Abstract
Recent studies have revealed that 10-20% of CD4+8- or CD4-8- thymocyte populations contain NK1.1+ T cell receptor (TCR)-alpha/beta+ cells. This subpopulation shows characteristics that are different from NK1.1- CD4+ or NK1.1- CD8+ T cells and seems to have developed in a manner different from NK1.1- T cells. Although extensive studies have been performed on the NK1.1+ TCR-alpha/beta+ thymocytes, the physiological role of the NK1.1+ TCR-alpha/beta+ thymocytes has been totally unclear. In the present study, we found that freshly isolated NK1.1+ TCR- alpha/beta+ thymocytes, but neither whole thymocytes nor lymph node T cells, directly killed CD4+8+ thymocytes from normal syngeneic or allogeneic mice by using a long-term cytotoxic assay in which flow cytometry was used to detect the cytotoxicity. However, only weak cytotoxicity was detected against thymocytes from lpr mice on which the Fas antigen that transduces signals for apoptosis into the cells is not expressed. Furthermore, the NK1.1+ TCR-alpha/beta+ thymocytes exhibited high cytotoxicity against T lymphoma targets transfected with fas genes as compared with the parental T lymphoma targets or target cells transfected with mutated fas genes, which lack the function of transducing signals. On the other hand, NK1.1+ effector thymocytes from gld mice that carry a point mutation in Fas ligand did not kill thymocyte targets from normal mice. The present findings, thus, consistently suggest that the NK1.1+ TCR-alpha/beta+ thymocytes kill a subpopulation among CD4+8+ thymocytes via Fas antigen and in this way regulate generation of T lineage cells in the thymus.
Full Text
The Full Text of this article is available as a PDF (1.0 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adachi M., Watanabe-Fukunaga R., Nagata S. Aberrant transcription caused by the insertion of an early transposable element in an intron of the Fas antigen gene of lpr mice. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1756–1760. doi: 10.1073/pnas.90.5.1756. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Agus D. B., Surh C. D., Sprent J. Reentry of T cells to the adult thymus is restricted to activated T cells. J Exp Med. 1991 May 1;173(5):1039–1046. doi: 10.1084/jem.173.5.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Allen R. D., Marshall J. D., Roths J. B., Sidman C. L. Differences defined by bone marrow transplantation suggest that lpr and gld are mutations of genes encoding an interacting pair of molecules. J Exp Med. 1990 Nov 1;172(5):1367–1375. doi: 10.1084/jem.172.5.1367. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Arase H., Arase-Fukushi N., Good R. A., Onoé K. Lymphokine-activated killer cell activity of CD4-CD8- TCR alpha beta + thymocytes. J Immunol. 1993 Jul 15;151(2):546–555. [PubMed] [Google Scholar]
- Arase H., Arase N., Nakagawa K., Good R. A., Onoé K. NK1.1+ CD4+ CD8- thymocytes with specific lymphokine secretion. Eur J Immunol. 1993 Jan;23(1):307–310. doi: 10.1002/eji.1830230151. [DOI] [PubMed] [Google Scholar]
- Arase H., Arase N., Ogasawara K., Good R. A., Onoé K. An NK1.1+ CD4+8- single-positive thymocyte subpopulation that expresses a highly skewed T-cell antigen receptor V beta family. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6506–6510. doi: 10.1073/pnas.89.14.6506. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ballas Z. K., Rasmussen W. NK1.1+ thymocytes. Adult murine CD4-, CD8- thymocytes contain an NK1.1+, CD3+, CD5hi, CD44hi, TCR-V beta 8+ subset. J Immunol. 1990 Aug 15;145(4):1039–1045. [PubMed] [Google Scholar]
- Bix M., Coles M., Raulet D. Positive selection of V beta 8+ CD4-8- thymocytes by class I molecules expressed by hematopoietic cells. J Exp Med. 1993 Sep 1;178(3):901–908. doi: 10.1084/jem.178.3.901. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bruce J., Symington F. W., McKearn T. J., Sprent J. A monoclonal antibody discriminating between subsets of T and B cells. J Immunol. 1981 Dec;127(6):2496–2501. [PubMed] [Google Scholar]
