14-9948-82

antibody from Invitrogen Antibodies

Targeting: ICOS AILIM, CD278

Provider product page for 14-9948-82

Immunoprecipitation

Immunohistochemistry

Flow cytometry

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Antibody data

Antibody Data
Antigen structure
References [20]
Comments [0]
Validations
Flow cytometry [1]
Other assay [14]

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Product number: 14-9948-82 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD278 (ICOS) Monoclonal Antibody (ISA-3), eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The ISA-3 monoclonal antibody reacts with human ICOS (Inducible COStimulatory molecule), also known as H4, CRP-1 and AILIM. ICOS is a T cell specific activation molecule and a third member of the CD28/CTLA-4 family. Human ICOS has a relative molecular mass of 55-60 kDa, composed of 27 kDa and 29 kDa chains. Human ICOS on activated T cells has potent costimulatory activity for T cell activation and is required for humoral immune responses, in particular for memory B cell and plasma cell generation. ICOS binds to its ligand, B7h/B7RP-1 expressed on activated APCs (antigen presenting cells) and on a number of inflamed peripheral tissues. Plate-bound ISA-3 is costimulatory for T cells and induces production of IL-4, IL-5, IL-10 and other cytokines, but not IL-2. ISA-3 has the same reactivity pattern and characteristics as F44. ISA-3 was generated against the human ICOS antigen. C398.4A, anti-mouse ICOS/H4
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: ISA-3
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: 4° C

ICOS protein structure - 14-9948-82 shown in red.

Supportive validation

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Experimental details: NULL

Supportive validation

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Experimental details: NULL

Supportive validation

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Experimental details: NULL

Supportive validation

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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 SARS-CoV-2 Infection Induces Durable, Functional Spike-Reactive CD4 + T Cells (A) Representative flow cytometry plots of ICOS and CD40L expression on antigen-experienced (non-CD45RA + CCR7 + ) CD4 + T cells 20 h after incubation of PBMCs from HC and CoV2 + individuals at V1 and V2 with vehicle or SARS-CoV-2 spike. (B) Number of antigen-experienced ICOS + CD40L + CD4 + T cells per 1x10 6 CD4 + T cells from HC and CoV2 + samples after incubation with vehicle (Veh.) or spike (S) at both time points (right) and calculated number of spike-responsive CD4 + T cells (number after incubation with spike minus number after incubation with vehicle) compared across time points (left). (C) Number of antigen-experienced CXCR5 + ICOS + CD40L + CD4 + T cells (cTfh) per 1 x 10 6 CD4 + T cells from HC and CoV2 + samples after incubation with Veh. or S at both time points (right) and calculated number of spike-responsive cTfh cells (number after incubation with spike minus number after incubation with vehicle) compared across time points (left). (D) Representative flow cytometry plots of sorted CD4 + naive (CD45RA + CCR7 + ), T CM (CD45RA - CCR7 + ), or T EM (CD45RA - CCR7 - ) T cells from HC and CoV2 + PBMCs after 5-6 days of co-culture with SARS-CoV-2 spike-protein-pulsed autologous monocytes and measuring proliferation by CPD dilution. (E) SARS-CoV-2 spike-specific expansion of sorted CD4 + naive T, T CM , and T EM cells from V1 (circles) and V2 (squares) reported as frequency of CXC

Supportive validation

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Experimental details: Figure 1 T FH cell compartment was altered during chronic HBV infection. (A, B) Representative flow cytometry plot indicates the percent frequencies of circulating T FH cells, PD-1 as well as ICOS expression in HC-vacc (n = 21) and CHB patients (n = 32) respectively. Throughout the flow cytometry plots, gates were placed based on single-color controls. (C, D) T FH cell subtypes were analyzed on the basis of expression of different cell surface molecules. Values in the quadrant designate the percent frequency of different T FH cell subsets. Cumulative data has been shown in scatter plot (HC-vacc: n = 21 and CHB: n = 32) and expressed as median. Each symbol on scatter plot represents an individual subject. Statistical analysis was performed using non-parametric, two-tailed Mann-Whitney U test. (E) Frequency of T FH cells in HBV patients with different ALT and HBV DNA levels, HBeAg-negative and positive patients was analyzed. Levels of ALT were divided as normal (range, 22-40; n=21) vs. raised (range, 41-193; n=15)). HBV DNA levels were divided as 10 4 (n = 8). Unpaired t-test was used for statistical significance.

