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mouse ifn-g, il-5, and il-17 immunospot elispot kits (Cellular Technology Ltd)

Name: mouse ifn-g, il-5, and il-17 immunospot elispot kits
Brand: Cellular Technology Ltd
Rating: 90 (1 reviews)

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Cellular Technology Ltd mouse ifn-g, il-5, and il-17 immunospot elispot kits
Mouse Ifn G, Il 5, And Il 17 Immunospot Elispot Kits, supplied by Cellular Technology Ltd, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse ifn-g, il-5, and il-17 immunospot elispot kits/product/Cellular Technology Ltd
Average 90 stars, based on 1 article reviews

mouse ifn-g, il-5, and il-17 immunospot elispot kits - by Bioz Stars, 2026-02

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Identification and characterization of T cell-restricted epitopes associated with AhpC C57G . C57BL/6 mice ( n = 12) were immunized on days 0, 21, and 35 with AhpC C57G . Spleens ( n = 4 per analyte) were harvested on day 42, and (a) IFN-γ-, (b) IL-5-, and (c) IL-17-secreting T cell responses against an AhpC C57G peptide library (15-mers overlapping by 10 amino acids) were quantitated by ELISpot. Black dots represent the means of assays conducted in duplicate for individual mice. SFC, spot-forming cells. (d) C57BL/6 mice ( n = 3) were immunized on days 0, 21, and 35 with AhpC C57G . Spleens harvested on day 42 were combined and used to prepare three equal pools of cells. The pools were either depleted of CD8 + cells or CD4 + cells or left untreated, and IFN-γ-secreting T cell responses against peptide 2 and peptide 12 were quantitated by ELISpot. Bars represent the means ± SD from three individual experiments conducted in duplicate. *, P < 0.05.

Journal: Infection and Immunity

Article Title: Development of Melioidosis Subunit Vaccines Using an Enzymatically Inactive Burkholderia pseudomallei AhpC

doi: 10.1128/iai.00222-22

Figure Lengend Snippet: Identification and characterization of T cell-restricted epitopes associated with AhpC C57G . C57BL/6 mice ( n = 12) were immunized on days 0, 21, and 35 with AhpC C57G . Spleens ( n = 4 per analyte) were harvested on day 42, and (a) IFN-γ-, (b) IL-5-, and (c) IL-17-secreting T cell responses against an AhpC C57G peptide library (15-mers overlapping by 10 amino acids) were quantitated by ELISpot. Black dots represent the means of assays conducted in duplicate for individual mice. SFC, spot-forming cells. (d) C57BL/6 mice ( n = 3) were immunized on days 0, 21, and 35 with AhpC C57G . Spleens harvested on day 42 were combined and used to prepare three equal pools of cells. The pools were either depleted of CD8 + cells or CD4 + cells or left untreated, and IFN-γ-secreting T cell responses against peptide 2 and peptide 12 were quantitated by ELISpot. Bars represent the means ± SD from three individual experiments conducted in duplicate. *, P < 0.05.

Article Snippet: Mouse IFN-γ, IL-5, and IL-17 ImmunoSpot ELISpot kits (Cellular Technology, Ltd.) were used per the manufacturer’s instructions.

Techniques: Enzyme-linked Immunospot

Characterization of cellular immune responses raised against AhpC C57G . C57BL/6 mice ( n = 4 per group) were immunized on days 0, 21, and 35 with CPS-CRM197 plus AhpC C57G . Spleens were harvested on day 42, and (a) IFN-γ-, (b) IL-5-, and (c) IL-17-secreting T cell responses against AhpC C57G , peptide 2, and peptide 12 were quantitated by ELISpot. White and black dots represent the means of assays conducted in duplicate for individual mice. Black bars represent geometric means for each group. *, ( P < 0.05).

Journal: Infection and Immunity

Article Title: Development of Melioidosis Subunit Vaccines Using an Enzymatically Inactive Burkholderia pseudomallei AhpC

doi: 10.1128/iai.00222-22

Figure Lengend Snippet: Characterization of cellular immune responses raised against AhpC C57G . C57BL/6 mice ( n = 4 per group) were immunized on days 0, 21, and 35 with CPS-CRM197 plus AhpC C57G . Spleens were harvested on day 42, and (a) IFN-γ-, (b) IL-5-, and (c) IL-17-secreting T cell responses against AhpC C57G , peptide 2, and peptide 12 were quantitated by ELISpot. White and black dots represent the means of assays conducted in duplicate for individual mice. Black bars represent geometric means for each group. *, ( P < 0.05).

