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Starting procedures for the isolation and purification of granulocyte chalone activities

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Summary

Several starting materials and procedures for the extraction and purification of granulocyte chalone activities were tested and evaluated. Among others, leukoadhesion of bovine blood granulocytes on nylon and cotton wool and direct extraction with polar organic solvents were found suitable. Following PVP-leukapheresis ascites fluids were collected from rats, purified by ultrafiltration and Sephadex G 25 chromatography to yield 2 inhibitors at Ve/Vo=2.1 and 2.6 and one stimulator at 2.0 by the in vitro3H-thymidine test. Fraction 2.1, which has met the criteria of a granulocyte chalone by the diffusion chamber and agar colony test, was found thermostabile and to contain several peptides. Yet evidence for the peptide nature of the inhibitor is not conclusive. Extracts from bovine blood granulocytes contained only the inhibitor at 2.1. Problems related to the in vitro test for chalone activity were discussed.

Zusammenfassung

Verschiedene Ausgangsmaterialien und Verfahren zur Extraktion und Reinigung von Granulozyten-Chalon-Aktivitäten wurden untersucht und kritisch beurteilt. Darunter erwiesen sich Leukoadhäsion von Rinderblut-Granulozyten an Nylon- und Baumwoll-Watte und direkte Extraktion mit polaren organischen Lösungsmitteln als lohnend. Durch PVP-Leukapherese gewonnene Ascites-Flüssigkeiten von Ratten lieferten nach Ultrafiltration und Sephadex G 25-Chromatographie im In-vitro-Test mit3H-Thymidin 2 Inhibitoren bei Ve/Vo=2,1 und 2,6 und einen Stimulator bei 2,0. Fraktion 2,1, die mit dem Diffusionskammer- und Agarkolonie-Test die Eigenschaften eines Granulozyten-Chalones erfüllt hat, war thermostabil und enthielt verschiedene Peptide. Doch ist die Peptid-Natur des Inhibitors nicht schlüssig bewiesen. Extrakte aus Rinderblut-Granulozyten enthielten nur den Inhibitor 2,1. Auf die Problematik des In-vitro-Tests zum Chalon-Nachweis wurde hingewiesen.

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References

  1. Boyum A.: Separation of leukocytes from blood and bone marrow.Scand. J. clin. Lab. Invest. Suppl.97, (1968).

  2. Bullough W. S.: Chalone control mechanisms.Life Sci. 16, 323 (1975).

    Google Scholar 

  3. Djerassi I., Kim J. S., Suvansri U., Mitrakul C. & Ciesielka W.: Continuous flow filtration-leukopheresis.Transfusion 12, 75 (1972).

    Google Scholar 

  4. Heit W., Kern P., Kubanek B. & Heimpel H.: Some factors influencing granulocytic colony formation in vitro by human white blood cells.Blood 44, 511 (1974).

    Google Scholar 

  5. Laerum O. D. & Maurer H. R.: Proliferation kinetics of myelopoietic cells and macrophages in diffusion chambers after treatment with granulocyte extracts (chalone).Virchows Arch. Abt. B14, 293 (1973).

    Google Scholar 

  6. Lajtha L. G.: Commentary on Chalone of the Granulocyte System by T. Rytömaa, and Granulopoiesis-Inhibiting Factor by W. R. Paukovits.Nat. Cancer Inst. Monogr. 38, 157 (1973).

    Google Scholar 

  7. Lewis J. P., Neal W. A., Welch E. T., Lewis W. G. III, DuBose C. M. Jr., Wright C. S. & Smith L. L.: The isolation of Erythropoiesis Regulatory Factors by an Electrofractionation Technique Combined with Selective Membrane Permeability.Proc. Soc. Exp. Biol. Med. 142, 293 (1973).

    Google Scholar 

  8. Lord B. I., Cercek L., Cercek B., Shah G. P., Dexter T. M. & Lajtha L. G.: Inhibitors of haemopoietic cell proliferation: Specificity of action within the haemopoietic system.Brit. J. Cancer 29, 168 (1974).

    Google Scholar 

  9. Lowry O. H., Rosenbrough N. J., Farr A. L. & Randall R. J.: Protein measurement with the Folin phenol reagent.J. biol. Chem. 193, 265 (1951).

