervisors and other researchers in the lab on the entire protocol including: preparation of recombinant antigens and DNA vaccines; generation and culturing of hybridomas; screening of hybridomas; characterization of antibodies produced by a clone of hybridoma cells. Promising mAbs will be characterized by flow analysis and immunohistochemistry on cells and organs from Atlantic cod. The experimental work will be done at IBV and at Rikshospitalet.
Relevance
Monoclonal antibodies are the main class of biologically produced drugs in the pharmaceutical industry selling for more than 200 billion US dollars a year, a market expected to exceed 500 billion USD by 2030.
Supervisors
Main supervisor is Finn-Eirik Johansen, and co-supervisor is Shuo-Wang Qiao (Medical Faculty). Finn-Eirik and Shuo-Wang work closely with PhD candidates and postdocs in the comparative immunology group in the lab and on bioinformatics. Contact: f.e.johansen@ibv.uio.no