Evidence for specific receptors for interleukin 3 on lymphokine- dependent cell lines established from long-term bone marrow cultures

• Published in: The Journal of immunology (1950) Vol. 132; no. 4; pp. 1872 - 1878
• Main Authors: Palaszynski, EW, Ihle, JN
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Am Assoc Immnol 01.04.1984; American Association of Immunologists
• Subjects: Analysis of the immune response. Humoral and cellular immunity; Animals; Binding, Competitive; Biological and medical sciences; Bone Marrow - immunology; Bone Marrow Cells; Cell interactions; Cell Line; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Immunobiology; Interleukin-3; Kinetics; Lymphokines - analysis; Lymphokines - metabolism; Lymphokines - physiology; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Rats; Receptors, Antigen, T-Cell - analysis; Receptors, Antigen, T-Cell - physiology; Receptors, Immunologic - analysis; Receptors, Immunologic - physiology; Receptors, Interleukin-2; Receptors, Interleukin-3; Cell culture; Rodentia; Lymphokine; Long term; Immunological method; Vertebrata; Specificity; Mammalia; Cell line; Mouse; Animal; Bone marrow; Biological receptor
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.132.4.1872

Utilizing 125I-labelled IL 3, an assay for specific IL 3 receptors on continuous cell lines was developed. The 32D-cl 23 and FDC-P1 cell lines examined were originally derived from long-term cultures of murine bone marrow supplemented with WEHI-3-conditioned media. Both of these cell lines have been shown to be absolutely dependent on IL 3, a component of WEHI-3-conditioned media, for growth in vitro. The binding of radiolabeled IL 3 to these cells was found to be specific, saturable, reversible, and time and temperature dependent. The specificity of IL 3 binding was demonstrated by the failure of a number of different lymphokines, hormones, and proteins to compete for the binding of radiolabeled IL 3. The IL 3-dependent cell lines uniquely express detectable receptors, whereas other cell lines of various types and origin known to be IL 3 independent had no detectable specific binding of IL 3. A Kd of 5.4 X 10(-11) M and a maximum binding capacity of 6.25 fmol per 1 X 10(6) cells was calculated for 32D-cl 23 from equilibrium binding studies. A similar analysis for FDC-P1 yielded a Kd of 1.7 X 10(-11) and a maximum binding capacity of 1.2 fmol per 1 X 10(6) cells. The data are consistent with the presence of 4000 to 5000 specific high-affinity binding sites/cell on 32D-cl 23, whereas FDC-P1 has 1500 to 2500 specific sites/cell. The Kd calculated for the FDC-P1 cell line from kinetic binding data was 2.5 X 10(-11) M, which agreed well with the Kd of 1.7 X 10(-11) M calculated from equilibrium data.