Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1992 May 1;175(5):1255–1269. doi: 10.1084/jem.175.5.1255

The extent of clonal structure in different lymphoid organs

PMCID: PMC2119216  PMID: 1569396

Abstract

To gain insight into the clonal organization of lymphoid organs, we studied the distribution in situ of donor-derived cells in near- physiological chimeras. We introduced RT7b fetal liver cells into nonirradiated congenic RT7a neonatal rats. The chimerism 6-20 wk after injection ranged from 0.3 to 20%. The numbers of cell clones simultaneously contributing to cell generation in a particular histological feature were deduced from the variance in donor cell distribution. In bone marrow and thymus, donor-derived lymphoid cells were found scattered among host cells, indicating a high mobility of cells. In bone marrow, donor cells were evenly distributed over the entire marrow, even at low chimerism. This indicates that leukopoiesis is maintained by the proliferation of many clones. In the thymus, the various lobules showed different quantities of donor-derived lymphoid cells. Mathematical analysis of these differences indicated that 17-18 cell division cycles occur in the cortex. In spleen, the distribution of donor-derived cells over the germinal centers indicated that 5 d after antigenic stimulation, germinal centers develop oligoclonally. The main conclusions of this work are that (a) bone marrow and thymus are highly polyclonal; (b) 17-18 divisions occur between prothymocyte and mature T cell; and (c) lymphoid cells disperse rapidly while proliferating and differentiating.

Full Text

The Full Text of this article is available as a PDF (3.1 MB).