Article Snippet: Mouse IFN-γ, IL-5, and IL-17 ImmunoSpot ELISpot kits (Cellular Technology, Ltd.) were used per the manufacturer’s instructions.

Techniques: Enzyme-linked Immunospot

PBMC from 45 donors was tested in ELISPOT assays without adding HCMV (antigen) or in the presence of UV-inactivated HCMV virions. The assay was performed as described in Materials and Methods with 250,000 PBMC per well and 50 μg/mL of HCMV antigen. ( A ) Representative well images of one donor for medium control and antigen-specific IFN-γ, IL-2, IL-4, and IL-17 assays are shown at the time point of peak secretion; ( B ) PBMC was stimulated with HCMV in ELISPOT assays for 24 h, 48 h, and 72 h. IFN-γ, IL-2, IL-4 and IL-17 responses exhibited as spot forming units (SFU) were recorded at each time point. ( n = 3) ( C ) Maximal number of IFN-γ spots was observed at 24 h. At later time points (72 h) the ELISPOT image was over developed.

Journal: Viruses

Article Title: Characterization of the HCMV-Specific CD4 T Cell Responses that Are Associated with Protective Immunity

doi: 10.3390/v7082828

Figure Lengend Snippet: PBMC from 45 donors was tested in ELISPOT assays without adding HCMV (antigen) or in the presence of UV-inactivated HCMV virions. The assay was performed as described in Materials and Methods with 250,000 PBMC per well and 50 μg/mL of HCMV antigen. ( A ) Representative well images of one donor for medium control and antigen-specific IFN-γ, IL-2, IL-4, and IL-17 assays are shown at the time point of peak secretion; ( B ) PBMC was stimulated with HCMV in ELISPOT assays for 24 h, 48 h, and 72 h. IFN-γ, IL-2, IL-4 and IL-17 responses exhibited as spot forming units (SFU) were recorded at each time point. ( n = 3) ( C ) Maximal number of IFN-γ spots was observed at 24 h. At later time points (72 h) the ELISPOT image was over developed.

Article Snippet: The 96-well ELISPOT assays were performed using human interferon-γ ImmunoSpot ® kit (CTL-HIFNG-1/5M), IL-2 ImmunoSpot ® kit (CTL-HIL2-1/5), IL-4 ImmunoSpot ® kit (HIL4-1/5) and IL-17 ImmunoSpot ® kit (CTL-HIL17-1/5) as well as the IFN-γ /IL-2 double-color enzymatic kit (CTL-HIFNGIL2-1/5) from Cellular Technology Ltd. Plates and all antibodies, tertiaries, diluents, and substrate are contained in the kits.

Techniques: Enzyme-linked Immunospot

The spot size distribution for different cytokines follow Log Normal distribution. The experimental size distribution of typical recall responses are shown as histograms for the specific cytokines (IFN-g, IL-2, IL-4, and IL-17) with the theoretical Log Normal distribution curve overlaid in red. To test for normality, Kolmogorov-Smirnov goodness of fit was used.

Journal: Viruses

Article Title: Characterization of the HCMV-Specific CD4 T Cell Responses that Are Associated with Protective Immunity

doi: 10.3390/v7082828

Figure Lengend Snippet: The spot size distribution for different cytokines follow Log Normal distribution. The experimental size distribution of typical recall responses are shown as histograms for the specific cytokines (IFN-g, IL-2, IL-4, and IL-17) with the theoretical Log Normal distribution curve overlaid in red. To test for normality, Kolmogorov-Smirnov goodness of fit was used.

