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. 1968 Oct 31;128(5):1099–1128. doi: 10.1084/jem.128.5.1099

CELLS INVOLVED IN THE IMMUNE RESPONSE

IV. THE RESPONSE OF NORMAL AND IMMUNE RABBIT BONE MARROW AND LYMPHOID TISSUE LYMPHOCYTES TO ANTIGENS IN VITRO

Sharwan K Singhal 1, Maxwell Richter 1
PMCID: PMC2138559  PMID: 4176224

Abstract

Cell suspensions of immune rabbit lymph nodes and spleen were capable of undergoing blastogenesis and mitosis and of incorporating tritiated thymidine when maintained in culture with the specific antigen in vitro. They did not respond to other, non-cross-reacting antigens. The blastogenic response obtained with immune lymph node cells could be correlated with the antibody synthesizing capacity of fragment cultures prepared from the same lymph nodes. Cell suspensions of immune bone marrow responded to non-cross-reacting antigens only whereas cell suspensions of immune thymus, sacculus rotundus, and appendix did not respond when exposed to any of the antigens tested. On the other hand, neither fragments nor cell suspensions prepared from lymph nodes, spleen, and thymus of normal, unimmunized rabbits responded with antibody formation and blastogenesis when exposed to any of the antigens. However, normal bone marrow cells responded with marked blastogenesis and tritiated thymidine uptake. The specificity of this in vitro bone marrow response was demonstrated by the fact that the injection of a protein antigen in vivo resulted in the loss of reactivity by the marrow cell to that particular antigen but not to the other, non-cross-reacting antigens. Furthermore, bone marrow cells of tolerant rabbits failed to respond to the specific antigen in vitro. It was also demonstrated that normal bone marrow cells incubated with antigen are capable of forming antibody which could be detected by the fluorescent antibody technique. This response of the bone marrow cells has been localized to the lymphocyte-rich fraction of the bone marrow. It is concluded that the bone marrow lymphocyte, by virtue of its capacity to react with blastogenesis and mitosis and with antibody formation upon initial exposure to the antigen, a capacity not possessed by lymphocytes of the other lymphoid organs, has a preeminent role in the sequence of cellular events culminating in antibody formation.

