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. 1972 Feb 29;135(3):675–697. doi: 10.1084/jem.135.3.675

IMMUNE RESPONSES IN VITRO

IV. SUPPRESSION OF PRIMARYγM, γG, ANDγA PLAQUE-FORMING CELL RESPONSES IN MOUSE SPLEEN CELL CULTURES BY CLASS-SPECIFIC ANTIBODY TO MOUSE IMMUNOGLOBULINS

Carl W Pierce 1, Susan M Solliday 1, Richard Asofsky 1
PMCID: PMC2139142  PMID: 4536706

Abstract

The suppressive effects of monospecific goat anti-mouse globulins on primary immunoglobulin class-specific plaque-forming cell responses in mouse spleen cell cultures were investigated. Anti-µ suppressed responses in all immunoglobulin classes, whereas anti-γ1 and anti-γ2 suppressed the γ1 and γ2 responses but not γM or γA responses, and anti-γA suppressed only γA responses. The mechanism of action of the anti-µ was studied in detail because of its suppression of responses in all immunoglobulin classes. The anti-µ was specific for µ-chain determinants; its activity was dose dependent, but was not mediated by killing cells with surface µ-chain determinants. Free γM but not γG myeloma proteins in solution effectively competed with µ-bearing cells for the anti-µ. An excess of anti-µ was necessary in the cultures for 48 hr to insure complete suppression of 5-day responses. However, after removal of excess anti-µ at 48 hr, responses could be stimulated by newly added antigen in cultures where incubation was prolonged to 7 days. Anti-µ was most effective when added at the initiation of cultures and had no suppressive effect when added at 48 hr. Excess antigen did not effectively compete with anti-µ for antigen receptors. Precursors of antibody-forming cells were shown to be the cell population where the suppressive activity of anti-µ was mediated. The experiments suggest that anti-µ combines with µ-chain determinants in antigen-specific receptors on the surfaces of antibody-forming cell precursors, prevents effective stimulation by antigen and subsequent antibody production. To explain suppression of responses in all Ig classes by anti-µ, several models were proposed. It is not possible to determine from the data whether stimulation of precursor cells with γG or γA receptors requires concommitant stimulation of separate cells with only γM receptors, or whether cells bearing γM receptors are precommitted to or differentiate into cells capable of synthesis of other Ig classes, or whether receptors of γM and another Ig class are present on some virgin precursors or the second Ig receptor appears after antigenic stimulation.

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Selected References

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  1. Ada G. L. Antigen binding cells in tolerance and immunity. Transplant Rev. 1970;5:105–129. doi: 10.1111/j.1600-065x.1970.tb00358.x. [DOI] [PubMed] [Google Scholar]
  2. Ada G. L., Byrt P. Specific inactivation of antigen-reactive cells with 125I-labelled antigen. Nature. 1969 Jun 28;222(5200):1291–1292. doi: 10.1038/2221291a0. [DOI] [PubMed] [Google Scholar]
  3. Avrameas S., Ternynck T. Biologically active water-insoluble protein polymers. I. Their use for isolation of antigens and antibodies. J Biol Chem. 1967 Apr 10;242(7):1651–1659. [PubMed] [Google Scholar]
  4. Bankhurst A. D., Warner N. L. Surface immunoglobulins on mouse lymphoid cells. J Immunol. 1971 Aug;107(2):368–373. [PubMed] [Google Scholar]
  5. Basten A., Miller J. F., Warner N. L., Pye J. Specific inactivation of thymus-derived (T) and non-thymus-derived (B) lymphocytes by 125I-labelled antigen. Nat New Biol. 1971 May 26;231(21):104–106. doi: 10.1038/newbio231104a0. [DOI] [PubMed] [Google Scholar]
