Interleukin‐2 Dependent Culture of Cytolytic T Cell Lines

Gillis, Steven; Watson, James D.

doi:10.1111/j.1600-065x.1981.tb00435.x

Cited by 96 publications

(47 citation statements)

References 85 publications

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“…The optimal DNA concentration (50 Ag template plus balance DNA) was pretested for each extract (Dobrzanski et al, 1988). Nuclear extracts (Dignam et al, 1983) were prepared from exponentially growing HeLa cells in suspension cultures or from primary human lymphocytes which were stimulated to exponential growth by phytohemagglutinin and interleukin 2 (Gillis and Watson, 1981). The RNA transcribed in vitro was extracted with phenol-chloroform, precipitated and directly analyzed by electrophoresis on 5% polyacrylamide gels in the presence of 7 M urea.…”

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confidence: 99%

DNA methylation in the Alu sequences of diploid and haploid primary human cells.

1993

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We have investigated DNA methylation in human Alu sequences, both in general and in specific Alu sequences associated with the genes for a1 globin, tissue plasminogen activator (tPA), adrenocorticotropic hormone (ACTH) and angiogenin. We studied DNAs from lymphocytes, granulocytes, brain, heart muscle and sperm, and from the human HeLa and KB cell lines by using cleavage with methylation-sensitive restriction enzymes combined with Southern blot hybridization and by using genomic sequencing. The results can be summarized as follows. (i) In differentiated primary human cells, Alu elements are often highly methylated even when they are in very 5'-CG-3'-rich regions. This finding is not consistent with the notion that hypermethylation would be a sufficient condition in itself for 5'-CG-3' sequences to undergo loss of 5-methyldeoxycytidine (5-mC) due to deamination and subsequent mutation. (ii) There are distinct differences in the levels of methylation in the specific Alu sequences. (iii) Alu elements in the DNA of haploid spermatozoa are much less methylated than in diploid cells. Preliminary data indicate that spermatozoa contain Alu-specific RNAs. (iv)The results of cell-free transcription experiments with Alu elements suggest that the in vitro transcription of Alu elements can be inhibited by 5'-CG-3' methylation. High levels of 5'-CG-3' methylation in Alu elements could contribute to their general transcriptional inactivity. (v) The patterns of methylation observed in the Alu elements and in the surrounding sequences are characterized by cell type specific interindividual concordance.

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confidence: 99%

DNA methylation in the Alu sequences of diploid and haploid primary human cells.

1993

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“…In addition, IL-2 has been used to derive functional T cell clones (12)(13)(14). These clones express relatively large amounts of both Ia and 4F2 antigens irrespective of their functional activity (15).…”

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confidence: 99%

IL-2-mediated T cell proliferation in humans is blocked by a monoclonal antibody directed against monomorphic determinants of HLA-DR antigens.

Moretta¹,

Accolla²,

Cerottini³

1982

The Journal of Experimental Medicine

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We have studied the effect on the interleukin (IL-) 2-dependent human T cell growth of two distinct monoclonal antibodies (Mab), D1-12 and 4F2, with specificity for common determinant of human Ia antigens and for a differentiation antigen expressed on all activated T cells, respectively. Strong inhibition of cell growth was found in cultures supplemented with the anti-Ia D1-12 Mab but not in cultures supplemented with 4F2 Mab. These results were obtained when either total mixed leukocyte culture (MLC) T cells or an MLC-derived T cell clone were used as indicator cell systems for IL-2 activity. The inhibition of cell growth appears to be mediated by a direct interaction of D1-12 Mab with the cells and not by a direct inactivation of the growth factor, as addition of the antibody to murine MLC T cells, which do not express the determinant defined by D1-12 Mab, resulted in no inhibition of their proliferation induced by the same source of human IL-2.

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“…The present study, however, provides some encouragement that systemic administration of IL-2 may be a feasible approach for the activation of T cells in vivo. Such an approach would be technically much simpler than injection of cytotoxic T cells grown in vitro in the presence of IL-2 as described by previous authors (Mills et al, 1980;Warren et al, 1979;Gillis & Watson, 1981) and may allow activation of lymphocytes at the site of tumour growth. Although the half-life of 20-25min for IL-2 in the circulation appears relatively short this does not appear to be due to inactivation by serum factors.…”

Section: Discussionmentioning

confidence: 99%

“…Addition of IL-2 from mitogen-stimulated lymphocytes to cultures of lymphocytes and tumour cells was shown to induce cytotoxic activity against syngeneic animal tumours (Mills & Paetkau, 1980;Warren et al, 1978Warren et al, , 1979Gillis & Watson, 1981) and against autologous human malignancies such as leukaemia (Zarling & Bach, 1979), melanoma (Lotze et al, 1980) and various carcinomas (Vose & Bonnard, 1982;Vose & Moore, 1981).…”

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confidence: 99%

Clearance rates and systemic effects of intravenously administered interleukin 2 (IL-2) containing preparations in human subjects

Bindon¹,

Czerniecki²,

Ruell³

et al. 1983

Br J Cancer

105

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Summary The present study was designed to examine the feasibility of in vivo administration of interleukin 2 (IL-2) to induce cytotoxic cell activity against tumours in human subjects. IL-2 was prepared from blood leukocytes stimulated with phytohaemagglutinin (PHA) and partially purified by membrane chromatography to exclude PHA. Administration of different amounts of IL-2 in vivo to 2 patients with melanoma revealed that the initial level of IL-2 in the circulation was related to the dose given and had a half-life of -22.5 minutes. The initial and subsequent levels of IL-2 were lower than that expected to occur from equilibration in plasma and extracellular fluid. This was not apparently due to inactivation by serum factors because fresh human serum had little effect in vitro on the induction of mitogenic or cytotoxic activity by IL-2. Spontaneous division of lymphocytes was increased following IL-2 administration and it is suggested that clearance of IL-2 in vivo may reflect, in part, absorption by activated lymphocytes in the circulation. Side effects noted shortly after administration of the partially-purified IL-2 preparations included transient pyrexia, hypoglycaemia, increased cortisol levels, lymphocytopenia and signs of mild intravascular coagulation. No long-term effects were noted. These initial results suggest that systemic injection of purified preparations of II-2 may be a feasible approach to induce cytotoxic T cells in vivo.

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Interleukin‐2 Dependent Culture of Cytolytic T Cell Lines

Cited by 96 publications

References 85 publications

DNA methylation in the Alu sequences of diploid and haploid primary human cells.

DNA methylation in the Alu sequences of diploid and haploid primary human cells.

IL-2-mediated T cell proliferation in humans is blocked by a monoclonal antibody directed against monomorphic determinants of HLA-DR antigens.

Clearance rates and systemic effects of intravenously administered interleukin 2 (IL-2) containing preparations in human subjects

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