Selected References

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

  1. Behringer R. R., Eldridge P. W., Dewey M. J. Stable genotypic composition of blood cells in allophenic mice derived from congenic C57BL/6 strains. Dev Biol. 1984 Jan;101(1):251–256. doi: 10.1016/0012-1606(84)90138-6. [DOI] [PubMed] [Google Scholar]
  2. Bell E. B., Sparshott S. M., Drayson M. T., Hunt S. V. The origin of T cells in permanently reconstituted old athymic nude rats. Analysis using chromosome or allotype markers. Immunology. 1989 Dec;68(4):547–556. [PMC free article] [PubMed] [Google Scholar]
  3. Bell E. B., Sparshott S. M. Interconversion of CD45R subsets of CD4 T cells in vivo. Nature. 1990 Nov 8;348(6297):163–166. doi: 10.1038/348163a0. [DOI] [PubMed] [Google Scholar]
  4. Benoist C., Mathis D. Positive selection of the T cell repertoire: where and when does it occur? Cell. 1989 Sep 22;58(6):1027–1033. doi: 10.1016/0092-8674(89)90501-1. [DOI] [PubMed] [Google Scholar]
  5. Capel B., Hawley R. G., Mintz B. Long- and short-lived murine hematopoietic stem cell clones individually identified with retroviral integration markers. Blood. 1990 Jun 15;75(12):2267–2270. [PubMed] [Google Scholar]
  6. Capel B., Hawley R., Covarrubias L., Hawley T., Mintz B. Clonal contributions of small numbers of retrovirally marked hematopoietic stem cells engrafted in unirradiated neonatal W/Wv mice. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4564–4568. doi: 10.1073/pnas.86.12.4564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deenen G. J., Hunt S. V., Opstelten D. A stathmokinetic study of B lymphocytopoiesis in rat bone marrow: proliferation of cells containing cytoplasmic mu-chains, terminal deoxynucleotidyl transferase and carrying HIS24 antigen. J Immunol. 1987 Aug 1;139(3):702–710. [PubMed] [Google Scholar]
  8. Deenen G. J., Van Balen I., Opstelten D. In rat B lymphocyte genesis sixty percent is lost from the bone marrow at the transition of nondividing pre-B cell to sIgM+ B lymphocyte, the stage of Ig light chain gene expression. Eur J Immunol. 1990 Mar;20(3):557–564. doi: 10.1002/eji.1830200315. [DOI] [PubMed] [Google Scholar]
  9. Dijkstra C. D., Döpp E. A., Joling P., Kraal G. The heterogeneity of mononuclear phagocytes in lymphoid organs: distinct macrophage subpopulations in the rat recognized by monoclonal antibodies ED1, ED2 and ED3. Immunology. 1985 Mar;54(3):589–599. [PMC free article] [PubMed] [Google Scholar]
  10. Ezine S., Weissman I. L., Rouse R. V. Bone marrow cells give rise to distinct cell clones within the thymus. Nature. 1984 Jun 14;309(5969):629–631. doi: 10.1038/309629a0. [DOI] [PubMed] [Google Scholar]
  11. Fink P. J., Bevan M. J., Weissman I. L. Thymic cytotoxic T lymphocytes are primed in vivo to minor histocompatibility antigens. J Exp Med. 1984 Feb 1;159(2):436–451. doi: 10.1084/jem.159.2.436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fleischman R. A., Mintz B. Development of adult bone marrow stem cells in H-2-compatible and -incompatible mouse fetuses. J Exp Med. 1984 Mar 1;159(3):731–745. doi: 10.1084/jem.159.3.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fry A. M., Jones L. A., Kruisbeek A. M., Matis L. A. Thymic requirement for clonal deletion during T cell development. Science. 1989 Nov 24;246(4933):1044–1046. doi: 10.1126/science.2511630. [DOI] [PubMed] [Google Scholar]
  14. Gregoire K. E., Goldschneider I., Barton R. W., Bollum F. J. Ontogeny of terminal deoxynucleotidyl transferase-positive cells in lymphohemopoietic tissues of rat and mouse. J Immunol. 1979 Sep;123(3):1347–1352. [PubMed] [Google Scholar]
  15. Guidos C. J., Danska J. S., Fathman C. G., Weissman I. L. T cell receptor-mediated negative selection of autoreactive T lymphocyte precursors occurs after commitment to the CD4 or CD8 lineages. J Exp Med. 1990 Sep 1;172(3):835–845. doi: 10.1084/jem.172.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gutman G. A., Weissman I. L. Lymphoid tissue architecture. Experimental analysis of the origin and distribution of T-cells and B-cells. Immunology. 1972 Oct;23(4):465–479. [PMC free article] [PubMed] [Google Scholar]
