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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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
Issue number1
Publication statusPublished - 1991

ASJC Scopus subject areas

  • Immunology


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