Biomedical Sciences ETDs

Publication Date

7-1-2013

Abstract

Obesity is defined as heightened fat accumulation leading to health impairments. It has been directly correlated to cardiovascular disease, type II diabetes mellitus, and cancer. Heightened cytokine levels are found in serum and adipose tissue of obese subjects, including TNFα (tumor necrosis factor alpha), IL-6 (interleukin-6), and MCP-1 (monocyte chemoattractant protein-1), being characterized as a chronic low-grade inflammatory disease. In this dissertation, I have generated a novel co-culture model between adipocytes and immune cells (derived from splenocytes) that mimics inflammation seen in obese adipose tissue. This co-culture model allows for distinct evaluation between secreted paracrine factors (indirect cultures) and these factors plus direct cell contact. Paracrine signaling from both cell types increased the release of IL-6 and MCP-1, with a concomitant decrease of TNFα, whereas direct physical contact exacerbated the effects. The anti-inflammatory cytokine IL-10 (interleukin-10) did not play a role in the decreased secretion of TNFα. A time course study showed that direct and indirect co-cultures exhibited differential secretion rates, demonstrating cytokine-specific regulatory mechanisms. To determine specific cellular cytokine contributions, directly cultured cells were separated and analyzed showing both adipocytes and immune cells contribute significantly to inflammation. Adipocytes express MCP-1 and IL-6, whereas immune cells, TNFα and IL-6. Additionally, TNFα is necessary for this augmentation of IL-6 and MCP-1 secretion in direct contact. By use of non-toxic levels of signaling pathway inhibitors, I verified that the changes in cytokine secretions are mediated by NF-κB (nuclear factor kappa B) and MAPKs (mitogen-activated protein kinases). Specifically, NF-κB is the major signaling cascade for TNFα production, IL-6 is regulated by NF-κB, JNK (c-Jun N-terminal kinase), and p38, whereas MCP-1 by NF-κB, JNK, and MEK1. Analysis of the cell adhesion molecules on adipocytes identified 42 molecules. Direct contact with immune cells produced an up-regulation of Cadherin-1, claudin-4, -8, and -11, and down-regulation of Integrin alpha 6 in adipocytes. In conclusion, these results illustrate that direct contact and "crosstalk" between adipocytes and immune cells is paramount for exacerbation of inflammation in obesity. These changes are mediated by specific signaling cascades and cell adhesion molecules, which are important novel targets for this disease.

Keywords

obesity, inflammation, cytokine, adipose tissue, immune cell, signaling, cell adhesion molecule

Document Type

Dissertation

Language

English

Degree Name

Biomedical Sciences

Level of Degree

Doctoral

Department Name

Biomedical Sciences Graduate Program

First Advisor

Orlando, Robert

First Committee Member (Chair)

Wandinger-Ness, Angela

Second Committee Member

Garver, William Sherman

Third Committee Member

Howdieshell, Thomas

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