Structured Review

Nycomed cd4 t cells
Suppression of CD4 +  CD25 –  T cells by flow cytometrically sorted CD4 +  CD25 hi  regulatory T cells from individuals with systemic lupus erythematosus (SLE). Fluorescence activated cell sorter-separated CD25 –  T cells were incubated at increasing ratios, from 1 : 1 to 4 : 1, with the 2% of CD4 +  T cells expressing the highest level of CD25, and stimulated with anti-CD3/anti-CD28-coated beads. The  x -axis shows the CD25 hi  population co-cultured with the indicated CD25 –  cells at ratios from 1 : 1 to 4 : 1. Percentage suppression was calculated using the equation 100-[(counts per minute CD25 – /CD25 hi  co-culture)/counts per minute CD25 –  alone] × (100). Bars at medians. There was no statistically significant difference between suppression at any ratios between individuals with SLE and controls: Kruskal–Wallis test with Dunn's multiple post-test comparison. (a) Suppression of CD25 –  cells from individuals with inactive lupus by autologous CD4 +  CD25 hi  T cells and CD4 +  CD25 hi  T cells from controls. (b) Suppression of CD25 –  cells from controls by autologous CD4 +  CD25 hi  T cells and CD4 +  CD25 hi  T cells from individuals with inactive lupus. (c) Suppression of CD25 –  cells from individuals with active lupus by autologous CD4 +  CD25 hi  T cells and CD4 +  CD25 hi  T cells from controls. (d) Suppression of CD25 –  cells from controls by autologous CD4 +  CD25 hi  T cells and CD4 +  CD25 hi  T cells from individuals with active lupus.
Cd4 T Cells, supplied by Nycomed, used in various techniques. Bioz Stars score: 92/100, based on 635 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/cd4 t cells/product/Nycomed
Average 92 stars, based on 635 article reviews
Price from $9.99 to $1999.99
cd4 t cells - by Bioz Stars, 2020-09
92/100 stars

Images

1) Product Images from "Natural regulatory T cells: number and function are normal in the majority of patients with lupus nephritis"

Article Title: Natural regulatory T cells: number and function are normal in the majority of patients with lupus nephritis

Journal: Clinical and Experimental Immunology

doi: 10.1111/j.1365-2249.2008.03665.x

Suppression of CD4 +  CD25 –  T cells by flow cytometrically sorted CD4 +  CD25 hi  regulatory T cells from individuals with systemic lupus erythematosus (SLE). Fluorescence activated cell sorter-separated CD25 –  T cells were incubated at increasing ratios, from 1 : 1 to 4 : 1, with the 2% of CD4 +  T cells expressing the highest level of CD25, and stimulated with anti-CD3/anti-CD28-coated beads. The  x -axis shows the CD25 hi  population co-cultured with the indicated CD25 –  cells at ratios from 1 : 1 to 4 : 1. Percentage suppression was calculated using the equation 100-[(counts per minute CD25 – /CD25 hi  co-culture)/counts per minute CD25 –  alone] × (100). Bars at medians. There was no statistically significant difference between suppression at any ratios between individuals with SLE and controls: Kruskal–Wallis test with Dunn's multiple post-test comparison. (a) Suppression of CD25 –  cells from individuals with inactive lupus by autologous CD4 +  CD25 hi  T cells and CD4 +  CD25 hi  T cells from controls. (b) Suppression of CD25 –  cells from controls by autologous CD4 +  CD25 hi  T cells and CD4 +  CD25 hi  T cells from individuals with inactive lupus. (c) Suppression of CD25 –  cells from individuals with active lupus by autologous CD4 +  CD25 hi  T cells and CD4 +  CD25 hi  T cells from controls. (d) Suppression of CD25 –  cells from controls by autologous CD4 +  CD25 hi  T cells and CD4 +  CD25 hi  T cells from individuals with active lupus.
Figure Legend Snippet: Suppression of CD4 + CD25 – T cells by flow cytometrically sorted CD4 + CD25 hi regulatory T cells from individuals with systemic lupus erythematosus (SLE). Fluorescence activated cell sorter-separated CD25 – T cells were incubated at increasing ratios, from 1 : 1 to 4 : 1, with the 2% of CD4 + T cells expressing the highest level of CD25, and stimulated with anti-CD3/anti-CD28-coated beads. The x -axis shows the CD25 hi population co-cultured with the indicated CD25 – cells at ratios from 1 : 1 to 4 : 1. Percentage suppression was calculated using the equation 100-[(counts per minute CD25 – /CD25 hi co-culture)/counts per minute CD25 – alone] × (100). Bars at medians. There was no statistically significant difference between suppression at any ratios between individuals with SLE and controls: Kruskal–Wallis test with Dunn's multiple post-test comparison. (a) Suppression of CD25 – cells from individuals with inactive lupus by autologous CD4 + CD25 hi T cells and CD4 + CD25 hi T cells from controls. (b) Suppression of CD25 – cells from controls by autologous CD4 + CD25 hi T cells and CD4 + CD25 hi T cells from individuals with inactive lupus. (c) Suppression of CD25 – cells from individuals with active lupus by autologous CD4 + CD25 hi T cells and CD4 + CD25 hi T cells from controls. (d) Suppression of CD25 – cells from controls by autologous CD4 + CD25 hi T cells and CD4 + CD25 hi T cells from individuals with active lupus.

