Biologic properties of homogeneous interleukin 3. I. Demonstration of WEHI-3 growth factor activity, mast cell growth factor activity, P cell-stimulating factor activity, colony-stimulating factor activity, and histamine-producing cell-stimulating factor activity

J. N. Ihle, J. Keller, S. Oroszlan, L. E. Henderson, T. D. Copeland, F. Fitch, Michael B. Prystowsky, E. Goldwasser, J. W. Schrader, E. Palaszynski, M. Dy, B. Lebel

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Abstract

Interleukin 3 (IL 3) was initially defined as a factor in conditioned media from concanavalin A-stimulated lymphocytes (Con A CM) that induces the enzyme 20-α-hydroxysteroid dehydrogenase (20αSDH) in cultures of nu/nu splenic lymphocytes. To determine the spectrum of additional 'biologic' activities, IL 3 was purified to homogeneity and its properties were assessed. The protein preparation was judged to be homogeneous IL 3 by the following criteria: 1) elution of a peak of IL 3 with a constant specific activity in the last step of purification, 2) presence of a single protein by SDS-PAGE analysis, 3) receptor-binding activity against IL 3-dependent cell lines, 4) a specific activity of ≃0.2 ng/ml required for 50% of maximal biologic activity, and 5) the presence of a single amino terminal sequence. With the use of this preparation of IL 3, the dose-response curves for 20αSDH induction were identical or similar to the dose-response curves of the activities of 1) WEHI-3 growth factor, 2) mast cell growth factor, 3) P cell-stimulating factor, and 4) histamine-producing cell-stimulating factor. In addition, homogeneous IL 3 had colony-stimulating factor activity although only approximately 2% of the total CSF activity found in Con A CM was associated with IL 3. The major peak of CSF activity could be resolved from IL 3 by DEAE column chromatography and lacked many of the biologic activities associated with IL 3.

Original languageEnglish (US)
Pages (from-to)282-287
Number of pages6
JournalJournal of Immunology
Volume131
Issue number1
StatePublished - 1983
Externally publishedYes

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Colony-Stimulating Factors
Stem Cell Factor
Interleukin-3
Granulocyte-Macrophage Colony-Stimulating Factor
Intercellular Signaling Peptides and Proteins
20-Hydroxysteroid Dehydrogenases
Lymphocytes
Conditioned Culture Medium
Concanavalin A
Interleukin-4
Chromatography
Polyacrylamide Gel Electrophoresis
Proteins

ASJC Scopus subject areas

  • Immunology

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Biologic properties of homogeneous interleukin 3. I. Demonstration of WEHI-3 growth factor activity, mast cell growth factor activity, P cell-stimulating factor activity, colony-stimulating factor activity, and histamine-producing cell-stimulating factor activity. / Ihle, J. N.; Keller, J.; Oroszlan, S.; Henderson, L. E.; Copeland, T. D.; Fitch, F.; Prystowsky, Michael B.; Goldwasser, E.; Schrader, J. W.; Palaszynski, E.; Dy, M.; Lebel, B.

In: Journal of Immunology, Vol. 131, No. 1, 1983, p. 282-287.

Research output: Contribution to journalArticle

Ihle, J. N. ; Keller, J. ; Oroszlan, S. ; Henderson, L. E. ; Copeland, T. D. ; Fitch, F. ; Prystowsky, Michael B. ; Goldwasser, E. ; Schrader, J. W. ; Palaszynski, E. ; Dy, M. ; Lebel, B. / Biologic properties of homogeneous interleukin 3. I. Demonstration of WEHI-3 growth factor activity, mast cell growth factor activity, P cell-stimulating factor activity, colony-stimulating factor activity, and histamine-producing cell-stimulating factor activity. In: Journal of Immunology. 1983 ; Vol. 131, No. 1. pp. 282-287.
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abstract = "Interleukin 3 (IL 3) was initially defined as a factor in conditioned media from concanavalin A-stimulated lymphocytes (Con A CM) that induces the enzyme 20-α-hydroxysteroid dehydrogenase (20αSDH) in cultures of nu/nu splenic lymphocytes. To determine the spectrum of additional 'biologic' activities, IL 3 was purified to homogeneity and its properties were assessed. The protein preparation was judged to be homogeneous IL 3 by the following criteria: 1) elution of a peak of IL 3 with a constant specific activity in the last step of purification, 2) presence of a single protein by SDS-PAGE analysis, 3) receptor-binding activity against IL 3-dependent cell lines, 4) a specific activity of ≃0.2 ng/ml required for 50{\%} of maximal biologic activity, and 5) the presence of a single amino terminal sequence. With the use of this preparation of IL 3, the dose-response curves for 20αSDH induction were identical or similar to the dose-response curves of the activities of 1) WEHI-3 growth factor, 2) mast cell growth factor, 3) P cell-stimulating factor, and 4) histamine-producing cell-stimulating factor. In addition, homogeneous IL 3 had colony-stimulating factor activity although only approximately 2{\%} of the total CSF activity found in Con A CM was associated with IL 3. The major peak of CSF activity could be resolved from IL 3 by DEAE column chromatography and lacked many of the biologic activities associated with IL 3.",
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T1 - Biologic properties of homogeneous interleukin 3. I. Demonstration of WEHI-3 growth factor activity, mast cell growth factor activity, P cell-stimulating factor activity, colony-stimulating factor activity, and histamine-producing cell-stimulating factor activity

AU - Ihle, J. N.

AU - Keller, J.

AU - Oroszlan, S.

AU - Henderson, L. E.

AU - Copeland, T. D.

AU - Fitch, F.

AU - Prystowsky, Michael B.

AU - Goldwasser, E.

AU - Schrader, J. W.

AU - Palaszynski, E.

AU - Dy, M.

AU - Lebel, B.

PY - 1983

Y1 - 1983

N2 - Interleukin 3 (IL 3) was initially defined as a factor in conditioned media from concanavalin A-stimulated lymphocytes (Con A CM) that induces the enzyme 20-α-hydroxysteroid dehydrogenase (20αSDH) in cultures of nu/nu splenic lymphocytes. To determine the spectrum of additional 'biologic' activities, IL 3 was purified to homogeneity and its properties were assessed. The protein preparation was judged to be homogeneous IL 3 by the following criteria: 1) elution of a peak of IL 3 with a constant specific activity in the last step of purification, 2) presence of a single protein by SDS-PAGE analysis, 3) receptor-binding activity against IL 3-dependent cell lines, 4) a specific activity of ≃0.2 ng/ml required for 50% of maximal biologic activity, and 5) the presence of a single amino terminal sequence. With the use of this preparation of IL 3, the dose-response curves for 20αSDH induction were identical or similar to the dose-response curves of the activities of 1) WEHI-3 growth factor, 2) mast cell growth factor, 3) P cell-stimulating factor, and 4) histamine-producing cell-stimulating factor. In addition, homogeneous IL 3 had colony-stimulating factor activity although only approximately 2% of the total CSF activity found in Con A CM was associated with IL 3. The major peak of CSF activity could be resolved from IL 3 by DEAE column chromatography and lacked many of the biologic activities associated with IL 3.

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