The role of crude saliva and purified salivary mucins in the inhibition of the Human Immunodeficiency Virus type 1

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dc.contributor.advisorMall, Anwar Sulemanen_ZA
dc.contributor.authorPeacocke, Julia Helenen_ZA
dc.date.accessioned2015-11-02T10:58:19Z
dc.date.available2015-11-02T10:58:19Z
dc.date.issued2011en_ZA
dc.descriptionIncludes bibliographical references.en_ZA
dc.description.abstractSub- Saharan Africa is the world's worst HIV-AIDS affected region. More interventions to manage this pandemic are urgently required. Transmission of the virus through an exchange of saliva is rarely known to occur. The role of saliva and its mucus in this inhibition requires clarification. This project sought to verify the previous findings in our laboratory (Habte et al. , 2006; Peacocked et al. (manuscript under revision)), that crude saliva from uninfected individuals together with its purified mucin components inhibited HIV-1 in an in vitro assay. Mucins from infected saliva did not show inhibition in this assay. Mucus was extracted in 6M guanidinium hydrochloride and a cocktail of protease inhibitors, pH 6.5. Sepharose CL-4B gel filtration separated MUC5B and MUC7 in saliva. Mucins were purified by density gradient ultra-centrifugation in caesium chloride. Western blotting and amino acid analysis determined the size, charge and identity of the mucins. The inhibitory activity of crude saliva, salivary mucin components MUC5B and MUC7 from HIV-negative and HIV-positive donors, was tested by their incubation in an in vitro HIV-neutralisation assay using a pseudovirus of Subtypes A and C, and infection of susceptible epithelial tumour cells (genetically modified TZM- bl cells). Glycosylation analysis by HPLC was also performed on each group. The presence of MUC5B and MUC7 in saliva was confirmed and it was shown that there was inter-individual variation in their amounts and electrophoretic behaviour between and within groups. Crude HIV-negative and HIV-positive saliva and its purified mucins MUC5B and MUC7 significantly inhibited the infection of HIV-1 in an in vitro pseudoviral assay in a dose-response manner. HPLC revealed two glycoforms of salivary MUC5B.en_ZA
dc.identifier.apacitationPeacocke, J. H. (2011). <i>The role of crude saliva and purified salivary mucins in the inhibition of the Human Immunodeficiency Virus type 1</i>. (Thesis). University of Cape Town ,Faculty of Health Sciences ,Department of Surgery. Retrieved from http://hdl.handle.net/11427/14613en_ZA
dc.identifier.chicagocitationPeacocke, Julia Helen. <i>"The role of crude saliva and purified salivary mucins in the inhibition of the Human Immunodeficiency Virus type 1."</i> Thesis., University of Cape Town ,Faculty of Health Sciences ,Department of Surgery, 2011. http://hdl.handle.net/11427/14613en_ZA
dc.identifier.citationPeacocke, J. 2011. The role of crude saliva and purified salivary mucins in the inhibition of the Human Immunodeficiency Virus type 1. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Peacocke, Julia Helen AB - Sub- Saharan Africa is the world's worst HIV-AIDS affected region. More interventions to manage this pandemic are urgently required. Transmission of the virus through an exchange of saliva is rarely known to occur. The role of saliva and its mucus in this inhibition requires clarification. This project sought to verify the previous findings in our laboratory (Habte et al. , 2006; Peacocked et al. (manuscript under revision)), that crude saliva from uninfected individuals together with its purified mucin components inhibited HIV-1 in an in vitro assay. Mucins from infected saliva did not show inhibition in this assay. Mucus was extracted in 6M guanidinium hydrochloride and a cocktail of protease inhibitors, pH 6.5. Sepharose CL-4B gel filtration separated MUC5B and MUC7 in saliva. Mucins were purified by density gradient ultra-centrifugation in caesium chloride. Western blotting and amino acid analysis determined the size, charge and identity of the mucins. The inhibitory activity of crude saliva, salivary mucin components MUC5B and MUC7 from HIV-negative and HIV-positive donors, was tested by their incubation in an in vitro HIV-neutralisation assay using a pseudovirus of Subtypes A and C, and infection of susceptible epithelial tumour cells (genetically modified TZM- bl cells). Glycosylation analysis by HPLC was also performed on each group. The presence of MUC5B and MUC7 in saliva was confirmed and it was shown that there was inter-individual variation in their amounts and electrophoretic behaviour between and within groups. Crude HIV-negative and HIV-positive saliva and its purified mucins MUC5B and MUC7 significantly inhibited the infection of HIV-1 in an in vitro pseudoviral assay in a dose-response manner. HPLC revealed two glycoforms of salivary MUC5B. DA - 2011 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 2011 T1 - The role of crude saliva and purified salivary mucins in the inhibition of the Human Immunodeficiency Virus type 1 TI - The role of crude saliva and purified salivary mucins in the inhibition of the Human Immunodeficiency Virus type 1 UR - http://hdl.handle.net/11427/14613 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/14613
dc.identifier.vancouvercitationPeacocke JH. The role of crude saliva and purified salivary mucins in the inhibition of the Human Immunodeficiency Virus type 1. [Thesis]. University of Cape Town ,Faculty of Health Sciences ,Department of Surgery, 2011 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/14613en_ZA
dc.language.isoengen_ZA
dc.publisher.departmentDepartment of Surgeryen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.subject.otherMedicineen_ZA
dc.titleThe role of crude saliva and purified salivary mucins in the inhibition of the Human Immunodeficiency Virus type 1en_ZA
dc.typeMaster Thesis
dc.type.qualificationlevelMasters
dc.type.qualificationnameMScen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceThesisen_ZA
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