Open AccessLong‐term cultures of human peripheral blood lymphocytes with recombinant human interleukin‐2 generate a population of virtually pure CD3 + CD16 ‐ CD56 ‐ large granular lymphocyte LAK cells
Author(s) -
ROUSSEL E.,
GERRARD J. M.,
GREENBERG A. H.
Publication year - 1990
Publication title -
clinical & experimental immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.329
H-Index - 135
eISSN - 1365-2249
pISSN - 0009-9104
DOI - 10.1111/j.1365-2249.1990.tb05463.x
Subject(s) - lymphokine activated killer cell , cd16 , population , interleukin 2 , cd3 , immunology , lymphokine , lymphocyte , biology , natural killer cell , cd8 , cytotoxic t cell , peripheral blood lymphocyte , t lymphocyte , microbiology and biotechnology , interleukin 21 , immune system , medicine , in vitro , biochemistry , environmental health
SUMMARY It has been reported that lymphocytes from peripheral blood (PBL) cultured with interleukin‐2 (IL‐2) produce predominantly CD16 + lymphokine‐activated killer (LAK) cells. We developed a two‐step method to generate LAK cells from human PBL in long‐term cultures (10–12 days) with recombinant human IL‐2 (rhIL‐2) and characterized the evolving LAK cell population by testing its phenotype and cytotoxic activity as a function of time. The starting PBL displayed some natural killer (NK) cytotoxicity but no LAK activity. At day 6 , the cells were a mixed population of about 80% CD3 + and 6% CD16 + cells. Little proliferation was evident but strong LAK activity was detected. After 10–12 days, major cell expansion had occurred and they were essentially a pure (>90%) CD3 + CD16 ‐ CD56 ‐ cell population large granular lymphocyte (LGL) by morphology that displayed strong non‐MHC‐restricted killing activity (> 200 lytic units). Over the same period of time, the CD16 + cells had almost completely regressed in these cultures. This preferential induction of CD + LAK cells was not an effect of IL‐2 concentration as 10 U/ml was as effective as 500 U/ml. Further characterization revealed a major population of CD4 + (60%) and CD8 + (30%) with a smaller fraction (<9%) of γδ + cells. These results indicate that a virtually pure CD3 + LAK cells population was produced with long‐term cultures of lymphocytes from peripheral blood in rhIL‐2, in which active proliferation of the CD3 + but not CD16 + cells occurred.