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. 1974 Jun 1;139(6):1553–1567. doi: 10.1084/jem.139.6.1553

CELLULAR INTERACTIONS IN THE PROLIFERATIVE RESPONSE OF HUMAN T AND B LYMPHOCYTES TO PHYTOMITOGENS AND ALLOGENEIC LYMPHOCYTES

Hans-Peter Lohrmann 1, Ligita Novikovs 1, Robert G Graw Jr 1
PMCID: PMC2139681  PMID: 4598019

Abstract

In vitro studies were performed to determine the proliferative responsiveness of human peripheral blood thymus-dependent (T) and thymus-independent (B) lymphocytes to phytomitogens and allogeneic lymphocytes. Recombination of T and B cells, with selective inhibition of proliferation of one of the two populations, was used to identify cellular interactions which may contribute to cell proliferation. The distinctive feature of human T lymphocytes to form rosettes with unsensitized sheep erythrocytes was utilized to separate human peripheral blood lymphocytes into highly purified resetting (T) and non-rosetting (B) cells. The proliferative response of these separated lymphocyte subpopulations to various stimulants was assessed from the uptake of tritiated thymidine into DNA. Phytohemagglutinin, concanavalin A, pokeweed mitogen, and allogeneic lymphocytes stimulated separated T cells, whereas no proliferation was observed with the T-cell-depleted B-cell population. This suggests that it is the human T cell which is activated directly by these stimulants. In the presence of T cells (proliferating or nonproliferating), B cells were capable of proliferation following stimulation with phytomitogens, but not in response to histocompatibility antigens. Thus, T-cell-mediated B-cell proliferation contributes to the overall lymphocyte response in phytomitogen-stimulated T + B cell mixtures, but not in human mixed leukocyte cultures. T-cell activation by allogeneic cells required the presence of monocytes; in contrast, the three tested phytomitogens stimulated T cells in the absence of monocytes. This indicates that direct interaction of mitogens with lymphocyte membrane receptors is sufficient to trigger T cells into proliferative response. However, monocytes considerably enhanced the proliferative response of T cells in a dose-dependent fashion; this monocyte-dependent mechanism of T-cell activation was predominant at lower concentrations of phytomitogens, and contributed relatively less at higher mitogen doses. Both, the direct, monocyte-independent, and the indirect, monocyte-dependent T-lymphocyte activation contribute to the total in vitro response of lymphocyte preparations to phytomitogens.