  6. Baur S., Vitetta E. S., Sherr C. J., Schenkein I., Uhr J. W. Isolation of heavy and light chains of immunoglobulin from the surfaces of lymphoid cells. J Immunol. 1971 Apr;106(4):1133–1135. [PubMed] [Google Scholar]
  7. Bert G., Massaro A. L., Di Cossano D. L., Maja M. Electrophoretic study of immunoglobulins and immunoglobulin sub-units on the surface of human peripheral blood lymphocytes. Immunology. 1969 Jul;17(1):1–6. [PMC free article] [PubMed] [Google Scholar]
  8. Biozzi G., Asofsky R., Lieberman R., Stiffel C., Mouton D., Benacerraf B. Serum concentrations and allotypes of immunoglobulins in two lines of mice genetically selected for "high" or "low" antibody synthesis. J Exp Med. 1970 Oct 1;132(4):752–764. doi: 10.1084/jem.132.4.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Boyse E. A., Itakura K., Stockert E., Iritani C. A., Miura M. Ly-C: a third locus specifying alloantigens expressed only on thymocytes and lymphocytes. Transplantation. 1971 Mar;11(3):351–353. [PubMed] [Google Scholar]
  10. Bretscher P. A., Cohn M. Minimal model for the mechanism of antibody induction and paralysis by antigen. Nature. 1968 Nov 2;220(5166):444–448. doi: 10.1038/220444a0. [DOI] [PubMed] [Google Scholar]
  11. Byrt P., Ada G. L. An in vitro reaction between labelled flagellin or haemocyanin and lymphocyte-like cells from normal animals. Immunology. 1969 Oct;17(4):503–516. [PMC free article] [PubMed] [Google Scholar]
  12. Chan E. L., Mishell R. I., Mitchell G. F. Cell interaction in an immune response in vitro: requirement for theta-carrying cells. Science. 1970 Dec 11;170(3963):1215–1217. doi: 10.1126/science.170.3963.1215. [DOI] [PubMed] [Google Scholar]
  13. Claman H. N., Chaperon E. A. Immunologic complementation between thymus and marrow cells--a model for the two-cell theory of immunocompetence. Transplant Rev. 1969;1:92–113. doi: 10.1111/j.1600-065x.1969.tb00137.x. [DOI] [PubMed] [Google Scholar]
  14. Cooper M. D., Raymond D. A., Peterson R. D., South M. A., Good R. A. The functions of the thymus system and the bursa system in the chicken. J Exp Med. 1966 Jan 1;123(1):75–102. doi: 10.1084/jem.123.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Cudkowicz G., Shearer G. M., Priore R. L. Cellular differentiation of the immune system of mice. V. Class differentiation in marrow precursors of plaque-forming cells. J Exp Med. 1969 Sep 1;130(3):481–491. doi: 10.1084/jem.130.3.481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Daguillard F., Richter M. Cells involved in the immune response. XII. The differing responses of normal rabbit lymphoid cells to phytohemagglutinin, goat anti-rabbit immunoglobulin antiserum and allogeneic and xenogeneic lymphocytes. J Exp Med. 1969 Nov 1;130(5):1187–1208. doi: 10.1084/jem.130.5.1187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Davie J. M., Paul W. E., Mage R. G., Goldman M. B. Membrane-associated immunoglobulin of rabbit peripheral blood lymphocytes: allelic exclusion at the b locus. Proc Natl Acad Sci U S A. 1971 Feb;68(2):430–434. doi: 10.1073/pnas.68.2.430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Davie J. M., Paul W. E. Receptors on immunocompetent cells. II. Specificity and nature of receptors on dinitrophenylated guinea pig albumin- 125 I-binding lymphocytes of normal guinea pigs. J Exp Med. 1971 Aug 1;134(2):495–516. doi: 10.1084/jem.134.2.495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Davie J. M., Rosenthal A. S., Paul W. E. Receptors on immunocompetent cells. 3. Specificity and nature of receptors on dinitrophenylated guinea pig albumin- 125 I-binding cells of immunized guinea pigs. J Exp Med. 1971 Aug 1;134(2):517–531. doi: 10.1084/jem.134.2.517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Davies A. J. The thymus and the cellular basis of immunity. Transplant Rev. 1969;1:43–91. doi: 10.1111/j.1600-065x.1969.tb00136.x. [DOI] [PubMed] [Google Scholar]
