Abstract
Antigen-specific B cells (ASC) were purified from other B cells by prior incubation with specific antigen followed by rosetting with erythrocytes conjugated with anti-mouse Ig and sedimenting on Ficoll- Isopaque. This procedure allowed the removal of most of the B cells, while those speicifc for the antigen used in incubation were retained. Relative to the B-cell content, ASC were enriched 64- to 132-fold. The method is highly specific in that B cells primed to two different antigens, turkey gamma globulin and sheep erythrocytes, could be separated from each other. The advantages of this indirect purification procedure over purification procedures which obtain ASC directly are the simplicity of obtaining the ASC and the ability of the ASC of respond to antigen without the addition of other cells.
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- Ault K. A., Unanue E. R. Events after the binding of antigen to lymphocytes: removal and regeneration of the antigen receptor. J Exp Med. 1974 May 1;139(5):1110–1124. doi: 10.1084/jem.139.5.1110. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bonner W. A., Hulett H. R., Sweet R. G., Herzenberg L. A. Fluorescence activated cell sorting. Rev Sci Instrum. 1972 Mar;43(3):404–409. doi: 10.1063/1.1685647. [DOI] [PubMed] [Google Scholar]
- Choi T. K., Sleight D. R., Nisonoff A. General method for isolation and recovery of B cells bearing specific receptors. J Exp Med. 1974 Mar 1;139(3):761–766. doi: 10.1084/jem.139.3.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davidson W. F., Parish C. R. A procedure for removing red cells and dead cells from lymphoid cell suspensions. J Immunol Methods. 1975 Jun;7(2-3):291–300. doi: 10.1016/0022-1759(75)90026-5. [DOI] [PubMed] [Google Scholar]
- Edelman G. M., Rutishauser U., Millette C. F. Cell fractionation and arrangement on fibers, beads, and surfaces. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2153–2157. doi: 10.1073/pnas.68.9.2153. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Falkoff R., Kettman J. Differential stimulation of precursor cells and carrier-specific thymus-derived cell activity in the in vivo reponse to heterologous erythrocytes in mice. J Immunol. 1972 Jan;108(1):54–58. [PubMed] [Google Scholar]
- Golub E. S., Mishell R. I., Weigle W. O., Dutton R. W. A modification of the hemolytic plaque assay for use with protein antigens. J Immunol. 1968 Jan;100(1):133–137. [PubMed] [Google Scholar]
- Haas W., Layton J. E. Separation of antigen-specific lymphocytes. I. Enrichment of antigen-binding cells. J Exp Med. 1975 May 1;141(5):1004–1014. doi: 10.1084/jem.141.5.1004. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haas W., Schrader J. W., Szenberg A. A new, simple method for the preparation of lymphocytes bearing specific receptors. Eur J Immunol. 1974 Aug;4(8):565–570. doi: 10.1002/eji.1830040809. [DOI] [PubMed] [Google Scholar]
- Haas W. Separation of antigen-specific lymphocytes. II. Enrichment of hapten-specific antibody-forming cell precursors. J Exp Med. 1975 May 1;141(5):1015–1029. doi: 10.1084/jem.141.5.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Julius M. H., Herzenberg L. A. Isolation of antigen-binding cells from unprimed mice: demonstration of antibody-forming cell precursor activity and correlation between precursor and secreted antibody avidities. J Exp Med. 1974 Oct 1;140(4):904–920. doi: 10.1084/jem.140.4.904. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Julius M. H., Masuda T., Herzenberg L. A. Demonstration that antigen-binding cells are precursors of antibody-producing cells after purification with a fluorescence-activated cell sorter. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1934–1938. doi: 10.1073/pnas.69.7.1934. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Katz D. H., Unanue E. R. Critical role of determinant presentation in the induction of specific responses in immunocompetent lymphocytes. J Exp Med. 1973 Apr 1;137(4):967–990. doi: 10.1084/jem.137.4.967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klinman N. R., Pickard A. R., Sigal N. H., Gearhart P. J., Metcalf E. S., Pierce S. K. Assessing B cell diversification by antigen receptor and precursor cell analysis. Ann Immunol (Paris) 1976 Jun-Jul;127(3-4):489–502. [PubMed] [Google Scholar]
- Klinman N. R., Press J. L. The B cell specificity repertoire: its relationship to definable subpopulations. Transplant Rev. 1975;24:41–83. doi: 10.1111/j.1600-065x.1975.tb00165.x. [DOI] [PubMed] [Google Scholar]
- Lawrence D. A., Spiegelberg H. L., Weigle W. O. 2,4-Dinitrophenyl receptors on mouse thymus and spleen cells. J Exp Med. 1973 Feb 1;137(2):470–482. doi: 10.1084/jem.137.2.470. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Louis J., Chiller J. M., Weigle W. O. Fate of antigen-binding cells in unresponsive and immune mice. J Exp Med. 1973 Feb 1;137(2):461–469. doi: 10.1084/jem.137.2.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Makinodan T., Albright J. F. Proliferative and differentiative manifestations of cellular immune potential. Prog Allergy. 1967;10:1–36. [PubMed] [Google Scholar]
- Mishell R. I., Dutton R. W. Immunization of dissociated spleen cell cultures from normal mice. J Exp Med. 1967 Sep 1;126(3):423–442. doi: 10.1084/jem.126.3.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nossal G. J., Layton J. E. Antigen-induced aggregation and modulation of receptors on hapten-specific B lymphocytes. J Exp Med. 1976 Mar 1;143(3):511–528. doi: 10.1084/jem.143.3.511. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nossal G. J., Pike B. L. Single cell studies on the antibody-forming potential of fractionated, hapten-specific B lymphocytes. Immunology. 1976 Feb;30(2):189–202. [PMC free article] [PubMed] [Google Scholar]
- Schlossman S. F., Hudson L. Specific purification of lymphocyte populations on a digestible immunoabsorbent. J Immunol. 1973 Jan;110(1):313–319. [PubMed] [Google Scholar]
- Schrader J. W. The mechanism of bone marrow-derived (B) lymphocyte activation. I. Early events in antigen-induced triggering in the presence of polymerized flagellin. Eur J Immunol. 1974 Jan;4(1):14–20. doi: 10.1002/eji.1830040105. [DOI] [PubMed] [Google Scholar]
- Scott D. W. Antifluorescein affinity columns. Isolation and immunocompetence of lymphocytes that bind fluoresceinated antigens in vivo or in vitro. J Exp Med. 1976 Jul 1;144(1):69–78. doi: 10.1084/jem.144.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stocker J. W. Estimation of hapten-specific antibody-forming cell precursors in microcultures. Immunology. 1976 Feb;30(2):181–187. [PMC free article] [PubMed] [Google Scholar]
- Stålenheim G., Sjöquist J. Protein A from Staphylococcus aureus. X. Complement-fixing activity of complexes between protein A and myeloma IgG or papain fragments of IgG. J Immunol. 1970 Oct;105(4):944–948. [PubMed] [Google Scholar]
- Wigzell H., Andersson B. Cell separation on antigen-coated columns. Elimination of high rate antibody-forming cells and immunological memory cells. J Exp Med. 1969 Jan 1;129(1):23–36. doi: 10.1084/jem.129.1.23. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wigzell H., Mäkelä O. Separation of normal and immune lymphoid cells by antigen-coated coated columns. Antigen-binding characteristics of membrane antibodies as analyzed by hapten-protein antigens. J Exp Med. 1970 Jul 1;132(1):110–126. doi: 10.1084/jem.132.1.110. [DOI] [PMC free article] [PubMed] [Google Scholar]