Techniques Used: Flow Cytometry, Fluorescence, Incubation, Expressing, Cell Culture, Co-Culture Assay

A higher proportion of CD4 +  T cells express CD25 in patients with active systemic lupus erythematosus (SLE), although the proportion of CD25 hi  remains similar to controls. Peripheral blood mononuclear cells were stained with anti-CD4-APC and an IgG1-PE isotype control antibody to establish levels of background staining. Lymphocytes were gated using forward-scatter/side-scatter characteristics. (a) Representative example of a PE-labelled isotype control and anti-CD25 staining from a control individual. (b) Staining pattern of an individual with active SLE with a particularly marked increase in the proportion of CD4 +  T cells expressing CD25 (1·4%). Note also the marked reduction in the proportion of lymphocytes that are CD4 +  (8.88%  versus  37% in a). (c) Comparing 18 controls, 12 patients with active SLE and six with inactive disease, there was a significant increase in the percentage of CD25 +  in those with active disease.  P
Figure Legend Snippet: A higher proportion of CD4 + T cells express CD25 in patients with active systemic lupus erythematosus (SLE), although the proportion of CD25 hi remains similar to controls. Peripheral blood mononuclear cells were stained with anti-CD4-APC and an IgG1-PE isotype control antibody to establish levels of background staining. Lymphocytes were gated using forward-scatter/side-scatter characteristics. (a) Representative example of a PE-labelled isotype control and anti-CD25 staining from a control individual. (b) Staining pattern of an individual with active SLE with a particularly marked increase in the proportion of CD4 + T cells expressing CD25 (1·4%). Note also the marked reduction in the proportion of lymphocytes that are CD4 + (8.88% versus 37% in a). (c) Comparing 18 controls, 12 patients with active SLE and six with inactive disease, there was a significant increase in the percentage of CD25 + in those with active disease. P

