Phenotypic and functional characterization of recombinant interleukin 2 (rIL 2)-induced activated killer cells: Analysis at the population and clonal levels

S. Ferrini, S. Miescher, M. R. Zocchi, V. von Fliedner, A. Moretta

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Abstract

In these studies we investigated the phenotypic and functional characteristics of human rIL 2-activated killer cells (LAK). By FACS sorting we separated PBL into Leu-11- and Leu-11+ cell fractions and cultured them for 4 days in 100 U/ml rIL 2. Under these culture conditions, cells of the Leu-11+ fraction acquired a stronger LAK activity against fresh autologous or allogeneic melanoma cells as compared with Leu-11- cells or unfractionated PBL. To better characterize the cells responsible for this cytolytic activity, we directly cloned Leu-11+ and Leu-11- FACS-sorted cells in the presence of 1% PHA, irradiated spleen feeder cells, and rIL 2. From 6 to 10% of the Leu-11+ cells and from 42 to 66% of the Leu-11- cells plated gave rise to clonal progenies that were tested simultaneously for cytolytic activity against fresh melanoma cells and NK-sensitive K562 target cells in a 4-hr 51Cr-release assay. Most of the Leu-11+ microcultures lysed fresh melanoma target cells (35 out of 38 and 26 out of 34 in two separate experiments), whereas only a few clones derived from the Leu-11- cell fraction had this capability (four out of 45 and one out of 41). All the clones lysing fresh melanoma cells also efficiently killed K562 target cells, whereas other clones lysing only K562 could be found among Leu-11+ and Leu-11- clones. Nine clones expressing LAK activity were tested for their reactivity against a panel of different tumor target cells. All clones were able to lyse a broad panel of target cells including NK-sensitive and NK-resistant cultured or noncultured human tumor target cells, as well as mouse tumor cell lines. Surface marker analysis of 14 clones displaying LAK activity, all derived from Leu-11+ cells, showed that they were all T3 (CD3)-, whereas 10 out of 14 expressed the T11 (CD2) antigen and only four were weakly stained by an anti-T8 (CD8) mAb. All 14 clones expressed the T40 (CD7) T cell marker and DR and LFA-1 antigens. Cytolysis inhibition experiments performed on a rIL 2-activated Leu-11+ population and on two LAK cell clones, both expressing T11 antigen, showed that anti-LFA-1 but not anti-T11 mAb could inhibit cytolysis of freshly derived tumor target cells.

Original languageEnglish
Pages (from-to)1297-1302
Number of pages6
JournalJournal of Immunology
Volume138
Issue number4
Publication statusPublished - 1987

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Interleukin-2
Clone Cells
Population
Melanoma
Lymphokine-Activated Killer Cells
Lymphocyte Function-Associated Antigen-1
K562 Cells
Natural Killer Cells
CD2 Antigens
Feeder Cells
Antigens
Neoplasms
Tumor Cell Line
Cultured Cells
Spleen
Cell Culture Techniques
T-Lymphocytes

ASJC Scopus subject areas

  • Immunology

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Phenotypic and functional characterization of recombinant interleukin 2 (rIL 2)-induced activated killer cells : Analysis at the population and clonal levels. / Ferrini, S.; Miescher, S.; Zocchi, M. R.; von Fliedner, V.; Moretta, A.

In: Journal of Immunology, Vol. 138, No. 4, 1987, p. 1297-1302.

Research output: Contribution to journalArticle

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AU - Ferrini, S.

AU - Miescher, S.

AU - Zocchi, M. R.

AU - von Fliedner, V.

AU - Moretta, A.

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