Abstract
It has been reported that lymphocytes from peripheral blood (PBL) cultured with interleukin-2 (IL-2) produce predominantly CD16+ lymphokine-activated killer (LAK) cells. We developed a two-step method to generate LAK cells from human PBL in long-term cultures (10-12 days) with recombinant human IL-2 (rhIL-2) and characterized the evolving LAK cell population by testing its phenotype and cytotoxic activity as a function of time. The starting PBL displayed some natural killer (NK) cytotoxicity but no LAK activity. At day 6, the cells were a mixed population of about 80% CD3+ and 6% CD16+ cells. Little proliferation was evident but strong LAK activity was detected. After 10-12 days, major cell expansion had occurred and they were essentially a pure (greater than 90%) CD3+ CD16- CD56- cell population large granular lymphocyte (LGL) by morphology that displayed strong non-MHC-restricted killing activity (greater than 200 lytic units). Over the same period of time, the CD16+ cells had almost completely regressed in these cultures. This preferential induction of CD+ LAK cells was not an effect of IL-2 concentration as 10 U/ml was as effective as 500 U/ml. Further characterization revealed a major population of CD4+ (60%) and CD8+ (30%) with a smaller fraction (less than 9%) of gamma delta + cells. These results indicate that a virtually pure CD3+ LAK cells population was produced with long-term cultures of lymphocytes from peripheral blood in rhIL-2, in which active proliferation of the CD3+ but not CD16+ cells occurred.
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- Anderson P., Caligiuri M., Ritz J., Schlossman S. F. CD3-negative natural killer cells express zeta TCR as part of a novel molecular complex. Nature. 1989 Sep 14;341(6238):159–162. doi: 10.1038/341159a0. [DOI] [PubMed] [Google Scholar]
- Biassoni R., Ferrini S., Prigione I., Moretta A., Long E. O. CD3-negative lymphokine-activated cytotoxic cells express the CD3 epsilon gene. J Immunol. 1988 Mar 1;140(5):1685–1689. [PubMed] [Google Scholar]
- Grimm E. A., Mazumder A., Zhang H. Z., Rosenberg S. A. Lymphokine-activated killer cell phenomenon. Lysis of natural killer-resistant fresh solid tumor cells by interleukin 2-activated autologous human peripheral blood lymphocytes. J Exp Med. 1982 Jun 1;155(6):1823–1841. doi: 10.1084/jem.155.6.1823. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grimm E. A., Ramsey K. M., Mazumder A., Wilson D. J., Djeu J. Y., Rosenberg S. A. Lymphokine-activated killer cell phenomenon. II. Precursor phenotype is serologically distinct from peripheral T lymphocytes, memory cytotoxic thymus-derived lymphocytes, and natural killer cells. J Exp Med. 1983 Mar 1;157(3):884–897. doi: 10.1084/jem.157.3.884. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grimm E. A., Robb R. J., Roth J. A., Neckers L. M., Lachman L. B., Wilson D. J., Rosenberg S. A. Lymphokine-activated killer cell phenomenon. III. Evidence that IL-2 is sufficient for direct activation of peripheral blood lymphocytes into lymphokine-activated killer cells. J Exp Med. 1983 Oct 1;158(4):1356–1361. doi: 10.1084/jem.158.4.1356. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hercend T., Reinherz E. L., Meuer S., Schlossman S. F., Ritz J. Phenotypic and functional heterogeneity of human cloned natural killer cell lines. Nature. 1983 Jan 13;301(5896):158–160. doi: 10.1038/301158a0. [DOI] [PubMed] [Google Scholar]
- Irle C., Beatty P. G., Mickelson E. M., Hansen J. A. Change in functional phenotype of cloned human alloreactive cytolytic T cells. Hum Immunol. 1984 Dec;11(4):183–191. doi: 10.1016/0198-8859(84)90058-2. [DOI] [PubMed] [Google Scholar]
- Ochoa A. C., Hasz D. E., Rezonzew R., Anderson P. M., Bach F. H. Lymphokine-activated killer activity in long-term cultures with anti-CD3 plus interleukin 2: identification and isolation of effector subsets. Cancer Res. 1989 Feb 15;49(4):963–968. [PubMed] [Google Scholar]