- Budd R. C., Miescher G. C., Howe R. C., Lees R. K., Bron C., MacDonald H. R. Developmentally regulated expression of T cell receptor beta chain variable domains in immature thymocytes. J Exp Med. 1987 Aug 1;166(2):577–582. doi: 10.1084/jem.166.2.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ceredig R., Lynch F., Newman P. Phenotypic properties, interleukin 2 production, and developmental origin of a "mature" subpopulation of Lyt-2- L3T4- mouse thymocytes. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8578–8582. doi: 10.1073/pnas.84.23.8578. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chu J. L., Drappa J., Parnassa A., Elkon K. B. The defect in Fas mRNA expression in MRL/lpr mice is associated with insertion of the retrotransposon, ETn. J Exp Med. 1993 Aug 1;178(2):723–730. doi: 10.1084/jem.178.2.723. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cohen P. L., Eisenberg R. A. Lpr and gld: single gene models of systemic autoimmunity and lymphoproliferative disease. Annu Rev Immunol. 1991;9:243–269. doi: 10.1146/annurev.iy.09.040191.001331. [DOI] [PubMed] [Google Scholar]
- Cohen P. L., Eisenberg R. A. The lpr and gld genes in systemic autoimmunity: life and death in the Fas lane. Immunol Today. 1992 Nov;13(11):427–428. doi: 10.1016/0167-5699(92)90066-G. [DOI] [PubMed] [Google Scholar]
- Davidson W. F., Holmes K. L., Roths J. B., Morse H. C., 3rd Immunologic abnormalities of mice bearing the gld mutation suggest a common pathway for murine nonmalignant lymphoproliferative disorders with autoimmunity. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1219–1223. doi: 10.1073/pnas.82.4.1219. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Drappa J., Brot N., Elkon K. B. The Fas protein is expressed at high levels on CD4+CD8+ thymocytes and activated mature lymphocytes in normal mice but not in the lupus-prone strain, MRL lpr/lpr. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10340–10344. doi: 10.1073/pnas.90.21.10340. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fowlkes B. J., Kruisbeek A. M., Ton-That H., Weston M. A., Coligan J. E., Schwartz R. H., Pardoll D. M. A novel population of T-cell receptor alpha beta-bearing thymocytes which predominantly expresses a single V beta gene family. Nature. 1987 Sep 17;329(6136):251–254. doi: 10.1038/329251a0. [DOI] [PubMed] [Google Scholar]
- Fukushi N., Arase H., Wang B., Ogasawara K., Gotohda T., Good R. A., Onoé K. Thymus: a direct target tissue in graft-versus-host reaction after allogeneic bone marrow transplantation that results in abrogation of induction of self-tolerance. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6301–6305. doi: 10.1073/pnas.87.16.6301. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hackett J., Jr, Tutt M., Lipscomb M., Bennett M., Koo G., Kumar V. Origin and differentiation of natural killer cells. II. Functional and morphologic studies of purified NK-1.1+ cells. J Immunol. 1986 Apr 15;136(8):3124–3131. [PubMed] [Google Scholar]
- Hara T., Miyajima A. Two distinct functional high affinity receptors for mouse interleukin-3 (IL-3). EMBO J. 1992 May;11(5):1875–1884. doi: 10.1002/j.1460-2075.1992.tb05239.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howe R. C., Pedrazzini T., MacDonald H. R. Functional responsiveness in vitro and in vivo of alpha/beta T cell receptors expressed by the B2A2 (J11d)- subset of CD4-8- thymocytes. Eur J Immunol. 1989 Jan;19(1):25–30. doi: 10.1002/eji.1830190105. [DOI] [PubMed] [Google Scholar]
- Kikly K., Dennert G. Evidence for extrathymic development of TNK cells. NK1+ CD3+ cells responsible for acute marrow graft rejection are present in thymus-deficient mice. J Immunol. 1992 Jul 15;149(2):403–412. [PubMed] [Google Scholar]
- Kobayashi S., Hirano T., Kakinuma M., Uede T. Transcriptional repression and differential splicing of Fas mRNA by early transposon (ETn) insertion in autoimmune lpr mice. Biochem Biophys Res Commun. 1993 Mar 15;191(2):617–624. doi: 10.1006/bbrc.1993.1262. [DOI] [PubMed] [Google Scholar]
- Koo G. C., Peppard J. R. Establishment of monoclonal anti-Nk-1.1 antibody. Hybridoma. 1984 Fall;3(3):301–303. doi: 10.1089/hyb.1984.3.301. [DOI] [PubMed] [Google Scholar]
