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fat-1 transgene expression prevents cell culture-induced loss of membrane n-3 fatty acids in activated CD4+ T-cells.

Abstract

In order to evaluate the effects of fatty acids on immune cell membrane structure and function, it is often necessary to maintain cells in culture. However, cell culture conditions typically reverse alterations in polyunsaturated fatty acid (PUFA) composition achieved by dietary lipid manipulation. Therefore, we hypothesized that T-cells from transgenic mice expressing the Caenorhabditis elegans n-3 desaturase (fat-1) gene would be resistant to the culture-induced loss of n-3 PUFA and, therefore, obviate the need to incorporate fatty acids or homologous serum into the medium. CD4+ T-cells were isolated from (i) control wild type (WT) mice fed a safflower oil-n-6 PUFA enriched diet (SAF) devoid of n-3 PUFA, (ii) fat-1 transgenic mice (enriched with endogenous n-3 PUFA) fed a SAF diet, or (iii) WT mice fed a fish oil (FO) based diet enriched in n-3 PUFA. T-cell phospholipids isolated from WT mice fed FO diet (enriched in n-3 PUFA) and fat-1 transgenic mice fed a SAF diet (enriched in n-6 PUFA) were both enriched in n-3 PUFA. As expected, the mol% levels of both n-3 and n-6 PUFA were decreased in cultures of CD4+ T-cells from FO-fed WT mice after 3d in culture. In contrast, the expression of n-3 desaturase prevented the culture-induced decrease of n-3 PUFA in CD4+ T-cells from the transgenic mice. Carboxyfluorescein succinidyl ester (CFSE) -labeled CD4+ T-cells from fat-1/SAF vs. WT/SAF mice stimulated with anti-CD3 and anti-CD28 for 3d, exhibited a reduced (P<0.05) number of cell divisions. We conclude that fat-1-containing CD4+ T-cells express a physiologically relevant, n-3 PUFA enriched, membrane fatty acid composition which is resistant to conventional cell culture-induced depletion.

Authors+Show Affiliations

Department of Nutrition and Food Science, Texas A&M University, College Station, TX 77843-2253, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

18977126

Citation

Fan, Yang-Yi, et al. "Fat-1 Transgene Expression Prevents Cell Culture-induced Loss of Membrane N-3 Fatty Acids in Activated CD4+ T-cells." Prostaglandins, Leukotrienes, and Essential Fatty Acids, vol. 79, no. 6, 2008, pp. 209-14.
Fan YY, Kim W, Callaway E, et al. Fat-1 transgene expression prevents cell culture-induced loss of membrane n-3 fatty acids in activated CD4+ T-cells. Prostaglandins Leukot Essent Fatty Acids. 2008;79(6):209-14.
Fan, Y. Y., Kim, W., Callaway, E., Smith, R., Jia, Q., Zhou, L., ... Chapkin, R. S. (2008). Fat-1 transgene expression prevents cell culture-induced loss of membrane n-3 fatty acids in activated CD4+ T-cells. Prostaglandins, Leukotrienes, and Essential Fatty Acids, 79(6), pp. 209-14. doi:10.1016/j.plefa.2008.09.017.
Fan YY, et al. Fat-1 Transgene Expression Prevents Cell Culture-induced Loss of Membrane N-3 Fatty Acids in Activated CD4+ T-cells. Prostaglandins Leukot Essent Fatty Acids. 2008;79(6):209-14. PubMed PMID: 18977126.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - fat-1 transgene expression prevents cell culture-induced loss of membrane n-3 fatty acids in activated CD4+ T-cells. AU - Fan,Yang-Yi, AU - Kim,Wooki, AU - Callaway,Evelyn, AU - Smith,Roger, AU - Jia,Qian, AU - Zhou,Lan, AU - McMurray,David N, AU - Chapkin,Robert S, Y1 - 2008/10/31/ PY - 2008/07/15/received PY - 2008/09/09/revised PY - 2008/09/11/accepted PY - 2008/11/4/pubmed PY - 2009/4/1/medline PY - 2008/11/4/entrez SP - 209 EP - 14 JF - Prostaglandins, leukotrienes, and essential fatty acids JO - Prostaglandins Leukot. Essent. Fatty Acids VL - 79 IS - 6 N2 - In order to evaluate the effects of fatty acids on immune cell membrane structure and function, it is often necessary to maintain cells in culture. However, cell culture conditions typically reverse alterations in polyunsaturated fatty acid (PUFA) composition achieved by dietary lipid manipulation. Therefore, we hypothesized that T-cells from transgenic mice expressing the Caenorhabditis elegans n-3 desaturase (fat-1) gene would be resistant to the culture-induced loss of n-3 PUFA and, therefore, obviate the need to incorporate fatty acids or homologous serum into the medium. CD4+ T-cells were isolated from (i) control wild type (WT) mice fed a safflower oil-n-6 PUFA enriched diet (SAF) devoid of n-3 PUFA, (ii) fat-1 transgenic mice (enriched with endogenous n-3 PUFA) fed a SAF diet, or (iii) WT mice fed a fish oil (FO) based diet enriched in n-3 PUFA. T-cell phospholipids isolated from WT mice fed FO diet (enriched in n-3 PUFA) and fat-1 transgenic mice fed a SAF diet (enriched in n-6 PUFA) were both enriched in n-3 PUFA. As expected, the mol% levels of both n-3 and n-6 PUFA were decreased in cultures of CD4+ T-cells from FO-fed WT mice after 3d in culture. In contrast, the expression of n-3 desaturase prevented the culture-induced decrease of n-3 PUFA in CD4+ T-cells from the transgenic mice. Carboxyfluorescein succinidyl ester (CFSE) -labeled CD4+ T-cells from fat-1/SAF vs. WT/SAF mice stimulated with anti-CD3 and anti-CD28 for 3d, exhibited a reduced (P<0.05) number of cell divisions. We conclude that fat-1-containing CD4+ T-cells express a physiologically relevant, n-3 PUFA enriched, membrane fatty acid composition which is resistant to conventional cell culture-induced depletion. SN - 0952-3278 UR - https://www.unboundmedicine.com/medline/citation/18977126/fat_1_transgene_expression_prevents_cell_culture_induced_loss_of_membrane_n_3_fatty_acids_in_activated_CD4+_T_cells_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0952-3278(08)00117-8 DB - PRIME DP - Unbound Medicine ER -