Establishment of Tac-negative, interleukin-2-dependent cytotoxic cell lines from large granular lymphocytes (LGL) of patients with expanded LGL populations

V. Pistoia, A. J. Carroll, E. F. Prasthofer, A. B. Tilden, K. S. Zuckerman, M. Ferrarini, C. E. Grossi

Research output: Contribution to journalArticle

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Abstract

Cell lines were established from purified large granular lymphocites (LGL) isolated from the peripheral blood of seven patients with phenotypically homogeneous LGL expansions. LGL were stimulated with phytohemagglutinin (PHA) or recombinant interleukin-2 (rIL-2) and further expanded in vitro in IL-2-containing media. The surface phenotype of LGL, as assessed by monoclonal antibody staining, was T3+ T8+ in five patients, T3- T8- in one, and T3+ T8- in another patient. The cells also expressed Leu 7, Leu 11, and/or OKM 1 markers in various proportions and were identifiable as LGL by their morphological and cytochemical features. The original surface phenotype of the unstimulated LGL was retained in the IL-2-dependent cell lines from each individual patient, i.e., T3+ T8+ cells originated T3+ T8+ cell lines and T3- T8- cells originated T3- T8- cell lines. Other markers, such as Leu 11 and OKM 1, were generally lost in culture. LGL proliferated in response to rIL-2 but did not express detectable IL-2 receptors, even after prolonged periods of culture. All cell lines from each individual patient had the same surface phenotype, and within the single lines, all of the cells expressed the same markers. Cell lines from two patients consistently displayed chromosomal abnormalities. Although different in the two patients, the abnormalities were identical in all of the lines from the same patient and detectable in most of the cells examined, suggesting a clonal origin for the abnormally expanded LGL populations. Freshly isolated LGL did not exert NK activity. However, the IL-2-dependent LGL lines acquired the ability to kill K562 target cells and to produce gamma interferon (γ-IFN). No direct correlation was observed between these two properties.

Original languageEnglish
Pages (from-to)457-466
Number of pages10
JournalJournal of Clinical Immunology
Volume6
Issue number6
DOIs
Publication statusPublished - Nov 1986

Fingerprint

Interleukin-2
Lymphocytes
Cell Line
CD8-Positive T-Lymphocytes
Population
Phenotype
K562 Cells
Interleukin-2 Receptors
Phytohemagglutinins
Chromosome Aberrations
Interferon-gamma
Monoclonal Antibodies
Staining and Labeling

Keywords

  • gamma interferon
  • interleukin-2
  • large granular lymphocytes
  • natural killer cells

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Establishment of Tac-negative, interleukin-2-dependent cytotoxic cell lines from large granular lymphocytes (LGL) of patients with expanded LGL populations. / Pistoia, V.; Carroll, A. J.; Prasthofer, E. F.; Tilden, A. B.; Zuckerman, K. S.; Ferrarini, M.; Grossi, C. E.

In: Journal of Clinical Immunology, Vol. 6, No. 6, 11.1986, p. 457-466.

Research output: Contribution to journalArticle

Pistoia, V. ; Carroll, A. J. ; Prasthofer, E. F. ; Tilden, A. B. ; Zuckerman, K. S. ; Ferrarini, M. ; Grossi, C. E. / Establishment of Tac-negative, interleukin-2-dependent cytotoxic cell lines from large granular lymphocytes (LGL) of patients with expanded LGL populations. In: Journal of Clinical Immunology. 1986 ; Vol. 6, No. 6. pp. 457-466.
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abstract = "Cell lines were established from purified large granular lymphocites (LGL) isolated from the peripheral blood of seven patients with phenotypically homogeneous LGL expansions. LGL were stimulated with phytohemagglutinin (PHA) or recombinant interleukin-2 (rIL-2) and further expanded in vitro in IL-2-containing media. The surface phenotype of LGL, as assessed by monoclonal antibody staining, was T3+ T8+ in five patients, T3- T8- in one, and T3+ T8- in another patient. The cells also expressed Leu 7, Leu 11, and/or OKM 1 markers in various proportions and were identifiable as LGL by their morphological and cytochemical features. The original surface phenotype of the unstimulated LGL was retained in the IL-2-dependent cell lines from each individual patient, i.e., T3+ T8+ cells originated T3+ T8+ cell lines and T3- T8- cells originated T3- T8- cell lines. Other markers, such as Leu 11 and OKM 1, were generally lost in culture. LGL proliferated in response to rIL-2 but did not express detectable IL-2 receptors, even after prolonged periods of culture. All cell lines from each individual patient had the same surface phenotype, and within the single lines, all of the cells expressed the same markers. Cell lines from two patients consistently displayed chromosomal abnormalities. Although different in the two patients, the abnormalities were identical in all of the lines from the same patient and detectable in most of the cells examined, suggesting a clonal origin for the abnormally expanded LGL populations. Freshly isolated LGL did not exert NK activity. However, the IL-2-dependent LGL lines acquired the ability to kill K562 target cells and to produce gamma interferon (γ-IFN). No direct correlation was observed between these two properties.",
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AU - Carroll, A. J.

AU - Prasthofer, E. F.

AU - Tilden, A. B.

AU - Zuckerman, K. S.

AU - Ferrarini, M.

AU - Grossi, C. E.

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AB - Cell lines were established from purified large granular lymphocites (LGL) isolated from the peripheral blood of seven patients with phenotypically homogeneous LGL expansions. LGL were stimulated with phytohemagglutinin (PHA) or recombinant interleukin-2 (rIL-2) and further expanded in vitro in IL-2-containing media. The surface phenotype of LGL, as assessed by monoclonal antibody staining, was T3+ T8+ in five patients, T3- T8- in one, and T3+ T8- in another patient. The cells also expressed Leu 7, Leu 11, and/or OKM 1 markers in various proportions and were identifiable as LGL by their morphological and cytochemical features. The original surface phenotype of the unstimulated LGL was retained in the IL-2-dependent cell lines from each individual patient, i.e., T3+ T8+ cells originated T3+ T8+ cell lines and T3- T8- cells originated T3- T8- cell lines. Other markers, such as Leu 11 and OKM 1, were generally lost in culture. LGL proliferated in response to rIL-2 but did not express detectable IL-2 receptors, even after prolonged periods of culture. All cell lines from each individual patient had the same surface phenotype, and within the single lines, all of the cells expressed the same markers. Cell lines from two patients consistently displayed chromosomal abnormalities. Although different in the two patients, the abnormalities were identical in all of the lines from the same patient and detectable in most of the cells examined, suggesting a clonal origin for the abnormally expanded LGL populations. Freshly isolated LGL did not exert NK activity. However, the IL-2-dependent LGL lines acquired the ability to kill K562 target cells and to produce gamma interferon (γ-IFN). No direct correlation was observed between these two properties.

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