- Kotzin B. L., Babcock S. K., Herron L. R. Deletion of potentially self-reactive T cell receptor specificities in L3T4-, Lyt-2- T cells of lpr mice. J Exp Med. 1988 Dec 1;168(6):2221–2229. doi: 10.1084/jem.168.6.2221. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kubo R. T., Born W., Kappler J. W., Marrack P., Pigeon M. Characterization of a monoclonal antibody which detects all murine alpha beta T cell receptors. J Immunol. 1989 Apr 15;142(8):2736–2742. [PubMed] [Google Scholar]
- Kubota H., Okazaki H., Onuma M., Kano S., Hattori M., Minato N. CD3+4-8- alpha beta T cell population with biased T cell receptor V gene usage. Presence in bone marrow and possible involvement of IL-3 for their extrathymic development. J Immunol. 1992 Aug 15;149(4):1143–1150. [PubMed] [Google Scholar]
- Landolfi M. M., Van Houten N., Russell J. Q., Scollay R., Parnes J. R., Budd R. C. CD2-CD4-CD8- lymph node T lymphocytes in MRL lpr/lpr mice are derived from a CD2+CD4+CD8+ thymic precursor. J Immunol. 1993 Jul 15;151(2):1086–1096. [PubMed] [Google Scholar]
- Leo O., Foo M., Sachs D. H., Samelson L. E., Bluestone J. A. Identification of a monoclonal antibody specific for a murine T3 polypeptide. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1374–1378. doi: 10.1073/pnas.84.5.1374. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levitsky H. I., Golumbek P. T., Pardoll D. M. The fate of CD4-8- T cell receptor-alpha beta+ thymocytes. J Immunol. 1991 Feb 15;146(4):1113–1117. [PubMed] [Google Scholar]
- Matsuzaki Y., Pannetien C., Kanagawa O., Gachelin G., Nakauchi H. Evidence for the existence of two parallel differentiation pathways in the thymus of MRL lpr/lpr mice. J Immunol. 1992 Aug 1;149(3):1069–1074. [PubMed] [Google Scholar]
- McConkey D. J., Hartzell P., Nicotera P., Orrenius S. Calcium-activated DNA fragmentation kills immature thymocytes. FASEB J. 1989 May;3(7):1843–1849. doi: 10.1096/fasebj.3.7.2497041. [DOI] [PubMed] [Google Scholar]
- Michon J. M., Caligiuri M. A., Hazanow S. M., Levine H., Schlossman S. F., Ritz J. Induction of natural killer effectors from human thymus with recombinant IL-2. J Immunol. 1988 May 15;140(10):3660–3667. [PubMed] [Google Scholar]
- Mountz J. D., Smith T. M., Toth K. S. Altered expression of self-reactive T cell receptor V beta regions in autoimmune mice. J Immunol. 1990 Mar 15;144(6):2159–2166. [PubMed] [Google Scholar]
- Niehues T., Gulwani-Akolkar B., Akolkar P. N., Tax W., Silver J. Unique phenotype and distinct TCR V beta repertoire in human peripheral blood alpha beta TCR+, CD4-, and CD8- double negative T cells. J Immunol. 1994 Feb 1;152(3):1072–1081. [PubMed] [Google Scholar]
- Ogasawara J., Watanabe-Fukunaga R., Adachi M., Matsuzawa A., Kasugai T., Kitamura Y., Itoh N., Suda T., Nagata S. Lethal effect of the anti-Fas antibody in mice. Nature. 1993 Aug 26;364(6440):806–809. doi: 10.1038/364806a0. [DOI] [PubMed] [Google Scholar]
- Ogasawara M., Iwabuchi K., Good R. A., Onoe K. Bone marrow cells from allogeneic bone marrow chimeras inhibit the generation of cytotoxic lymphocyte responses against both donor and recipient cells. J Immunol. 1988 Nov 15;141(10):3306–3312. [PubMed] [Google Scholar]
- Ohteki T., Okuyama R., Seki S., Abo T., Sugiura K., Kusumi A., Ohmori T., Watanabe H., Kumagai K. Age-dependent increase of extrathymic T cells in the liver and their appearance in the periphery of older mice. J Immunol. 1992 Sep 1;149(5):1562–1570. [PubMed] [Google Scholar]
- Palathumpat V., Dejbakhsh-Jones S., Holm B., Wang H., Liang O., Strober S. Studies of CD4- CD8- alpha beta bone marrow T cells with suppressor activity. J Immunol. 1992 Jan 15;148(2):373–380. [PubMed] [Google Scholar]
- Porcelli S., Morita C. T., Brenner M. B. CD1b restricts the response of human CD4-8- T lymphocytes to a microbial antigen. Nature. 1992 Dec 10;360(6404):593–597. doi: 10.1038/360593a0. [DOI] [PubMed] [Google Scholar]