Techniques Used: Staining, Expressing

Phenotypic comparison of CD25 subsets. Active lupus is associated with phenotypic changes in the CD25 –  subset. Peripheral blood mononuclear cells were triple-stained for CD4, CD25 and expression of markers associated with CD4 +  CD25 hi  T regs  or T cell activation. Subsets were gated using CD25 expression level and the proportion of each subset expressing the marker determined by comparison with staining with the isotype control antibody. Values shown are median percentage positive, with figures in brackets the 25th and 75th percentiles. Eleven controls, 11 with active systemic lupus erythematosus, five with inactive disease. (a) CD4 +  T cells expressing CD25 at levels above the upper limit of CD25 expression by CD4 –  lymphocytes. (b) The 2% CD4 +  T cells expressing the highest CD25 levels (definition of CD25 hi  used for FACS separation). (c) CD25 int . (d) CD25 – . CCR7 from 10 controls, nine patients with active and five with inactive disease. ×:  P -value comparing controls and patients with inactive lupus; ♦,  P -value comparing controls and patients with active lupus, †:  P -value comparing patients with active and inactive lupus. Statistical comparison was made using the Kruskal–Wallis test with Dunn's post-test. FACS, fluorescence activated cell sorter; HLA-DR, human leucocyte antigen D-related.
Figure Legend Snippet: Phenotypic comparison of CD25 subsets. Active lupus is associated with phenotypic changes in the CD25 – subset. Peripheral blood mononuclear cells were triple-stained for CD4, CD25 and expression of markers associated with CD4 + CD25 hi T regs or T cell activation. Subsets were gated using CD25 expression level and the proportion of each subset expressing the marker determined by comparison with staining with the isotype control antibody. Values shown are median percentage positive, with figures in brackets the 25th and 75th percentiles. Eleven controls, 11 with active systemic lupus erythematosus, five with inactive disease. (a) CD4 + T cells expressing CD25 at levels above the upper limit of CD25 expression by CD4 – lymphocytes. (b) The 2% CD4 + T cells expressing the highest CD25 levels (definition of CD25 hi used for FACS separation). (c) CD25 int . (d) CD25 – . CCR7 from 10 controls, nine patients with active and five with inactive disease. ×: P -value comparing controls and patients with inactive lupus; ♦, P -value comparing controls and patients with active lupus, †: P -value comparing patients with active and inactive lupus. Statistical comparison was made using the Kruskal–Wallis test with Dunn's post-test. FACS, fluorescence activated cell sorter; HLA-DR, human leucocyte antigen D-related.

Techniques Used: Staining, Expressing, Activation Assay, Marker, FACS, Fluorescence

2) Product Images from "Interleukin-33 promoting Th1 lymphocyte differentiation dependents on IL-12"

Article Title: Interleukin-33 promoting Th1 lymphocyte differentiation dependents on IL-12

Journal: Immunobiology

doi: 10.1016/j.imbio.2015.11.013

IL-33 enhances Th1 cell polarization  via  ST2 and IL-12. CD4 +  T cells from WT, ST2 −/−  mice (A) or human cord blood (B) were stimulated with anti-CD3 Abs with or without IL-33 (10 ng/ml) or IL-12(10 ng/ml) for 72 h. Supernatants IFN-γ and IL-4 concentrations were measured by ELISA. (C) Human CD4 +  T cells were stimulated as above and intracellular cytokines were determined using FACScan. Data are presented as mean ± SD,  n  = 5 mice/group, and are representative of three independent experiments.  ∗ p
Figure Legend Snippet: IL-33 enhances Th1 cell polarization via ST2 and IL-12. CD4 + T cells from WT, ST2 −/− mice (A) or human cord blood (B) were stimulated with anti-CD3 Abs with or without IL-33 (10 ng/ml) or IL-12(10 ng/ml) for 72 h. Supernatants IFN-γ and IL-4 concentrations were measured by ELISA. (C) Human CD4 + T cells were stimulated as above and intracellular cytokines were determined using FACScan. Data are presented as mean ± SD, n  = 5 mice/group, and are representative of three independent experiments. ∗ p