Techniques:

HCMV induced recall response is produced by CD4 cells for all cytokines. Unseparated PBMC, CD8 depleted PBMC, and CD4 depleted PBMC (2.5 × 10 5 cells each) were tested in human ELISPOT assays to detect the secretion of IFN-γ, IL-2, IL-4, and IL-17 at their appropriate time points as described in . CD4 and CD8 depletions were carried out by magnetic bead separations. ( A ) FACS analysis of the unseparated, CD8-depleted and CD4-depleted PBMC populations is shown; ( B ) Representative well images from one donor for the unseparated PBMC, CD8-depleted PBMC, and CD4-depleted PBMC with IFN-γ, IL-2, IL-4, and IL-17 recall responses, following activation with HCMV antigen for 24 h, 24 h, 48 h, and 72 h respectively is shown; ( C ) The mean and SD (Standard deviation) of cytokine spot forming units (SFU) from 2.5 × 10 5 unseparated PBMC (shown as solid bars), CD8-depleted PBMC (hatched bars), and CD4-depleted PBMC (empty bars) each are shown (n = 6).

Journal: Viruses

Article Title: Characterization of the HCMV-Specific CD4 T Cell Responses that Are Associated with Protective Immunity

doi: 10.3390/v7082828

Figure Lengend Snippet: HCMV induced recall response is produced by CD4 cells for all cytokines. Unseparated PBMC, CD8 depleted PBMC, and CD4 depleted PBMC (2.5 × 10 5 cells each) were tested in human ELISPOT assays to detect the secretion of IFN-γ, IL-2, IL-4, and IL-17 at their appropriate time points as described in . CD4 and CD8 depletions were carried out by magnetic bead separations. ( A ) FACS analysis of the unseparated, CD8-depleted and CD4-depleted PBMC populations is shown; ( B ) Representative well images from one donor for the unseparated PBMC, CD8-depleted PBMC, and CD4-depleted PBMC with IFN-γ, IL-2, IL-4, and IL-17 recall responses, following activation with HCMV antigen for 24 h, 24 h, 48 h, and 72 h respectively is shown; ( C ) The mean and SD (Standard deviation) of cytokine spot forming units (SFU) from 2.5 × 10 5 unseparated PBMC (shown as solid bars), CD8-depleted PBMC (hatched bars), and CD4-depleted PBMC (empty bars) each are shown (n = 6).

Techniques: Produced, Enzyme-linked Immunospot, Activation Assay, Standard Deviation

ELISPOT responses for IFN-γ, IL-2, IL-4, and IL-17. PBMC from 45 donors were plated at 250,000 cells/well and 50 μg/mL of HCMV antigen was added to induce a recall response for INF-γ (blue bars), IL-2 (red bars), IL-4 (green bars), and IL-17 (purple bars) from CD4 cells. SFU for each cytokine was established. SFU greater than 100 spots per well are not shown here. Raw data is available as part of Table S1.

Journal: Viruses

Article Title: Characterization of the HCMV-Specific CD4 T Cell Responses that Are Associated with Protective Immunity

doi: 10.3390/v7082828

Figure Lengend Snippet: ELISPOT responses for IFN-γ, IL-2, IL-4, and IL-17. PBMC from 45 donors were plated at 250,000 cells/well and 50 μg/mL of HCMV antigen was added to induce a recall response for INF-γ (blue bars), IL-2 (red bars), IL-4 (green bars), and IL-17 (purple bars) from CD4 cells. SFU for each cytokine was established. SFU greater than 100 spots per well are not shown here. Raw data is available as part of Table S1.

Techniques: Enzyme-linked Immunospot

Cell number dependence of ELISPOT formation. PBMC were plated at 100,000 250,000, and 500,000 cells/well and 50 μg/mL of HCMV antigen was added to induce a recall response for INF-γ, IL-2, IL-4, and IL-17 from CD4 cells. SFU at the different PBMC numbers plated were established. Mean values and the regression coefficient (R 2 ) from four donors is shown.

Journal: Viruses

Article Title: Characterization of the HCMV-Specific CD4 T Cell Responses that Are Associated with Protective Immunity

doi: 10.3390/v7082828

Figure Lengend Snippet: Cell number dependence of ELISPOT formation. PBMC were plated at 100,000 250,000, and 500,000 cells/well and 50 μg/mL of HCMV antigen was added to induce a recall response for INF-γ, IL-2, IL-4, and IL-17 from CD4 cells. SFU at the different PBMC numbers plated were established. Mean values and the regression coefficient (R 2 ) from four donors is shown.

Techniques: Enzyme-linked Immunospot

Establishing optimal antigen dose for HCMV-specific CD4 cell stimulation. PBMC were plated at 250,000 cells per well and was stimulated with different concentrations of HCMV antigen starting from 100 μg/mL to 0.01 μg/mL. SFU recorded from IFN-γ, IL-2, IL-4, and IL-17 ELISPOT assays are shown for each of these antigen concentrations.

