Title page for ETD etd-0107102-160706

Document Type thesis

Author Name Farfan Arribas, Diego Jose

URN etd-0107102-160706

Title DNA Vaccines Against HIV-1: Augmenting Immunogenicity of gp120

Degree MS

Department Biology & Biotechnology

Advisors
David S. Adams, PhD, Advisor
Shan Lu, MD, PhD, Co-Advisor
Joseph C. Bagshaw, Committee Member

Keywords
Gene Therapy
Immunology
Vaccine
HIV
AIDS

Date of Presentation/Defense 2002-01-07

Availability unrestricted

Abstract
There is currently no protective vaccine against HIV. It is known that a high mutation rate and the existence of many subspecies or clades generated by point mutations or recombination events, are at least partly responsible for the ability of the virus to escape immune responses elicited by classical vaccines. Protein subunit vaccines may not be effective due to this pronounced viral mutability. An immune evasion
mechanism has been postulated in which variable domains occlude conserved epitopes crucial for infectivity. The use of DNA vaccines appeared as a favorable approach.
Here, a DNA vaccine approach is presented in which the DNA constructs have been engineered to circumvent the aforementioned problems by 1) introducing elements to enhance expression, such as a heterologous promoter, a heterologous signal sequence and intron sequences, 2) by optimizing codon usage, and 3) by vaccinating with antigens
that have a modified glycosylation pattern which will make them more immunogenic.
The results indicated that deglycosylation of different clades of gp120 did not affect the
protein conformation, and ‘in vitro’ expression levels were good. Antigen codon optimization dramatically increased antibody production. In the animals vaccinated with non-codon-optimized constructs, the presence of an intron and a heterologous signal sequence was required to achieve a good antibody response. Therefore, antigen engineering is required to obtain a powerful immune response against HIV-1 gp120.

Files
arribas.pdf

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Document Type	thesis
Author Name	Farfan Arribas, Diego Jose
URN	etd-0107102-160706
Title	DNA Vaccines Against HIV-1: Augmenting Immunogenicity of gp120
Degree	MS
Department	Biology & Biotechnology
Advisors	David S. Adams, PhD, Advisor Shan Lu, MD, PhD, Co-Advisor Joseph C. Bagshaw, Committee Member
Keywords	Gene Therapy Immunology Vaccine HIV AIDS
Date of Presentation/Defense	2002-01-07
Availability	unrestricted
Abstract There is currently no protective vaccine against HIV. It is known that a high mutation rate and the existence of many subspecies or clades generated by point mutations or recombination events, are at least partly responsible for the ability of the virus to escape immune responses elicited by classical vaccines. Protein subunit vaccines may not be effective due to this pronounced viral mutability. An immune evasion mechanism has been postulated in which variable domains occlude conserved epitopes crucial for infectivity. The use of DNA vaccines appeared as a favorable approach. Here, a DNA vaccine approach is presented in which the DNA constructs have been engineered to circumvent the aforementioned problems by 1) introducing elements to enhance expression, such as a heterologous promoter, a heterologous signal sequence and intron sequences, 2) by optimizing codon usage, and 3) by vaccinating with antigens that have a modified glycosylation pattern which will make them more immunogenic. The results indicated that deglycosylation of different clades of gp120 did not affect the protein conformation, and ‘in vitro’ expression levels were good. Antigen codon optimization dramatically increased antibody production. In the animals vaccinated with non-codon-optimized constructs, the presence of an intron and a heterologous signal sequence was required to achieve a good antibody response. Therefore, antigen engineering is required to obtain a powerful immune response against HIV-1 gp120.
Files	arribas.pdf