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Selected References

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  1. BACH F. H., HIRSCHHORN K. THE IN VITRO IMMUNE RESPONSE OF PERIPHERAL BLOOD LYMPHOCYTES. Semin Hematol. 1965 Jan;2:68–89. [PubMed] [Google Scholar]
  2. BACH F., HIRSCHHORN K. LYMPHOCYTE INTERACTION: A POTENTIAL HISTOCOMPATIBILITY TEST IN VITRO. Science. 1964 Feb 21;143(3608):813–814. doi: 10.1126/science.143.3608.813. [DOI] [PubMed] [Google Scholar]
  3. BAIN B., LOWENSTEIN L. GENETIC STUDIES ON THE MIXED LEUKOCYTE REACTION. Science. 1964 Sep 18;145(3638):1315–1316. doi: 10.1126/science.145.3638.1315. [DOI] [PubMed] [Google Scholar]
  4. BAIN B., VAS M. R., LOWENSTEIN L. THE DEVELOPMENT OF LARGE IMMATURE MONONUCLEAR CELLS IN MIXED LEUKOCYTE CULTURES. Blood. 1964 Jan;23:108–116. [PubMed] [Google Scholar]
  5. BOYDEN S. V. The adsorption of proteins on erythrocytes treated with tannic acid and subsequent hemagglutination by antiprotein sera. J Exp Med. 1951 Feb;93(2):107–120. doi: 10.1084/jem.93.2.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Byers V. S., Sercarz E. E. The X-Y-Z scheme of immunocyte maturation. IV. The exhaustion of memory cells. J Exp Med. 1968 Feb 1;127(2):307–325. doi: 10.1084/jem.127.2.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. CHAPMAN N. D., PARKHOUSE R. M., DUTTON R. W. ANTIGEN STIMULATED PROLIFERATION IN LYMPHOID AND MYELOID TISSUES FROM IMMUNIZED RABBITS. Proc Soc Exp Biol Med. 1964 Dec;117:708–711. doi: 10.3181/00379727-117-29674. [DOI] [PubMed] [Google Scholar]
  8. CONGDON C. C. Experimental treatment of total-body irradiation injury: a brief review. Blood. 1957 Aug;12(8):746–754. [PubMed] [Google Scholar]
  9. COWLING D. C., QUAGLINO D. EFFECT OF SOME ANTIGENS ON LEUCOCYTE CULTURES. J Pathol Bacteriol. 1965 Jan;89:63–70. doi: 10.1002/path.1700890108. [DOI] [PubMed] [Google Scholar]
  10. Chalmers D. G., Coulson A. S., Evans C., Yealland S. Immunologically stimulated human peripheral blood lymphocytes in vitro. II. Mixed lymphocyte cultures with related and unrelated donors. Int Arch Allergy Appl Immunol. 1966;30(2):177–189. doi: 10.1159/000229804. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Claman H. N., Chaperon E. A., Triplett R. F. Thymus-marrow cell combinations. Synergism in antibody production. Proc Soc Exp Biol Med. 1966 Aug-Sep;122(4):1167–1171. doi: 10.3181/00379727-122-31353. [DOI] [PubMed] [Google Scholar]
  13. Cooper M. D., Gabrielsen A. E., Good R. A. Role of the thymus and other central lymphoid tissues in immunological disease. Annu Rev Med. 1967;18:113–138. doi: 10.1146/annurev.me.18.020167.000553. [DOI] [PubMed] [Google Scholar]
  14. Coulson A. S., Chalmers D. G. Response of human blood lymphocytes to tuberculin PPD in tissue culture. Immunology. 1967 Apr;12(4):417–429. [PMC free article] [PubMed] [Google Scholar]
  15. DUTTON R. W., EADY J. D. AN IN VITRO SYSTEM FOR THE STUDY OF THE MECHANISM OF ANTIGENIC STIMULATION IN THE SECONDARY RESPONSE. Immunology. 1964 Jan;7:40–53. [PMC free article] [PubMed] [Google Scholar]
  16. Davies A. J., Leuchars E., Wallis V., Koller P. C. The mitotic response of thymus-derived cells to antigenic stimulus. Transplantation. 1966 Jul;4(4):438–451. doi: 10.1097/00007890-196607000-00008. [DOI] [PubMed] [Google Scholar]
  17. Dowden S. J., Sercarz E. E. The X-Y-Z scheme of immunocyte maturation. II. The effect of antigen on spontaneous escape from immune paralysis. J Immunol. 1967 Apr;98(4):827–835. [PubMed] [Google Scholar]
  18. Dutton R. W. In vitro studies of immunological responses of lymphoid cells. Adv Immunol. 1967;6:253–336. doi: 10.1016/s0065-2776(08)60524-6. [DOI] [PubMed] [Google Scholar]
