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Differences in iron requirements by concanavalin A-treated and anti-CD3-treated murine splenic lymphocytes

Published online by Cambridge University Press:  09 March 2007

Solo R. Kuvibidila ,
Maria Velez ,
Lolie Yu ,
Raj P. Warrier  and
B. Surendra Baliga
Solo R. Kuvibidila*
Affiliation:
Louisiana State University, Department of Pediatrics, Divisions of Hematology/Oncology, 1542 Tulane Avenue, New Orleans, LA 70112, USA
Maria Velez
Affiliation:
Louisiana State University, Department of Pediatrics, Divisions of Hematology/Oncology, 1542 Tulane Avenue, New Orleans, LA 70112, USA
Lolie Yu
Affiliation:
Louisiana State University, Department of Pediatrics, Divisions of Hematology/Oncology, 1542 Tulane Avenue, New Orleans, LA 70112, USA
Raj P. Warrier
Affiliation:
Louisiana State University, Department of Pediatrics, Divisions of Hematology/Oncology, 1542 Tulane Avenue, New Orleans, LA 70112, USA
B. Surendra Baliga
Affiliation:
University of South Alabama College of Medicine, Department of Pediatrics, 2451 Fillingim Street, Mobile AL 36617, USA
*
*Corresponding author: Dr Solo R. Kuvibidila, fax +1 504 568 3078, email [email protected]
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Abstract

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Fe availability is critical for optimal lymphocyte proliferation; however, the minimum required levels are unknown. Such information is valuable when assessing in vitro immune responses in Fe-deficient subjects, because serum (Fe) added to the culture medium may replete lymphocytes. To address this issue, splenic lymphocytes obtained from seventeen 3-month-old C57BL/6 mice were incubated without and with 1 mg/l concanavalin A or 50 μg/l anti-CD3 antibody in media that contained between 0·113 and 9·74 μmol Fe/l. Fe was provided by either fetal calf serum (FCS, 0–100 ml/l), newborn calf serum (NBCS, 0–100 ml/l), or NBCS (10 ml/l) plus ferric ammonium citrate. As expected, the rate of DNA synthesis increased with Fe levels (P<0·01). Maximum DNA synthesis was obtained with 2·26 μmol Fe/l (50 ml FCS/l) for concanavalin A and 0·895 μmol/l (20 ml FCS/l) for anti-CD3-treated cells. In serum-free media (0·113 μmol Fe/l), the proliferative responses to concanavalin A were below the background, while they rose 5·5-fold in anti-CD3-treated cells (P<0·05). In apotransferrin-supplemented media (0·13 μmol Fe/l), the proliferative responses to concanavalin A and anti-CD3 antibody were 18·6 and 71 %, respectively, of that obtained with 4·66 μmol Fe/l (100 ml FCS/l). Interleukin 2 secretion also followed the same trend as lymphocyte proliferation. Since differences between both mitogens persisted after FCS was substituted with NBCS, we can rule out an effect on ribonucleotide reductase activity, or by other serum growth factors. We speculate an Fe effect at an early step of T-cell activation. Data suggest that the minimum Fe concentration required for lymphocyte proliferation varies with the mitogen.

Keywords

IronLymphocyte proliferationConcanavalin AAnti-C antibodyInterleukin 2
Type
Research Article
Information
British Journal of Nutrition , Volume 88 , Issue 1 , July 2002 , pp. 67 - 72
Copyright
Copyright © The Nutrition Society 2002

