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. 1970 Jun 1;131(6):1093–1108. doi: 10.1084/jem.131.6.1093

IMMUNOLOGICAL MEMORY IN MICE

I. PHYSICAL SEPARATION AND PARTIAL CHARACTERIZATION OF MEMORY CELLS FOR DIFFERENT IMMUNOGLOBULIN CLASSES FROM EACH OTHER AND FROM ANTIBODY-PRODUCING CELLS

Johanna L'age-Stehr 1, Leonard A Herzenberg 1
PMCID: PMC2138841  PMID: 4192567

Abstract

Plaque forming cells (PFC) of different immunoglobulin classes producing antibodies against sheep erythrocytes were separated according to their buoyant densities by means of equilibrium centrifugation in a stepwise BSA gradient. In the period of 7–10 days after immunization γM PFC are markedly enriched in fractions of low density and relatively depleted in fractions of high density. The distribution of total γG PFC shows less enrichment in the lower density fractions and less depletion in the higher density fractions. The density profile for γG2a PFC is even flatter, with a significant difference (depletion) relative to the unseparated spleen cells only in the highest density fraction. The density gradient distributions of cells able to transfer an adoptive immune response of the various immunoglobulin classes are markedly different from the PFC distribution. Cells obtained 7–10 days after immunization able to transfer an IgM response are present in the same proportions across the density gradient, whereas memory cells for γG2a obtained at this time are markedly enriched in fractions of low density and virtually depleted from high density fractions. With increasing time after primary immunization, the γG2a memory cells increase progressively in density and by 6 weeks the higher and lower density fractions have the same proportions of γG2a memory cells. The total γG (mainly γG1) memory cells by 7–10 days show slight enrichment in low density fractions and no depletion in high density fractions. The conclusions were reached that (a) memory for γG1 develops earlier than memory for γG2a and (b) that memory for anti-SRBC antibodies of different classes is carried in separate cells. When gradient fractions enriched for PFC and memory cells for all classes were completely depleted of PFC using glass bead columns, the ability of this fraction to transfer memory for all classes was not diminished. This shows that memory cells are not identical with cells secreting antibodies.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BERNIER G. M., CEBRA J. J. POLYPEPTIDE CHAINS OF HUMAN GAMMA-GLOBULIN: CELLULAR LOCALIZATION BY FLUORESCENT ANTIBODY. Science. 1964 Jun 26;144(3626):1590–1591. doi: 10.1126/science.144.3626.1590. [DOI] [PubMed] [Google Scholar]
  2. BORUN E. R., FIGUEROA W. G., PERRY S. M. The distribution of Fe59 tagged human erythrocytes in centrifuged specimens as a function of cell age. J Clin Invest. 1957 May;36(5):676–679. doi: 10.1172/JCI103468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Claflin A. J., Smithies O. Antibody-producing cells in division. Science. 1967 Sep 29;157(3796):1561–1562. [PubMed] [Google Scholar]
  4. Claman H. N., Chaperon E. A., Triplett R. F. Immunocompetence of transferred thymus-marrow cell combinations. J Immunol. 1966 Dec;97(6):828–832. [PubMed] [Google Scholar]
  5. Cunningham A. J. Studies on the cellular basis of IgM immunological memory. Immunology. 1969 May;16(5):621–632. [PMC free article] [PubMed] [Google Scholar]
  6. Dresser D. W., Wortis D. H. Use of an antiglobulin serum to detect cells producing antibody with low haemolytic efficiency. Nature. 1965 Nov 27;208(5013):859–861. doi: 10.1038/208859a0. [DOI] [PubMed] [Google Scholar]
  7. EISEN H. N., SISKIND G. W. VARIATIONS IN AFFINITIES OF ANTIBODIES DURING THE IMMUNE RESPONSE. Biochemistry. 1964 Jul;3:996–1008. doi: 10.1021/bi00895a027. [DOI] [PubMed] [Google Scholar]
  8. Green I., Vassalli P., Nussenzweig V., Benacerraf B. Specificity of the antibodies produced by single cells following immunization with antigens bearing two types of antigenic determinants. J Exp Med. 1967 Mar 1;125(3):511–526. doi: 10.1084/jem.125.3.511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Haskill J. S. Density distribution analysis of antigen sensitive cells in the rat. Nature. 1967 Dec 23;216(5121):1229–1231. doi: 10.1038/2161229a0. [DOI] [PubMed] [Google Scholar]
  10. Haskill J. S., Legge D. G., Shortman K. Density distribution analysis of cells forming 19S hemolytic antibody in the rat. J Immunol. 1969 Mar;102(3):703–712. [PubMed] [Google Scholar]
  11. Klein J., Herzenberg L. A. Congenic mouse strains with different immunoglobulin allotypes. I. Breeding scheme, histocompatibility tests, and kinetics of gamma G2a-globulin production by transferred cells for C3H.SW and its congenic partner CWB/5. Transplantation. 1967 Nov;5(6):1484–1495. doi: 10.1097/00007890-196711000-00013. [DOI] [PubMed] [Google Scholar]