  21. Doria G., Martinozzi M., Agarossi G., Di Pietro S. In vitro primary immune response resulting from the interaction between bone marrow-derived and thymus cells. Experientia. 1970 Apr 15;26(4):410–411. doi: 10.1007/BF01896919. [DOI] [PubMed] [Google Scholar]
  22. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  23. Feldmann M., Diener E. Reversible blocking effect of anti-mouse immunoglobulin serum on the induction of immunity and tolerance in vitro. Nat New Biol. 1971 Jun 9;231(23):183–184. doi: 10.1038/newbio231183a0. [DOI] [PubMed] [Google Scholar]
  24. Greaves M. F. The expression of immunoglobulin determinants on the surface of antigen-binding lymphoid cells in mice. I. An analysis of light and heavy chain restrictions on individual cells. Eur J Immunol. 1971 Jun;1(3):186–194. doi: 10.1002/eji.1830010308. [DOI] [PubMed] [Google Scholar]
  25. Hartmann K. U. Induction of a hemolysin response in vitro. II. Influence of the thymus-derived cells during the development of the antibody-producing cells. J Exp Med. 1971 Jun 1;133(6):1325–1333. doi: 10.1084/jem.133.6.1325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Hartmann K. U. Induction of a hemolysin response in vitro. Interaction of cells of bone marrow origin and thymic origin. J Exp Med. 1970 Dec 1;132(6):1267–1278. doi: 10.1084/jem.132.6.1267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Hartmann K. U., Reeg S., Mehner C. The induction of haemolysin producing cells in vitro: inhibition by antiglobulin antisera. Immunology. 1971 Jan;20(1):29–36. [PMC free article] [PubMed] [Google Scholar]
  28. Haskill J. S., Byrt P., Marbrook J. In vitro and in vivo studies of the immune response to sheep erythrocytes using partially purified cell preparations. J Exp Med. 1970 Jan 1;131(1):57–76. doi: 10.1084/jem.131.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Hirst J. A., Dutton R. W. Cell components in the immune response. 3. Neonatal thymectomy: restoration in culture. Cell Immunol. 1970 Jul;1(2):190–195. doi: 10.1016/0008-8749(70)90006-7. [DOI] [PubMed] [Google Scholar]
  30. JERNE N. K., NORDIN A. A. Plaque formation in agar by single antibody-producing cells. Science. 1963 Apr 26;140(3565):405–405. [PubMed] [Google Scholar]
  31. Johansson B., Klein E. Cell surface localized IgM-kappa immunoglobulin reactivity in a case of chronic lymphocytic leukaemia. Clin Exp Immunol. 1970 Mar;6(3):421–428. [PMC free article] [PubMed] [Google Scholar]
  32. Klein E., Eskeland T., Inoue M., Strom R., Johansson B. Surface immunoglobulin-moieties on lymphoid cells. Exp Cell Res. 1970 Sep;62(1):133–148. doi: 10.1016/0014-4827(79)90515-9. [DOI] [PubMed] [Google Scholar]
  33. Lawton A. R., 3rd, Asofsky R., Hylton M. B., Cooper M. D. Suppression of immunoglobulin class synthesis in mice. I. Effects of treatment with antibody to -chain. J Exp Med. 1972 Feb 1;135(2):277–297. doi: 10.1084/jem.135.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Lesley J., Dutton R. W. Antigen receptor molecules: inhibition by antiserum against kappa light chains. Science. 1970 Jul 31;169(3944):487–488. doi: 10.1126/science.169.3944.487. [DOI] [PubMed] [Google Scholar]
  35. Mann D., Granger H., Fahey J. L. Use of insoluble antibody for quantitative determination of small amounts of immunoglobulin. J Immunol. 1969 Mar;102(3):618–624. [PubMed] [Google Scholar]
