Background: Activation-induced apoptosis is believed to limit cell proliferation and eliminate the high number of activated cells during an immune response. Methods: Activation-induced apoptosis was investigated in splenic T cells isolated from young (6 months) and old (24 months) male Fischer 344 rats. The cells were incubated with anti-CD3, concanavalin A or staphylococcal enterotoxin B (primary stimulus) for 72 h, followed by restimulation with anti-CD3 or concanavalin A (secondary stimulus). Apoptosis was assessed by DNA fragmentation assay and DNA gel electrophoresis. The expression of the apoptotic marker CD95 was analyzed by flow cytometry and the relative levels of CD95 ligand, Bcl-2 and Bax protein were measured by immunoblotting. Results: It was shown that DNA fragmentation was very low in the unstimulated T cells from both young and old rats. However, the level of DNA fragmentation was 45–55% greater in the activated T cells from old rats than in the activated T cells from young rats. The increase in DNA fragmentation was paralleled by an increase in the proportion of cells expressing the CD95 molecule. The proportion of CD95+ cells was approximately 40% higher in T cells from old rats than in T cells from young rats. In addition, it was found that the expression of CD95 ligand and Bax increased and Bcl-2 decreased in the activated T cells from old rats compared to the activated T cells from young rats. Conclusion: Our data suggest that the increase in sensitivity of T cells to apoptosis with age may contribute to age-associated immune dysfunction and disorders.

Keywords:
T cell, Apoptosis, CD95, CD95L, Bax, Bcl-2, Rat, Aging
1.
Kabelitz D, Janssen O: Antigen-induced death of T-lymphocytes. Front Biosci 1997;2:d61–d77.
2.
Perandones CE, Illera VA, Peckham D, Stunz LL, Ashman RF: Regulation of apoptosis in vitro in mature murine spleen T cells. J Immunol 1993;151:3521–3529.
3.
Critchfield JM, Lenardo MJ: Antigen-induced programmed T cell death as a new approach to immune therapy. Clin Immunol Immunopathol 1995;75:13–19.
4.
Cohen JJ: Apoptosis: Mechanisms of life and death in the immune system. J Allergy Clin Immunol 1999;103:548–554.
5.
Potestio M, Caruso C, Gervasi F, Scialabba G, D’Anna C, Di Lorenzo G, Balistreri CR, Candore G, Romano GC: Apoptosis and ageing. Mech Ageing Dev 1998;102:221–237.
6.
Mogil RJ, Radvanyi L, Gonzalez-Quintial R, Miller R, Mills G, Theofilopoulos AN, Green DR: Fas (CD95) participates in peripheral T cell deletion and associated apoptosis in vivo. Int Immunol 1995;7:1451–1458.
7.
Smith CA, Williams GT, Kingston R, Jenkinson EJ, Owen JJ: Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures. Nature 1989;337:181–184.
8.
Shi YF, Bissonnette RP, Parfrey N, Szalay M, Kubo RT, Green DR: In vivo administration of monoclonal antibodies to the CD3 T cell receptor complex induces cell death (apoptosis) in immature thymocytes. J Immunol 1991;146:3340–3346.
9.
Jenkinson EJ, Kingston R, Smith CA, Williams GT, Owen JJ: Antigen-induced apoptosis in developing T cells: A mechanism for negative selection of the T cell receptor repertoire. Eur J Immunol 1989;19:2175–2187.
10.
Radvanyi LG, Shi Y, Mills GB, Miller RG: Cell cycle progression out of G1 sensitizes primary-cultured nontransformed T cells to TCR-mediated apoptosis. Cell Immunol 1996;170:260–273.
11.
Radvanyi LG, Shi Y, Vaziri H, Sharma A, Dhala R, Mills GB, Miller RG: CD28 costimulation inhibits TCR-induced apoptosis during a primary T cell response. J Immunol 1996;156:1788–1798.
12.
