Regular article
The spontaneous apoptotic cell death of normal human lymphocytes in vitro: The release of, and immunoproliferative response to, nucleosomes in vitro

https://doi.org/10.1016/0090-1229(91)90108-MGet rights and content

Abstract

Nucleosomes released spontaneously in short-term culture from murine spleen cells have a significant immunoproliferative effect in vitro, including the stimulation of anti-DNA antibody responses. The present studies show that during short-term cultures, tonsil lymphoid cells also undergo spontaneous apoptosis revealed morphologically (electron microscopy) by the appearance of changes in nuclear chromatin, typical of apoptosis and similar to morphologic changes of apoptosis in cultured normal splenic lymphocytes. This process is followed by the release, in the >30-kDa cell free supernatant fraction, of core histones (H2A, H2B, H3, H4) and low molecular weight DNA (approx 160 bp) constituents of nucleosomes. The >30-kDa tonsil lymphocyte cell-free supernatant material containing the constituents of core nucleosomes, as well as the >30-kDa supernatant fractions of tonsil cell lysates harvested at the same time, had a significant immunoproliferative effect on human or murine lymphocytes, increasing both DNA and immunoglobulin synthesis (protein A plaque-forming cells). Thus the release of immunoproliferative nucleosomes form dying human lymphoid cells provides an autocrine lymphocyte stimulatory network which may be important in immunoproliferative disorders and in normal cell turnover. Apoptosis in vivo may also provide a potential source for the circulating nucleosomal DNA identified in plasma in some systemic lupus erythematosus patients as well as contributing to increased polyclonal B lymphocyte stimulation and autoantibody responses in this disorder.

References (24)

  • J.O. Thomas et al.

    The major core histone antigenic determinants in SLE are in the trypsin-sensitive regions

    FEBS Lett.

    (1984)
  • P.J. Boender et al.

    Nucleosomal fragments in serum may directly reflect cell-mediated cytotoxic activity in vivo

    Clin. Immunol. Immunopathol.

    (1989)
  • J.F.R. Kerr et al.

    Apoptosis: A basic biological phenomenon with wide-ranging implications in tissue kinetics

    Br. J. Cancer

    (1972)
  • J.F.R. Kerr et al.

    The digestion of cellular fragments within phagolysosomes in carcinoma cells

    J. Pathol.

    (1972)
  • J. Searle et al.

    Necrosis and apoptosis: Distinct modes of cell death with fundamentally different significance

    Pathol. Annu.

    (1982)
  • R.C. Duke et al.

    Endogenous endonuclease-induced DNA fragmentation: An early event in cell-mediated cytolysis

  • D.A. Bell et al.

    Immunogenic DNA related factors: Nucleosomes spontaneously released from normal nmurine lymphoid cells stimulate proliferation and immunoglobulin synthesis of normal mouse lymphocytes

    J. Clin. Invest.

    (1990)
  • M.J. Atkinson et al.

    A naturally occurring polyclonal B cell activator of normal and autoantibody responses

    J. Immunol.

    (1985)
  • E. Grokowicz et al.

    A plaque assay for all cells secreting Ig of a given type or class

    Eur. J. Immunol.

    (1976)
  • D.G. Chung et al.

    Intermolecular histone H4 interactions in core nucleosomes

    Biochemistry

    (1986)
  • K.W. Giles et al.

    An improved diphenylamine method fo the estimation of deoxyribonucleic acid

    Nature

    (1965)
  • J.O. Thomas et al.

    An octamer of histones in chromatin and free in solution

  • Cited by (109)

    • Cell death in the pathogenesis of systemic lupus erythematosus and lupus nephritis

      2017, Clinical Immunology
      Citation Excerpt :

      In SLE, accelerated primary necrosis or accelerated apoptosis resulting in secondary necrosis, defective clearance of necrotic cells, and post-translational modifications during necrosis may play an important role in the development of LN. As previously mentioned, accelerated apoptosis leading to secondary necrosis has been observed in SLE patients and mice and promotes autoimmunity and renal damage [41,46,47,49,53–55,152,153]. Necrosis is frequently observed in subjects with LN and is associated with enhanced serological activity and proteinuria compared to LN patients that do not display necrosis [154].

    • Differential clearance mechanisms, neutrophil extracellular trap degradation and phagocytosis, are operative in systemic lupus erythematosus patients with distinct autoantibody specificities

      2015, Immunology Letters
      Citation Excerpt :

      However, at present there is no general unifying theory that leads to the basis for this distinctive autoantibody profile. Impaired phagocytosis in SLE delays apoptotic and microbial clearance thus exposing microbial and cellular debris to the immune cells leading to autoantibody production [1–5]. In addition to delayed apoptotic and microbial clearance, an interesting and newly identified process, NETosis or neutrphil extracellular trap (NET) formation has also been implicated as an alternative source of immunogens in SLE [5].

    • Pullulan-histone antibody nanoconjugates for the removal of chromatin fragments from systemic circulation

      2013, Biomaterials
      Citation Excerpt :

      It has been estimated that 1011–1012 cells, primarily of haematogenous origin, die in the adult human body daily due to normal physiology, and a similar number is regenerated through mitosis to maintain homeostasis [2]. It is now well established that the chromatin fragments (Cfs) enter the systemic circulation even though most of the apoptotic bodies are cleared by the macrophages [3,4]. It is also reported that if enhanced cell death occurs due to various conditions such as autoimmune diseases, inflammatory diseases, malignant tumors, chemotherapy etc the phagocytic systems can be overloaded or impaired leading to higher levels of circulating chromosomes [4,5].

    View all citing articles on Scopus

    Supported by a grant from the Medical Research Council of Canada.

    View full text