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n-3 Polyunsaturated Fatty Acids Suppress the Localization and Activation of Signaling Proteins at the Immunological Synapse in Murine CD4⁺ T Cells by Affecting Lipid Raft Formation¹

Wooki Kim^*, Yang-Yi Fan^*, Rola Barhoumi^,, Roger Smith, David N. McMurray^*,,,¶ and Robert S. Chapkin^2,*,,

^* Faculty of Nutrition, Center for Environmental and Rural Health, Department of Veterinary Integrative Biosciences, Department of Veterinary Pathology, Texas A&M University, and ^¶ Department of Microbial & Molecular Pathogenesis, Texas A&M University System Health Science Center, College Station, TX 77843

The molecular properties of immunosuppressive n-3 polyunsaturated fatty acids (PUFA) have not been fully elucidated. Using CD4⁺ T cells from wild-type control and fat-1 transgenic mice (enriched in n-3 PUFA), we show that membrane raft accumulation assessed by Laurdan (6-dodecanoyl-2-dimethyl aminonaphthalene) labeling was enhanced in fat-1 cells following immunological synapse (IS) formation by CD3-specific Ab expressing hybridoma cells. However, the localization of protein kinase C, phospholipase C-1, and F-actin into the IS was suppressed. In addition, both the phosphorylation status of phospholipase C-1 at the IS and cell proliferation as assessed by CFSE labeling and [³H]thymidine incorporation were suppressed in fat-1 cells. These data imply that lipid rafts may be targets for the development of dietary agents for the treatment of autoimmune and chronic inflammatory diseases.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work was supported by Grants DK071707, CA59034, and P30ES09106 from the National Institutes of Health (NIH). Confocal and multiphoton microscopy was supported with funding by Grant 1 S10 RR22532-01 from National Center for Research Resources-NIH Shared Instrumentation, performed at the College of Veterinary Medicine & Biomedical Sciences Image Analysis Laboratory, Texas A&M University, College Station, TX.

² Address correspondence and reprint requests to Dr. Robert S. Chapkin, Faculty of Nutrition, 321 Kleberg Center, MS 2253, Texas A&M University, College Station, TX 77843-2253. E-mail address: r-chapkin{at}tamu.edu

³ Abbreviations used in this paper: PUFA, polyunsaturated fatty acid; IS, immunological synapse; DHA, docosahexaenoic acid; PKC, protein kinase C; MFI, mean fluorescence intensity; GP, generalized polarization; LAT, linker for activation of T cells; PLC, phospholipase C; CTx, cholera toxin; RRI, relative relocation index; WT, wild type.

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