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

Blood. 1995 Dec 15;86(12):4568-78.

Abstract

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

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Division / drug effects
  • Cell Line
  • Dosage Compensation, Genetic
  • Erythroid Precursor Cells / drug effects*
  • Female
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology*
  • Humans
  • Interleukin-2 / pharmacology*
  • Kinetics
  • Leukemia, Erythroblastic, Acute / pathology*
  • Lymphoma / pathology
  • Male
  • Mice
  • Molecular Sequence Data
  • Protein Conformation
  • Receptors, Interleukin-2 / analysis
  • Receptors, Interleukin-2 / chemistry
  • Receptors, Interleukin-2 / genetics
  • Receptors, Interleukin-2 / physiology*
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / metabolism
  • Severe Combined Immunodeficiency / genetics
  • Severe Combined Immunodeficiency / metabolism
  • Severe Combined Immunodeficiency / pathology
  • Structure-Activity Relationship
  • Thymus Neoplasms / pathology
  • Transfection
  • Tumor Cells, Cultured
  • X Chromosome

Substances

  • Interleukin-2
  • Receptors, Interleukin-2
  • Recombinant Fusion Proteins
  • Granulocyte-Macrophage Colony-Stimulating Factor