Radvanyi LG, Mills GB, Miller RG: Religation of the T cell receptor after primary activation of mature T cells inhibits proliferation and induces apoptotic cell death. J Immunol 1993;150:5704–5715.
13.
Yuan J: Transducing signals of life and death. Curr Opin Cell Biol 1997;9:247–251.
14.
Klas C, Debatin KM, Jonker RR, Krammer PH: Activation interferes with the APO-1 pathway in mature human T cells. Int Immunol 1993;5:625–630.
15.
Alderson MR, Tough TW, Braddy S, Davis-Smith T, Roux E, Shooley K, Miller RE, Lynch DH: Regulation of apoptosis and T cell activation by Fas-specific mAb. Int Immunol 1994;6:1799–1806.
16.
Vignaux F, Vivier E, Malissen B, Depraetere V, Nagata S, Golstein P: TCR/CD3 coupling to Fas-based cytotoxicity. J Exp Med 1995;181:781–786.
17.
Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD: The release of cytochrome c from mitochondria: A primary site for Bcl-2 regulation of apoptosis (see comments). Science 1997;275:1132–1136.
18.
Vaux DL: CED-4 – the third horseman of apoptosis (comment). Cell 1997;90:389–390.
19.
Vaux DL: Immunopathology of apoptosis – introduction and overview. Springer Semin Immunopathol 1998;19:271–278.
20.
Reed JC: Double identity for proteins of the Bcl-2 family. Nature 1997;387:773–776.
21.
Miller RA: Aging and immune function. Int Rev Cytol 1991;124:187–215.
22.
Pahlavani MA, Richardson A: The effect of age on the expression of interleukin-2. Mech Ageing Dev 1996;89:125–154.
23.
Pawelec G, Remarque E, Barnett Y, Solana R: T cells and aging. Front Biosci 1998;3:d59–d99.
24.
Pahlavani MA: T cell signaling: Effect of age. Front Biosci 1998;3:D1120–D1133.
25.
Zhou T, Edwards CK III, Mountz JD: Prevention of age-related T cell apoptosis defect in CD2-fas-transgenic mice (see comments). J Exp Med 1995;182:129–137.
26.
Coleman GL, Barthold W, Osbaldiston GW, Foster SJ, Jonas AM: Pathological changes during aging in barrier-reared Fischer 344 male rats. J Gerontol 1977;32:258–278.
27.
Pahlavani MA, Richardson A: Age-related decrease in the naive (OX22+) T cells in F344 rats. Mech Ageing Dev 1994;74:171–176.
28.
Herrmann M, Lorenz HM, Voll R, Grunke M, Woith W, Kalden JR: A rapid and simple method for the isolation of apoptotic DNA fragments. Nucleic Acids Res 1994;22:5506–5507.
29.
Duke RC, Cohen JJ: Morphological and biochemical assays of apoptosis; in Coligan JE, Kruisbeek AM, Margulies DH, Shevac EM, Strober W (eds): Current Protocols in Immunology. New York, Wiley, 1992, pp 3.17.1–3.17.16.
30.
Ferlini C, Di Cesare S, Rainaldi G, Malorni W, Samoggia P, Biselli R, Fattorossi A: Flow cytometric analysis of the early phases of apoptosis by cellular and nuclear techniques. Cytometry 1996;24:106–115.
31.
Pahlavani MA, Harris MD, Richardson A: Activation of p21ras/MAPK signal transduction molecules decreases with age in mitogen-stimulated T cells from rats. Cell Immunol 1998;185:39–48.
32.
Duke RC, Chervenak R, Cohen JJ: Endogenous endonuclease-induced DNA fragmentation: An early event in cell-mediated cytolysis. Proc Natl Acad Sci USA 1983;80:6361–6365.
33.
Cohen JJ, Duke RC: Glucocorticoid activation of a calcium-dependent endonuclease in thymocyte nuclei leads to cell death. J Immunol 1984;132:38–42.
34.
Arends MJ, Morris RG, Wyllie AH: Apoptosis. The role of the endonuclease. Am J Pathol 1990;136:593–608.