  19. GENGOZIAN N., MAKINODAN T., SHEKARCHI I. C. Transplantation of antibody-forming cells in lethally irradiated mice. J Immunol. 1961 Feb;86:113–122. [PubMed] [Google Scholar]
  20. GOWANS J. L. The fate of parental strain small lymphocytes in F1 hybrid rats. Ann N Y Acad Sci. 1962 Oct 24;99:432–455. doi: 10.1111/j.1749-6632.1962.tb45326.x. [DOI] [PubMed] [Google Scholar]
  21. Girard J. P., Rose N. R., Kunz M. L., Kobayashi S., Arbesman C. E. In vitro lymphocyte transformation in atopic patients: induced by antigens. J Allergy. 1967 Feb;39(2):65–81. doi: 10.1016/0021-8707(67)90113-x. [DOI] [PubMed] [Google Scholar]
  22. Gowans J. L., Uhr J. W. The carriage of immunological memory by small lymphocytes in the rat. J Exp Med. 1966 Nov 1;124(5):1017–1030. doi: 10.1084/jem.124.5.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. HARRIS J. E., FORD C. E. CELLULAR TRAFFIC OF THE THYMUS: EXPERIMENTS WITH CHROMOSOME MARKERS. EVIDENCE THAT THE THYMUS PLAYS AN INSTRUCTIONAL PART. Nature. 1964 Feb 29;201:884–885. doi: 10.1038/201884a0. [DOI] [PubMed] [Google Scholar]
  24. HIRSCHHORN K., BACH F., KOLODNY R. L., FIRSCHEIN I. L., HASHEM N. IMMUNE RESPONSE AND MITOSIS OF HUMAN PERIPHERAL BLOOD LYMPHOCYTES IN VITRO. Science. 1963 Nov 29;142(3596):1185–1187. doi: 10.1126/science.142.3596.1185. [DOI] [PubMed] [Google Scholar]
  25. Juhasz A., Richter M. The secondary antibody response in tissue culture. II. Differences in the course of antibody synthesis to diphtheria toxoid and rat serum albumin. Can J Biochem. 1965 Jun;43(6):803–809. doi: 10.1139/o65-090. [DOI] [PubMed] [Google Scholar]
  26. Juhasz A., Rose B. The secondary antibody response in tissue culture. I. Observations on prolonged antibody synthesis by lymph node fragments in media of different composition. Can J Biochem. 1965 Jun;43(6):787–801. doi: 10.1139/o65-089. [DOI] [PubMed] [Google Scholar]
  27. Loewi G., Temple A., Vischer T. L. The immunological significance in the guinea-pig of in vitro transformation of lymph node, spleen and peripheral blood lymphocytes. Immunology. 1968 Feb;14(2):257–264. [PMC free article] [PubMed] [Google Scholar]
  28. MANNICK J. A., LOCHTE H. L., Jr, ASHLEY C. A., THOMAS E. D., FERREBEE J. W. Autografts of bone marrow in dogs after lethal total-body radiation. Blood. 1960 Feb;15:255–266. [PubMed] [Google Scholar]
  29. McGregor D. D., McCullagh P. J., Gowans J. L. The role of lymphocytes in antibody formation. I. Restoration of the haemolysin response in x-irradiated rats with lymphocytes from normal and immunologically tolerant donors. Proc R Soc Lond B Biol Sci. 1967 Sep 12;168(1012):229–243. doi: 10.1098/rspb.1967.0063. [DOI] [PubMed] [Google Scholar]
  30. Micklem H. S., Ford C. E., Evans E. P., Gray J. Interrelationships of myeloid and lymphoid cells: studies with chromosome-marked cells transfused into lethally irradiated mice. Proc R Soc Lond B Biol Sci. 1966 Jul 19;165(998):78–102. doi: 10.1098/rspb.1966.0059. [DOI] [PubMed] [Google Scholar]
  31. Miller J. F., Mitchell G. F. The thymus and the precursors of antigen reactive cells. Nature. 1967 Nov 18;216(5116):659–663. doi: 10.1038/216659a0. [DOI] [PubMed] [Google Scholar]
  32. Miller J. F., Osoba D. Current concepts of the immunological function of the thymus. Physiol Rev. 1967 Jul;47(3):437–520. doi: 10.1152/physrev.1967.47.3.437. [DOI] [PubMed] [Google Scholar]
  33. Mills J. A. The immunologic significance of antigen induced lymphocyte transformation in vitro. J Immunol. 1966 Aug;97(2):239–247. [PubMed] [Google Scholar]
  34. Mitchell G. F., Miller J. F. Immunological activity of thymus and thoracic-duct lymphocytes. Proc Natl Acad Sci U S A. 1968 Jan;59(1):296–303. doi: 10.1073/pnas.59.1.296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Morrison J. H., Toepfer J. R. Survival of lethally X-irradiated rats after treatment with isogenic marrow lymphocytes. Am J Physiol. 1967 Oct;213(4):923–927. doi: 10.1152/ajplegacy.1967.213.4.923. [DOI] [PubMed] [Google Scholar]