  17. HARRIS J. E., FORD C. E., BARNES D. W., EVANS E. P. EVIDENCE FROM PARABIOSIS FOR AN AFFERENT STREAM OF CELLS. Nature. 1964 Feb 29;201:886–887. doi: 10.1038/201886a0. [DOI] [PubMed] [Google Scholar]
  18. Harrison D. E., Lerner C., Hoppe P. C., Carlson G. A., Alling D. Large numbers of primitive stem cells are active simultaneously in aggregated embryo chimeric mice. Blood. 1987 Mar;69(3):773–777. [PubMed] [Google Scholar]
  19. Hengartner H., Odermatt B., Schneider R., Schreyer M., Wälle G., MacDonald H. R., Zinkernagel R. M. Deletion of self-reactive T cells before entry into the thymus medulla. Nature. 1988 Nov 24;336(6197):388–390. doi: 10.1038/336388a0. [DOI] [PubMed] [Google Scholar]
  20. Hermans M. H., Hartsuiker H., Opstelten D. An in situ study of B-lymphocytopoiesis in rat bone marrow. Topographical arrangement of terminal deoxynucleotidyl transferase-positive cells and pre-B cells. J Immunol. 1989 Jan 1;142(1):67–73. [PubMed] [Google Scholar]
  21. Hermans M. H., Opstelten D. In situ visualization of hemopoietic cell subsets and stromal elements in rat and mouse bone marrow by immunostaining of frozen sections. J Histochem Cytochem. 1991 Dec;39(12):1627–1634. doi: 10.1177/39.12.1940317. [DOI] [PubMed] [Google Scholar]
  22. Hirokawa K., Utsuyama M., Sado T. Immunohistological analysis of immigration of thymocyte-precursors into the thymus: evidence for immigration of peripheral T cells into the thymic medulla. Cell Immunol. 1989 Mar;119(1):160–170. doi: 10.1016/0008-8749(89)90232-3. [DOI] [PubMed] [Google Scholar]
  23. Hunt S. V., Fowler M. H. A repopulation assay for B and T lymphocyte stem cells employing radiation chimaeras. Cell Tissue Kinet. 1981 Jul;14(4):445–464. doi: 10.1111/j.1365-2184.1981.tb00551.x. [DOI] [PubMed] [Google Scholar]
  24. Hwang W. S., Ho T. Y., Luk S. C., Simon G. T. Ultrastructure of the rat thymus. A transmission, scanning electron microscope, and morphometric study. Lab Invest. 1974 Nov;31(5):473–487. [PubMed] [Google Scholar]
  25. Jacob J., Kassir R., Kelsoe G. In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. I. The architecture and dynamics of responding cell populations. J Exp Med. 1991 May 1;173(5):1165–1175. doi: 10.1084/jem.173.5.1165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Jacobsen K., Osmond D. G. Microenvironmental organization and stromal cell associations of B lymphocyte precursor cells in mouse bone marrow. Eur J Immunol. 1990 Nov;20(11):2395–2404. doi: 10.1002/eji.1830201106. [DOI] [PubMed] [Google Scholar]
  27. Jordan C. T., Lemischka I. R. Clonal and systemic analysis of long-term hematopoiesis in the mouse. Genes Dev. 1990 Feb;4(2):220–232. doi: 10.1101/gad.4.2.220. [DOI] [PubMed] [Google Scholar]
  28. Jordan C. T., McKearn J. P., Lemischka I. R. Cellular and developmental properties of fetal hematopoietic stem cells. Cell. 1990 Jun 15;61(6):953–963. doi: 10.1016/0092-8674(90)90061-i. [DOI] [PubMed] [Google Scholar]
  29. Kampinga J., Kroese F. G., Pol G. H., Opstelten D., Seijen H. G., Boot J. H., Roser B., Nieuwenhuis P., Aspinall R. RT7-defined alloantigens in rats are part of the leucocyte common antigen family. Scand J Immunol. 1990 Jun;31(6):699–710. doi: 10.1111/j.1365-3083.1990.tb02821.x. [DOI] [PubMed] [Google Scholar]
  30. Keller G., Paige C., Gilboa E., Wagner E. F. Expression of a foreign gene in myeloid and lymphoid cells derived from multipotent haematopoietic precursors. Nature. 1985 Nov 14;318(6042):149–154. doi: 10.1038/318149a0. [DOI] [PubMed] [Google Scholar]
