PMID: 6981047Jan 1, 1982Paper

Regulatory factors of lymphocyte-lymphocyte interaction. II. Characterization of human mitogenic factor derived from the culture supernatant of a mouse-human hybridoma

Microbiology and Immunology
M KawanoM Hanaoka

Abstract

Cell hybridization of phytohemagglutinin (PHA)-stimulated human peripheral blood lymphocyte (PBL) with murine lymphoma (EL-4) provided three hybridomas (MHH-16, MHH-20, and MHH-22) which spontaneously produced human mitogenic factor (MF). MHH-16 was serially subcloned by limiting dilution procedures, which resulted in maintaining two subclones producing human MF spontaneously for more than one year (PQL-3 and PQL-5 subcloned lines). Human MF MHH-MF) derived from supernatants of PQL-5 line cultures had a molecular weight (m.w.) of about 26,000-30,000 daltons (the major peak) with a minor peak with an m.w. of 15,000 daltons on Sephadex G-100 chromatography, and at a high concentration of NaCl (1 M), the activity of the 26,000-30,000-m.w. fraction became weak and that of the 15,000-m.w. fraction became predominant. MHH-MF had an isoelectric point of Ph 5.0-6.5. On DEAE-cellulose chromatography, MHH-MF was eluted at a fairly low salt concentration (sodium phosphate buffer 0.02 M, pH 8.0, NaCl 10 mM). After periodate treatment of this MHH-MF, the mitogenic activity almost disappeared. MHH-MF was relatively unstable to heating at 56 c for 20 min. In the presence of tunicamycin (0.3 microgram/ml), an inhibitor of N-linked glycosylatio...Continue Reading

References

Aug 3, 1978·Nature·S KontiainenM Feldmann
May 3, 1972·Nature: New Biology·A Schimpl, E Wecker
Nov 1, 1974·The Journal of Experimental Medicine·R E RocklinJ R David
Jan 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·P LonaiG Hämmerling
Oct 1, 1980·The Journal of Experimental Medicine·L HarwellJ Kappler

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