Supportive validation

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Experimental details: Figure 1. Robust haemagglutinin (HA)-specific CD4 + T cells' response to seasonal influenza vaccination. ( A ) Overview of cohort characteristics, vaccination strains, and immune variables measured before (d0), 7 days (d7), and 42 days (d42) after seasonal influenza vaccination. ( B ) IgG responses to HA proteins from vaccine influenza strains measured by Luminex. Dashed line indicates limit of detection in Luminex assay. ( C ) Log 2 fold change versus -log10 false discovery rate (FDR)-adjusted p-value of Flu HA Luminex, Cytokine Luminex, B cell and CD4 + T cell immunophenotyping parameters before and after vaccination at indicated time points for cohort 1. Dashed line represents the p-value cut-off at FDR-adjusted p=0.05. ( D ) Representative flow cytometry plots of HA 257-276 HLADR*0701 staining on CD4 + CD45RA - cells, gate corresponds to HA-specific TET + T cells, and ( E ) the frequency of HA-specific TET + T cells among all CD4 + CD45RA - cells on d0 compared to d7. ( F ) Representative ICOS staining and ( G ) the percentage of ICOS + cells among HA-specific TET + T cells on d0 and d7. ( H ) CXCR5 and PD1 staining on HA-specific TET + T cells on d0 and d7, gate corresponds to 'TET + cTfh' T cells, and ( I ) the percentage of circulating T follicular helper (cTfh) cells among HA-specific TET + T cells on d0 and d7 for each cohort. In parts ( D-I ), cohort 1 n = 16; cohort 2 n = 19. Paired p-values determined using Wilcoxon signed-rank test. Figure 1--figure supplement 1.

Supportive validation

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Experimental details: Inducible costimulator (ICOS) signaling induces C-X-C motif chemokine receptor 3 (CXCR3) expression in vivo and in vitro. ( A ) Representative flow cytometric dot plots of the frequencies of CXCR3 + CD4 + T cells and CXCR3 + CD4 + CD25 + FOXP3 + Tregs within patients with stable chronic obstructive pulmonary disease (SCOPD). Comparisons of the percentage of CXCR3 + T cells within CD4 + T cells ( B ) and CD4 + CD25 + FOXP3 + Tregs ( C ) in the peripheral blood from 15 healthy controls (HC), 11 healthy smokers (HS), 22 patients with SCOPD (SCOPD), and 14 patients with acute exacerbation of COPD (AECOPD). ( D ) Representative gating of CXCR3 + ICOS + CD4 + T cells and CXCR3 + ICOS - CD4 + T cells and the corresponding histograms (n = 22). ( E ) Naive CD4 + T cells obtained from the peripheral blood of healthy people were cultured with plate-bound anti-CD3/28 mAbs in the presence of IL-2 with or without anti-ICOS mAb (ISA-3) for four days. The frequency of CXCR3 + CD4 + T cells was analyzed with flow cytometry. Comparable results were obtained from six independent experiments. ( F ) The percentage of CXCR3 + CD4 + T cells in the peripheral blood and corresponding BALF of four patients with SCOPD. *p

Supportive validation

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Experimental details: Figure 3. A GLA-SE adjuvanted vaccine promotes cTfh expansion. (A and B) Flow cytometric contour plots of (A) PD-1 and CXCR5 on total CD45RA - CD4 + CD3 + cells and (B) ICOS and CD38 on the population gated in A on peripheral blood cells from individuals 7 d after the third P27A vaccination. Alum group, n = 7; GLA-SE group, n = 8. (C) Fold change of CD38 + ICOS + CXCR5 + PD-1 + cTfh cells 7 d after the third P27A vaccination (frequency of cTfh cells at day 63/baseline). Alum group, n = 7; GLA-SE, group n = 8. P = 0.027. The P value is from an unpaired Student's t test. (D) Correlation of anti-P27A IgG antibody titer 28 d after the third vaccination against fold change in CD38 + ICOS + CXCR5 + PD-1 + cTfh cells 7 d after the third vaccination. Rho = 0.73; P = 0.003 by Spearman's correlation (Rho = coefficient); n = 15. (E) Scatterplot of RNA-sequencing data comparing all genes expressed in CD38 + ICOS + CXCR5 + PD-1 + cTfh cells 7 d after the third P27A vaccination in either Alum- (x axis) or GLA-SE- (y axis) vaccinated groups. Alum group, n = 7; GLA-SE group, n = 8. (F-I) Bar plots showing log-normalized reads from RNA-sequencing data of CD40LG . P = 0.593 (F), IL21 P = 0.846 (G), CXCR3 P = 0.606 (H), and CCR6 P = 0.977 (I) for transcripts in ICOS in CD38 + ICOS + CXCR5 + PD-1 + cTfh cells 7 d after the third P27A vaccination; Alum group, n = 7; GLA-SE group, n = 8. P values were calculated using DESeq2 with a false discovery rate corr