FluoroSpot Path: SARS‑CoV‑2 (S+NMO) Human IFN‑γ/Granzyme B/IL‑2

FSP-013602-P12-1

Publications: 1

Documents

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Paves your way to functional analysis of a specific research question.

Components

Plate	Pre-coated FluoroSpot plates (mAbs 1-D1K, MT34B6, and MT2A91/2C95)
Detection mAbs	Anti-IFN-γ mAb (7-B6-1), BAM
	Anti-Granzyme B (MT28), biotin
	Anti-IL-2 mAb (MT8G10), WASP
Fluorophore conjugates	Anti-BAM mAb, 490
	SA-550
	Anti-WASP mAb, 640
Buffer/Solution	FluoroSpot enhancer
Positive control	Anti-CD3 mAb (CD3-2)
Co-stimulus	Anti-CD28 mAb (CD28-A)
Peptide pools	SARS-CoV-2 (S), human
	SARS-CoV-2 (NMO), human

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Product specifications

Intended use

This FluoroSpot Path kit is intended for the enumeration of cells secreting human IFN-γ, granzyme B, and IL-2 in response to SARS-CoV-2 (S+NMO) peptides using the FluoroSpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

Peptide pool composition
The SARS-CoV-2 S defined peptide pool contains 100 peptides from the human SARS-CoV-2 virus. The peptides are derived from the spike (S) protein.

The SARS-CoV-2 NMO defined peptide pool contains 101 peptides from the human SARS-CoV-2 virus. The peptides are derived from the nucleoprotein (N), membrane protein (M), and open reading frame (ORF) proteins (O): ORF1, nsp3, ORF-3a, ORF-7a, and ORF8.

Published HLA types covered by the SARS-CoV-2 NMO defined peptide pool are listed in the datasheets.

Recommendation

Product details

Product	FluoroSpot Path: SARS-CoV-2 (S+NMO) Human IFN-γ/Granzyme B/IL-2
Application	FluoroSpot
Analyte	IFN-γ, Granzyme B, IL-2
Reactivity	Human
Specificity	This kit is based on matched pairs of mAbs specific for human IFN-γ, granzyme B, and IL-2.

Shipping and Storage

Shipping	Shipped at ambient temperature.
Storage	Store antibodies, fluorophore conjugates, and FluoroSpot enhancer at 4-8 °C upon receipt. Peptide pools should be stored at -20 °C or below. Plates may be kept at room temperature.
Shelf life	At least 12 months from date of receipt.

Performance

Human IFN-γ/Granzyme B/IL-2 FluoroSpot. PBMC were prepared from an individual after receiving two doses of SARS-CoV-2 vaccine and after subsequent SARS-CoV-2 infection. PBMC were incubated 48 h in the absence or presence of the SARS-CoV-2 S defined peptide pool or NMO defined peptide pool. All cells were co-stimulated with anti-CD28 mAb. Single cytokine secreting cells as well as double- and triple-secreting cells were determined. Analysis was done with using Mabtech IRIS.

Documents

Datasheet/Protocol SDS

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Third vaccine dose important for T cell response against Omicron

Three doses mRNA vaccine had better T cell responses than recipients of two doses. Mabtech IRIS and FluoroSpot Flex: Human IFN-γ/TNF-α/IL-2/Granzyme B were used to assess T cell immunity in mRNA vaccine recipients.

Highlighted research, Published December 16, 2021

SARS-CoV-2-specific immune responses in patients with autoimmune diseases

Patients with rheumatic diseases showed T cell responses comparable to healthy control. Patients with MS and after mRNA vaccination show different memory B cell IgG responses. SARS-CoV-2 specific immune responses in autoimmunity was analyzed using FluoroSpot, ELISpot Path, and Mabtech IRIS.

Highlighted research, Published December 2, 2021

Importance of studying SARS-CoV-2-specific memory B cells

When studying antibodies in circulation, remember that memory B cells might hide out of sight. In this study, detection of SARS-CoV-2-specific T and memory B cells 6 and 15 months after infection.

Highlighted research, Published November 24, 2021

Polyfunctional T cells with Mabtech IRIS

Study of development and persistence of virus-specific cellular and humoral immune responses, during the acute phase and two follow-ups after COVID-19 disease in hospitalized patients. Mabtech’s FluoroSpot Plus: Human IFN-γ/IL-2/TNF-α were analyzed with Mabtech IRIS.

Highlighted research, Published June 30, 2020

T cell immunity in COVID-19

Much emphasis has been put on antibody responses, but also T cells are important in the context of COVID-19. In this study, the authors characterized SARS-CoV-2-specific T cell responses by using an extensive FACS panel, peptide stimulations, tetramer staining, and Mabtech IFN-γ ELISpot.

Highlighted research, Published May 30, 2020

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Highlighted research, Published May 11, 2020

Longevity of immunity against SARS-CoV-2

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Publications (1)

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Analyte information

IFN-γ

Analyte description	Interferon-γ (IFN-γ) is the only type II interferon. This proinflammatory cytokine is secreted by activated T cells and NK cells. It activates macrophages and endothelial cells and regulates immune responses by affecting APCs, T cells, and B cells. Production of IFN-γ by helper T cells and cytotoxic T cells is a hallmark of the Th1-type phenotype. Thus, high-level production of IFN-γ is typically associated with effective host defense against intracellular pathogens.
Alternative names	Interferon-γ, Interferon-gamma, IFN-γ, IFN-gamma, IFN-g, IFNg, IFG, IFI, IMD69
Cell type	T cell, Tc, Th1, NK cell
Gene ID	3458

Granzyme B

Analyte description	Granzyme B is a serine protease released by the cytoplasmic granules within cytotoxic T cells and NK cells. Granzyme B will act together with perforin and other cytotoxic mediators to induce target cell death by apoptosis.
Alternative names	Granzyme B, GzmB, GrB, GrzB, C11, CCPI, CGL-1, CGL1, CSP-B, CSPB, CTLA1, CTSGL1, HLP, SECT
Cell type	T cell, Tc, Th1, NK cell
Gene ID	3002

IL-2

Analyte description	Interleukin 2 (IL-2) is predominantly produced by activated T cells. IL-2 promotes the proliferation, differentiation, and survival of antigen-activated T cells and NK cells. After antigen stimulation, the cytokine also promotes the differentiation of T cells into effector and memory T cells. Therefore IL-2 plays an important role in the body's response to infection. IL-2 is also key to tolerance, as it promotes the differentiation of certain immature T cells into regulatory T cells in the thymus, ultimately preventing autoimmune diseases.
Alternative names	Interleukin-2, Interleukin 2, IL-2, IL2, T cell growth factor, TCGF, lymphokine
Cell type	T cell, Th1, Th2, Th17, Tfh
Gene ID	3558