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The bovine and human respiratory syncytial viruses (RSV)s can cause severe lower respiratory tract infections. The surface glycoprotein, G protein, ofRSV is primarily thought to be an attachment protein and is an important protective antigen. Previous studies have shown the possibility ofthe G protein or regions within it playing a role in the pathogenesis ofRSV through modulation ofthe host immune system. In this report, the role ofthe G protein in the pathogenesis ofRSV in vivo was investigated. Infection of calves with recombinant (r) bovine (B) RSV expressing different forms of the G protein showed that the soluble form ofthe G protein (Gs) is essential for successful replication in the lungs. In addition, although rBRSV expressing only the membraneanchored G protein (Gm) was attenuated in the lungs and did not induce any pulmonary pathology, mucosal vaccination with this virus was able to protect calves against subsequent BRSV challenge. Studies using rBRSVs were also carried out in vitro by observing rBRSV infection of bovine airway epithelial cell cultures (BAEC). Previous work with recombinant vaccinia virus (rVV) has shown that the presence ofRSV G protein enhances replication ofrVV in the lungs of mice compared to control rVVs. To investigate which region(s) ofthe G protein may be responsible forJhis effect; mice were infected with rVV expressing the HRSV G protein containing deletions of specific regions. The replication ofrVV expressing the G protein lacking residues 193-205 (rVVG~193-105) was attenuated in the lungs of mice compared to rVVGwt. In addition rVVG~193-205 infection of mice induced lower amouptsofIFN-y in the broncho-alveolar lavage (BAL) compared to rVVGwt. The results described indicate that further investigation ofthe innate immune response induced following infection by different forms of RSV-G wiII contribute to a better understanding of how the adaptive immune response to RSV is influenced by the G protein.

Multiple myeloma (MM) is a haematological malignancy characterised by the proliferation of clonal plasma cells in the bone marrow, usually leading to the secretion of a monoclonal immunoglobulin termed the M-protein. Multiple myeloma is a complex disease, and despite great advances in both detection and treatment of the disease, there remains a vast amount of interest in improving patient outcomes. In the era of novel agents and clinical trials, patients with MM are living longer and there is a requirement for frequent monitoring to assess treatment response and signs of relapse. Despite these improvements in treatment, even those patients achieving deep responses are still relapsing due to the presence of very low levels of residual disease, termed minimal residual disease (MRD). Increasingly sensitive techniques are needed to detect this MRD and, as MM is a bone marrow based disease, the majority of techniques focus on bone marrow based assays. However, as bone marrow sampling is invasive, painful and unpleasant for patients it is important to investigate whether blood-based assays could be equally or more informative of disease status. This project therefore aimed to obtain evidence as to whether blood-based assays can act as a safe and effective marker for disease in the bone marrow of treated multiple myeloma patients. Firstly, a bone marrow flow cytometry assay was validated to act as the gold standard technique. Next, three blood-based assays were investigated: circulating tumour plasma cell measurement, heavy/light chain measurement and mass spectrometry evaluation. This study shows the significance of blood-based assays at different disease stages in both transplant and nontransplant treated patients. These results lead to the suggestion of a testing algorithm focused on the addition of heavy/light chain measurement which can be pre-emptively used to influence decisions regarding bone marrow analysis and treatment, ultimately improving the patient experience.

Rheumatoid Arthritis (RA) is a chronic, systemic autoimmune disease, primarily affecting the small joints of the hands, wrists and feet. Disease onset is insidious with classical symptoms including pain, morning stiffness, and persistent synovitis; accompanied by fatigue, malaise and weight loss. RA patients also suffer from a number of extra-articular manifestations and additional comorbidities, including rheumatoid nodules and scleritis; as well as an increased risk of cardiovascular disease (CVD), lymphoma and infection. Ultimately, RA is a devastatingly progressive disease, if a diagnosis and an effective therapeutic regimen is not established quickly. Identifying when a therapy is proving ineffective can be difficult, however, and is reliant upon accurate formal joint counts, CRP measurements and patient global assessments. These are potentially insensitive means of assessing remission and may result in delayed changes to therapeutic regimens. The identification of new biomarkers to improve the predictive value of these evaluation models would therefore greatly benefit patient care. Based upon current evidence, it has been hypothesized that RA patients who fail to make a clinical improvement following the initiation of csDMARD and bDMARD therapeutic regimens are identifiable by their profile of the following CD4+ T-cell populations: follicular helper, memory helper, antigen-specific and naturally-occurring regulatory CD4+ T-cells. As such, flow cytometric assays have developed for the identification and enumeration of these respective CD4+ T-cell populations; the relative and absolute counts of which were investigated within a cohort of healthy controls, with normal distributions established. Subsequently, we have identified distinct alterations to the CD4+ T-cell compartments of newly-diagnosed, treatment-naïve and longstanding RA patients, with clear associations with patient clinical outcomes observed. Of notable significance, the relative distribution of CCR4- Th2-like follicular helper CD4+ T-cells within newly-diagnosed, treatment-naïve RA patients at the time of diagnosis, has proven highly predictive of therapeutic efficacy; with changes in the relative distribution of Th17-like follicular helper and Th17 memory helper CD4+ T-cells also associated with changes to disease activity over time. Furthermore, consistent with previous studies, the CMV status of newly-diagnosed, treatment-naïve and longstanding RA patients, is associated with an aggravated clinical course; with changes in CMV-specific CCR4+ Th17 cell responses related to changes in disease activity 12-months after the initiation of a csDMARD therapeutic regimen. No such relationships could, however, be discerned within the naturally-occurring regulatory CD4+ T-cell compartment, limiting the utility of employing naturally-occurring regulatory CD4+ T-cell measurements in therapeutic efficacy monitoring. Nevertheless, these preliminary results indicate that the addition of follicular helper, memory helper and CMV-specific CD4+ T-cell responses to current remission assessment practices, could improve the identification of RA therapy non-responders, enabling the swift adjustment of RA patient therapeutic regimens and, consequently, improving disease management.