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

Mechanism by which IL-33 enhances Th1 cell development  in vitro . CD4 +  T cells from WT or MyD88 −/−  mice were stimulated with anti-CD3 Abs in the presence or absence of IL-33, IL-12 or a combination of the cytokines. The supernatants were collected for cytokine measurement by ELISA after 72 h culture (A) and cells collected for the detection of T-bet and GATA3 (B), st2 and IL-12R (C, D) by qPCR. Data are presented as mean ± SD,  n  = 5 pooled mice/group and are representative of three independent experiments;  ∗ p
Figure Legend Snippet: Mechanism by which IL-33 enhances Th1 cell development in vitro . CD4 + T cells from WT or MyD88 −/− mice were stimulated with anti-CD3 Abs in the presence or absence of IL-33, IL-12 or a combination of the cytokines. The supernatants were collected for cytokine measurement by ELISA after 72 h culture (A) and cells collected for the detection of T-bet and GATA3 (B), st2 and IL-12R (C, D) by qPCR. Data are presented as mean ± SD, n  = 5 pooled mice/group and are representative of three independent experiments; ∗ p

Techniques Used: In Vitro, Mouse Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

3) Product Images from "Relationship between functional ability in older people, immune system status, and intensity of response to CMV"

Article Title: Relationship between functional ability in older people, immune system status, and intensity of response to CMV

Journal: Age

doi: 10.1007/s11357-011-9240-6

CD69 expression and proliferative capacity of CD4+ and CD8+ depending on the functional capacity. Whole blood from the BI groups of elders (group 0 + 1,  n  = 12 and group 2 + 3,  n  = 14) was stimulated for 18 h and expression of CD69 in CD4+ and CD8+ T cells was evaluated by flow cytometry. Proliferative capacity of CD4+ and CD8+ T cells subsets was also evaluated in the two groups (group 0 + 1,  n  = 19 and group 2 + 3,  n  = 16) by labeling the PBMC with CFSE. Cells were stained and 1 × 10 5  cells were acquired per experiment.  a  Representative dot plots showing the frequency of CD69 expression in CD4+ and CD8+ subset from elderly with different functional status. Cells were stimulated using anti-CD3 (10 ng/mL). Percentage of positive cells in each subpopulation in this representative experiment is expressed in the  upper right corner  and summarized results from all donors (median and IR) were also expressed in dot plots.  b  Proliferative capacity of CD4+ and CD8+ T cells subsets in response to anti-CD3. PBMC were labeled with CFSE (1.5 μM) and cultured in presence of anti-CD3 (10 ng/mL) for 5 days. Percentage of dividing CD4+ and CD8+ T cells is represented.  Bars  represent results from the grouped elders (mean ± SEM).  c  Expression of CD69 into de CD4+ and CD8+ T cell subset was analyzed in the same way as in Fig. 5a in response to a CMV supernatant (10 4  PFU/mL).  d  Proliferative capacity of CD4+ and CD8+ T-cell subsets in response to the CMV supernatant.  Bars  represent resulted from the grouped elders (mean ± SEM). The Student’s  t  test (when data were normally distributed) and Mann–Whitney non-parametric (when data were not normally distributed) methods were used to compare frequencies between groups.  p  values are depicted in the panels
Figure Legend Snippet: CD69 expression and proliferative capacity of CD4+ and CD8+ depending on the functional capacity. Whole blood from the BI groups of elders (group 0 + 1, n  = 12 and group 2 + 3, n  = 14) was stimulated for 18 h and expression of CD69 in CD4+ and CD8+ T cells was evaluated by flow cytometry. Proliferative capacity of CD4+ and CD8+ T cells subsets was also evaluated in the two groups (group 0 + 1, n  = 19 and group 2 + 3, n  = 16) by labeling the PBMC with CFSE. Cells were stained and 1 × 10 5  cells were acquired per experiment. a Representative dot plots showing the frequency of CD69 expression in CD4+ and CD8+ subset from elderly with different functional status. Cells were stimulated using anti-CD3 (10 ng/mL). Percentage of positive cells in each subpopulation in this representative experiment is expressed in the upper right corner and summarized results from all donors (median and IR) were also expressed in dot plots. b Proliferative capacity of CD4+ and CD8+ T cells subsets in response to anti-CD3. PBMC were labeled with CFSE (1.5 μM) and cultured in presence of anti-CD3 (10 ng/mL) for 5 days. Percentage of dividing CD4+ and CD8+ T cells is represented. Bars represent results from the grouped elders (mean ± SEM). c Expression of CD69 into de CD4+ and CD8+ T cell subset was analyzed in the same way as in Fig. 5a in response to a CMV supernatant (10 4  PFU/mL). d Proliferative capacity of CD4+ and CD8+ T-cell subsets in response to the CMV supernatant. Bars represent resulted from the grouped elders (mean ± SEM). The Student’s  t test (when data were normally distributed) and Mann–Whitney non-parametric (when data were not normally distributed) methods were used to compare frequencies between groups. p values are depicted in the panels

