Abstract.
The germinal center (GC) develops after antigen stimulation and is thought to occur at the site of various immune responses. We observed apoptotic cells within the GC using in situ end labeling (TUNEL), small amount DNA ladder assay, and RT-PCR analysis of Bcl-2 mRNA expression. Apoptosis was detected within GCs at all phases of the GC reaction by both TUNEL and DNA ladder assays. The number of TUNEL+ nuclei within the GC did not increase over the course of the GC reaction. However, the density of DNA in the ladder assay was higher in later-phase GCs. Bcl-2 mRNA expression was detected within GCs during the early phases of the GC reaction. These results indicate that accumulation of apoptotic cells and rescue from apoptosis occur within chicken GCs. In the present paper, the reasons for the accumulation of apoptotic cells will be discussed.
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References
Arakawa H, Furusawa S, Ekino S, Yamagishi H (1996) Immunoglobulin gene hyperconversion ongoing in chicken splenic germinal centers. EMBO J 15:2540–2546
Arakawa H, Kuma K, Yasuda M, Furusawa S, Ekino S, Yamagishi H (1998) Oligoclonal development of B cells bearing discrete Ig chains in chicken single germinal centers. J Immunol 160:4232–4241
Bhan AK, Nadler LM, Stashenko P, McCluskey RT, Schlossman SF (1981) Stages of B cell differentiation in human lymphoid tissue. J Exp Med 154:737–749
Buja LM, Eigenbrodt ML, Eigenbrodt EH (1993) Apoptosis and necrosis. Basic types and mechanisms of cell death. Arch Pathol Lab Med 117:1208–1214
Casamayor-Palleja M, Feuillard J, Ball J, Drew M, MacLennan ICM (1996) Centrocytes rapidly adopt a memory B cell phenotype on co-culture with autologous germinal centre T cell-enriched preparations. Int Immunol 8:737–744
Cooper MD, Chen C-LH, Bucy RP, Thompson CB (1991) Avian T cell ontogeny. Adv Immunol 50:87–117
Cory S (1995) Regulation of lymphocyte survival by the bcl-2 gene family. Annu Rev Immunol 13:513–543
Eguchi Y, Ewert DL, Tsujimoto Y (1992) Isolation and characterization of the chicken bcl-2 gene: expression in variety of tissues including lymphoid and neuronal organs in adult and embryo. Nucleic Acids Res 20:4187–4192
Ford JE, McHeyzer-Williams MG, Lieber MR (1994) Analysis of individual immunoglobulin λ light chain genes amplified from single cells is inconsistent with variable region gene conversion in germinal-center B cell somatic mutation. Eur J Immunol 24:1816–1822
Furusawa S, Okitsi-Negishi S, Yoshino K, Mizoguchi M, Noguchi Y (1987) Anti-idiotypic antibody as a mirror image of the paratope of the original antibody. Int Arch Allergy Appl Immunol 84:263–270
Gracia de Vinuesa C, Cooka MC, Balla J, Drewa M, Sunnersa Y, Cascalhob M, Wablb M, Klausc GGB, MacLennan ICM (2000) Germinal center without T cells. J Exp Med 191:485–493
Grandien A, Modigliani Y, Freitas A, Andersson J, Coutinho A (1994) Positive and negative selection of antibody repertoires during B-cell differentiation. Immunol Rev 137:54–89
Hardie DL, Johnson GD, Khan M, MacLennan ICM (1993) Quantitative analysis of molecules which distinguish functional compartments within germinal centers. Eur J Immunol 23:997–1004
Houssaint E, Diez E, Pink JRL (1987) Ontogeny and tissue distribution of the chicken Bu-1a antigen. Immunology 62:463–470
Kasagi N, Gomyo Y, Shirai H, Tsujitani S, Ito H (1994) Apoptotic cell death in human gastric carcinoma: analysis by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling. Jpn J Cancer Res 85:939–945
Kondo T, Arakawa H, Kitao H, Hirota Y, Yamagishi H (1993) Signal joint of immunoglobulin Vλ1-Jλ and novel joints of chimeric V pseudogenes on extrachromosomal circular DNA from chicken bursa. Eur J Immunol 23:245–249
Leanderson T, Källeberg E, Gray D (1992) Expansion, selection and mutation of antigen-specific B cells in germinal centers. Immunol Rev 126:47–61