  12. Miller J. F., Mitchell G. F. Cell to cell interaction in the immune response. I. Hemolysin-forming cells in neonatally thymectomized mice reconstituted with thymus or thoracic duct lymphocytes. J Exp Med. 1968 Oct 1;128(4):801–820. doi: 10.1084/jem.128.4.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Mitchell G. F., Miller J. F. Cell to cell interaction in the immune response. II. The source of hemolysin-forming cells in irradiated mice given bone marrow and thymus or thoracic duct lymphocytes. J Exp Med. 1968 Oct 1;128(4):821–837. doi: 10.1084/jem.128.4.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Möller G. Regulation of cellular antibody synthesis. Cellular 7S production and longevity of 7S antigen-sensitive cells in the absence of antibody feedback. J Exp Med. 1968 Feb 1;127(2):291–306. doi: 10.1084/jem.127.2.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. NOSSAL G. J., MAKELA O. Kinetic studies on the incidence of cells appearing to form two antibodies. J Immunol. 1962 May;88:604–612. [PubMed] [Google Scholar]
  17. PUCK T. T., CIECIURA S. J., ROBINSON A. Genetics of somatic mammalian cells. III. Long-term cultivation of euploid cells from human and animal subjects. J Exp Med. 1958 Dec 1;108(6):945–956. doi: 10.1084/jem.108.6.945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pernis B., Chiappino G., Kelus A. S., Gell P. G. Cellular localization of immunoglobulins with different allotypic specificities in rabbit lymphoid tissues. J Exp Med. 1965 Nov 1;122(5):853–876. doi: 10.1084/jem.122.5.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Plotz P. H., Talal N. Fractionation of splenic antibody-forming cells on glass bead columns. J Immunol. 1967 Dec;99(6):1236–1242. [PubMed] [Google Scholar]
  20. Raidt D. J., Mishell R. I., Dutton R. W. Cellular events in the immune response : analysis and in vitro response of mouse spleen cell populations separated by differential flotation in albumin gradients. J Exp Med. 1968 Oct 1;128(4):681–698. doi: 10.1084/jem.128.4.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. STAATS J. STANDARDIZED NOMENCLATURE FOR INBRED STRAINS OF MICE. THIRD LISTING. Cancer Res. 1964 Feb;24:147–168. [PubMed] [Google Scholar]
  22. Sercarz E. E., Byers V. S. The X-Y-Z scheme of immunocyte maturation. 3. Early IgM memory and the nature of the memory cell. J Immunol. 1967 Apr;98(4):836–843. [PubMed] [Google Scholar]
  23. Shearer G. M., Cudkowicz G., Priore R. L. Cellular differentiation of the immune system of mice. II. Frequency of unipotent splenic antigen-sensitive units after immunization with sheep erythrocytes. J Exp Med. 1969 Jan 1;129(1):185–199. doi: 10.1084/jem.129.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shortman K. The separation of different cell classes from lymphoid organs. II. The purification and analysis of lymphocyte populations by equilibrium density gradient centrifugation. Aust J Exp Biol Med Sci. 1968 Aug;46(4):375–396. doi: 10.1038/icb.1968.32. [DOI] [PubMed] [Google Scholar]
  25. Sterzl J., Ríha I. Detection of cells producing 7S antibodies by the plaque technique. Nature. 1965 Nov 27;208(5013):858–859. doi: 10.1038/208858a0. [DOI] [PubMed] [Google Scholar]
  26. Szenberg A., Cunningham A. J. DNA synthesis in the development of antibody-forming cells during the early stages of the immune response. Nature. 1968 Feb 24;217(5130):747–748. doi: 10.1038/217747a0. [DOI] [PubMed] [Google Scholar]
  27. Tannenberg W. J., Malaviya A. N. The life cycle of antibody-forming cells. I. The generation time of 19S hemolytic plaque-forming cells during the primary and secondary responses. J Exp Med. 1968 Nov 1;128(5):895–925. doi: 10.1084/jem.128.5.895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Uhr J. W., Möller G. Regulatory effect of antibody on the immune response. Adv Immunol. 1968;8:81–127. doi: 10.1016/s0065-2776(08)60465-4. [DOI] [PubMed] [Google Scholar]
  29. Vazquez J. J., Makinodan T. Cytokinetic events following antigenic stimulation. Fed Proc. 1966 Nov-Dec;25(6):1727–1733. [PubMed] [Google Scholar]
  30. Wanebo H. J., Gallmeier W. M., Boyse E. A., Old L. J. Paraproteinemia and reticulum cell sarcoma in an inbred mouse strain. Science. 1966 Nov 18;154(3751):901–903. doi: 10.1126/science.154.3751.901. [DOI] [PubMed] [Google Scholar]
  31. Weiler E., Melletz E. W., Breuninger-Peck E. Facilitation of immune hemolysis by an interaction between red cell-sensitizing antibody and gamma-globulin allotype antibody. Proc Natl Acad Sci U S A. 1965 Nov;54(5):1310–1317. doi: 10.1073/pnas.54.5.1310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wortis H. H., Dresser D. W., Anderson H. R. Antibody production studied by means of the localized haemolysis in gel (LHG) assay. 3. Mouse cells producing five different classes of antibody. Immunology. 1969 Jul;17(1):93–110. [PMC free article] [PubMed] [Google Scholar]

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