Techniques Used: Expressing, Functional Assay, Flow Cytometry, Cytometry, Labeling, Staining, Cell Culture, MANN-WHITNEY

Immune phenotype in peripheral blood from elderly according to their BI group. Elders were stratified according to their BI (group 0 BI = 100, group 1 BI = 95–80, group 2 BI = 75–40, and group 3 BI = 35–0). The number of donors in each group was group 0 = 24, group 1 = 26, group 2 = 27, and group 3 = 23. Whole blood from elderly individuals was stained with different antibody combinations and analyzed by flow cytometry (10 5  cells acquired in each experiment). Outlier values were represented by  circles  and extreme values by  stars , calculated by adding 1.5 and 3 times the IR to the 75th percentile, respectively. The ANOVA test (when data were normally distributed) and Kruskal–Wallis non-parametric methods (when data were not normally distributed) were used to compare frequencies between groups.  p  values are depicted in the panels.  a  Percentages of CD16+56+ and CD19+ cells with respect to the total CD45+ cells were compared between the four groups of elderly. Staining was performed with “Multiset CD3-FITC/CD16+56-PE/CD45-PerCP/CD19-APC” and frequencies of CD16+56 and CD19+ cells in gated CD45+ subsets were analyzed.  b  Percentages of CD4+ and CD8+ cells were analyzed with respect to the total CD45+CD3+ and were compared between the four groups. Staining was performed with anti-CD3-FITC, anti-CD4-APC, and anti-CD8-PerCP to gate CD4+ and CD8+ T cells.  c  Percentages of CD4+ T cells expressing CD8 and expressing NKG2D in peripheral blood from elderly. Whole blood was stained with anti-CD3-FITC, anti-NKG2D-PE, anti-CD8-PerCP, and anti-CD4-APC. Frequencies of NKG2D+ cells in gated CD3+CD4+ lymphocytes were quantified
Figure Legend Snippet: Immune phenotype in peripheral blood from elderly according to their BI group. Elders were stratified according to their BI (group 0 BI = 100, group 1 BI = 95–80, group 2 BI = 75–40, and group 3 BI = 35–0). The number of donors in each group was group 0 = 24, group 1 = 26, group 2 = 27, and group 3 = 23. Whole blood from elderly individuals was stained with different antibody combinations and analyzed by flow cytometry (10 5 cells acquired in each experiment). Outlier values were represented by circles and extreme values by stars , calculated by adding 1.5 and 3 times the IR to the 75th percentile, respectively. The ANOVA test (when data were normally distributed) and Kruskal–Wallis non-parametric methods (when data were not normally distributed) were used to compare frequencies between groups. p values are depicted in the panels. a Percentages of CD16+56+ and CD19+ cells with respect to the total CD45+ cells were compared between the four groups of elderly. Staining was performed with “Multiset CD3-FITC/CD16+56-PE/CD45-PerCP/CD19-APC” and frequencies of CD16+56 and CD19+ cells in gated CD45+ subsets were analyzed. b Percentages of CD4+ and CD8+ cells were analyzed with respect to the total CD45+CD3+ and were compared between the four groups. Staining was performed with anti-CD3-FITC, anti-CD4-APC, and anti-CD8-PerCP to gate CD4+ and CD8+ T cells. c Percentages of CD4+ T cells expressing CD8 and expressing NKG2D in peripheral blood from elderly. Whole blood was stained with anti-CD3-FITC, anti-NKG2D-PE, anti-CD8-PerCP, and anti-CD4-APC. Frequencies of NKG2D+ cells in gated CD3+CD4+ lymphocytes were quantified

