Abstract
The relationship of natural killer (NK) cells to T cells has been a longstanding enigma. It is well established that development of NK cells does not require a thymus (because they are present in athymic mice), recombination activating genes (because they are present in RAG-1 and RAG-2 knockout mice) or DNA-dependent protein kinase (because they are present in severe combined immunodeficiency [SCID] mice) (1,2). NK cells do not express CD3 or TCR on their surface and they are classically defined as CD3–CD56+ (in humans) or CD3–NK1.1+ (in mice) (3–5). However, there has been some suggestion that NK cells might share a common progenitor with T cells. This has been derived from the observations that some NK cells contain a truncated mRNA for TCR-β (4), that activated NK contain cytoplasmic CD3ε and that fetal NK cells also contain cytoplasmic CD3ε (3–7). Several studies demonstrated that fetal thymi contain progenitors that might develop into T cells or NK cells depending on whether they mature within a thymic or extrathymic microenvironment, respectively (7). The discovery of a novel lymphocyte subset that expresses markers for both T cells and NK cells, the so-called T/NK lymphocytes, raised further questions about the ontogenic relationship of NK to T cells (8,9). More recently, Sanchez et al. (7) showed that human fetal thymi contain a bipotential progenitor that could develop along either of the T or NK maturation pathways. This led us and others to determine whether NK cells can be demonstrated and cloned from fetal thymi (10,11). Surprisingly, we found that NK1.1 is among the earliest lymphohematopoietic genes to be transcribed as its mRNA is demonstrable in d 9 fetuses (exact organ distribution not yet determined) and in the earliest of thymic anlages (d 11 of gestation) (11). Although T/NK cells can be demonstrated in fetal thymic organ cultures (FTOC) established from day 14 fetal thy mi, the majority of NK1.1+ cells obtained from fetal thymi appear to be the classical, CD3– NK1.1+ cells (10,11).
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References
Dorshkind K., Pollack S. B., Bosma M. J., and Phillips R. A. (1985) Natural killer (NK) cells are present in mice with severe combined immunodeficiency (scid). J. Immunol. 134, 3798–3801.
Mombaerts P., Iacomini J., Johnson R. S., Herrup K., Tonegawa S., and Papaioannou V. E. (1992) RAG-1-deficient mice have no mature B and T lymphocytes. Cell 68, 869–877.
Whiteside T. L. and Herberman R. B. (1994) Role of human natural killer cells in health and disease. Clin. Diag. Lab. Immunol. 1, 125–133.
Lanier L. L., Phillips J. H., Hackett Jr J., Tutt M., Kumar V., and Terhorst C. (1986) Natural killer cells: definition of a cell type rather than a function. J. Immunol. 137, 2735–2739.
Trinchieri G. (1989) Biology of natural killer cells. Adv. Immunol. 47, 187–376.
Lanier L. L., Chang C., Spits H., and Phillips J. H. (1992) Expression of cytoplasmic CD3 epsilon proteins in activated human adult natural killer (NK) cells and CD3 gamma, delta, epsilon complexes in fetal NK cells. Implications for the relationship of NK and T lymphocytes. J. Immunol. 149, 1876–1880.
Sanchez M. J., Muench M. O., Roncarolo M. G., Lanier L. L., and Phillips J. H. (1994) Identification of a common T/natural killer cell progenitor in human fetal thymus. J. Exp. Med. 180, 569–576.
Bendelac A., Rivera M. N., Park S. H., and Roark J. H. (1997) Mouse CD1-specific NK 1 T cells: development, specificity and function. Ann. Rev. Immunol. 15, 535–562.
Ballas Z. K. and Rasmussen W. (1990) NK1. 1+ thymocytes: adult murine CD4- CD8- thymocytes contain an NK1. 1+, CD3+, CD5 high, CD44 high, TCR-Vb8+ subset. J. Immunol. 145, 1039–1045.
Brooks C. G., Georgiou A., and Jordan R. K. (1993) The majority of immature fetal thymocytes can be induced to proliferate to IL-2 and differentiate into cells indistinguishable from mature natural killer cells. J. Immunol. 151, 6645–6656.
Ballas, Z. K., Rasmussen W. L., Alber C. A., and Sandor M. (1997) Ontogeny of Thymic NK1. 1+ cells. J. Immunol. 159, 1174–1181.
Robey E. and Fowlkes B. J. (1994) Selective events in T cell development. Ann. Rev. Immunol. 12, 675–705.
Spits H., Lanier L. L., and Phillips J. H. (1995) Development of human T and natural killer cells. Blood 85, 2654–2670.
Rodewald H. R., Moingeon P., Lucich J. L., Dosiou C., Lopes P., and Reinherz E. L. (1992) A population of early fetal thymocytes expressing FcγRII/III contains precursors of T lymphocytes and natural killer cells. Cell 69, 139–150.
Jenkinson E. J. and Owen J. J. (1990) T cell differentiation in thymus organ culture. Semin. Immunol. 2, 51–58.
Carlyle J. R., Michie A. M., Furlonger C., Nakano T., Lenardo M. J., Paige C. J., and Zuniga-Pflucker J. C. (1997) Identification of a novel developmental stage marking lineage commitment of progenitor thymocytes. J. Exp. Med. 186, 173–182.
Ernst B., Surh C. D., and Sprent J. (1995) Thymic selection and cell division. J. Exp. Med. 182, 961–971.
Kumamoto T., Inaba M., Toki J., Adachi Y., Imamura H., and Ikehara S. (1995) Cytotoxic effects of irradiation and deoxyguanosine on fetal thymus. Immunobiology 192, 365–381.
Kingston R., Jenkinson E. J., and Owen J. J. (1985) A single stem cell can recolonize an embryonic thymus, producing phenotypically distinct T cell populations. Nature 317, 811–813.
Puzanov I. J., Bennett M., and Kumar V. (1996) IL-15 can substitute for the marrow microenvironment in the differentiation of natural killer cells. J. Immunol. 157, 4282–4285.
Williams N. S., Moore T. A., Schatzle J. D., Puzanov I. J., Sivakumar P. V., Zlotnik A., Bennett M., and Kumar V. (1997) Generation of lytic natural killer 1. 1+, Ly49- cells from multipotential murine bone marrow progenitors in a stroma-free culture: definition of cytokine requirements and developmental intermediates. J. Exp. Med. 186, 1609–1614
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Ballas, Z.K., Rasmussen, W.L. (1999). Murine Natural Killer Cell Cloning from Fetal Thymic Organ Cultures. In: Campbell, K.S., Colonna, M. (eds) Natural Killer Cell Protocols. Methods in Molecular Biology, vol 121. Humana Press. https://doi.org/10.1385/1-59259-044-6:39
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DOI: https://doi.org/10.1385/1-59259-044-6:39
Publisher Name: Humana Press
Print ISBN: 978-0-89603-683-3
Online ISBN: 978-1-59259-044-5
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