  36. Mason S., Warner N. L. The immunoglobulin nature of the antigen recognition site on cells mediating transplantation immunity and delayed hypersentivity. J Immunol. 1970 Mar;104(3):762–765. [PubMed] [Google Scholar]
  37. McConnell I., Munro A., Gurner B. W., Coombs R. R. Studies on actively allergized cells. I. The cyto-dynamics and morphology of rosete-forming lymph node cells in mice and inhibition of rosette-formation with antibody to mouse immunoglobulins. Int Arch Allergy Appl Immunol. 1969;35(3):209–227. [PubMed] [Google Scholar]
  38. Miller H. C., Cudkowicz G. Density gradient separation of marrow cells restricted for antibody class. Science. 1971 Mar 5;171(3974):913–915. doi: 10.1126/science.171.3974.913. [DOI] [PubMed] [Google Scholar]
  39. Miller J. F., Mitchell G. F. Thymus and antigen-reactive cells. Transplant Rev. 1969;1:3–42. doi: 10.1111/j.1600-065x.1969.tb00135.x. [DOI] [PubMed] [Google Scholar]
  40. Mosier D. E. A requirement for two cell types for antibody formation in vitro. Science. 1967 Dec 22;158(3808):1573–1575. doi: 10.1126/science.158.3808.1573. [DOI] [PubMed] [Google Scholar]
  41. Mosier D. E., Fitch F. W., Rowley D. A., Davies A. J. Cellular deficit in thymectomized mice. Nature. 1970 Jan 17;225(5229):276–277. doi: 10.1038/225276a0. [DOI] [PubMed] [Google Scholar]
  42. Munro A., Hunter P. In vitro reconstitution of the immune response of thymus-deprived mice to sheep red blood cells. Nature. 1970 Jan 17;225(5229):277–278. doi: 10.1038/225277a0. [DOI] [PubMed] [Google Scholar]
  43. Mäkelä O., Cross A. M. The diversity and specialization of immunocytes. Prog Allergy. 1970;14:145–207. doi: 10.1159/000289379. [DOI] [PubMed] [Google Scholar]
  44. Naor D., Sulitzneau D. Binding of radioiodinated bovine serum albumin to mouse spleen cells. Nature. 1967 May 13;214(5089):687–688. doi: 10.1038/214687a0. [DOI] [PubMed] [Google Scholar]
  45. Nordin A. A., Cosenza H., Sell S. Immunoglobulin classes of antibody-forming cells in mice. II. Class restriction of plaque-forming cells demonstrated by replica plating. J Immunol. 1970 Feb;104(2):495–501. [PubMed] [Google Scholar]
  46. Oppenheim J. J., Rogentine G. N., Terry W. D. The transformation of human lymphocytes by monkey antisera to human immunoglobulins. Immunology. 1969 Jan;16(1):123–138. [PMC free article] [PubMed] [Google Scholar]
  47. Paul W. E. Functional specificity of antigen-binding receptors of lymphocytes. Transplant Rev. 1970;5:130–166. doi: 10.1111/j.1600-065x.1970.tb00359.x. [DOI] [PubMed] [Google Scholar]
  48. Pernis B., Forni L., Amante L. Immunoglobulin spots on the surface of rabbit lymphocytes. J Exp Med. 1970 Nov;132(5):1001–1018. doi: 10.1084/jem.132.5.1001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Pierce C. W. Immune responses in vitro. I. Cellular requirements for the immune response by nonprimed and primed spleen cells in vitro. J Exp Med. 1969 Aug 1;130(2):345–364. doi: 10.1084/jem.130.2.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Pierce C. W., Johnson B. M., Gershon H. E., Asofsky R. Immune responses in vitro. 3. Development of primary gamma-M, gamma-G, and gamma-A plaque-forming cell responses in mouse spleen cell cultures stimulated with heterologous erythrocytes. J Exp Med. 1971 Aug 1;134(2):395–416. doi: 10.1084/jem.134.2.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Pierce C. W., Solliday S. M., Asofsky R. Immune responses in vitro. V. Suppression of M, G, and A plaque-forming cell responses in cultures of primed mouse spleen cells by class-specific antibody to mouse immunoglobulins. J Exp Med. 1972 Mar 1;135(3):698–710. doi: 10.1084/jem.135.3.698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Plotz P. H., Talal N., Asofsky R. Assignment of direct and facilitated hemolytic plaques in mice to specific immunoglobulin classes. J Immunol. 1968 Apr;100(4):744–751. [PubMed] [Google Scholar]