Techniques Used: Staining, Flow Cytometry, Cytometry, Expressing

TREC content in CD4+ T cells, its correlation with the NAÏVE subset and Ki-67 quantification.  a  The TREC content was measured in CD4+ T cells from elders belonging to group 0 + 1 ( n  = 14) and group 2 + 3 ( n  = 15). CD4+ population was isolated by magnetic bead separation and the TREC copy number was determined by real-time PCR. Experiments were conducted in duplicate and  bars  represented results from the grouped elders (mean ± SEM).  b  Relationship between TREC content and NAÏVE (CD4+CCR7+CD45RA+) subset in the four groups of elderly was analyzed. The correlations,  p  value, and coefficient of correlation were calculated by using the non-parametric Spearman test and are listed in the  upper left hand corner .  c  The quantification of Ki-67 was performed in CD4+ and CD8+ T cells from elders belonging to group 0 + 1 ( n  = 10) and group 2 + 3 ( n  = 10). CD4+ and CD8+ populations were isolated by magnetic bead separation and the Ki-67 quantification was determined by intracellular staining and flow cytometry. Representative dot plots show the frequency of Ki-67 expression in CD4+ and CD8+ subsets from elderly with different functional status. Percentage of positive cells in each subpopulation in these representative experiments are expressed in the  upper right corner . Appropriate isotype control mAbs were used for marker settings.  d  Histograms summarize the percentage of positive cells for Ki-67 (mean ± SEM). The Student’s  t  test method was used to compare frequencies between groups
Figure Legend Snippet: TREC content in CD4+ T cells, its correlation with the NAÏVE subset and Ki-67 quantification. a The TREC content was measured in CD4+ T cells from elders belonging to group 0 + 1 ( n  = 14) and group 2 + 3 ( n  = 15). CD4+ population was isolated by magnetic bead separation and the TREC copy number was determined by real-time PCR. Experiments were conducted in duplicate and bars represented results from the grouped elders (mean ± SEM). b Relationship between TREC content and NAÏVE (CD4+CCR7+CD45RA+) subset in the four groups of elderly was analyzed. The correlations, p value, and coefficient of correlation were calculated by using the non-parametric Spearman test and are listed in the upper left hand corner . c The quantification of Ki-67 was performed in CD4+ and CD8+ T cells from elders belonging to group 0 + 1 ( n  = 10) and group 2 + 3 ( n  = 10). CD4+ and CD8+ populations were isolated by magnetic bead separation and the Ki-67 quantification was determined by intracellular staining and flow cytometry. Representative dot plots show the frequency of Ki-67 expression in CD4+ and CD8+ subsets from elderly with different functional status. Percentage of positive cells in each subpopulation in these representative experiments are expressed in the upper right corner . Appropriate isotype control mAbs were used for marker settings. d Histograms summarize the percentage of positive cells for Ki-67 (mean ± SEM). The Student’s  t test method was used to compare frequencies between groups

Techniques Used: Isolation, Real-time Polymerase Chain Reaction, Staining, Flow Cytometry, Cytometry, Expressing, Functional Assay, Marker