References

Abbas, AK, Lichtman, AH & Pober, JS (1997 a) T cell maturation in the thymus. In Cellular and Molecular Immunology, pp. 171–193 [Abbas, AK, Lichman, AH and Pober, JS, editors]. Philadelphia, PA: W.B. Saunder Company.Google Scholar
Abbas, AK, Lichtman, AH & Pober, JS (1997 b) Maturation of B lymphocytes and expression of immunoglobulin genes. In Cellular and Molecular Immunology, pp. 66–95 [Abbas, AK, Lichtman, AH and Pober, JS, editors]. Philadelphia, PA: W.B. Saunder Company.Google Scholar
Abbas, AK, Lichtman, AH & Pober, JS (1997 c) T cell antigen recognition and activation. In Cellular and Molecular Immunology, pp. 139–170 [Abbas, AK, Lichtman, AH and Pober, JS, editors]. Philadelphia, PA: W.B. Saunder Company.Google Scholar
Alcantara, O, Obeid, L, Hannun, Y, Ponka, P & Boldt, DH (1994) Regulation of protein kinase C (PKC) expression by iron: effect of different iron compounds on PKC-beta and PKC-alpha gene expression and the role of the 5'-flanking region of the PKC-beta gene in the response to ferric transferrin. Blood 84, 3510–3517.CrossRefGoogle ScholarPubMed
Brock, HJ (1981) The effect of iron and transferrin on the response of serum-free cultures of mouse lymphocytes to concanavalin and lipopolysaccharides. Immunology 43, 387–392.Google Scholar
Brock, HJ (1992) Iron and the immune system. In Iron and Human Disease, pp. 161–178 [Lauffer, RB, editor]. Boca Raton, FL: CRC Press.Google Scholar
Cazzola, M, Bergamaschi, G, Dezza, L & Arosio, P (1990) Manipulation of cellular iron metabolism for modulating normal and malignant cell proliferation: achievements and prospects. Blood 75, 1903–1919.CrossRefGoogle Scholar
Furukawa, T, Naitoh, Y, Kohno, H, Tokinaga, R & Taketani, S (1992) Iron deprivation decreases ribonucleotide reductase and DNA synthesis. Life Sciences 50, 2059–2065.CrossRefGoogle ScholarPubMed
Galan, P, Thibault, H, Preziosi, P & Herceberg, S (1992) Interleukin-2 production in iron-deficient children. Biological Trace Element Research 32, 421–426.CrossRefGoogle ScholarPubMed
Gao, J & Richardson, DR (2001) The potential of iron chelators of the pyridoxal isonicotinoyl hydrazone class as effective antiproliferative agents. IV: the mechanisms involved in inhibiting cell-cycle progression. Blood 98, 842–850.CrossRefGoogle ScholarPubMed
Golding, S & Young, SP (1995) Iron requirements of human lymphocytes: relative contribution of intra- and extra-cellular iron. Scandinavian Journal of Immunology 41, 229–236.CrossRefGoogle ScholarPubMed
Gross, RL, Reid, JVO, Newberne, PM, Burgess, B, Marston, R & Hift, W (1975) Depressed cell-mediated immunity in megaloblastic anemia due to folic acid deficiency. American Journal of Clinical Nutrition 28, 225–232.CrossRefGoogle ScholarPubMed
Haq, RU, Wereley, JP & Chitambar, CR (1995) Induction of apoptosis by iron deprivation in human leukemic CCRF-CEM cells. Experimental Hematology 23, 428–432.Google ScholarPubMed
Hoffbrand, AV, Ganeshaguru, K, Hooton, JWL & Tattersall, MHN (1976) Effect of iron deficiency and desferrioxamine on DNA synthesis. British Journal of Haematology 33, 517–526.CrossRefGoogle ScholarPubMed
Kuvibidila, SR, Baliga, BS, Warrier, RP & Suskind, RM (1998) Iron deficiency reduces the hydrolysis of cell membrane phosphatidyl-inositol 4,5 bisphosphate during splenic lymphocyte activation in C57BL/6 mice. Journal of Nutrition 128, 1077–1083.CrossRefGoogle ScholarPubMed
Kuvibidila, SR, Kitchens, D & Baliga, BS (1999) In vivo and in vitro iron deficiency reduces protein kinase C activity and translocation in murine splenic and purified T cells. Journal of Cellular Biochemistry 74, 468–478.3.0.CO;2-G>CrossRefGoogle ScholarPubMed
Lederman, HM, Cohen, A, Lee, JWW, Freedman, MH & Gelfand, EW (1984) Deferoxamine: a reversible S-phase inhibitor of human lymphocyte proliferation. Blood 64, 748–753.CrossRefGoogle ScholarPubMed
Mainou-Fowler, T & Brock, JH (1985) Effect of iron deficiency on the response of mouse lymphocytes to concanavalin A: the importance of transferrin-bound iron. Immunology 54, 325–332.Google ScholarPubMed
Munro, HB (1993) Differences among group means: one-way analysis of variance. In Statistical Methods for Health Care Research, 2nd ed., pp. 99–128 [Munro, HB and Page, EB, editors]. Philadelphia, PA: J.B. Lippincott Company.Google Scholar
Nagase, F, Abo, T, Hiramatsu, K, Suzuki, S, Du, J & Nakashima, I (1998) Induction of apoptosis and tyrosine phosphorylation of cellular proteins in T cells and non-T cells by stimulation with concanavalin A. Microbiology and Immunology 42, 567–574.CrossRefGoogle Scholar
Omar, FO & Blakley, BR (1994) The effects of iron deficiency and iron overload on cell-mediated immunity in the mouse. British Journal of Nutrition 72, 899–909.CrossRefGoogle Scholar
Phillips, JL, Bodt, DH & Harper, J (1987) Iron-transferrin-induced increase in protein kinase C activity in CCRF-CEM cells. Journal of Cellular Physiology 132, 349–353.CrossRefGoogle ScholarPubMed
Scaccabarozzi, A, Arosio, P, Weiss, G, Valenti, L, Dongiovanni, P, Fracanzani, AL, Mattioli, M, Levi, S, Fiorelli, G & Fargion, S (2000) Relationship between TNF-α and iron metabolism in differentiating human monocytic THP-1 cells. British Journal of Haematology 110, 978–984.CrossRefGoogle ScholarPubMed
Thelander, L, Graslund, A & Thelander, M (1983) Continual presence of oxygen and iron required for mammalian ribonucleotide reduction. Possible regulation mechanism. Biochemical and Biophysical Research Communications 110, 859–865.CrossRefGoogle ScholarPubMed
Thibault, H, Galan, P, Selz, F, Preziosi, P, Olivier, C, Badoual, J & Hercberg, S (1993) The immune response in iron-deficient young children: effect of iron supplementation on cell-mediated immunity. European Journal of Pediatrics 152, 120–124.CrossRefGoogle ScholarPubMed