  53. Rabellino E., Colon S., Grey H. M., Unanue E. R. Immunoglobulins on the surface of lymphocytes. I. Distribution and quantitation. J Exp Med. 1971 Jan 1;133(1):156–167. doi: 10.1084/jem.133.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Raff M. C., Sternberg M., Taylor R. B. Immunoglobulin determinants on the surface of mouse lymphoid cells. Nature. 1970 Feb 7;225(5232):553–554. doi: 10.1038/225553a0. [DOI] [PubMed] [Google Scholar]
  55. Raff M. C. Two distinct populations of peripheral lymphocytes in mice distinguishable by immunofluorescence. Immunology. 1970 Oct;19(4):637–650. [PMC free article] [PubMed] [Google Scholar]
  56. Raff M. C., Wortis H. H. Thymus dependence of theta-bearing cells in the peripheral lymphoid tissues of mice. Immunology. 1970 Jun;18(6):931–942. [PMC free article] [PubMed] [Google Scholar]
  57. Reif A. E., Allen J. M. Mouse thymic iso-antigens. Nature. 1966 Jan 29;209(5022):521–523. doi: 10.1038/209521b0. [DOI] [PubMed] [Google Scholar]
  58. SELL S., GELL P. G. STUDIES ON RABBIT LYMPHOCYTES IN VITRO. I. STIMULATION OF BLAST TRANSFORMATION WITH AN ANTIALLOTYPE SERUM. J Exp Med. 1965 Aug 1;122:423–440. doi: 10.1084/jem.122.2.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Siskind G. W., Benacerraf B. Cell selection by antigen in the immune response. Adv Immunol. 1969;10:1–50. doi: 10.1016/s0065-2776(08)60414-9. [DOI] [PubMed] [Google Scholar]
  60. Talmage D. W., Radovich J., Hemmingsen H. Cell interaction in antibody synthesis. Adv Immunol. 1970;12:271–282. doi: 10.1016/s0065-2776(08)60171-6. [DOI] [PubMed] [Google Scholar]
  61. Taylor R. B. Cellular cooperation in the antibody response of mice to two serum albumins: specific function of thymus cells. Transplant Rev. 1969;1:114–149. doi: 10.1111/j.1600-065x.1969.tb00138.x. [DOI] [PubMed] [Google Scholar]
  62. Truffa-Bachi P., Wofsy L. Specific separation of cells on affinity columns. Proc Natl Acad Sci U S A. 1970 Jul;66(3):685–692. doi: 10.1073/pnas.66.3.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Unanue E. R., Grey H. M., Rabellino E., Campbell P., Schmidtke J. Immunoglobulins on the surface of lymphocytes. II. The bone marrow as the main source of lymphocytes with detectable surface-bound immunoglobulin. J Exp Med. 1971 Jun 1;133(6):1188–1198. doi: 10.1084/jem.133.6.1188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Vitetta E. S., Baur S., Uhr J. W. Cell surface immunoglobulin. II. Isolation and characterization of immunoglobulin from mouse splenic lymphocytes. J Exp Med. 1971 Jul 1;134(1):242–264. doi: 10.1084/jem.134.1.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Walters C. S., Wigzell H. Demonstration of heavy and light chain antigenic determinants on the cell-bound receptor for antigen. Similarities between membrane-attached and humoral antibodies produced by the same cell. J Exp Med. 1970 Dec 1;132(6):1233–1249. doi: 10.1084/jem.132.6.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Warner N. L., Byrt P., Ada G. L. Blocking of the lymphocyte antigen receptor site with anti-immunoglobulin sera in vitro. Nature. 1970 Jun 6;226(5249):942–943. doi: 10.1038/226942a0. [DOI] [PubMed] [Google Scholar]
  67. 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]
  68. 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]
  69. Wigzell H. Specific fractionation of immunocompetent cells. Transplant Rev. 1970;5:76–104. doi: 10.1111/j.1600-065x.1970.tb00357.x. [DOI] [PubMed] [Google Scholar]

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