A solid-phase enzyme-linked immunospot (ELISPOT) assay for enumeration of specific antibody-secreting cells

Czerkinsky, Cécil; Nilsson, Lars‐Åke; Nygren, Håkan; Ouchterlony, Örjan; Tarkowski, Andrej

doi:10.1016/0022-1759(83)90308-3

Cited by 959 publications

(497 citation statements)

References 8 publications

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“…ELISPOT assays 10,11 are often used to measure the T-cell responses in different vaccine immunogenicity and efficacy studies. T cells isolated from the blood are stimulated with different antigens in order to induce cytokine synthesis and/or secretion, such as IFN-g, which is used especially in monitoring of CD8-specific CTL or CD4-specific Th 1 responses.…”

Section: Elispot Assaymentioning

confidence: 99%

Validation overview of bio-analytical methods

2005

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Section: Elispot Assaymentioning

confidence: 99%

Validation overview of bio-analytical methods

2005

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“…The inter-assay variation, calculated from control cells which were frozen and then thawed and assayed on four different occasions, was for IFN-g 31% (mean > 100 spots) and for IL-4 38% (mean < 10 spots). To compensate for this known variation of the ELISPOT assay [28], all cells were assayed in quadruplicates.…”

Section: Stimulation Of Lymphocytes and Enumeration Of Ifn-g-and Il-4mentioning

confidence: 99%

Th1-like responses to peptides from peripheral nerve myelin proteins in patients with polyneuropathy associated with monoclonal gammopathy

Ekerfelt¹,

Ernerudh²,

Vrethem³

1997

Clinical and Experimental Immunology

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SUMMARYPolyneuropathy associated with monoclonal gammopathy is usually regarded as an immune-mediated disorder with autoantibody activity against myelin glycoproteins. The pathogenic mechanisms are, however, not fully understood. Several reports have indicated an additional involvement of T cells. We investigated the occurrence of myelin-specific T cells as well as their functional characteristics. Cytokine responses generated after stimulation of peripheral blood mononuclear cells with selected peptides of myelin proteins P0 and P2 were investigated with an ELISPOT method. Three P0 peptides caused significantly elevated levels of interferon-gamma (IFN-g)-secreting cells in patients with polyneuropathy associated with monoclonal gammopathy (n ¼ 8) compared with controls (n ¼ 8), whereas none of the peptides caused elevated levels of IL-4-secreting cells. Patients with nonimmunological types of neuropathy (n ¼ 4) did not reveal any cytokine responses to myelin peptides. Our results indicate that a Th1-like response against myelin proteins occurs in polyneuropathy associated with monoclonal gammopathy.

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“…This assay can detect < 20 pg IFN-. For detection of IFN--secreting cells, a modification of the ELISPOT technique [10] was employed, as described in detail elsewhere [11]. Briefly, cell suspensions were plated in 96-well microplates that had previously been coated with mouse anti-human IFN-MoAbs.…”

Section: Detection Of Interferon-gammamentioning

confidence: 99%

Histaminergic regulation of interferon-gamma (IFN-γ) production by human natural killer (NK) cells

Asea¹,

Hansson²,

Czerkinsky³

et al. 1996

Clinical and Experimental Immunology

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SUMMARYMonocytes, recovered from human peripheral blood by counter-current centrifugal elutriation, effectively inhibit the production of IFN-°by CD3 ÿ /56 NK cells in response to IL-2. This study aimed at defining the nature of the inhibitory signal, particularly the importance of monocyte-derived reactive metabolites of oxygen. It was found that monocytes recovered from patients with chronic granulomatous disease (CGD), a condition characterized by deficient NADPH-oxidase activity of phagocytes, did not inhibit IFN-°production by NK cells. Further, catalase, a scavenger of hydrogen peroxide, completely reversed the inhibitory signal, whereas scavengers of the superoxide anion, hypohalous acids, the hydroxyl radical, or nitric oxide synthesis inhibitors such as L-NMMA were ineffective. Inhibition of IFN-°production was operating on a pre-translational level, as indicated by the inability of enriched NK cells to accumulate IFN-°mRNA in the presence of elutriated monocytes. Hydrogen peroxide, at micromolar concentrations, reconstituted the inhibition of IFN-°production when added to enriched NK cells. Histamine, a biogenic amine which inhibits the generation of reactive oxygen metabolites in monocytes, abrogated the inhibition of IFN-°production in NK cells; by this mechanism, histamine strongly synergized with IL-2 to induce IFN-°in mixtures of NK cells and monocytes. The synergizing effect of histamine was specifically mediated by H 2 -type histamine receptors. We conclude that: (i) the induction of IFN-°mRNA in NK cells is effectively down-regulated by products of the oxidative metabolism of monocytes; and (ii) histamine effectively enhances IFN-°production by preventing monocyte-induced oxidative damage to NK cells.

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A solid-phase enzyme-linked immunospot (ELISPOT) assay for enumeration of specific antibody-secreting cells

Cited by 959 publications

References 8 publications

Validation overview of bio-analytical methods

Validation overview of bio-analytical methods

Th1-like responses to peptides from peripheral nerve myelin proteins in patients with polyneuropathy associated with monoclonal gammopathy

Histaminergic regulation of interferon-gamma (IFN-γ) production by human natural killer (NK) cells

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