Sequential TNF and TGF-β regulation of expansion and induction of cytotoxicity in long-term cultures of lymphokine-activated killer cells

J. Koberda, D. Przepiorka, R. P. Moser, Elizabeth A Grimm

Research output: Contribution to journalArticle

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Abstract

Therapeutic use of lymphokine-activated killer (LAK) cells frequently requires higher numbers of effector cells than can be produced in short-term cultures. Extended stimulation of these cells by interleukin-2 (IL-2) past 2 weeks leads to a decrease in cytolytic activity and cell proliferation. To determine how known regulatory cytokines are involved in the mechanism responsible for loss of IL-2 responsiveness, adherent LAK (A-LAK), nonadherent LAK (NA-LAK), monocyte-depleted (MD-LAK), and unmanipulated LAK (UN-LAK) cells derived from human peripheral blood were stimulated with high- dose IL-2 for 4 weeks, and cytokine production was measured serially. Despite continued supplementation with IL-2, cell number plaeaued at 2 weeks with a 2.5-3.0 log increase in A-LAK cultures and a 1.0 log increase in NA-LAK, MD- LAK, and UN-LAK cultures. Cytolytic activity had decreased significantly in all four culture systems after only 14 days of stimulation with IL-2 as assessed by the chromium release assay using K562, Daudi, and RP-mel tumor cell lines as targets, and LAK activity was barely detectable after 28 days of stimulation. Bioactive tumor necrosis factor (TNF) was present in concentrations of 15-55 U/ml during the first week of culture and at less than 10 U/ml thereafter. Bioactive transforming growth factor-β (TGF-β) was detected at 1-36 U/ml from 5 to 14 days of culture and decreased thereafter. Immunoreactive TGF-β2 was still present at concentrations of 20-90 pg/ml after 21 days of culture. Cytokine profiles were similar in all four culture systems, although TNF levels were higher and TGF levels lower in the A-LAK cultures. Supplementation of UN-LAK cultures with TNF failed to reverse the decrease in cell proliferation and cytolytic activity in the long-term system, but IL-2 responsiveness was partially restored when cultures were supplemented with a neutralizing anti-TGF antibody whether or not TNF and lymphotoxin (LT) were also added. These results indicate that production of TGF, a negative regulatory cytokine, is mainly responsible for the loss of IL-2 responsiveness in the long-term A-LAK, NA-LAK, MD-LAK, and UN-LAK cultures, although the inability of TGF-neutralizing antibody to completely reverse the inhibition suggests that other negative regulatory cytokines not investigated here may also have a role.

Original languageEnglish (US)
Pages (from-to)139-145
Number of pages7
JournalLymphokine and Cytokine Research
Volume13
Issue number2
StatePublished - 1994

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Lymphokine-Activated Killer Cells
Transforming Growth Factors
Lymphokines
Interleukin-2
Tumor Necrosis Factor-alpha
Cytokines
Neutralizing Antibodies
Activated Killer Monocytes
Cell Count
Cell Proliferation
Lymphotoxin-alpha
Therapeutic Uses
Chromium
Tumor Cell Line
Interleukin-4
Anti-Idiotypic Antibodies

ASJC Scopus subject areas

  • Immunology
  • Hematology

Cite this

Sequential TNF and TGF-β regulation of expansion and induction of cytotoxicity in long-term cultures of lymphokine-activated killer cells. / Koberda, J.; Przepiorka, D.; Moser, R. P.; Grimm, Elizabeth A.

In: Lymphokine and Cytokine Research, Vol. 13, No. 2, 1994, p. 139-145.

Research output: Contribution to journalArticle

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