N-acetylmuramyl-L-alanyl-D-isoglutamine stimulates the immune response of B cells. Use of fast protein liquid chromatography to purify a novel monokine involved in this effect.
The immune response of murine B cells, which requires the presence of various cytokines, was stimulated by N-acetylmuramyl-L-alanyl-D-isoglutamine. A novel monokine involved in this effect has been partially purified from the conditioned medium of peritoneal macrophages by using affinity chromatography on Blue-Sepharose and high-performance liquid chromatography. Hydrophobic interaction chromatography was highly efficient in removing almost all protein contaminants. The resulting biologically active material exhibited heterogeneity in ion-exchange chromatography, and apparent isoelectric points between 5.2 and 5.5 on chromatofocusing. Its apparent molecular weight, as determined by size-exclusion filtration, was about 35,000. Preliminary results on the kinetics of appearance of intracellular biological activity suggested that the monokine could also be obtained from cytoplasmic extracts.
Souvannavong
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Institut de Biochimie, Université de Paris-Sud, Orsay, France.
Adam
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D000119:Acetylmuramyl-Alanyl-Isoglutamine / Q000302:isolation & purification* / Q000494:pharmacology; D000276:Adjuvants, Immunologic; D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D004912:Erythrocytes / Q000276:immunology; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D007525:Isoelectric Focusing; D051379:Mice; D008815:Mice, Inbred Strains
D000276:Adjuvants, Immunologic; D007202:Indicators and Reagents; D000119:Acetylmuramyl-Alanyl-Isoglutamine