  31. Keller G., Snodgrass R. Life span of multipotential hematopoietic stem cells in vivo. J Exp Med. 1990 May 1;171(5):1407–1418. doi: 10.1084/jem.171.5.1407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kincade P. W., Lee G., Pietrangeli C. E., Hayashi S., Gimble J. M. Cells and molecules that regulate B lymphopoiesis in bone marrow. Annu Rev Immunol. 1989;7:111–143. doi: 10.1146/annurev.iy.07.040189.000551. [DOI] [PubMed] [Google Scholar]
  33. Kroese F. G., Opstelten D., Wubbena A. S., Deenen G. J., Aten J., Schwander E. H., de Leij L., Nieuwenhuis P. Monoclonal antibodies to rat B lymphocyte (sub-)populations. Adv Exp Med Biol. 1985;186:81–89. doi: 10.1007/978-1-4613-2463-8_10. [DOI] [PubMed] [Google Scholar]
  34. Kroese F. G., Wubbena A. S., Joling P., Nieuwenhuis P. T-lymphocytes in rat lymphoid follicles are a subset of T helper cells. Adv Exp Med Biol. 1985;186:443–449. doi: 10.1007/978-1-4613-2463-8_54. [DOI] [PubMed] [Google Scholar]
  35. Kroese F. G., Wubbena A. S., Opstelten D., Deenen G. J., Schwander E. H., De Leij L., Vos H., Poppema S., Volberda J., Nieuwenhuis P. B lymphocyte differentiation in the rat: production and characterization of monoclonal antibodies to B lineage-associated antigens. Eur J Immunol. 1987 Jul;17(7):921–928. doi: 10.1002/eji.1830170705. [DOI] [PubMed] [Google Scholar]
  36. Kroese F. G., Wubbena A. S., Seijen H. G., Nieuwenhuis P. Germinal centers develop oligoclonally. Eur J Immunol. 1987 Jul;17(7):1069–1072. doi: 10.1002/eji.1830170726. [DOI] [PubMed] [Google Scholar]
  37. Kyewski B. A. Seeding of thymic microenvironments defined by distinct thymocyte-stromal cell interactions is developmentally controlled. J Exp Med. 1987 Aug 1;166(2):520–538. doi: 10.1084/jem.166.2.520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Lemischka I. R., Raulet D. H., Mulligan R. C. Developmental potential and dynamic behavior of hematopoietic stem cells. Cell. 1986 Jun 20;45(6):917–927. doi: 10.1016/0092-8674(86)90566-0. [DOI] [PubMed] [Google Scholar]
  39. Lubaroff D. M. An alloantigenic marker on rat thymus and thymus-derived cells. Transplant Proc. 1973 Mar;5(1):115–118. [PubMed] [Google Scholar]
  40. MacLennan I. C., Liu Y. J., Oldfield S., Zhang J., Lane P. J. The evolution of B-cell clones. Curr Top Microbiol Immunol. 1990;159:37–63. doi: 10.1007/978-3-642-75244-5_3. [DOI] [PubMed] [Google Scholar]
  41. Micklem H. S., Ford C. E., Evans E. P., Ogden D. A., Papworth D. S. Competitive in vivo proliferation of foetal and adult haematopoietic cells in lethally irradiated mice. J Cell Physiol. 1972 Apr;79(2):293–298. doi: 10.1002/jcp.1040790214. [DOI] [PubMed] [Google Scholar]
  42. Micklem H. S., Lennon J. E., Ansell J. D., Gray R. A. Numbers and dispersion of repopulating hematopoietic cell clones in radiation chimeras as functions of injected cell dose. Exp Hematol. 1987 Mar;15(3):251–257. [PubMed] [Google Scholar]
  43. Moore M. A., Owen J. J. Experimental studies on the development of the thymus. J Exp Med. 1967 Oct 1;126(4):715–726. doi: 10.1084/jem.126.4.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Opstelten D., Deenen G. J., Rozing J., Hunt S. V. B lymphocyte-associated antigens on terminal deoxynucleotidyl transferase-positive cells and pre-B cells in bone marrow of the rat. J Immunol. 1986 Jul 1;137(1):76–84. [PubMed] [Google Scholar]
  45. Paterson D. J., Green J. R., Jefferies W. A., Puklavec M., Williams A. F. The MRC OX-44 antigen marks a functionally relevant subset among rat thymocytes. J Exp Med. 1987 Jan 1;165(1):1–13. doi: 10.1084/jem.165.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Pellas T. C., Weiss L. Migration pathways of recirculating murine B cells and CD4+ and CD8+ T lymphocytes. Am J Anat. 1990 Apr;187(4):355–373. doi: 10.1002/aja.1001870405. [DOI] [PubMed] [Google Scholar]
  47. Penit C. In vivo thymocyte maturation. BUdR labeling of cycling thymocytes and phenotypic analysis of their progeny support the single lineage model. J Immunol. 1986 Oct 1;137(7):2115–2121. [PubMed] [Google Scholar]