Journal: Viruses

Article Title: Characterization of the HCMV-Specific CD4 T Cell Responses that Are Associated with Protective Immunity

doi: 10.3390/v7082828

Figure Lengend Snippet: Establishing optimal antigen dose for HCMV-specific CD4 cell stimulation. PBMC were plated at 250,000 cells per well and was stimulated with different concentrations of HCMV antigen starting from 100 μg/mL to 0.01 μg/mL. SFU recorded from IFN-γ, IL-2, IL-4, and IL-17 ELISPOT assays are shown for each of these antigen concentrations.

Techniques: Cell Stimulation, Enzyme-linked Immunospot

Establishing cytokine signature of CD4 cells. PBMC from 45 healthy donors were tested for HCMV-induced cytokine production of IFN-γ, IL-2, IL-4, and IL-17. ( A ) The percentage of donors that generated a recall response to the HCMV antigen for each of the cytokines is shown. ( B ) The percentage of individual donors representing the various CD4 effector lineages is shown.

Journal: Viruses

Article Title: Characterization of the HCMV-Specific CD4 T Cell Responses that Are Associated with Protective Immunity

doi: 10.3390/v7082828

Figure Lengend Snippet: Establishing cytokine signature of CD4 cells. PBMC from 45 healthy donors were tested for HCMV-induced cytokine production of IFN-γ, IL-2, IL-4, and IL-17. ( A ) The percentage of donors that generated a recall response to the HCMV antigen for each of the cytokines is shown. ( B ) The percentage of individual donors representing the various CD4 effector lineages is shown.

Techniques: Generated

Longitudinal studies of donors show similarity in responses over multiple years. PBMC from six different donors were isolated at different time points over years. Some donors were tested multiple times within 1 year (Donor ID 13, 41, 44), others within 3 years (ID 43, 45) and one donor was tested over 5 years (ID 42). The dates at which PBMC was isolated is specified in the figure for each donor. PBMC isolated from each donor at the different time points, at 250,000 per well, was activated with 50 μg/mL HCMV antigen and the cytokine response for IFN-γ, IL-2, IL-4, and IL-17 was determined. Mean and SD of SFU from three repetitive wells are shown.

Journal: Viruses

Article Title: Characterization of the HCMV-Specific CD4 T Cell Responses that Are Associated with Protective Immunity

doi: 10.3390/v7082828

Figure Lengend Snippet: Longitudinal studies of donors show similarity in responses over multiple years. PBMC from six different donors were isolated at different time points over years. Some donors were tested multiple times within 1 year (Donor ID 13, 41, 44), others within 3 years (ID 43, 45) and one donor was tested over 5 years (ID 42). The dates at which PBMC was isolated is specified in the figure for each donor. PBMC isolated from each donor at the different time points, at 250,000 per well, was activated with 50 μg/mL HCMV antigen and the cytokine response for IFN-γ, IL-2, IL-4, and IL-17 was determined. Mean and SD of SFU from three repetitive wells are shown.

Techniques: Isolation

Restoration of functionality of exhausted CD4 cells with IL-7. PBMC from 40 donors were tested in an ELISPOT assay with 50 μg/mL of HCMV antigen, with and without IL-7 added to the culture. Cytokine recall response for IFN-γ, IL-2, IL-4, and IL-17 was recorded as SFU. PBMC were plated at 250,000 cells and the concentration of IL-7 was 30 ng/mL. The SFU with and without IL-7 for each of the cytokines is shown.

Journal: Viruses

Article Title: Characterization of the HCMV-Specific CD4 T Cell Responses that Are Associated with Protective Immunity

doi: 10.3390/v7082828

Figure Lengend Snippet: Restoration of functionality of exhausted CD4 cells with IL-7. PBMC from 40 donors were tested in an ELISPOT assay with 50 μg/mL of HCMV antigen, with and without IL-7 added to the culture. Cytokine recall response for IFN-γ, IL-2, IL-4, and IL-17 was recorded as SFU. PBMC were plated at 250,000 cells and the concentration of IL-7 was 30 ng/mL. The SFU with and without IL-7 for each of the cytokines is shown.

Techniques: Enzyme-linked Immunospot, Concentration Assay

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