Evaluation of the expression of IFN-γ in lymphocytes using a monoclonal antibody and flow cytometry

Caruso, Arnaldo; Terlenghi, Luigina; Scalzini, Alfredo; Verardi, Rosanna; Foresti, I; Pollara, P.; Bonfanti, Carlo; Ravizzola, G; Manca, N.; Turano, A

doi:10.1016/0022-1759(88)90379-1

Cited by 9 publications

(4 citation statements)

References 11 publications

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“…Recently, several assays have been reported that determine the IFN-y response at the singlecell level after polyclonal in vitroTcel1 activation. One such assay is the recently described assay for detection of intracellular [6-111 and/or surface-expressed IFN-y [12]. However, these assays do not measure the number of cells which actually secrete IFN-y.…”

Section: Introductionmentioning

confidence: 99%

Detection of intracellular expression and secretion of interferon‐γ at the single‐cell level after activation of human T cells with tetanus toxoid in vitro

Andersson²,

et al. 1990

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Activation of T cells results in intracellular expression and secretion of cytokines such as interferon (IFN)-gamma. Here we have used three different assays for determination of IFN-gamma in tetanus toxoid- or mitogen-activated human T cell cultures. Two of these assays [intracytoplasmic immunofluorescence and enzyme-linked immuno spot assay (ELISPOT)] determined the expression and secretion of IFN-gamma at the single-cell level while the third assay enzyme-linked immunosorbent assay (ELISA) measured IFN-gamma secreted into the culture supernatant. Comparison of all three tests revealed a good correlation between the ELISPOT assay and the ELISA, whereas expression of intracellular IFN-gamma showed a qualitative but not a quantitative correlation with the latter. Both the immunospot assay and the immunofluorescence may be used to detect approximate numbers of specific T cells even when present at low frequencies. With the use of the immunospot assay antigen-specific T cells could be detected even in the absence of detectable IFN-gamma in the culture supernatants. However, the ELISA assay should be more convenient for screening large clinical material.

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Section: Introductionmentioning

confidence: 99%

Detection of intracellular expression and secretion of interferon‐γ at the single‐cell level after activation of human T cells with tetanus toxoid in vitro

Andersson²,

et al. 1990

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“…In an earlier report, Caruso et al (15) had shown that human lymphocytes activated by PHA and viruses express cytoplasmic and surface IFN-y. In our own work, we have also used semi-quantitative PCR to measure the kinetic expression of IFN-yspecific mRNA in PHA-activated lymphoblasts.…”

Section: Binding Specificities Of Anti-ifn-y (Adi-1 and Adi-23) Monocmentioning

confidence: 98%

Characterization of anti‐canine cytokine monoclonal antibodies specific for IFN‐γ: Effect of anti‐IFN‐γ on renal transplant rejection

et al. 1994

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Two murine monoclonal antibodies specific for IFN-gamma, ADI-1, and ADI-23 (both IgG1 kappa), were generated in BALB/c mice. The ADI-1 exhibited a higher avidity for canine rIFN-gamma than for nIFN-gamma and human rIFN-gamma. In contrast, the ADI-23 showed equal avidity for the three IFN-gamma preparations. The anti-canine IFN-gamma mAb did not bind to mouse and rat rIFN-gamma. The ADI-1, and ADI-23 mAb were also tested for binding to human rTFN-alpha and, contrary to our expectations, it was found that ADI-23 showed significant binding to human rTFN-alpha and rIFN-gamma, in contrast to ADI-1. Both anti-canine IFN-gamma mAb stained 48-h PHA-induced dog lymphoblasts. A two-site mAb ELISA was developed, which was linear in the range of 7-500 ng of canine rIFN-gamma, which indicated that the two mAb detected non-overlapping epitopes on the canine rIFN-gamma molecule. We studied the effect of ADI-1 on the prolongation of canine renal allografts. Recipients of kidney allografts, that were treated with ADI-1 by continuous arterial infusion, were prolonged to 22 and 25 days, compared to 9 and 13 days for animals given the IgG1 isotype control.

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“…MAb IGMB-14 unlike previous anti-IFN-y MAbs, recognizes in Western blot two polypeptides of MW 2S,000 and 56,000 present in affinity purified IFN-y preparations, without reacting with the IFNy subspecies whose molecular weight is already known (namely MW 25 ,000 and 20,000); it is also able to recognize the new molecular weight IFN-y subspecies on the surface of circulating lymphocytes (13). The assay was performed as earlier described (10,13). …”

Section: Ifn -Y Assaymentioning

confidence: 99%

Four Different Markers for Activated Cell Mediated Immunity Compared with the Clinical Stages of HIV Infection

et al. 1990

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Summary Activated cell-mediated immunity, as defined by four markers, has been studied in HIV-infected patients, and compared with the clinical stages of HIV infection. All the values obtained with such markers (interferon-y expressed on lymphocytes, soluble CD8, soluble IL-2 receptor, and serum neopterin) were observed to increase with the progressive stages of HIV infection, with the highest figures occurring in stage IV-C patients. Since the replication of HIV depend strictly on the stimulation of infected cells, T-cell activation in vitro might contribute to a further spread of HIV-1 in infected subjects. Thus permanent or multiple stimulation of the cellular immune system could be an important cofactor in the progression of this syndrome.

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Evaluation of the expression of IFN-γ in lymphocytes using a monoclonal antibody and flow cytometry

Cited by 9 publications

References 11 publications

Detection of intracellular expression and secretion of interferon‐γ at the single‐cell level after activation of human T cells with tetanus toxoid in vitro

Detection of intracellular expression and secretion of interferon‐γ at the single‐cell level after activation of human T cells with tetanus toxoid in vitro

Characterization of anti‐canine cytokine monoclonal antibodies specific for IFN‐γ: Effect of anti‐IFN‐γ on renal transplant rejection

Four Different Markers for Activated Cell Mediated Immunity Compared with the Clinical Stages of HIV Infection

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