Abstract
PRIMARY antigenic exposure results in an initial antibody response and the T cell-dependent induction of B-cell memory1,2. Memory B-cell differentiation is characterized by somatic hypermutation in antibody variable region genes (V) and selection of B cells expressing high-affinity variants of this antigen receptor3–5. Despite our current understanding of B-cell memory6–8, the origin of memory B cells and the regulation of their differentiation remain elusive. This is largely due to the difficulties in observing and purifying this minor component of the immunized spleen. Further, molecular characterization of memory B cells requires hybridoma formation which restricts analyses to only those clones capable of fusion and does not allow isolation of cells in a normal physiological state. We have therefore developed a unique system which allows isolation and unambiguous enumeration of lgG+1 memory B cells, based on six-parameter flow cytometry, secretion of antibody in clonal cultures and analysis of clonally expressed V genes using the polymerase chain reaction9. Here we report that single IgG+1 antigen-binding B cells from an early secondary immune response proliferate in lipopolysaccharide-driven microcultures and produce antigen-specific IgG1 antibodies. Individual B-cell clones in these cultures express somatically mutated heavy chain V genes, confirming their designation as memory B cells. Although isolated memory B cells undergo extensive proliferation in vitro, V gene sequence analysis of their individual progeny shows that further hypermutation does not occur.
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References
MacLennan, I. C. M., Liu, Y. J., Oldfield, S., Zhang, J. & Lane, P. J. L. Curr. Topics Microbiol. Immun. 159, 37–63 (1990).
Gray, D. & Leanderson, T. Curr. Topics Microbiol. Immun. 159, 1–17 (1990).
Weigert, M., Cesari, I. M., Yonkovich, S. J. & Cohn, M. Nature 228, 1045–1048 (1970).
Berek, C. & Milstein, C. Immunol. Rev. 105, 5–26 (1988).
Kocks, C. & Rajewsky, K. A. Rev. Immun. 7, 537–559 (1989).
Schittek, B. & Rajewsky, K. Nature 346, 749–751 (1990).
Gray, D. & Skarvall, H. Nature 336, 70–73 (1988).
Linton, P-J., Decker, D. J. & Klinman, N. R. Cell 59, 1049–1059 (1989).
Saiki, R. A. et al. Science 239, 487–491 (1988).
Imanishi, T. & Mäkelä, O. Eur. J. Immun. 3, 323–330 (1973).
Reth, M., Hammerling, G. J. & Rajewsky, K. Eur. J. Immun. 8, 393–400 (1978).
Bothwell, A. L. M. et al. Cell 24, 625–637 (1981).
Allen, D. et al. Immunol. Rev. 96, 5–22 (1987).
Rajewsky, K., Förster, I. & Cumano, A. Science 238, 1088–1094 (1987).
Cumano, A. & Rajewsky, K. EMBO J. 5, 2459–2468 (1986).
Pike, B. L., Vaux, D. L., Clarke-Lewis, I., Schrader, J. W. & Nossal, G. J. V. Proc. natn. Acad. Sci. U.S.A. 79, 6350–6354 (1982).
McHeyzer-Williams, M. G. Eur. J. Immun. 19, 2025–2030 (1989).
Hayakawa, K., Ishii, R., Katsuhiko, Y., Kishimoto, T. & Hardy, R. R. Proc. natn. Acad. Sci. U.S.A. 84, 1379–1383 (1987).
McKean, D. et al. Proc. natn. Acad. Sci. U.S.A. 81, 3180–3184 (1984).
Siekevitz, M., Kocks, C., Rajewsky, K. & Dildrop, R. Cell 48, 757–770 (1987).
Manser, T. Immunol. Today 11, 305–308 (1990).
Lebecque, S. G. & Gearhart, P. J. J. exp. Med. (in the press).
Levy, N. S., Malipiero, U. V., Lebecque, S. G. & Gearhart, P. J. J. exp. Med. 169, 2007–2019 (1989).
Tao, W. & Bothwell, A. L. M. J. Immun. 145, 3216–3222 (1990).
Lalor, P. A. & Morahan, G. Eur. J. Immun. 20, 485–492 (1990).
Lalor, P. A., Stall, A. M., Adams, S. & Herzenberg, L. A. Eur. J. Immun. 19, 501–506 (1989).
Chomczynski, P. & Sacchi, N. Analyt. Biochem. 162, 156–159 (1987).
Sambrook, J., Fritsch, E. F. & Maniatis, T. in Molecular Cloning- A Laboratory Manual (Cold Spring Harbor Laboratory Press, New York, 1989).
Sanger, F., Miklen, S. & Coulson, A. R. Proc. natn. Acad. Sci. U.S.A. 74, 5463–5467 (1977).
Winship, P. R. Nucleic Acids Res. 17, 1266 (1989).
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McHeyzer-Williams, M., Nossal, G. & Lalor, P. Molecular characterization of single memory B cells. Nature 350, 502–505 (1991). https://doi.org/10.1038/350502a0
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DOI: https://doi.org/10.1038/350502a0
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