Distinction between γc detection and function in YT lymphoid cells and in the granulocyte-macrophage colony-stimulating factor-responsive human myeloid cell line, Tf-1

Nancy L. Farner, Stephan D. Voss, Thomas P. Leary, Jacek Gan, John Hakimi, Gerald Evans, Grace Ju, Paul M. Sondel

Research output: Contribution to journalArticle

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Abstract

Peripheral blood monocytes respond to interleukin-2 (IL-2) and express the γcommon (γc) subunit of the IL-2 receptor (IL-2R) complex. However, the role of IL-2 in myeloid development has recently become of interest for several reasons, including the effect γc mutations may or may not have on myeloid development in patients with XSCID. Many studies of IL-2 function in the myeloid cell lineage have been performed on a murine background. To study γc expression and function in human myeloid precursors, we introduced the human myelomonocytic cell line, Tf-1, with a retroviral vector containing the human IL-2Rβ subunit to create functional human intermediate IL-2R consisting of βγc dimers. We have characterized this transfected variant of Tf-1 (Tf-1β) with regard to its response to IL-2. Unlike the parental Tf-1 cell line that is deficient in both IL-2Rα and IL-2Rβ expression, the Tf-1β transfectant binds and responds to IL-2 through intermediate-affinity IL-2Rs. Scatchard analyses indicate the number of intermediate-affinity receptors on Tf-1β is similar to the number found on the well-characterized YT cell line. However, detection of γc on Tf-1β cells is dramatically less than on YT cells by Western blot analysis and is undetectable by flow cytometric studies and surface iodinations. The γc component on YT cells is readily detected by all three methods. We conclude from these studies that the intermediateaffinity IL-2Rs on the Tf-1 cell line behave differently than those on YT cells with respect to γc detection. Either the γc molecule itself is different, or the cellular environment in which it functions is altered. Elucidation of γc function on this cell line will allow for its use as a model in which other cytokines using γc (including IL-2, IL-4, and IL-15) can be studied on the same cellular background.

Original languageEnglish (US)
Pages (from-to)4568-4578
Number of pages11
JournalBlood
Volume86
Issue number12
StatePublished - Dec 15 1995

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Myeloid Cells
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-2 Receptors
Interleukin-2
Cells
Lymphocytes
Cell Line
X-Linked Combined Immunodeficiency Diseases
Interleukin-15
Halogenation
Cell Lineage
Interleukin-4
Dimers
Monocytes
Blood
Western Blotting
Cytokines
Mutation
Molecules

ASJC Scopus subject areas

  • Hematology

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Distinction between γc detection and function in YT lymphoid cells and in the granulocyte-macrophage colony-stimulating factor-responsive human myeloid cell line, Tf-1. / Farner, Nancy L.; Voss, Stephan D.; Leary, Thomas P.; Gan, Jacek; Hakimi, John; Evans, Gerald; Ju, Grace; Sondel, Paul M.

In: Blood, Vol. 86, No. 12, 15.12.1995, p. 4568-4578.

Research output: Contribution to journalArticle

Farner, Nancy L. ; Voss, Stephan D. ; Leary, Thomas P. ; Gan, Jacek ; Hakimi, John ; Evans, Gerald ; Ju, Grace ; Sondel, Paul M. / Distinction between γc detection and function in YT lymphoid cells and in the granulocyte-macrophage colony-stimulating factor-responsive human myeloid cell line, Tf-1. In: Blood. 1995 ; Vol. 86, No. 12. pp. 4568-4578.
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abstract = "Peripheral blood monocytes respond to interleukin-2 (IL-2) and express the γcommon (γc) subunit of the IL-2 receptor (IL-2R) complex. However, the role of IL-2 in myeloid development has recently become of interest for several reasons, including the effect γc mutations may or may not have on myeloid development in patients with XSCID. Many studies of IL-2 function in the myeloid cell lineage have been performed on a murine background. To study γc expression and function in human myeloid precursors, we introduced the human myelomonocytic cell line, Tf-1, with a retroviral vector containing the human IL-2Rβ subunit to create functional human intermediate IL-2R consisting of βγc dimers. We have characterized this transfected variant of Tf-1 (Tf-1β) with regard to its response to IL-2. Unlike the parental Tf-1 cell line that is deficient in both IL-2Rα and IL-2Rβ expression, the Tf-1β transfectant binds and responds to IL-2 through intermediate-affinity IL-2Rs. Scatchard analyses indicate the number of intermediate-affinity receptors on Tf-1β is similar to the number found on the well-characterized YT cell line. However, detection of γc on Tf-1β cells is dramatically less than on YT cells by Western blot analysis and is undetectable by flow cytometric studies and surface iodinations. The γc component on YT cells is readily detected by all three methods. We conclude from these studies that the intermediateaffinity IL-2Rs on the Tf-1 cell line behave differently than those on YT cells with respect to γc detection. Either the γc molecule itself is different, or the cellular environment in which it functions is altered. Elucidation of γc function on this cell line will allow for its use as a model in which other cytokines using γc (including IL-2, IL-4, and IL-15) can be studied on the same cellular background.",
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AU - Farner, Nancy L.

AU - Voss, Stephan D.

AU - Leary, Thomas P.

AU - Gan, Jacek

AU - Hakimi, John

AU - Evans, Gerald

AU - Ju, Grace

AU - Sondel, Paul M.

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