Liu Y-J, Arpin C (1997) Germinal center development. Immunol Rev 156:111–126
Liu Y-J, Joshua DE, Williams GT, Smith CA, Gordon J, MacLennan ICM (1989) Mechanism of antigen-driven selection in germinal centers. Nature 342:929–931
Liu Y-J, Mason DY, Johnson GD, Abbot S, Gregory CD, Hardie DL, Gordon J, MacLennan ICM (1991a) Germinal center cells express Bcl-2 protein after activation by signals which prevent their entry into apoptosis. Eur J Immunol 21:1905–1910
Liu Y-J, Zhang J, Lane PJL, Chan EY-T, MacLennan ICM (1991b) Sites of specific B cell activation in primary and secondary responses to T cell-dependent and T cell-independent antigens. Eur J Immunol 21:2951–2962
Martinez-Valdez H, Guret C, de Bouteiller O, Fugier I, Banchereau J, Liu Y-J (1996) Human germinal center B cells express the apoptosis-inducing genes, Fas, c-myc, P53, and Bax but not survival gene bcl-2. J Exp Med 183:971–977
Motyka B, Reynolds JD (1991) Apoptosis is associated with the extensive B cell death in the sheep ileal Peyer's patch and the chicken bursa of Fabricius: a possible role in B cell selection. Eur J Immunol 21:1951–1958
Nakamura M, Yagi H, Kayaba S, Ishii T, Gotoh T, Ohtsu S, Itoh T (1996) Death of germinal center B cells without DNA fragmentation. Eur J Immunol 26:1211–1216
Pulendran B, Kannourakis G, Nouri S, Smith KGC, Nossal GJV (1995) Soluble antigen can cause enhanced apoptosis of germinal-centre B cells. Nature 375:331–334
Shokat KM, Goodnow CC (1995) Antigen-induced B-cell death and elimination during germinal-centre immune responses. Nature 375:334–338
Smith JP, Lister AM, Tew JG, Szakal AK (1991) Kinetics of the tingible body macrophage response in mouse germinal center development and its depression with age. Anat Rec 229:511–520
Smith KGC, Light A, O'Reilly LA, Ang S, Strasser A, Tarlinton D (2000) bcl-2 transgene expression inhibits apoptosis in the germinal center and reveals differences in the selection of memory B cells and bone marrow antibody-forming cells. J Exp Med 191:475–484
Smithyman AM (1977) A simple procedure for the isolation of germinal centres from chicken spleen. Dev Comp Immunol 1:263–270
Surh CD, Sprent J (1994) T-cell apoptosis detected in situ during positive and negative selection in the thymus. Nature 372:100–103
Tarlinton DM, Smith KGC (1997) Apoptosis and the B cell response to antigen. Int Rev Immunol 15:53–71
Tsiagbe VK, Inghirami G, Thorbecke GJ (1996) The physiology of germinal centers. Crit Rev Immunol 16:381–421
Yasuda M, Taura Y, Yokomizo Y, Ekino S (1998) A comparative study of germinal center: fowls and mammals. Comp Immunol Microbiol Infect Dis 21:179–189
Yasuda M, Kajiwara E, Ekino S, Taura Y, Hirota Y, Horiuchi H, Matsuda H, Furusawa S (2003) Immunobiology of chicken germinal center: I. Changes in surface Ig class expression in the chicken splenic germinal center after antigenic stimulation. Dev Comp Immunol 27:159–166
Zhang W, Bardwell PD, Woo CJ, Poltoratsky V, Scharf MD, Martin A (2001) Clonal instability of V-region hypermutation in the Ramos Burkitt's lymphoma cell line. Int Immunol 13:1175–1184
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This work was supported by Grants-in-Aid for Scientific Research (Nos. 11670322 and 10306017) from the Ministry of Education, Science, Sport and Culture, and the Ministry of Agriculture, Forestry and Fisheries of Japan (Special Scientific Research and Pioneering Research Project in Biotechnology), as well as from the Bio-oriented Technology Research Advancement Institution (BRAIN)
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Yasuda, M., Horiuch, H., Matsuda, H. et al. Immunobiology of chicken germinal center: II. Accumulation of apoptotic cells within the germinal center. Cell Tissue Res 314, 215–221 (2003). https://doi.org/10.1007/s00441-003-0790-5
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DOI: https://doi.org/10.1007/s00441-003-0790-5