An investigation of FcRL4 positive B cells in the rheumatoid synovium and The development of a cultured ELISPOT assay for the detection of low frequency t cell responses

Lom, Hannah (2013). An investigation of FcRL4 positive B cells in the rheumatoid synovium and The development of a cultured ELISPOT assay for the detection of low frequency t cell responses. University of Birmingham. M.Res.

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Abstract

FcRL4 is a transmembrane protein expressed on a subset of memory B cells. Using immunofluorescent microscopy and qPCR we have investigated the prevalence of FcRL4+ B cells in the synovial tissue, synovial fluid and peripheral blood of Rheumatoid Arthritis patients. FcRL4+ B cells were not identified in synovial tissue sections by immunofluorescent microscopy due to weak staining. qPCR suggests FcRL4 expression is higher in synovial tissue of established RA patients compared to healthy controls. FcRL4 expression is also elevated in the synovial fluid and peripheral blood of RA patients.

HPV-16 is found in ~60% of cases of head and neck squamous cell carcinoma (HNSCC). As a step towards the analysis of HPV-specific T cell responses in tumours of HNSCC patients we have optimised a cultured IFN-γ ELISPOT assay using Epstein Barr virus (EBV) peptide CLG as a model. We found lowest backgrounds when cells were cultured in serum-free AIM-V medium, and ELISPOT plates are blocked with 10% albumin from bovine serum (BSA)or 10%autologous plasma. Culturing PBMCs with peptide prior to performing the ELISPOT assay significantly amplifies the antigen-specific T cell response detected compared to standard ELISPOT assays performed overnight on PBMCs ex vivo.

Type of Work:

Thesis (Masters by Research > M.Res.)

Award Type:

Masters by Research > M.Res.

Supervisor(s):