    Google Scholar 

  10. MacVittie T. J. & McCarthy K. F.: Inhibition of Granulopoiesis in Diffusion Chambers by a Granulocyte Chalone.Exp. Hemat. 2, 182 (1974).

    Google Scholar 

  11. Maurer H. R. & Laerum O. D.: Granulocyte chalone extraction and in vitro testing. Internat. Symp. Chalone Control. Mechan., Lane End, Bucks, England 1973, abstract p. 14.

  12. Maurer H. R., Weiß G. & Laerum O. D.: Evalution of a short term in vitro test of granulocyte chalone activity.Virchows Arch. Abt. B Cell Path.20, 229 (1976).

    Google Scholar 

  13. Müller-Berat N., Laerum O. D. & Maurer H. R.: Chalone inhibition of the committed stem cell to granulopoiesis in vitro. 6th Meeting Europ. Study Group for Cell Proliferation, Moscow 1973, abstract p. 41.

  14. Nakao M., Nakayama T. & Kankura T.: A new method for separation of human blood components.Nature New Biol. 246, 94 (1973).

    Google Scholar 

  15. Pataki G.: Techniques of thin-layer chromatography in amino acid and peptide chemistry.Ann Arbor Sci. Publ. Inc. pp. 90 & 93 (1968).

  16. Paukovits W. R.: Control of granulocyte production: Separation and chemical identification of a specific inhibitor (chalone).Cell Tiss. Kinet. 4, 539 (1971).

    Google Scholar 

  17. Paukovits W. R.: Granulopoiesis-Inhibiting Factor: Demonstration and Preliminary Chemical and Biological Characterization of a Specific Polypeptide (Chalone).Nat. Cancer Inst. Monogr. 38, 147 (1973).

    Google Scholar 

  18. Paukovits W. R. & Paukovits J. B.: Mechanism of action of granulopoiesis inhibiting factor (chalone). I. Evidence for a receptor protein on bone marrow cells.Exp. Pathol. 10, 348 (1975).

    Google Scholar 

  19. Paukovits W. R.: Chemistry of granulocyte chalone. 8th Saratoga Conf. Mol. Biol. Pathol., Saratoga Springs, N.Y., USA, Aug. 1975.

    Google Scholar 

  20. Price G. B., McCulloch E. A. & Till J. E.: A new human low molecular weight granulocyte colony stimulating activity.Blood 42, 341 (1973).

    Google Scholar 

  21. Rytömaa T. & Kiviniemi K.: Control of granulocyte production I. Chalone and antichalone, two specific humoral regulators.Cell Tissue Kinet. 1, 329 (1968).

    Google Scholar 

  22. Rytömaa T. & Kiviniemi K.: Control of granulocyte production. II. Mode of action of chalone and antichalone.Cell Tissue Kinet. 1, 341 (1968).

    Google Scholar 

  23. Rytömaa T.: Chalone of the Granulocyte System,Nat. Cancer Inst. Monogr. 38, 143 (1973).

    Google Scholar 

  24. Simard A., Corneille L., Deschamps Y. & Verly W. G.: Inhibition of cell proliferation in the livers of hepatectomized rats by a rabbit hepatic chalone.Proc. Nat. Acad. Sci. USA71, 1763 (1974).

    Google Scholar 

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Author notes

  1. H. Rainer Maurer

    Present address: Institut für Pharmazie der Freien Universität Berlin, Königin-Luise-Straße 2+4, 1000, Berlin 33, Germany

Authors and Affiliations

  1. Max-Planck-Institut für Virus-Forschung, Tübingen

    H. Rainer Maurer, Gerhard Weiß & Ole Didrik Laerum

  2. Institutt for Patologi, Universitetet i Oslo, Rikshospitalet, Oslo, Norwegen

    H. Rainer Maurer, Gerhard Weiß & Ole Didrik Laerum

Authors
  1. H. Rainer Maurer
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  2. Gerhard Weiß
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  3. Ole Didrik Laerum
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Supported by the Deutsche Forschungsgemeinschaft (Ma 505/4).

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Maurer, H.R., Weiß, G. & Laerum, O.D. Starting procedures for the isolation and purification of granulocyte chalone activities. Blut 33, 161–170 (1976). https://doi.org/10.1007/BF00995997

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