Clunk, Marilee
(2017)
Siglec-10, CD1B, and Palladin are differentially expressed between Human Immunodeficiency Virus+ (HIV) progressors and nonprogressors.
Master's Thesis, University of Pittsburgh.
(Unpublished)
Abstract
HIV plasma viremia and CD4+ lymphocyte counts dictate the classification of HIV+ disease progression to Acquired Immunodeficiency Syndrome (AIDS), i.e. progressor (PR) vs. non-progressor (NP). In the absence of antiretroviral therapy, NP CD4+ counts remain stable and plasma viremia is substantially reduced. Recent data affirms that NPs exhibit decreased trans-infection efficiency due to reduced cellular cholesterol levels in antigen presenting cells (APCs) through cholesterol efflux protein, ABCA-1. Because defective trans infection in NPs was also observed prior to seroconversion, it is believed that nonprogression is influenced by a genetic trait. Further transcriptome analysis of Multicenter AIDS Cohort Study (MACS) immature dendritic cells (iDCs) reveals elevated expression profiles for Siglec-10, CD1B, and Palladin (PALLD) in NPs compared to PRs. While the explicit mechanism remains elusive, our aim is to elucidate the genetic origin of NP viremic control. I have hypothesized that upregulated expression of Siglec-10, CD1b, and Palladin in HIV+ MACS NPs is responsible for control of HIV progression by attenuating trans infection between iDCs and CD4 lymphocytes through unrelated molecular and immunological mechanisms.
MACS CD14+ monocytes were cultured with IL-4 and GM-CSF to yield iDCs. Protein expression of Siglec-10, CD1B, and PALLD in seronegative (SN), PR, and NP iDCs has been assessed using flow cytometry. Quantitative PCR (qPCR) analysis of MACS iDCs RNA is used for corroboration of gene expression data from the transcriptome findings. TaqMan® SNP genotyping assays examined if there is a link between NP disease state and single base-pair variations within DNA.
Flow cytometry shows no significant difference in Siglec-10, CD1b, or PALLD protein expression levels between PR and NP donors. SyBr green qPCR primers for resulted in artefactual amplicons for control and experimental melt curves. IDT Taq-Man® primers were subsequently designed. Analysis of qPCR data shows significant differences in CD1b and PALLD expression levels between NPs and PRs (p < 0.05). Haplotype analysis of CD1b SNPs between NP, PR, and SN individuals via TaqMan Genotyping was not statistically significant.
Public Health Significance: Identifying the underlying genetic factors that drive HIV-1 disease progression could reveal potential therapeutic targets with the long term aims of improving HIV health outcomes.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
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| ETD Committee: |
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| Date: |
25 September 2017 |
| Date Type: |
Publication |
| Defense Date: |
19 June 2017 |
| Approval Date: |
25 September 2017 |
| Submission Date: |
5 June 2017 |
| Access Restriction: |
No restriction; Release the ETD for access worldwide immediately. |
| Number of Pages: |
58 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Public Health > Infectious Diseases and Microbiology |
| Degree: |
MS - Master of Science |
| Thesis Type: |
Master's Thesis |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
Human Immunodeficiency Virus, HIV, HIV disease progression, Progressor, Nonprogressor, AIDS, Siglec-10, CD1b, Palladin |
| Date Deposited: |
25 Sep 2017 14:18 |
| Last Modified: |
25 Sep 2017 14:18 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/32360 |
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