Purification of immunomodulatory factors in human peripheral blood leukocytes

J Chromatogr. 1990 Jul 20:512:139-47. doi: 10.1016/s0021-9673(01)89480-2.

Abstract

Procedure is described for purifying low-molecular-weight factors with antigen-aspecific properties from a dialysate of human leukocyte extract. It includes gel chromatography on Sephadex G-25 and G-15, ion-exchange chromatography, reversed-phase high-performance liquid chromatography (HPLC) on a C18 hydrophobic column and gel permeation HPLC. The immunosuppressive factor (mol.wt. 800-1000) was purified to near homogeneity. It is probably of peptidic nature, although it is pronase resistant. The enhancer factor (mol.wt. 300-600) is eluted from chromatographic columns together with a hypoxanthine-like substance. Nevertheless, the biological activity cannot be attributed to the purine derivative. Identification of this amplifier activity is still lacking.

MeSH terms

  • Adjuvants, Immunologic / analysis
  • Adjuvants, Immunologic / isolation & purification*
  • Amino Acids / analysis
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Humans
  • Immunosuppressive Agents / analysis
  • Immunosuppressive Agents / isolation & purification*
  • In Vitro Techniques
  • Leukocytes / chemistry*
  • Leukocytes / immunology
  • Lymphocyte Activation / drug effects
  • Molecular Weight
  • Phytohemagglutinins / pharmacology
  • Spectrophotometry, Ultraviolet

Substances

  • Adjuvants, Immunologic
  • Amino Acids
  • Immunosuppressive Agents
  • Phytohemagglutinins