Decreased Proviral DNA Loads and Changes in Innate Immune System-Related Transcription Factor Expression in Simian Immunodeficiency Virus-Infected Rhesus Macaques after Blocking the alpha4beta7 Gut-Homing Integrin Pubblico

Abstract

Abstract
Decreased Proviral DNA Loads and Changes in Innate Immune System-Related
Transcription Factor Expression in Simian Immunodeficiency Virus-Infected Rhesus
Macaques after Blocking the alpha4beta7 Gut-Homing Integrin
By Mohammad Yahya Zaidi



Administration of a novel recombinant rhesus monoclonal antibody (mAb) with specificity to
the alpha4beta7 gut-homing integrin prior to and 28 days after infection with the Simian
Immunodeficiency Virus (SIV) resulted in a significant decrease in colorectal and jejunal
proviral DNA loads of anti-alpha4beta7 treated rhesus macaques compared to control SIV-infected
rhesus macaques. The anti-alpha4beta7 monoclonal antibody administration also affected innate
immune system-related transcription factor expression, with significant changes in expression
of the transcription factors related to Th17 cell activity and Regulatory T cell activity (Tregs)
compared to the control SIV-infected rhesus macaques. Changes in innate immune system-
related plasma cytokine levels were also seen in the anti-alpha4beta7 mAb treated monkeys
compared to the control SIV-infected rhesus macaques. The inhibition of MIP-3alpha synthesis by
gut tissues complemented other data collected in the lab which suggested that the anti-alpha4beta7
mAb blocks trafficking of CD4+ T cells as well as inhibits the mobilization of other innate
immune system cell lineages to the gut, thereby protecting the gut tissue and modulating
acute SIV infection.

Table of Contents

Table of Contents

Introduction ..................................................................................................................................... 1
Hypothesis ....................................................................................................................................... 4
Materials and Methods .................................................................................................................... 5
Animals and administration of recombinant anti-alpha4beta7 monoclonal antibody ............................... 5
Virus used for infection and mononuclear cell isolation from blood and biopsy tissues ............ 6
Mononuclear cell isolation from the blood and biopsy tissues ................................................... 6
Isolation of Cellular DNA .............................................................................................................. 8
Calculation of the Pro-viral Load in PBMC, colorectal and jejunal cells ...................................... 9
Isolation of Cellular RNA and Reverse Transcription of RNA ..................................................... 10
Quantitative mRNA analysis ...................................................................................................... 11
Table 1 .................................................................................................................................... 12
Enzyme-linked Immunosorbent Assay (ELISA) Analysis of Plasma Levels of IL-10, IL-7, IL-15,
TGF-beta and IFN-alpha ...................................................................................................................... 12
Table 2 .................................................................................................................................... 14
Results ............................................................................................................................................ 15
Effect of in vivo anti-α4β7 mAb infusion on proviral DNA load in PBMCs and mononuclear cells
from jejunal and colorectal biopsies .......................................................................................... 15
Figure 1. ................................................................................................................................. 15
Figure 2. ................................................................................................................................. 17
Figure 3. ................................................................................................................................. 18
The Kinetics of Various Transcription Factor and Cytokine mRNA in jejunal biopsies during
acute SIVmac239 infection in anti-alpha4/beta7 treated and control monkeys. ..................... 20
Figure 4. ................................................................................................................................. 21
Figure 5. ................................................................................................................................. 25
Effect of in vivo anti-α4β7 mAb infusion on plasma levels of IL-10, IL-7, IL-15, TGF-beta, and IFN-alpha: ................................................................................................................................................ 29
Figure 6. ................................................................................................................................. 30
Discussion ...................................................................................................................................... 32
Appendix ........................................................................................................................................ 45
Supplemental Figure 1........................................................................................................... 45
Supplemental Figure 2 ........................................................................................................... 47
Supplemental Figure 3 ........................................................................................................... 47

Supplemental Figure 4 ........................................................................................................... 48
Supplemental Figure 5 ........................................................................................................... 49
Supplemental Figure 6 ........................................................................................................... 50
References ……………………………………………………………………………………………………………………………....51

About this Master's Thesis

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School	Laney Graduate School
Department	Biological and Biomedical Sciences
Degree	MS
Submission	Master's Thesis
Language	English
Research Field	Health Sciences, Immunology Biology, Virology
Parola chiave	SIV a4b7 Gut-Homing Integrin
Committee Chair / Thesis Advisor	Ansari, Aftab A, Emory University
Committee Members	Gerardo, Nicole Marie, Emory University Stokes, Darrell R, Emory University Starnes, Amanda, Emory University

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