Distribution of CD4+ and CD8+ T cells into naïve, central memory, effector memory and effector memory RA, and distribution of EM and EMRA in CD4+ and CD8+ T cells into subsets defined by CD28 and CD27 expression. Expression of CD45RA, CCR7, CD27, and CD28 was analyzed by flow cytometry in isolated CD4+ and CD8+ T cells from the four BI groups of elders. a Schematic model of the T-cell differentiation subsets accordingly to CD45RA and CCR7 expression. Whole blood was stained with anti-CD45RA-FICT, anti-CD8-PE, anti-CD4-PerCP, and anti-CCR7-APC, and 10 5 cells were acquired in each experiment. b Histograms represent percentage of cells in each subset (NAÏVE, CM, EM, and EMRA) in the four groups of elderly accordingly to their functional status (group 0— white bars , group 1— light gray bars , group 2— dark gray bars , group 3— black bars ). Significant differences between subsets are indicated (ANOVA or Kruskal–Wallis non-parametric method). Each bar in the histograms represented the mean ± SEM. c Representative dot plots of the subsets defined by CD27 and CD28 expression for individuals in each group. EM T cells can be divided into EM1 (CD27+CD28+), EM2 (CD27+CD28null, only in CD8+ T cells), EM3 (CD27nullCD28null), and EM4 (CD27nullCD28+). Similarly, EMRA can be divided into pE1 (CD27+CD28+) and pE2 (CD27+CD28null, only in CD8 T cells) and E (CD27nullCD28null). d Percentage of cells in each subset in the four groups of elderly accordingly to their functional status (group 0— white bars , group 1— light gray bars , group 2— dark gray bars , group 3— black bars ). Bars in the histograms represented the mean ± SEM
Figure Legend Snippet: Distribution of CD4+ and CD8+ T cells into naïve, central memory, effector memory and effector memory RA, and distribution of EM and EMRA in CD4+ and CD8+ T cells into subsets defined by CD28 and CD27 expression. Expression of CD45RA, CCR7, CD27, and CD28 was analyzed by flow cytometry in isolated CD4+ and CD8+ T cells from the four BI groups of elders. a Schematic model of the T-cell differentiation subsets accordingly to CD45RA and CCR7 expression. Whole blood was stained with anti-CD45RA-FICT, anti-CD8-PE, anti-CD4-PerCP, and anti-CCR7-APC, and 10 5 cells were acquired in each experiment. b Histograms represent percentage of cells in each subset (NAÏVE, CM, EM, and EMRA) in the four groups of elderly accordingly to their functional status (group 0— white bars , group 1— light gray bars , group 2— dark gray bars , group 3— black bars ). Significant differences between subsets are indicated (ANOVA or Kruskal–Wallis non-parametric method). Each bar in the histograms represented the mean ± SEM. c Representative dot plots of the subsets defined by CD27 and CD28 expression for individuals in each group. EM T cells can be divided into EM1 (CD27+CD28+), EM2 (CD27+CD28null, only in CD8+ T cells), EM3 (CD27nullCD28null), and EM4 (CD27nullCD28+). Similarly, EMRA can be divided into pE1 (CD27+CD28+) and pE2 (CD27+CD28null, only in CD8 T cells) and E (CD27nullCD28null). d Percentage of cells in each subset in the four groups of elderly accordingly to their functional status (group 0— white bars , group 1— light gray bars , group 2— dark gray bars , group 3— black bars ). Bars in the histograms represented the mean ± SEM

Techniques Used: Expressing, Flow Cytometry, Cytometry, Isolation, Cell Differentiation, Staining, Functional Assay

4) Product Images from "5-Azacytidine treatment sensitizes tumor cells to T-cell mediated cytotoxicity and modulates NK cells in patients with myeloid malignancies"

Article Title: 5-Azacytidine treatment sensitizes tumor cells to T-cell mediated cytotoxicity and modulates NK cells in patients with myeloid malignancies

