Leucoagglutinin induces differential proliferation of lymphocyte subsets

S. P. Goedegebuure, R. J. van de Griend, C. P.M. Ronteltap, R. L.H. Bolhuis

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Abstract

In this study we have established culture conditions that allow the preferential and rapid expansion of either T cell receptor (TCR)+/CD3+16- T lymphocytes or TCR-/CD3-16- natural killer (NK) cells, or the non-selective outgrowth of both subsets. Optimal proliferation of lymphocytes was obtained using a combination of irradiated allogeneic peripheral blood lymphocytes (PBL) and irradiated Epstein Barr virus (EBV) transformed lymphoblastoid B cell lines (B-LCL). Addition of 1 μg/ml leucoagglutinin to the culture medium induced a preferential outgrowth of TCR-/CD3-16- T lymphocytes. The proportion of TCR-/CD3-16- NK cells was decreased to 5% or less, although still a 2000-fold multiplication of TCR-/CD3-16- NK cells was obtained at day 13. Without leucoagglutinin a 1000-fold increase of about 70% pure TCR-/CD3-16- NK cells was obtained at day 13. Intermediate concentrations of leucoagglutinin (0.1-0.3 μg/ml) resulted in a non-selective expansion of both NK cells and T cells. Irrespective whether leucoagglutinin was added or not, the number of TCR+/CD3+8+ lymphocytes increased more rapidly relative to the TCR+/CD3+4+ lymphocytes resulting in an increased TCR+/CD3+8+ population size. Also under limiting dilution conditions leucoagglutinin increased the frequency of proliferating cells. In contrast to the preferential outgrowth of TCR+/CD3+8+ lymphocytes in bulk cultures, approximately 80% of the clones generated was TCR+/CD3+4+, demonstrating a growth promoting effect of TCR+/CD3+4+ lymphocytes on TCR+/CD3+8+ lymphocytes in PBL bulk cultures.

Original languageEnglish
Pages (from-to)73-83
Number of pages11
JournalBiotherapy
Volume1
Issue number1-2
DOIs
StatePublished - Mar 1989
Externally publishedYes

Keywords

  • TCR/CD3 NK cells
  • TCR/CD3 T lymphocytes
  • leucoagglutinin
  • proliferation

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    Goedegebuure, S. P., van de Griend, R. J., Ronteltap, C. P. M., & Bolhuis, R. L. H. (1989). Leucoagglutinin induces differential proliferation of lymphocyte subsets. Biotherapy, 1(1-2), 73-83. https://doi.org/10.1007/BF02170137