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Hypercholesterolemia Induced Immune Response and Inflammation on Progression of Atherosclerosis in Apob tm2Sgy Ldlr tm1Her/J Mice

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Lipids

Abstract

The effect of hypercholesterolemia induced immune response and inflammation on progression of atherosclerosis in ApoB tm25gy LDLr tm1Her mice, expressing only ApoB100 and deficient in the low density lipoprotein (LDL) receptor, thus closely resembling human cholesterol transport is not well defined. Atherosclerosis was induced by a high cholesterol diet and its progression was studied at 8, 14 and 20 weeks. Antibody response was determined by ELISA. Lymphocytes in spleen and aortic expression of inflammatory markers were studied by flow cytometry, and immunohistochemistry respectively. A rapid increase in plasma LDL levels in the first 8 weeks was followed by the exponential development of atherosclerosis between 8 and 14 weeks. Progression of the disease was accompanied by an accumulation of macrophages and increased expression of IL17 and IFN-γ in the aorta. Hypercholesterolemia resulted in increased immune response to modified lipids and aortic inflammation, with an expansion of Th17 cells in the spleen. Progression of atherosclerosis showed a positive correlation (r = 0.84, P < 0.001) with Th17 cells and a negative correlation with Treg cells (r = 0.83, P < 0.001). IgM antibodies to Ox-LDL and Th17 cells in spleen showed greatest association with disease development. Our results suggest that anti Ox-LDL IgM antibodies, Th17 cells could be developed as a potential marker to study disease progression and to study the effect of therapeutic regulation of inflammation.

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Abbreviations

Apo B:

Apolipoprotein B

Apo E:

Apolipoprotein E

Con A:

Concanavalin A

EVG:

Elastica van Gieson

GAPDH:

Glyceraldehyde-3 phosphate dehydrogenase

HDL:

High-density lipoprotein

HFD:

High fat diet

HBSS:

Hanks' balanced salt solution

IL-17:

Interleukin-17

IL-10:

Interleukin-10

IFN-γ:

Interferon-γ

LDL:

Low-density lipoprotein

LDL-R:

Low-density lipoprotein receptor

NBF:

Neutral buffered phenol

Ox-LDL:

Oxidized low-density lipoprotein

ORO:

Oil red O

PMA:

Phorbol myristate acetate

RT-PCR:

Real-time polymerase chain reaction

RPMI:

Rosewell Park Memorial Institute

TC:

Total cholesterol

Th1:

T-helper 1

TGF-β:

Transforming growth factor

TNF-α:

Tumor necrosis factor α

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Acknowledgments

We gratefully acknowledge the support of the trustees of Thrombosis Research Institute, London and Bangalore and the Tata Social Welfare Trust, India (TSWT/IG/SNB/JP/Sdm). The study was supported by Department of Biotechnology, Ministry of Science and Technology, Government of India (BT/01/CDE/08/07) and Garry Weston foundation UK.

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Authors and Affiliations

  1. Research Scholar Manipal University at Molecular Immunology Unit, Thrombosis Research Institute, Bangalore, India

    Lakshmi Narasimha Rao & Thiruvelselvan Ponnusamy

  2. Molecular Immunology Unit, Thrombosis Research Institute, Narayana Hrudayalaya, 258/A, Bommasandra Industrial Area, Anekal Taluk, Bangalore, 560099, India

    Sheena Philip & Lakshmi Mundkur

  3. Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur, 784 028, Assam, India

    Rupak Mukhopadhyay

  4. Thrombosis Research Institute, Bangalore, India

    Vijay V. Kakkar

  5. Thrombosis Research Institute, London, UK

    Vijay V. Kakkar

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  1. Lakshmi Narasimha Rao
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Rao, L.N., Ponnusamy, T., Philip, S. et al. Hypercholesterolemia Induced Immune Response and Inflammation on Progression of Atherosclerosis in Apob tm2Sgy Ldlr tm1Her/J Mice. Lipids 50, 785–797 (2015). https://doi.org/10.1007/s11745-015-4046-4

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