A convenient and reliable IL‐2 bioassay using frozen CTLL‐2 to improve the detection of helper T lymphocyte precursors

Weston, Lyanne; Geczy, Andrew F.; Farrell, C.

doi:10.1046/j.1440-1711.1998.00733.x

Cited by 12 publications

(6 citation statements)

References 6 publications

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“…We confirmed our hypothesis about the binding of peptide (I) to IL 2Rα by the experiments with the use of the CTLL cell line that permanently expresses IL 2R and can exist only in the presence of IL 2 [17]. Peptide (I) stimulated growth of the CTLL cells which were preliminarily incubated without IL 2 for 24 h within the concentration range from 1.6 to 200 μg/mL (Fig.…”

Section: Resultssupporting

confidence: 82%

Regulation of activity of the immune cells by modified peptide fragments of Human IL-2

Onoprienko

Mikhaleva

Voytenkov³

et al. 2012

Russ J Bioorg Chem

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Synthetic peptides corresponding to the 59-72 (I), 60-72 (II) and 61-72 (III) sequences of human interleukin 2 with their N(alpha) acetylated and C(alpha) methylated termini were shown to exhibit pronounced hepatoprotective properties. These peptides neutralized hepatotoxic effects of such agents as tetrachloromethane and galactosamine in experiments in vivo. The peptide action revealed as normalization of duration of the thiopental narcosis of experimental animals and the level of hepatospecific enzymes in their blood. The effects of peptides (I)-(III) proved to be similar to that of prednisolone (the well-known anti-inflammatory agent), whereas the bestatine cytotoxic dipeptide had no hepatoprotecting effect. The target of the hepatoprotective activity of the peptides was shown to be the preliminary activated macrophages. We proposed that this activity of the peptides was associated with their interaction with the a-subunit of the interleukin 2 receptor (IL-2Ralpha), because the X-Ray analysis pointed to this region as one of binding sites of IL-2 with IL-2Ralpha. Experiments on the influence of the most active (59-72)-peptide on growth of the IL-2 dependent cell line (CTLL) confirmed this proposal. The 3H-labeled peptide corresponding to the 59-72 sequence ofthe human IL-2 was shown to bind to the CTLL cels. We assumed that the binding of this peptide was specific and occurred precisely with IL-2Ra and virtually determined the binding constant. Its value (1.41 x 10(-6) M) was comparable with that of the interaction of IL-2 with IL-2Ralpha (approximately 10(-7) M).

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Section: Resultssupporting

confidence: 82%

Regulation of activity of the immune cells by modified peptide fragments of Human IL-2

Onoprienko

Mikhaleva

Voytenkov³

et al. 2012

Russ J Bioorg Chem

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“…CTLL‐2 cells were cryopreserved 3 days after subculturing, in culture medium without IL‐2. These cryopreserved cells were assessed in dose–response assays and their sensitivity was judged acceptable if the cells were capable of detecting 0.01 U rIL‐2 15 . CTLL‐2 cells were thawed using 20% FCS in RPMI, washed, counted and adjusted to a concentration of 1 × 10 3 cells/mL, in media without IL‐2.…”

Section: Methodsmentioning

confidence: 99%

Interleukin‐2‐producing helper T lymphocyte precursor frequency and rejection: A longitudinal cellular study after cardiac transplantation

et al. 2000

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Summary Interleukin-2-producing helper T lymphocyte precursors (HTLp) in the recipient recognize donor alloantigen expressed by the transplanted organ. The frequency of these reactive cells in the peripheral blood was determined and correlated with rejection episodes. Endomyocardial biopsy is generally used to quantify cardiac allograft rejection and guide immunotherapy. While non-invasive techniques have been investigated, none of these has demonstrated sufficient sensitivity or specificity to replace myocardial biopsy. Twelve cardiac transplant recipients were assessed over a 1 year period using limiting dilution analysis, to determine the frequency of HTLp in response to cadaver donor splenocytes. The IL-2-dependent mouse cell line CTLL-2 was used to measure the IL-2 present and the precursor frequency was calculated using maximum likelihood estimation. In the months immediately post transplantation, six of the 12 recipients displayed an association with increases in HTLp frequency, which preceded histologically detectable rejection. The second six recipients had fewer rejection episodes and achieved 'acceptance' of their graft sooner. Once 'acceptance' was achieved, the association between IL-2 HTLp frequency and rejection was no longer apparent. The ability to identify two groups of patients on the basis of IL-2 HTLp frequency clearly highlights the heterogenous nature of the response to the graft and this emphasizes the need to monitor other cytokines, which may influence the functioning of effector T cells.

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“…40 Briefly, 1 Â 10 4 CTLL cells per well/50 ml were plated with 50 ml of varying concentrations of rec IL-2 and test supernatants. CTLL proliferation was determined 24 h later by [ 3 H]-thymidine incorporation (for 5 h) and measured by liquid scintillation counting (Hewlett Packard, Palo Alto, CA).…”

Section: Cytotoxic T Cell Line Assay To Determine the Biological Actimentioning

confidence: 99%

Cytokine-armed vaccinia virus infects the mesothelioma tumor microenvironment to overcome immune tolerance and mediate tumor resolution

Jackaman

Nelson

2010

Cancer Gene Ther

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Intratumoral (i.t.) administration of cytokine genes expressed by viral vectors represents a rational approach that induces cytokine secretion at the site they are needed, and i.t. vaccinia virus (VV) has shown promise in mesothelioma patients. However, we and others have shown that the mesothelioma tumor microenvironment includes macrophages, dendritic cells (DCs), and T cells. Therefore, we investigated which of these cell types are infected after exposure to VV or Fowlpox virus (FPV)-cytokine gene constructs. In vitro studies showed that mesothelioma tumor cells were resistant to FPV infection yet highly permissive to infection by VV vectors resulting in significant cytokine production and impaired proliferation. Macrophages secreted low levels of cytokine suggestive of resistance to overt infection. DCs transiently secreted virally derived cytokines, but did not mature during VV infection. VV inhibition of T cell proliferation was rescued by the interleukin (IL)-2 and IL-12 VV constructs. In vivo studies of murine mesotheliomas showed that i.t. injection of the parent VV could not hinder tumor progression. In contrast, the VV-cytokine constructs induced profound tumor regression. These data suggest that i.t. VV-cytokine gene constructs retard tumor growth by infecting mesothelioma cells and targeting the immune system through tumor-infiltrating DC and T cells.

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A convenient and reliable IL‐2 bioassay using frozen CTLL‐2 to improve the detection of helper T lymphocyte precursors

Cited by 12 publications

References 6 publications

Regulation of activity of the immune cells by modified peptide fragments of Human IL-2

Regulation of activity of the immune cells by modified peptide fragments of Human IL-2

Interleukin‐2‐producing helper T lymphocyte precursor frequency and rejection: A longitudinal cellular study after cardiac transplantation

Cytokine-armed vaccinia virus infects the mesothelioma tumor microenvironment to overcome immune tolerance and mediate tumor resolution

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