Madlala, Paradise Zamokuhle.Ntshangase, Wenzile Senorita.2023-09-202023-09-2020232023https://researchspace.ukzn.ac.za/handle/10413/22288Masters Degree. University of KwaZulu-Natal, Durban.Background: Human immunodeficiency virus type 1 (HIV-1) ribonucleic acid (RNA) is characteristically lower in the central nervous system (CNS) than in plasma of antiretroviral treatment naïve patients. Paradoxically, there is higher HIV-1 viral load in the cerebral spinal fluid (CSF) than plasma of treatment naïve patients co-infected with tuberculous meningitis (TBM). The mechanisms that govern high viral replication in the CNS of TBM co-infected antiretroviral therapy naïve patients remain to be determined. Methodology: The study population comprised of 17 TBM and 3 non-TBM participants selected from an HIV-1 positive and TBM co-infected cohort. The HIV-1 viral RNA was reversed transcribed into complementary deoxyribonucleic acid (cDNA) thus the U3/R region of 3’ long terminal repeat (LTR) was amplified from CSF and plasma RNA by KAPA HiFi HotStart PCR Kits (ThermoFisher Scientific, Invitrogen™, USA). The patients CSF and plasma derived LTR were subsequently cloned into a pGL3 plasmid and further transfected in Jurkat and Astrocyte cell lines to assess the LTR transcriptional activity using Bright-Glo™ Luciferase Assay System (Promega, Madison, WI, USA). Results: CSF derived LTR had a significantly (p<0.0001) higher basal and Tat induced transcriptional activity compared to plasma derived LTR in Astrocyte (SVG) cell line. Similarly, CSF derived LTR had significantly higher (p=0.0024) Tat induced transcriptional activity compared to plasma derived LTR in Jurkat cell lines. LTR sequences containing an Adenine (A) at position 5 of the Sp1III binding site were associated with significantly high basal (p<0.0001) and Tat induced (p=0.0002) transcriptional activity compared to the LTR sequences containing a Thymine (T) at the same position when it was assessed in SVG cell. A similar case was observed in Jurkat cell lines. Consistently, CSF LTR sequences containing an A at position 5 of the Sp1III transcription binding site were associated with significantly higher HIV-1 viral load compared to LTR sequences containing a T at the same position (p=0.0093). Conclusion: Our data clearly show that CSF derived LTR from TBM co-infected individuals exhibit significantly higher transcriptional. Particularly, sequences containing the A5T mutation are significantly associated with higher LTR transcriptional activity and viral load.enCentral Nervous System (CNS).Long Terminal Repeat (LTR).Human Immunodeficiency Virus Type 1 (HIV-1).Tuberculous Meningitis (TBM).Co-Infection.Thesis