Production of a monoclonal antibody useful in the molecular characterization of murine T-cell-replacing factor/B-cell growth factor II.

Abstract

T-cell-replacing factor (TRF) is a T-cell-derived factor required for terminal differentiation of activated B cells to immunoglobulin-secreting cells. Previous studies have shown that a murine T-cell hybrid (B151K12) produces factor(s) that (i) induce immunoglobulin secretion by the B-cell leukemia line BCL1 and secondary anti-2,4-dinitrophenyl IgG synthesis in vitro by dinitrophenyl-primed B cells (TRF activity) and (ii) cause proliferation of the BCL1 cells [B-cell growth factor II (BCGF-II) activity]. Both activities appear to be associated with the same molecule. Here we report the production of a monoclonal antibody to murine TRF. The monoclonal antibody, designated TB13, strongly inhibited both TRF and BCGF-II activities and absorbed TRF- as well as BCGF-II-active molecules produced by B151K12 and by Xenopus oocytes that had been injected with mRNA transcribed from plasmid pSP6K-mTRF23. Inhibition was linearly dependent on the concentration of both TB13 and TRF. Monoclonal antibody TB13 did not inhibit the activities of B-cell stimulatory factor 1, interleukin 1, interleukin 2, or interleukin 3. TRF activity in dissolved samples of immunoprecipitates obtained with TB13 was recovered after NaDodSO4/PAGE, in the fractions corresponding to a protein band at Mr 46,000. Our results indicate that monoclonal antibody TB13 recognizes a molecule that has both TRF and BCGF-II activities.

• Authors: N Harada, T Takahashi, M Matsumoto, T Kinashi, J Ohara, Y Kikuchi, N Koyama, E Severinson, Y Yaoita, T Honjo
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 84 Issue 13 Pg. 4581-5 (Jul 1987) ISSN: 0027-8424 [Print] United States
• PMID: 2440025 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• Epitopes
• Growth Substances
• Interleukin-5
• Lymphokines
• Recombinant Proteins
• Interleukin-4

Topics

• Animals
• Antibodies, Monoclonal (immunology)
• Antibody Specificity
• B-Lymphocytes (drug effects, immunology)
• Cell Line
• Chromatography, Affinity
• Epitopes (immunology)
• Female
• Growth Substances (immunology, pharmacology)
• Interleukin-4
• Interleukin-5
• Lymphokines (immunology, pharmacology)
• Mice
• Mice, Inbred BALB C
• Mice, Inbred C3H
• Molecular Weight
• Rats
• Rats, Inbred Strains
• Recombinant Proteins (pharmacology)