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Selected References

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  1. ASKONAS B. A., RHODES J. M. IMMUNOGENICITY OF ANTIGEN-CONTAINING RIBONUCLEIC ACID PREPARATIONS FROM MACROPHAGES. Nature. 1965 Jan 30;205:470–474. doi: 10.1038/205470a0. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Alter B. J., Bach F. H. Lymphocyte reactivity in vitro. I. Cellular reconstitution of purified lymphocyte response. Cell Immunol. 1970 Jul;1(2):207–218. doi: 10.1016/0008-8749(70)90008-0. [DOI] [PubMed] [Google Scholar]
  4. Andersson J., Edelman G. M., Möller G., Sjöberg O. Activation of B lymphocytes by locally concentrated concanavalin A. Eur J Immunol. 1972 Jun;2(3):233–235. doi: 10.1002/eji.1830020307. [DOI] [PubMed] [Google Scholar]
  5. Andersson J., Möller G., Sjöberg O. B lymphocytes can be stimulated by concanavalin A in the presence of humoral factors released by T cells. Eur J Immunol. 1972 Feb;2(1):99–101. doi: 10.1002/eji.1830020119. [DOI] [PubMed] [Google Scholar]
  6. Andersson J., Möller G., Sjöberg O. Selective induction of DNA synthesis in T and B lymphocytes. Cell Immunol. 1972 Aug;4(4):381–393. doi: 10.1016/0008-8749(72)90040-8. [DOI] [PubMed] [Google Scholar]
  7. Andersson L. C., Nordling S., Häyry P. Proliferation of B and T cells in mixed lymphocyte cultures. J Exp Med. 1973 Jul 1;138(1):324–329. doi: 10.1084/jem.138.1.324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Babianskas M. A., Kotkes F. N., Moshkov S. N. Blastransformatsiia limfotsitov, indutsirovannaia monosloem makrofagov, obrabotannykh razlichnymi mitogenami. Biull Eksp Biol Med. 1972 Sep;73(9):79–82. [PubMed] [Google Scholar]
  9. COHN Z. A., BENSON B. THE DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. MORPHOLOGY, CYTOCHEMISTRY, AND BIOCHEMISTRY. J Exp Med. 1965 Jan 1;121:153–170. doi: 10.1084/jem.121.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Elfenbein G. J., Harrison M. R., Green I. Demonstration of proliferation by bone marrow-derived lymphocytes of guinea pigs, mice and rabbits in response to mitogen stimulation in vitro. J Immunol. 1973 May;110(5):1334–1339. [PubMed] [Google Scholar]
  12. Folch H., Yoshinaga M., Waksman B. H. Regulation of lymphocyte responses in vitro. 3. Inhibition by adherent cells of the T-lymphocyte response to phytohemagglutinin. J Immunol. 1973 Mar;110(3):835–839. [PubMed] [Google Scholar]
  13. Gery I., Krüger J., Spiesel S. Z. Stimulation of B-lymphocytes by endotoxin. Reactions of thymus-deprived mice and karyotypic analysis of dividing cells in mice bearing T 6 T 6 thymus grafts. J Immunol. 1972 Apr;108(4):1088–1091. [PubMed] [Google Scholar]
  14. Gery I., Waksman B. H. Potentiation of the T-lymphocyte response to mitogens. II. The cellular source of potentiating mediator(s). J Exp Med. 1972 Jul 1;136(1):143–155. doi: 10.1084/jem.136.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gordon J. Role of monocytes in the mixed leukocyte culture reaction. Proc Soc Exp Biol Med. 1968 Jan;127(1):30–33. doi: 10.3181/00379727-127-32613. [DOI] [PubMed] [Google Scholar]
  16. Greaves M. F., Bauminger S. Activation of T and B lymphocytes by insoluble phytomitogens. Nat New Biol. 1972 Jan 19;235(55):67–70. doi: 10.1038/newbio235067a0. [DOI] [PubMed] [Google Scholar]
  17. Greaves M. F., Bauminger S., Janossy G. Lymphocyte activation. 3. Binding sites for phytomitogens on lymphocyte subpopulations. Clin Exp Immunol. 1972 Mar;10(3):537–554. [PMC free article] [PubMed] [Google Scholar]
  18. Greaves M. F., Brown G. Purification of human T and B lymphocytes. J Immunol. 1974 Jan;112(1):420–423. [PubMed] [Google Scholar]
  19. Hersh E. M., Harris J. E. Macrophage-lymphocyte interaction in the antigen-induced blastogenic response of human peripheral blood leukocytes. J Immunol. 1968 Jun;100(6):1184–1194. [PubMed] [Google Scholar]
  20. Janossy G., Greaves M. F. Lymphocyte activation. I. Response of T and B lymphocytes to phytomitogens. Clin Exp Immunol. 1971 Oct;9(4):483–498. [PMC free article] [PubMed] [Google Scholar]
  21. Janossy G., Greaves M. F. Lymphocyte activation. II. discriminating stimulation of lymphocyte subpopulations by phytomitogens and heterologous antilymphocyte sera. Clin Exp Immunol. 1972 Mar;10(3):525–536. [PMC free article] [PubMed] [Google Scholar]
  22. Johnston J. M., Wilson D. B. Origin of immunoreactive lymphocytes in rats. Cell Immunol. 1970 Oct;1(4):430–444. doi: 10.1016/0008-8749(70)90019-5. [DOI] [PubMed] [Google Scholar]
  23. Jondal M., Holm G., Wigzell H. Surface markers on human T and B lymphocytes. I. A large population of lymphocytes forming nonimmune rosettes with sheep red blood cells. J Exp Med. 1972 Aug 1;136(2):207–215. doi: 10.1084/jem.136.2.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jones G. Lymphocyte activation. I. Expression of theta, H-2 and immunoglobulin determinants on lymphocytes stimulated by phytohaemagglutinin, pokeweed mitogen, concanavalin A or histocompatibility antigen. Clin Exp Immunol. 1972 Nov;12(3):391–402. [PMC free article] [PubMed] [Google Scholar]
  25. Jones G. Reversible binding of phytomitogens to lymphocytes. Cell Immunol. 1973 Dec;9(3):393–404. doi: 10.1016/0008-8749(73)90054-3. [DOI] [PubMed] [Google Scholar]
