Date of Award

Spring 5-8-2024

Author's School

College of Arts & Sciences

Author's Program

Biology

Degree Name

Bachelor of Arts (A.B.)

Restricted/Unrestricted

Unrestricted

Abstract

Human T-cell leukemia virus type-1 (HTLV-1) is an oncogenic retrovirus that causes multiple disorders, including adult T-cell leukemia and HTLV-1-associated myelopathy/tropical spastic paraparesis. HTLV-1 retroviral integrase binds to the regulatory B’56γ subunit of the host cell Protein Phosphatase 2A (PP2A). Integrase contains a highly conserved LxxIxE motif that is essential for binding, which increases integration efficiency and facilitates HTLV-1 hijack of host cell machinery. We aim to understand how mutations introduced in the highly conserved binding site can affect viral particle production and infectivity. We transfected 729B human lymphoblastoid cells and 293T cells with mutant and wildtype virus. Mutations L213A, I216A, and E218A within the LxxIxE motif interfered with integrase-PP2A binding and the D122N mutation eliminated integrase catalytic activity, serving as a negative control. Single cell clones were obtained from mutant and wildtype virus transfected cells. Virus production for each clone was determined using a p19 ELISA assay. The clones with higher and similar p19 production were selected for infectivity assays and will be used to infect humanized mice for our in vivo study. The binding site mutation decreased viral replication in 729B-ACH cell clones and decreased infectivity in both cell lines. We anticipate that humanized mice will exhibit slower disease progression or even no HTLV-1 infection and disease when infected with the mutant viruses. This study provides preliminary evidence for treatments that target integrase-PP2A binding since binding site mutations decreased HTLV-1 viral replication and infectivity.

Mentor

Lee Ratner, MD, PhD

Additional Advisors

Xiaogang Cheng, Nehla Banu

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