- Porcelli S., Yockey C. E., Brenner M. B., Balk S. P. Analysis of T cell antigen receptor (TCR) expression by human peripheral blood CD4-8- alpha/beta T cells demonstrates preferential use of several V beta genes and an invariant TCR alpha chain. J Exp Med. 1993 Jul 1;178(1):1–16. doi: 10.1084/jem.178.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rouvier E., Luciani M. F., Golstein P. Fas involvement in Ca(2+)-independent T cell-mediated cytotoxicity. J Exp Med. 1993 Jan 1;177(1):195–200. doi: 10.1084/jem.177.1.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmidt R. E., Murray C., Daley J. F., Schlossman S. F., Ritz J. A subset of natural killer cells in peripheral blood displays a mature T cell phenotype. J Exp Med. 1986 Jul 1;164(1):351–356. doi: 10.1084/jem.164.1.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Singer P. A., Balderas R. S., McEvilly R. J., Bobardt M., Theofilopoulos A. N. Tolerance-related V beta clonal deletions in normal CD4-8-, TCR-alpha/beta + and abnormal lpr and gld cell populations. J Exp Med. 1989 Dec 1;170(6):1869–1877. doi: 10.1084/jem.170.6.1869. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Slezak S. E., Horan P. K. Cell-mediated cytotoxicity. A highly sensitive and informative flow cytometric assay. J Immunol Methods. 1989 Feb 24;117(2):205–214. doi: 10.1016/0022-1759(89)90142-7. [DOI] [PubMed] [Google Scholar]
- Smith C. A., Williams G. T., Kingston R., Jenkinson E. J., Owen J. J. Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures. Nature. 1989 Jan 12;337(6203):181–184. doi: 10.1038/337181a0. [DOI] [PubMed] [Google Scholar]
- Suda T., Takahashi T., Golstein P., Nagata S. Molecular cloning and expression of the Fas ligand, a novel member of the tumor necrosis factor family. Cell. 1993 Dec 17;75(6):1169–1178. doi: 10.1016/0092-8674(93)90326-l. [DOI] [PubMed] [Google Scholar]
- Suda T., Zlotnik A. Origin, differentiation, and repertoire selection of CD3+CD4-CD8- thymocytes bearing either alpha beta or gamma delta T cell receptors. J Immunol. 1993 Jan 15;150(2):447–455. [PubMed] [Google Scholar]
- Sykes M. Unusual T cell populations in adult murine bone marrow. Prevalence of CD3+CD4-CD8- and alpha beta TCR+NK1.1+ cells. J Immunol. 1990 Nov 15;145(10):3209–3215. [PubMed] [Google Scholar]
- Takahama Y., Kosugi A., Singer A. Phenotype, ontogeny, and repertoire of CD4-CD8- T cell receptor alpha beta + thymocytes. Variable influence of self-antigens on T cell receptor V beta usage. J Immunol. 1991 Feb 15;146(4):1134–1141. [PubMed] [Google Scholar]
- Takahashi T., Tanaka M., Brannan C. I., Jenkins N. A., Copeland N. G., Suda T., Nagata S. Generalized lymphoproliferative disease in mice, caused by a point mutation in the Fas ligand. Cell. 1994 Mar 25;76(6):969–976. doi: 10.1016/0092-8674(94)90375-1. [DOI] [PubMed] [Google Scholar]
- Tutt M. M., Kuziel W. A., Hackett J., Jr, Bennett M., Tucker P. W., Kumar V. Murine natural killer cells do not express functional transcripts of the alpha-, beta-, or gamma-chain genes of the T cell receptor. J Immunol. 1986 Nov 1;137(9):2998–3001. [PubMed] [Google Scholar]
- Watanabe-Fukunaga R., Brannan C. I., Copeland N. G., Jenkins N. A., Nagata S. Lymphoproliferation disorder in mice explained by defects in Fas antigen that mediates apoptosis. Nature. 1992 Mar 26;356(6367):314–317. doi: 10.1038/356314a0. [DOI] [PubMed] [Google Scholar]
- Watanabe-Fukunaga R., Brannan C. I., Itoh N., Yonehara S., Copeland N. G., Jenkins N. A., Nagata S. The cDNA structure, expression, and chromosomal assignment of the mouse Fas antigen. J Immunol. 1992 Feb 15;148(4):1274–1279. [PubMed] [Google Scholar]
- Yonehara S., Ishii A., Yonehara M. A cell-killing monoclonal antibody (anti-Fas) to a cell surface antigen co-downregulated with the receptor of tumor necrosis factor. J Exp Med. 1989 May 1;169(5):1747–1756. doi: 10.1084/jem.169.5.1747. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhou T., Bluethmann H., Eldridge J., Berry K., Mountz J. D. Origin of CD4-CD8-B220+ T cells in MRL-lpr/lpr mice. Clues from a T cell receptor beta transgenic mouse. J Immunol. 1993 Apr 15;150(8 Pt 1):3651–3667. [PubMed] [Google Scholar]
- Zlotnik A., Godfrey D. I., Fischer M., Suda T. Cytokine production by mature and immature CD4-CD8- T cells. Alpha beta-T cell receptor+ CD4-CD8- T cells produce IL-4. J Immunol. 1992 Aug 15;149(4):1211–1215. [PubMed] [Google Scholar]