The Centre for Experimental Medicine & Rheumatology (EMR) comprises an integrated, multidisciplinary research team of Clinicians, Scientists, Biostatisticians and Bioinformaticians led by Professor Costantino Pitzalis.
EMR research aims to better understand disease pathogenesis in order to develop novel therapeutics and maximise therapeutic utility of existing therapies for a number of autoimmune conditions including RA, SLE and Sjogren’s Syndrome.
The Centre runs a strong basic science programme in the field of immunology & inflammation linked to a clinical translational programme in the following 4 research areas: (1) Breach of Tolerance and Autoimmunity, (2) Development and resolution of inflammation, (3) Mechanisms of Tissue Damage and Repair and (4) Developing new therapies through basic research (e.g. tissue specific targeting) and clinical trials.
The Centre is based at the William Harvey Research Institute (WHRI) working closely with partner Centres and Institutes on the Charterhouse Square campus.
To assist and provide scientific and technical support for a research project investigating the role of IKZF1, IKZF3 and other targets in B cell homeostasis in SLE, Sjogren’s syndrome and rheumatoid arthritis (RA).
Main Duties & Responsibilities
This will involve a number of experimental procedures listed below. It is understood that the researcher will need supervision and training to learn some such procedures.
- Perform immunohistochemistry of multiple therapeutic targets in synovial biopsy tissue and salivary gland tissue. Quantify histological staining using slide scanner and digital imaging analysis tools.
- Perform double or triple staining using fluorescence microscopy and/or confocal microscopy to localise protein expression with cell types for B/T cells and subsets, plasma cells, tissue macrophages.
- Cell culture and in vitro differentiation of primary human cells
- Purify PBMC from human blood samples and perform magnetic bead isolation of B cells and other cell types from blood
- Perform cell culture of primary human cells and stimulation with cytokines. Some experiments may include layering of cultured cells on stromal feeder cell lines for long-term culture, therefore it is essential that the candidate has good cell culture and advanced flow cytometry skills.
- Complex flow cytometry
- 10-colour flow cytometry of cultured cells using BD Fortessa and cell sorting using BD Aria.
- Imagestream X2 analysis of NF-kB p65 translocation
- Genetic and transcriptomic analysis
- Isolate RNA from cells for RNA-seq analysis and perform gene expression analysis using real-time PCR
- Isolate DNA from cells for genotyping
- Serum analysis
- Purification of serum from blood samples for ELISA and multiplex cytokine assays (e.g. Luminex)
- Lentiviral silencing and other in vitro cell manipulation experiments
- Perform in vitro gain and loss of function experiments in human cell lines and/or primary human cells using recombinant molecules, lentiviruses, siRNA and shRNA. The researcher is expected to generate some of these reagents in house; therefore he/she will be expected to procure the human/ murine cells, clone, design and generate siRNAs, shRNAs and viruses. Some molecules will require mutagenesis.
- Generate and modify plasmids and reagents required for expression and production of lentiviruses
- Stably transfect cell lines and measure transfection efficiency using flow cytometry or western blot
In addition, the researcher will be expected to:
- Perform all in vitro and in vivo experiments required in this project, analyse results, report to the group leader and to other colleagues, and write the resulting paper(s).
- Contribute intellectually and practically to the group, including supervision of juniors, help colleagues as required, actively participate in the weekly journal club and data sessions.
- Contribute to the overall scientific endeavour of the Centre, and to take responsibility for areas of several of the current projects, as demand requires.
- Work as part of the Institute’s research team, being mutually supportive and covering duties as necessary during colleagues’ absences and at times of additional pressure, as directed.
The above list of responsibilities is not exhaustive, and the jobholder may be required to undertake other duties commensurate with the level of the role, as reasonable requested by their line manager.
This job description sets out the duties of the post at the time it was drawn up. Such duties may vary from time to time without changing the general character of the duties or level of the responsibility entailed. Such variations are a common occurrence and cannot in themselves justify a reconsideration of the grading of the post.