- Olabuenaga S. E., Brooks C. G., Gillis S., Henney C. S. Interleukin 2 is not sufficient for the continuous growth of cloned NK-like cytotoxic cell lines. J Immunol. 1983 Nov;131(5):2386–2391. [PubMed] [Google Scholar]
- Ortaldo J. R., Mason A., Overton R. Lymphokine-activated killer cells. Analysis of progenitors and effectors. J Exp Med. 1986 Oct 1;164(4):1193–1205. doi: 10.1084/jem.164.4.1193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Patel S. S., Thiele D. L., Lipsky P. E. Major histocompatibility complex-unrestricted cytolytic activity of human T cells. Analysis of precursor frequency and effector phenotype. J Immunol. 1987 Dec 1;139(11):3886–3895. [PubMed] [Google Scholar]
- Pawelec G., Kahle P., Wernet P. Specificity spectrum and cell surface markers of mono- and multi-functional mixed leukocyte culture-derived T cell clones in man. Eur J Immunol. 1982 Jul;12(7):607–615. doi: 10.1002/eji.1830120714. [DOI] [PubMed] [Google Scholar]
- Perussia B., Ramoni C., Anegon I., Cuturi M. C., Faust J., Trinchieri G. Preferential proliferation of natural killer cells among peripheral blood mononuclear cells cocultured with B lymphoblastoid cell lines. Nat Immun Cell Growth Regul. 1987;6(4):171–188. [PubMed] [Google Scholar]
- Phillips J. H., Lanier L. L. Dissection of the lymphokine-activated killer phenomenon. Relative contribution of peripheral blood natural killer cells and T lymphocytes to cytolysis. J Exp Med. 1986 Sep 1;164(3):814–825. doi: 10.1084/jem.164.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pohajdak B., Gomez J. L., Wilkins J. A., Greenberg A. H. Tumor-activated NK cells trigger monocyte oxidative metabolism. J Immunol. 1984 Nov;133(5):2430–2436. [PubMed] [Google Scholar]
- Punturieri A., Velotti F., Piccoli M., Herberman R. B., Frati L., Santoni A. Overnight incubation of mouse spleen cells in recombinant IL-2 generates cytotoxic cells with NK characteristics from precursors enriched with or devoid of LGL. Clin Exp Immunol. 1989 Jan;75(1):155–160. [PMC free article] [PubMed] [Google Scholar]
- Scott C. F., Jr, Lambert J. M., Kalish R. S., Morimoto C., Schlossman S. F. Human T cells can be directed to lyse tumor targets through the alternative activation/T11-E rosette receptor pathway. J Immunol. 1988 Jan 1;140(1):8–14. [PubMed] [Google Scholar]
- Shaw S., Luce G. E. The lymphocyte function-associated antigen (LFA)-1 and CD2/LFA-3 pathways of antigen-independent human T cell adhesion. J Immunol. 1987 Aug 15;139(4):1037–1045. [PubMed] [Google Scholar]
- Trinchieri G., Matsumoto-Kobayashi M., Clark S. C., Seehra J., London L., Perussia B. Response of resting human peripheral blood natural killer cells to interleukin 2. J Exp Med. 1984 Oct 1;160(4):1147–1169. doi: 10.1084/jem.160.4.1147. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weil-Hillman G., Fisch P., Prieve A. F., Sosman J. A., Hank J. A., Sondel P. M. Lymphokine-activated killer activity induced by in vivo interleukin 2 therapy: predominant role for lymphocytes with increased expression of CD2 and leu19 antigens but negative expression of CD16 antigens. Cancer Res. 1989 Jul 1;49(13):3680–3688. [PubMed] [Google Scholar]
- Wencel-Drake J. D., Plow E. F., Zimmerman T. S., Painter R. G., Ginsberg M. H. Immunofluorescent localization of adhesive glycoproteins in resting and thrombin-stimulated platelets. Am J Pathol. 1984 May;115(2):156–164. [PMC free article] [PubMed] [Google Scholar]
- Zalman L. S., Brothers M. A., Chiu F. J., Müller-Eberhard H. J. Mechanism of cytotoxicity of human large granular lymphocytes: relationship of the cytotoxic lymphocyte protein to the ninth component (C9) of human complement. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5262–5266. doi: 10.1073/pnas.83.14.5262. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zalman L. S., Brothers M. A., Müller-Eberhard H. J. Self-protection of cytotoxic lymphocytes: a soluble form of homologous restriction factor in cytoplasmic granules. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4827–4831. doi: 10.1073/pnas.85.13.4827. [DOI] [PMC free article] [PubMed] [Google Scholar]