  48. Rossant J., Vijh K. M., Grossi C. E., Cooper M. D. Clonal origin of haematopoietic colonies in the postnatal mouse liver. Nature. 1986 Feb 6;319(6053):507–511. doi: 10.1038/319507a0. [DOI] [PubMed] [Google Scholar]
  49. Sainte-Marie G. Tridimensional reconstruction of the rat thymus. Anat Rec. 1974 Aug;179(4):517–526. doi: 10.1002/ar.1091790412. [DOI] [PubMed] [Google Scholar]
  50. Schwartz R. H. Acquisition of immunologic self-tolerance. Cell. 1989 Jun 30;57(7):1073–1081. doi: 10.1016/0092-8674(89)90044-5. [DOI] [PubMed] [Google Scholar]
  51. Scollay R. G., Butcher E. C., Weissman I. L. Thymus cell migration. Quantitative aspects of cellular traffic from the thymus to the periphery in mice. Eur J Immunol. 1980 Mar;10(3):210–218. doi: 10.1002/eji.1830100310. [DOI] [PubMed] [Google Scholar]
  52. Shortman K., Egerton M., Spangrude G. J., Scollay R. The generation and fate of thymocytes. Semin Immunol. 1990 Jan;2(1):3–12. [PubMed] [Google Scholar]
  53. Shortman K., Vremec D., Egerton M. The kinetics of T cell antigen receptor expression by subgroups of CD4+8+ thymocytes: delineation of CD4+8+3(2+) thymocytes as post-selection intermediates leading to mature T cells. J Exp Med. 1991 Feb 1;173(2):323–332. doi: 10.1084/jem.173.2.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Smith L. G., Weissman I. L., Heimfeld S. Clonal analysis of hematopoietic stem-cell differentiation in vivo. Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2788–2792. doi: 10.1073/pnas.88.7.2788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Sunderland C. A., McMaster W. R., Williams A. F. Purification with monoclonal antibody of a predominant leukocyte-common antigen and glycoprotein from rat thymocytes. Eur J Immunol. 1979 Feb;9(2):155–159. doi: 10.1002/eji.1830090212. [DOI] [PubMed] [Google Scholar]
  56. Thomas D. B., Smith C. M., Sumpster J. M. The proliferation of transplanted haematopoietic cells derived from bone marrow and fetal liver. J Anat. 1976 Sep;122(Pt 1):15–22. [PMC free article] [PubMed] [Google Scholar]
  57. Verfaillie C. M., McCarthy J. B., McGlave P. B. Differentiation of primitive human multipotent hematopoietic progenitors into single lineage clonogenic progenitors is accompanied by alterations in their interaction with fibronectin. J Exp Med. 1991 Sep 1;174(3):693–703. doi: 10.1084/jem.174.3.693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Weinberg W. C., Howard J. C., Iannaccone P. M. Histological demonstration of mosaicism in a series of chimeric rats produced between congenic strains. Science. 1985 Feb 1;227(4686):524–527. doi: 10.1126/science.3966159. [DOI] [PubMed] [Google Scholar]
  59. Wekerle H., Ketelsen U. P., Ernst M. Thymic nurse cells. Lymphoepithelial cell complexes in murine thymuses: morphological and serological characterization. J Exp Med. 1980 Apr 1;151(4):925–944. doi: 10.1084/jem.151.4.925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Westermann J., Ronneberg S., Fritz F. J., Pabst R. Proliferation of lymphocyte subsets in the adult rat: a comparison of different lymphoid organs. Eur J Immunol. 1989 Jun;19(6):1087–1093. doi: 10.1002/eji.1830190619. [DOI] [PubMed] [Google Scholar]
  61. Williams A. F., Galfrè G., Milstein C. Analysis of cell surfaces by xenogeneic myeloma-hybrid antibodies: differentiation antigens of rat lymphocytes. Cell. 1977 Nov;12(3):663–673. doi: 10.1016/0092-8674(77)90266-5. [DOI] [PubMed] [Google Scholar]
  62. van Ewijk W., van Soest P. L., van den Engh G. J. Fluorescence analysis and anatomic distribution of mouse T lymphocyte subsets defined by monoclonal antibodies to the antigens Thy-1, Lyt-1, Lyt-2, and T-200. J Immunol. 1981 Dec;127(6):2594–2604. [PubMed] [Google Scholar]
  63. von Boehmer H. Developmental biology of T cells in T cell-receptor transgenic mice. Annu Rev Immunol. 1990;8:531–556. doi: 10.1146/annurev.iy.08.040190.002531. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES