Interleukin 2 receptor expression by K-562 cells

Author

William C. Kouns

Date of Award

12-1987

Degree Type

Thesis

Degree Name

Master of Science

Major

Comparative and Experimental Medicine

Major Professor

Albert T. Ichiki

Committee Members

Roger Carroll, John Tyler, Clinton Lothrop

Abstract

The K-562 cell line was derived from cells in the pleural effusion of a patient with chronic myelogenous leukemia. The cells have been classified as pluripotent stem cells with an arrest of differentiation. A number of sublines have been derived from various serial passages of the original line. These sublines differ in karyotype, as well as in the expression of varous cell surface determinants. The F-1 subline was shown to express several lymphocytic determinants. It was observed that this subline was reactive with the monoclonal antibody (MoAb) anti-IL-2R1, which recognizes the interleukin 2 receptor (IL2R). The IL2R is expressed by activated T- and B-cells, as well as some monocytes. This study provides further evidence of IL2R expression by K-562 subline F-1 cells. Serological studies demonstrated that the cells (approximately 80%) constitutively express the IL2R epitope recognized by anti-IL-2R1. The IL2R epitopes recognized by anti-Tac, 767/B6, and anti-IL2R are variably expressed at low levels by the cells. Phytohemagglutinin and phorbol myristate acetate, which up-regulate IL2R expression on T-cells, do not up-regulate IL2R expression by K-562 cells. This data suggests that the K-562 IL2R differs in conformation or in the expression of various epitopes from normal activated T-cell IL2R, as well as differing in the mechanisms controlling the regulation of IL2R expression. Immunoblotting and immunoprecipitation experiments were unable to fully characterize the receptor as expressed by the K-562 cells. The cells were capable of specifically binding IL2, as demonstrated when rIL2 was shown to compete with ¹²⁵IL2 for binding. The stimulation of the cells with IL2 results in the enhanced expression of the B4 and 0KT4a binding antigens. As IL2 binding to high affinity IL2R results in the proliferation of activated helper and cytotoxic T-cells, it is noteworthy that IL2 stimulation of K-562 cells results in the enhanced expression of the T-cell helper/inducer antigen recognized by 0KT4a. The enhanced expression of these lymphocytic determinants further suggests that the IL2 may have been endocytosed by the K-562 cells via functional IL2R. Hence, the serological and binding data strongly suggest that the K-562 subline F-1 cells do express IL2R.

Recommended Citation

Kouns, William C., "Interleukin 2 receptor expression by K-562 cells. " Master's Thesis, University of Tennessee, 1987.
https://trace.tennessee.edu/utk_gradthes/13511