Progress in Vaccine Development for HCV Infection

Hepatitis C virus (HCV) is a blood-transmitted disease that spreads among 3% of the world’s population causing seriously increasing mortality rates. The HCV prevalence in Egypt in October 2008 was 14.7% and declined to 6.3% in the survey carried out in October 2015. Nowadays, the new direct-acting antivirals (DAAs) show amazing results especially with regard to HCV genotype 1, but there is still a great necessity to produce a vaccine to avoid this viral infection. Additionally, neutralizing anti-HCV antibodies could be utilized in combination with DAAs empowering their effect. A powerful candidate HCV vaccine should create comprehensively cross-receptive T cells CD4 and CD8 and effectively neutralizing antibodies to successfully clear the virus. The current clinical trials for HCV vaccines comprise synthetic peptides, DNA-based vaccines, or recombinant protein vaccines. Several preclinical vaccine studies are under research including cell culture-derived HCV (HCVcc), HCV-like particles, and recombinant adenoviral vaccines. This mini-review will discuss the prevalence of HCV worldwide and in Egypt. We will present the recent progress in basic research and preclinical and clinical studies for HCV vaccine. Finally, it will present the phenomena of spontaneous clearance of HCV without treatment as a model for study of HCV vaccine development

The expression of HLA class I molecules and complement regulatory proteins in ovarian cancer

Over recent decades, translational ovarian cancer research has been impeded by its underappreciated molecular heterogeneity and five-year survival has remained poor. One strategy for addressing this problem is to search for molecular biomarkers that can better inform the development and targeting of novel treatments. The aim of this thesis was to construct and validate a tissue microarray of ovarian cancer cases and to survey the expression and prognostic capabilities of immunological molecular markers: specifically HLA class I and the membrane bound complement regulatory proteins CD46, CD55 and CD59. These are central to the efficacy of certain immunotherapies and while they have been shown to have prognostic power in breast and colorectal cancer, they have been investigated less in ovarian cancer. Five copies of a tissue microarray representing 339 cases of ovarian cancer which presented to Derby City General Hospital between 1982 and 1997 were made. The array was stained for CK7, CK20, CA125, CEA, p53 and Bcl-2 following a standard immunohistochemical protocol. A linked clinical database was adapted and assessed for data consistency and subsequently used to analyse the prognostic and clinicopathological associations of the expression data. The array was then stained for HLA class I, B2microglobulin and CD59 using commercial antibodies and for CD55 and CD46 using in-house antibodies. Retained expression of HLA class I molecules independently predicted improved prognosis. High expression of CD55 and CD59 were associated with worse prognosis, though not independently of other factors. CD55 expression was more widespread than previously appreciated. This thesis describes the discovery of a new independent marker of prognosis which suggests that immunoediting occurs in ovarian cancer, describes the distribution of markers known to have a negative impact on immunotherapy in ovarian cancer in a large series for the first time and documents the production of a valuable resource for future studies

Human Surfactant Protein D as an Innate Immune Surveillance Molecule

This thesis was submitted for the award of Doctor of Philosophy and was awarded by Brunel University LondonSurfactant protein D (SP-D) belongs to the family called Collectins (collagen-containing lectins). It has a primary structure characterised by an N-terminal cysteine-rich region, triple-helical collagen region composed of Gly-X-Y repeats (where X and Y can be any amino acid), an a-helical coiled-coil neck region, and a C-terminal carbohydrate recognition domain (CRD). The primary subunit oligomerizes to form a trimeric structure that can further acquire a cruciform organization and multimers (dodecamers). SP-D is a potent innate immune molecule whose presence in the lungs as well as at a range mucosal surfaces and extrapulmonary tissues allows immune surveillance against pathogens, apoptotic/necrotic cells, allergens, and cancer cells. A recombinant fragment of human SP-D (rfhSP-D) composed of 8 Gly-X-Y repeats, neck and CRD region, expressed in E. coli, is well known to act against a range of pathogen and allergen challenge in vitro, in vivo and ex vivo. In this thesis, we have examined the interaction between rfhSP-D and two subtypes (pH1N1 and H3N2) of Influenza A Virus (IAV) (Chapter 3). rfhSP-D interacted with haemagglutinin, neuraminidase and matrix protein 1; inhibited their infectivity against lung epithelia cell lines; and suppressed the cytokine storm induced by the viral challenge. The protective role of rfhSP-D against IAV as an entry inhibitor was further validated by the use of pseudotyped lentiviruses containing haemagglutinin and neuraminidase of the two IAV subtypes. Continuing with the theme of innate immune surveillance by SP-D against lung, pancreatic, ovarian, and prostate cancers, we extended on recent studies to breast cancer using HER2 over-expressing (SKBR3), triple-positive (BT474) and triple-negative (BT20) breast cancer cell lines (Chapter 4). rfhSP-D induced apoptosis at 24 h in SKBR3 and BT747 cells (but not in BT20) via intrinsic apoptosis pathway. However, this protective effect of rfhSP-D was fully negated by the presence of hyaluronic acid, a major extracellular matrix component within the tumour microenvironment (Chapter 5). This thesis highlights the therapeutic potential of rfhSP-D in influenza A Virus infection as well as breast cancer and merits pre-clinical trials in murine models

