In vitro proliferation and cloning of CD3-CD161 cells from human thymocyte precursors

Maria Cristina Mingari, Alessandro Poggi, Roberto Biassoni, Rosanna Bellomo, Ermanno Ciccone, Nicoletta Pella, Luigia Morelli, Simonetta Verdiani, Alessandro Moretta, Lorenzo Moretta

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Abstract

Purified CD3-4- thymocytes were obtained by depletion of CD3+ and CD4+ cells from fresh thymocyte suspensions. 5-15% of these cells were found to express CD16 antigen, while other natural killer (NK) cell markers were virtually absent. Double fluorescence analysis revealed that 20-40% of thymic CD16+ cells coexpressed CD1, while approximately half were cyCD3+. When cultured in the presence of peripheral blood lymphocytes and H9 leukemia cell line as a source of irradiated feeder cells and interleukin 2 (IL-2), CD3-4- thymocytes underwent extensive proliferation. In addition, after 1-2 wk of culture, 30-50% of these cells were found to express CD16 surface antigen. Cloning under limiting dilution conditions of either CD3-4- or CD3-4-16- thymocytes in the presence of irradiated H9 cells resulted in large proportions (∼50%) of CD16+ clones. On the basis of the expression of surface CD16 and/or cyCD3 antigen, clones could be grouped in the following subsets: CD16+ cyCD3+ ; CD16+ cyCD3+; CD16-cyCD3+ ; and CD16-cyCD3-. All clones expressed CD56 surface antigen, displayed a strong cytolytic activity against NK sensitive (K562) and NK-resistant (M14) target cells, and produced IFN-γ and tumor necrosis factor, but not IL-2. Similar to peripheral NK cells, thymic CD16+ cells expressed transcripts for CD16 and for CD3 ε (Biassoni, R., S. Ferrini, I. Prigione, A. Moretta, and E.O. Long. 1988. J. Im̀munol 140:1685.) and ζ chains (Anderson, P., M. Caligiuri, J. Ritz, and S.F. Schlossman. 1989. Nature [Land.]. 341:159). Therefore, it appears that cells that are phenotypically and functionally similar to CD3- CD16+ NK cells may arise from immature thymocytes.

Original languageEnglish
Pages (from-to)21-26
Number of pages6
JournalJournal of Experimental Medicine
Volume174
Issue number1
Publication statusPublished - 1991

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Thymocytes
Organism Cloning
Natural Killer Cells
IgG Receptors
Clone Cells
Surface Antigens
Interleukin-2
CD56 Antigens
Feeder Cells
In Vitro Techniques
Suspensions
Leukemia
Tumor Necrosis Factor-alpha
Fluorescence
Lymphocytes
Antigens
Cell Line

ASJC Scopus subject areas

  • Immunology

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In vitro proliferation and cloning of CD3-CD161 cells from human thymocyte precursors. / Mingari, Maria Cristina; Poggi, Alessandro; Biassoni, Roberto; Bellomo, Rosanna; Ciccone, Ermanno; Pella, Nicoletta; Morelli, Luigia; Verdiani, Simonetta; Moretta, Alessandro; Moretta, Lorenzo.

In: Journal of Experimental Medicine, Vol. 174, No. 1, 1991, p. 21-26.

Research output: Contribution to journalArticle

Mingari, MC, Poggi, A, Biassoni, R, Bellomo, R, Ciccone, E, Pella, N, Morelli, L, Verdiani, S, Moretta, A & Moretta, L 1991, 'In vitro proliferation and cloning of CD3-CD161 cells from human thymocyte precursors', Journal of Experimental Medicine, vol. 174, no. 1, pp. 21-26.
Mingari, Maria Cristina ; Poggi, Alessandro ; Biassoni, Roberto ; Bellomo, Rosanna ; Ciccone, Ermanno ; Pella, Nicoletta ; Morelli, Luigia ; Verdiani, Simonetta ; Moretta, Alessandro ; Moretta, Lorenzo. / In vitro proliferation and cloning of CD3-CD161 cells from human thymocyte precursors. In: Journal of Experimental Medicine. 1991 ; Vol. 174, No. 1. pp. 21-26.
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abstract = "Purified CD3-4- thymocytes were obtained by depletion of CD3+ and CD4+ cells from fresh thymocyte suspensions. 5-15{\%} of these cells were found to express CD16 antigen, while other natural killer (NK) cell markers were virtually absent. Double fluorescence analysis revealed that 20-40{\%} of thymic CD16+ cells coexpressed CD1, while approximately half were cyCD3+. When cultured in the presence of peripheral blood lymphocytes and H9 leukemia cell line as a source of irradiated feeder cells and interleukin 2 (IL-2), CD3-4- thymocytes underwent extensive proliferation. In addition, after 1-2 wk of culture, 30-50{\%} of these cells were found to express CD16 surface antigen. Cloning under limiting dilution conditions of either CD3-4- or CD3-4-16- thymocytes in the presence of irradiated H9 cells resulted in large proportions (∼50{\%}) of CD16+ clones. On the basis of the expression of surface CD16 and/or cyCD3 antigen, clones could be grouped in the following subsets: CD16+ cyCD3+ ; CD16+ cyCD3+; CD16-cyCD3+ ; and CD16-cyCD3-. All clones expressed CD56 surface antigen, displayed a strong cytolytic activity against NK sensitive (K562) and NK-resistant (M14) target cells, and produced IFN-γ and tumor necrosis factor, but not IL-2. Similar to peripheral NK cells, thymic CD16+ cells expressed transcripts for CD16 and for CD3 ε (Biassoni, R., S. Ferrini, I. Prigione, A. Moretta, and E.O. Long. 1988. J. Im̀munol 140:1685.) and ζ chains (Anderson, P., M. Caligiuri, J. Ritz, and S.F. Schlossman. 1989. Nature [Land.]. 341:159). Therefore, it appears that cells that are phenotypically and functionally similar to CD3- CD16+ NK cells may arise from immature thymocytes.",
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AU - Ciccone, Ermanno

AU - Pella, Nicoletta

AU - Morelli, Luigia

AU - Verdiani, Simonetta

AU - Moretta, Alessandro

AU - Moretta, Lorenzo

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