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Isolation and Activation of Murine Lymphocytes
08:08

Isolation and Activation of Murine Lymphocytes

Published on: October 30, 2016

Modulation of human lymphocyte mitogen responsiveness and interleukin-2 production by polymorphonuclear leukocytes

M Lyte1

  • 1Department of Biological Sciences, Mankato State University, MN 56002.

Journal of Clinical & Laboratory Immunology
|June 1, 1990
PubMed

Insights

Polymorphonuclear leukocytes (PMN) significantly enhance human lymphocyte proliferation and IL-2 production in response to mitogens like PHA and PWM. This enhancement is critically dependent on the lymphocyte:PMN ratio and serum source.

Area of Science:

  • Immunology
  • Cell Biology

Background:

  • Human peripheral blood lymphocytes (L) respond to mitogens like phytohemagglutinin (PHA) and pokeweed mitogen (PWM).
  • The role of polymorphonuclear leukocytes (PMN) in modulating lymphocyte responses is debated, with some studies suggesting suppression.

Purpose of the Study:

  • To investigate the effect of autologous polymorphonuclear leukocytes (PMN) on human lymphocyte proliferation and IL-2 production stimulated by PHA and PWM.
  • To determine the influence of lymphocyte:PMN ratios and serum supplementation on these interactions.

Main Methods:

  • Human peripheral blood lymphocytes were cultured with PHA or PWM at various sub-optimal and optimal concentrations.
  • Cultures were supplemented with autologous PMN at different lymphocyte:PMN ratios.
  • Lymphocyte proliferation was assessed by 3H-thymidine incorporation.
  • IL-2 production was measured for PHA-stimulated cultures.

Main Results:

  • PMN supplementation resulted in up to 25,000-fold increases in mitogen-stimulated lymphocyte proliferation compared to control cultures.
  • Enhanced IL-2 production was observed in PHA-stimulated cultures with PMN.
  • The degree of enhancement was dependent on the lymphocyte:PMN ratio and the source of serum (autologous vs. xenogeneic).

Conclusions:

  • Cell ratio is a critical factor in studying lymphocyte-PMN interactions.
  • Serum supplementation significantly influences the observed effects of PMN on lymphocyte reactivity.
  • Previous findings of PMN-induced suppression or no effect may need re-evaluation due to variations in experimental conditions, particularly cell ratios.

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