Multiple myeloma : investigation of the expression of light chain isotypes on the surface of peripheral blood lymphocytes and bone marrow mononuclear cells during different stages of the disease

Multiple myeloma (MM) is a relatively common haematological neoplasm composed of antibody-secreting cells. Currently, prolonged complete remissions are rarely obtained and there is little prospect of cure as on clinical presentation there is often irreversible pathological damage. Treatment of myeloma aims to reduce the tumour cell mass if possible, or to at least retard its growth. Current therapy involves the agents melphalan or cyclophosphamide, singly or combined with prednisone, and has resulted in a median survial for patients responding to treatment in excess of three years (Durie & Salmon, 1982), compared with a median survial of 3.5 months in 1950 (Feinleib & MacMahon). Despite the advances made in understanding the pathophysiology of myeloma in the last 20 years, it is difficult to correlate clinical observations and laboratory data with tumour mass staging (Durie & Salmon, 1975) as although such staging systems have some prognostic importance, survival duration within comparable tumour mass groups varies greatly (Cooper et al, 1982)

The secretory antigens of Schistosoma mansoni and their involvement in the immune elimination of worms from rats.

SIGLEAvailable from British Library Document Supply Centre-DSC:DXN003593 / BLDSC - British Library Document Supply CentreGBUnited Kingdo

Past Endemic Malaria and Adaptive Responses in the Fens and Marshlands of Eastern England

Changes in climate have increased concerns over the return of temperate malaria to the United Kingdom. Hence, studies of ancient disease are becoming more relevant for future health predictions in areas which are under threat of disease re-emergence. Conditions were likely ideal for Plasmodium vivax malaria from at least the Roman period, and recent research on Anglo-Saxon Fen populations has suggested an indigenous malarial presence. The primary aim of this project was to investigate the presence of English malaria in archaeological Fen cemetery populations from the Roman, Anglo-Saxon, and later medieval periods (c. AD 40-1600), using biomolecular analyses of human bone, and palaeopathological analyses via extant published data. A further aim was to investigate Masters’ (1987) hypothesis concerning preferential survival of non-collagenous proteins (NCPs) within archaeological bone. Indirect evidence for malaria was sought by reassessing archaeological reports for osteological evidence of the genetic anaemia β thalassaemia at 13 cemetery sites (five Roman, seven Anglo-Saxon, one late medieval) closely associated with the Fens and marshlands of Lincolnshire and Cambridgeshire. A palaeodemographic comparison of 30 Fen (five Roman, 21 Anglo-Saxon, four late medieval) and 31 non-Fen (nine Roman, 18 Anglo-Saxon, four late medieval) cemetery populations was also undertaken to assess any impact of vivax malaria on mortality. Osteological evidence does not support the presence of past thalassaemia, with palaeodemographic analysis suggesting an acquired, rather than genetic immunity in the Fens. Possible evidence emerged for ‘healthy adaptation’ to the increased stresses of Fenland life, and one population provided tentative evidence of intrauterine growth restriction, a condition strongly linked to endemic P. vivax. Direct evidence was sought by attempting to extract and test anti-malarial antibodies from human bone samples from 13 Fen-associated cemetery sites (five Roman, seven Anglo-Saxon, one late medieval), encompassing 24 individuals. Bone preservation was assessed in over 200 samples to provide a baseline for sample selection for biomolecular analysis. Analysis of bone samples proved unsupportive of Masters’ (1987) hypothesis. However, a range of endogenous proteins and a possible pathogenic disease marker were revealed, as was a correlation between bone preservation and NCP content. Evaluation of extraction protocols failed to yield antibodies which, if present, were consistently masked by collagen. Consequently, a novel antibody extraction technique has been developed. If successful, this could lead to a replicable technique of ancient, reactive antibody isolation, which would offer an invaluable new tool in biomolecular palaeopathology