35.
McConkey DJ, Orrenius S, Jondal M: Cellular signalling in programmed cell death (apoptosis). Immunol Today 1990;11:120–121.
36.
Salmon M, Pilling D, Borthwick NJ, Viner N, Janossy G, Bacon PA, Akbar AN: The progressive differentiation of primed T cells is associated with an increasing susceptibility to apoptosis. Eur J Immunol 1994;24:892–899.
37.
Jones LA, Chin LT, Longo DL, Kruisbeek AM: Peripheral clonal elimination of functional T cells. Science 1990;250:1726–1729.
38.
Kawabe Y, Ochi A: Programmed cell death and extrathymic reduction of Vbeta8+ CD4+ T cells in mice tolerant to Staphylococcus aureus enterotoxin B (see comments). Nature 1991;349:245–258.
39.
Owen-Schaub LB, Yonehara S, Crump WL III, Grimm EA: DNA fragmentation and cell death is selectively triggered in activated human lymphocytes by Fas antigen engagement. Cell Immunol 1992;140:197–205.
40.
Akbar AN, Borthwick N, Salmon M, Gombert W, Bofill M, Shamsadeen N, Pilling D, Pett S, Grundy JE, Janossy G: The significance of low bcl-2 expression by CD45RO T cells in normal individuals and patients with acute viral infections. The role of apoptosis in T cell memory. J Exp Med 1993;178:427–438.
41.
Yoshino T, Kondo EI, Cao L, Takahashi K, Hayashi K, Nomura S, Akagi T: Inverse expression of bcl-2 protein and Fas antigen in lymphoblasts in peripheral lymph nodes and activated peripheral blood T and B lymphocytes. Blood 1994;83:1856–1861.
42.
Chrest FJ, Buchholz MA, Kim YH, Kwon TK, Nordin AA: Anti-CD3-induced apoptosis in T-cells from young and old mice. Cytometry 1995;20:33–42.
43.
Lechner H, Amort M, Steger MM, Maczek C, Grubeck-Loebenstein B: Regulation of CD95 (APO-1) expression and the induction of apoptosis in human T cells: Changes in old age. Int Arch Allergy Immunol 1996;110:238–243.
44.
Phelouzat MA, Arbogast A, Laforge T, Quadri RA, Proust JJ: Excessive apoptosis of mature T lymphocytes is a characteristic feature of human immune senescence. Mech Ageing Dev 1996;88:25–38.
45.
Roberts AD, Orme IM: CD95 expression in aged mice infected with tuberculosis. Infect Immun 1998;66:5036–5040.
46.
Herndon FJ, Hsu HC, Mountz JD: Increased apoptosis of CD45RO– T cells with aging. Mech Ageing Dev 1997;94:123–134.
47.
Miyawaki T, Uehara T, Nibu R, Tsuji T, Yachie A, Yonehara S, Taniguchi N: Differential expression of apoptosis-related Fas antigen on lymphocyte subpopulations in human peripheral blood. J Immunol 1992;149:3753–3758.
48.
Kishimoto H, Sprent J: Strong TCR ligation without costimulation causes rapid onset of Fas-dependent apoptosis of naïve murine CD4+ T cells. J Immunol 1999;163:1817–1826.
49.
Desbarats J, Wade T, Wade WF, Newell MK: Dichotomy between naive and memory CD4+ T cell responses to Fas engagement. Proc Natl Acad Sci USA 1999;96:8104–8109.
50.
Buttke TM, Sandstrom PA: Oxidative stress as a mediator of apoptosis. Immunol Today 1994;15:7–10.
51.
Slater AF, Nobel SI, Orrenius S: The role of intracellular oxidants in apoptosis. Biochim Biophys Acta 1995;1271:59–62.
52.
King CM, Barnett YA: Oxidative stress and human ageing. Biochem Soc Trans 1995;23:375S–378S.
© 2000 S. Karger AG, Basel
2000
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.