  26. Jones G. The number of reactive cells in mouse lymphocyte cultures stimulated by phytohemagglutinin, concanavalin A or histocompatibility antigen. J Immunol. 1973 Sep;111(3):914–920. [PubMed] [Google Scholar]
  27. Lay W. H., Mendes N. F., Bianco C., Nussenzweig V. Binding of sheep red blood cells to a large population of human lymphocytes. Nature. 1971 Apr 23;230(5295):531–532. doi: 10.1038/230531a0. [DOI] [PubMed] [Google Scholar]
  28. Levis W. R., Robbins J. H. Effect of glass-adherent cells on the blastogenic response of 'purified' lymphocytes to phytohemagglutinin. Exp Cell Res. 1970 Jul;61(1):153–158. doi: 10.1016/0014-4827(70)90269-7. [DOI] [PubMed] [Google Scholar]
  29. Lohrmann H. P., Graw C. M., Graw R. G., Jr Stimulated lymphocyte cultures: responder recruitment and cell cycle kinetics. J Exp Med. 1974 May 1;139(5):1037–1048. doi: 10.1084/jem.139.5.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mackler B. F., Amkraut A. A., Malley A. Cellular receptors for PHA and antigen-induced transformation of peripheral blood and thoracic duct lymphocytes. Cell Immunol. 1972 Jan;3(1):138–143. doi: 10.1016/0008-8749(72)90234-1. [DOI] [PubMed] [Google Scholar]
  31. Meuwissen H. J., Van Alten P. A., Good R. A. Decreased lymphoid cell multiplication in the post-thymectomy state. Transplantation. 1969 Jan;7(1):1–11. doi: 10.1097/00007890-196901000-00001. [DOI] [PubMed] [Google Scholar]
  32. Mosier D. E. Cell interactions in the primary immune response in vitro: a requirement for specific cell clusters. J Exp Med. 1969 Feb 1;129(2):351–362. doi: 10.1084/jem.129.2.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Oppenheim J. J., Leventhal B. G., Hersh E. M. The transformation of column-purified lymphocytes with nonspecific and specific antigenic stimuli. J Immunol. 1968 Aug;101(2):262–267. [PubMed] [Google Scholar]
  34. Plate J. M., McKenzie I. F. "B"-cell stimulation of allogeneic T-cell proliferation in mixed lymphocyte cultures. Nat New Biol. 1973 Oct 24;245(147):247–249. doi: 10.1038/newbio245247a0. [DOI] [PubMed] [Google Scholar]
  35. Ramseier H. T cells in recognition of histocompatibility antigens. Nature. 1973 Dec 7;246(5432):351–352. doi: 10.1038/246351a0. [DOI] [PubMed] [Google Scholar]
  36. Rode H. N., Gordon J. The mixed leukocyte culture: a three component system. J Immunol. 1970 Jun;104(6):1453–1457. [PubMed] [Google Scholar]
  37. Schiff R. I., Buckley R. H., Gilbertsen R. B., Metzgar R. S. Membrane receptors and in vitro responsiveness of lymphocytes in human immunodeficiency. J Immunol. 1974 Jan;112(1):376–386. [PubMed] [Google Scholar]
  38. Schwarz M. R. Transformation of rat small lymphocytes with pokeweed mitogen (PWM). Anat Rec. 1968 Jan;160(1):47–58. doi: 10.1002/ar.1091600106. [DOI] [PubMed] [Google Scholar]
  39. Seeger R. C., Oppenheim J. J. Synergistic interaction of macrophages and lymphocytes in antigen-induced transformation of lymphocytes. J Exp Med. 1970 Jul 1;132(1):44–65. doi: 10.1084/jem.132.1.44. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Stobo J. D., Rosenthal A. S., Paul W. E. Functional heterogeneity of murine lymphoid cells. I. Responsiveness to and surface binding of concanavalin A and phytohemagglutinin. J Immunol. 1972 Jan;108(1):1–17. [PubMed] [Google Scholar]
  41. Twomey J. J., Sharkey O., Jr, Brown J. A., Laughter A. H., Jordan P. H., Jr Cellular requirements for the mitotic response in allogeneic mixed leukocyte cultures. J Immunol. 1970 Apr;104(4):845–853. [PubMed] [Google Scholar]
  42. Vischer T. L. Immunoglobulin-like surface molecules and theta antigen during the specific and non-specific stimulation of mouse spleen cells in vitro. Clin Exp Immunol. 1972 Aug;11(4):523–534. [PMC free article] [PubMed] [Google Scholar]
  43. WALKER R. I., FOWLER I. GRANULOCYTE INHIBITION OF HUMAN PERIPHERAL BLOOD LYMPHOCYTE GROWTH IN VITRO. Exp Cell Res. 1965 May;38:379–385. doi: 10.1016/0014-4827(65)90411-8. [DOI] [PubMed] [Google Scholar]
  44. Wagner H. The correlation between the proliferative and the cytotoxic responses of mouse lymphocytes to allogeneic cells in vitro. J Immunol. 1972 Sep;109(3):630–637. [PubMed] [Google Scholar]
  45. Weber W. T. Direct evidence for the response of B and T cells to pokeweed mitogen. Cell Immunol. 1973 Dec;9(3):482–487. doi: 10.1016/0008-8749(73)90064-6. [DOI] [PubMed] [Google Scholar]
  46. Weiner M. S., Bianco C., Nussenzweig V. Enhanced binding of neuraminidase-treated sheep erythrocytes to human T lymphocytes. Blood. 1973 Dec;42(6):939–946. [PubMed] [Google Scholar]
  47. Wilson D. B., Blyth JL NOWELL P. C. Quantitative studies on the mixed lymphocyte interaction in rats. 3. Kinetics of the response. J Exp Med. 1968 Nov 1;128(5):1157–1181. doi: 10.1084/jem.128.5.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wybran J., Carr M. C., Fudenberg H. H. The human rosette-forming cell as a marker of a population of thymus-derived cells. J Clin Invest. 1972 Oct;51(10):2537–2543. doi: 10.1172/JCI107069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wybran J., Chantler S., Fudenberg H. H. Isolation of normal T cells in chronic lymphatic leukaemia. Lancet. 1973 Jan 20;1(7795):126–129. doi: 10.1016/s0140-6736(73)90196-7. [DOI] [PubMed] [Google Scholar]

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