  36. Nossal G. J. Mechanisms of antibody production. Annu Rev Med. 1967;18:81–96. doi: 10.1146/annurev.me.18.020167.000501. [DOI] [PubMed] [Google Scholar]
  37. O'Brien T. F., Michaelides M. C., Coons A. H. STUDIES ON ANTIBODY PRODUCTION : VI. THE COURSE, SENSITIVITY, AND HISTOLOGY OF THE SECONDARY RESPONSE IN VITRO. J Exp Med. 1963 Jun 1;117(6):1053–1062. doi: 10.1084/jem.117.6.1053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Oppenheim J. J., Whang J., Frei E., 3rd The effect of skin homograft rejection on recipient and donor mixed leukocyte cultures. J Exp Med. 1965 Oct 1;122(4):651–664. doi: 10.1084/jem.122.4.651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. ROBBINS J. H. TISSUE CULTURE STUDIES OF THE HUMAN LYMPHOCYTE. Science. 1964 Dec 25;146(3652):1648–1654. doi: 10.1126/science.146.3652.1648. [DOI] [PubMed] [Google Scholar]
  40. Richter M., Juhasz A., Drechsler B., Myers J. Synthesis of antibody in vitro by lymph node fragments using absorbable gelatin sponges. Nature. 1966 May 7;210(5036):645–645. doi: 10.1038/210645a0. [DOI] [PubMed] [Google Scholar]
  41. Richter M., Naspitz C. K. The in vitro blastogenic response of lymphocytes of ragweed-sensitive individuals. J Allergy. 1968 Mar;41(3):140–151. doi: 10.1016/0021-8707(68)90054-3. [DOI] [PubMed] [Google Scholar]
  42. Richter M., Naspitz C. K. The variation in response of human peripheral lymphocytes to phytohemagglutinin in vitro. Int Arch Allergy Appl Immunol. 1967;32(3):288–293. doi: 10.1159/000229939. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Singhal K., Richter M. Cells involved in the immune response. I. The response of normal rabbit bone marrow cells to antigens in vitro. Int Arch Allergy Appl Immunol. 1968;33(5):493–500. doi: 10.1159/000230064. [DOI] [PubMed] [Google Scholar]
  45. Sterzl J., Jílek M. Number of antibody-forming cells in primary and secondary reactions after administration of antigen. Nature. 1967 Dec 23;216(5121):1233–1235. doi: 10.1038/2161233a0. [DOI] [PubMed] [Google Scholar]
  46. Stone S. H. Cellular commitments to immune responses. Annu Rev Microbiol. 1967;21:181–204. doi: 10.1146/annurev.mi.21.100167.001145. [DOI] [PubMed] [Google Scholar]
  47. TIMBURY M. C. INHIBITION OF VIRAL REPLICATION BY ANTICELLULAR SERUM. Virology. 1963 Aug;20:629–635. doi: 10.1016/0042-6822(63)90288-5. [DOI] [PubMed] [Google Scholar]
  48. TRENTIN J. J. Effect of x-ray dose on mortality and skin transplantability in mice receiving F1 hybrid marrow. Proc Soc Exp Biol Med. 1956 Oct;93(1):98–100. doi: 10.3181/00379727-93-22674. [DOI] [PubMed] [Google Scholar]
  49. Vischer T. L., Stastny P. Time of appearance and distribution of cells capable of secondary immune response following primary immunization. Immunology. 1967 Jun;12(6):675–687. [PMC free article] [PubMed] [Google Scholar]
  50. Wilson D. B., Billingham R. E. Lymphocytes and transplantation immunity. Adv Immunol. 1967;7:189–273. doi: 10.1016/s0065-2776(08)60129-7. [DOI] [PubMed] [Google Scholar]
  51. Wilson D. B. Quantitative studies on the mixed lymphocyte interaction in rats. I. Conditions and parameters of response. J Exp Med. 1967 Oct 1;126(4):625–654. doi: 10.1084/jem.126.4.625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Zeitz S. J., Van Arsdel P. P., Jr, McClure D. K. Specific response of human lymphocytes to pollen antigen in tissue culture. J Allergy. 1966 Dec;38(6):321–329. doi: 10.1016/0021-8707(66)90030-x. [DOI] [PubMed] [Google Scholar]

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