Essential fatty acids and iron are involved at distinct stages of the proliferative cycle but not in the activation of human T cells.
In the absence of serum, optimal lymphocyte proliferation is obtained when cultures are supplemented with transferrin and an essential fatty acid (EFA). In order to study the effects of iron in conjunction with EFA on T-cell proliferation, we have utilized a chemically defined serum-free culture system to achieve better control of the variables involved. This system includes three different serum-free media (SFM) that differ in total iron content and source of iron: (i) transferrin-free medium containing a high concentration (500 microns) of a soluble iron salt in the form of ferric citrate (Fe-SFM); (ii) iron-saturated human transferrin (5 micrograms/ml) (T-SFM); and (iii) iron-free medium (SFM(-Fe)) without any apparent source of iron. None of these SFM supported proliferation of T cells stimulated by the combination of phorbol 12,13-dibutyrate/ionomycin or phytohemagglutinin. Restoration of the proliferative response was only observed following supplementation of the iron-containing media with linoleic acid (complexed to bovine serum albumin (LA/BSA)). In cultures containing LA/BSA, the addition of iron alone in the absence of transferrin (Fe-SFM) resulted in similar responses to the transferrin-containing medium (T-SFM). Low levels of RNA synthesis in mitogen-stimulated T cells could be demonstrated in the presence or absence of iron and the addition of LA/BSA resulted in marked enhancement of RNA synthesis, regardless of the availability of iron. Cell cycle analysis showed that 91-94% of the cells cultured in SFM were arrested in G0/G1. These cells could progress through the cell cycle following the addition of LA/BSA, but only in the iron-containing media. Unlike DNA or RNA synthesis, activation of T cells could be demonstrated in SFM with or without iron as shown by the normal induction of c-fos and early growth response gene mRNA, normal expression of IL2 and transferrin receptors, and normal IL2 production, despite the arrest of cells in G0/G1. These results suggest that although human T-cell growth is iron and EFA dependent, the early events of T-cell activation are both iron and EFA independent.
Department of Pediatrics, National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado 80206.
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Kovar
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Domenico
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Gelfand
D002453:Cell Cycle; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis; D005786:Gene Expression Regulation; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D007501:Iron / Q000502:physiology*; D019787:Linoleic Acid; D008041:Linoleic Acids / Q000502:physiology*; D008213:Lymphocyte Activation; D012313:RNA / Q000096:biosynthesis; D015375:Receptors, Interleukin-2 / Q000502:physiology; D011990:Receptors, Transferrin / Q000502:physiology; D013601:T-Lymphocytes / Q000276:immunology*
D016895:Culture Media, Serum-Free; D007376:Interleukin-2; D008041:Linoleic Acids; D015375:Receptors, Interleukin-2; D011990:Receptors, Transferrin; D012313:RNA; D004247:DNA; D019787:Linoleic Acid; D007501:Iron