SACM - United Kingdom
Permanent URI for this collectionhttps://drepo.sdl.edu.sa/handle/20.500.14154/9667
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Item Restricted Intracellular and extracellular immunisation for HIV(The University of Manchester, 2024-11) Alatiq, Abdulrahman; Klapper, Paul; Valley, Pamela; Faqih, LaylaThe prevalence of HIV infection remains a serious challenge for public health. HIV infection results in considerable mortality and morbidity, as well as imposing a substantial financial burden on healthcare systems globally. Despite the availability of effective antiretroviral treatments to control infection, an effective protective vaccine remains elusive. This project aimed to develop a practical, affordable, and effective combined vaccine and treatment by combining intracellular antibodies and extracellular immunisation to prevent or reduce HIV viraemia. Specifically, an approach in which DNA-encoding N49P7 broadly neutralising antibodies against the HIV-1 envelope glycoprotein 120 is delivered to cells using recombinant baculovirus or lipid nanoparticle (LNP) transfection. Extracellular antibodies were anticipated to prevent gp120 from attaching to cells, and intracellular antibodies were intended to inhibit the genesis of virions. The expression of N49P7, either as a human IgG antibody or as human Fab, was attempted. The expression of the N49P7 IgG antibody was not achieved in HEK 293 cells despite employing three delivery models for transfections: recombinant baculovirus, MC3-based LNP, or lipofectamine 2000 LNP. While transfection was successfully achieved (monitored via an eGFP gene included within the plasmid design), functional antibody was not achieved. The most likely explanation for the failure was thought to be the use of dual promoters within the expression cassette. Redesign of the plasmid to create a bicistronic vector including the N49P7 Fab region and signal peptide sequences of murine IgG and IL-2 allowed successful expression of N49P7 Fab through recombinant baculovirus or lipofectamine 2000 reagent transfection of HEK 293 cells. However, the expressed N49P7 Fab region was predominantly accumulated intracellularly. A further redesign was implemented to incorporate homogenous signal peptides H7 and L1 into the N49P7 Fab gene, which significantly enhanced the secretion of the antibody fragment. This design maintained functional intracellular and extracellular antibody activity. Lower cellular cytotoxicity was seen with recombinant baculovirus transfection compared to lipofectamine 2000 LNP mediated transfection although both were equally efficient. The selection of an optimum method for transfection will form part of future investigations progressing to animal model testing. This proof of principle study showed that recombinant baculovirus or lipofectamine 2000-mediated transfection systems allowed efficient N49P7 Fab expression both intracellularly and extracellularly in mammalian cells, suggesting that this approach indicates potential for providing a vaccine against HIV infection in addition to a therapeutic intervention for those who already have HIV infection.13 0Item Restricted An exploration of the knowledge and attitudes of Saudi occupational therapists (OTs) regarding people living with HIV\AIDS in Saudi Arabia(Saudi Digital Library, 2023-01-20) Bin Numay, Abdulelah; Sakellariou, DikaiosIntroduction: Relatively 38 million people are living with the acquired immunodeficiency syndrome (AIDS) in addition to the virus which causes AIDS, the human immunodeficiency virus (HIV). AIDS-related complications were responsible for the deaths of 35 million people globally between the beginning of the pandemic and the end of 2020, with the disease's prevalence continuing to rise across the Middle East and North Africa. Due to an early misunderstanding of HIV transmission patterns, many Muslims incorrectly think that only "sinners" may get HIV/AIDS. Consequently, stigma and prejudice are seen as the greatest challenges for persons with HIV/AIDS in Saudi Arabia. Therefore, the purpose of this research is to get an understanding of and investigate the experience and knowledge of Saudi Arabian Occupational Therapists. Methodology: The researcher selected the framework qualitative design as the research's methodology to collect data from six participants, 4 male and 2 female Occupational Therapy practitioners. Semi-structured audio recorded interviews were conducted, recordings were transcribed digitally for data analysis. Then, the qualitative data were thematically analysed. Result: The analysis concluded four main themes and eight subthemes. Proceeded with the participants' general perception of HIV/AIDS, then move on to their knowledge development in relation to the virus, their clinical practise in the context of working with a person living with HIV/AIDS, and finally, the socio-cultural obstacles and future prospects from the perspective of the participants. Conclusion: Participants mentioned they lacked significant understanding about HIV/AIDS and the occupational therapy role with HIV+ patients. Their collective understanding of HIV/AIDS was derived from the theory they studied, movies, television programmes, chats with others, and books or articles, owing their lack of understanding to the incomplete HIV/AIDS education incorporated in the curriculum. In addition, while they would not mind working with an HIV- positive client, HIV/AIDS related discrimination in the study population was presented by altering the treatment approach, take extra contact precautions, minimize interaction between HIV positive patients with other patients9 0