Journal: Blood Cancer Journal

doi: 10.1038/bcj.2014.14

Cancer testis antigen (CTA)-specific T cells in the peripheral blood of patients. Detection of CTA- or viral-specific T cells in PBMCs by MHC multimers, expressed in percentage of CD8 T cells. ( a ) The sum of CTA-specific T cells as measured in peripheral blood at different time points during treatment. Eight patients were tested, results from the six patients with detectable responses are shown. First cycle represents a sample obtained before treatment. The following responses were found for each patient: AZA 1 (SART-3 WLE , SART-3 QIR , Sp17 ILD ), AZA 2 (MAGE-A2 LVH , MAGE-A2 KMV , TAG-1 SLG ), AZA 4 (MAGE-A2 LVH , MAGE-A2 LVQ , NY-ESO-1 QLS ), AZA 5 (MAGE-A1 EAD ), AZA 12 (MAGE-A2 LVH , MAGE-A2 KMV , CDCA1 KLA , TAG-1 SLG , NY-ESO-1 SLL , MAGE-A1 EAD ) and AZA 16 (MAGE-A2 LVH , MAGE-A2 KMV , GnTV VLP , TAG-1 SLG ). ( b ) The frequency of individual CTA-specific T cells detected after an in vitro peptide pre-stimulation was performed at different time points during treatment. ( c ) The sum of virus-specific T cells detected over the course of treatment. The following responses were detected: AZA 1 (EBV RLR , EBV RLR , FLU ILR ), AZA 2 (EBV GLC , FLU ILR ), AZA 4 (EBV GLC , EBV YVL , FLU GIL ), AZA 5 (FLU BP-VSD ), AZA 12 (CMV VTE , CMV YSE , CMV NLV , FLU GIL ), AZA 14 (CMV YSE , CMV VTE , FLU BP-VSD ), AZA 16 (CMV NLV , EBV GLC ) and AZA 17 (CMV YSE , CMV VTE , FLU BP-VSD ). MHC-multimer-specific T cells are given in percentage of CD8 cells. Significance is indicated by * P
Figure Legend Snippet: Cancer testis antigen (CTA)-specific T cells in the peripheral blood of patients. Detection of CTA- or viral-specific T cells in PBMCs by MHC multimers, expressed in percentage of CD8 T cells. ( a ) The sum of CTA-specific T cells as measured in peripheral blood at different time points during treatment. Eight patients were tested, results from the six patients with detectable responses are shown. First cycle represents a sample obtained before treatment. The following responses were found for each patient: AZA 1 (SART-3 WLE , SART-3 QIR , Sp17 ILD ), AZA 2 (MAGE-A2 LVH , MAGE-A2 KMV , TAG-1 SLG ), AZA 4 (MAGE-A2 LVH , MAGE-A2 LVQ , NY-ESO-1 QLS ), AZA 5 (MAGE-A1 EAD ), AZA 12 (MAGE-A2 LVH , MAGE-A2 KMV , CDCA1 KLA , TAG-1 SLG , NY-ESO-1 SLL , MAGE-A1 EAD ) and AZA 16 (MAGE-A2 LVH , MAGE-A2 KMV , GnTV VLP , TAG-1 SLG ). ( b ) The frequency of individual CTA-specific T cells detected after an in vitro peptide pre-stimulation was performed at different time points during treatment. ( c ) The sum of virus-specific T cells detected over the course of treatment. The following responses were detected: AZA 1 (EBV RLR , EBV RLR , FLU ILR ), AZA 2 (EBV GLC , FLU ILR ), AZA 4 (EBV GLC , EBV YVL , FLU GIL ), AZA 5 (FLU BP-VSD ), AZA 12 (CMV VTE , CMV YSE , CMV NLV , FLU GIL ), AZA 14 (CMV YSE , CMV VTE , FLU BP-VSD ), AZA 16 (CMV NLV , EBV GLC ) and AZA 17 (CMV YSE , CMV VTE , FLU BP-VSD ). MHC-multimer-specific T cells are given in percentage of CD8 cells. Significance is indicated by * P

Techniques Used: In Vitro, Gas Chromatography

5) Product Images from "Molecular mechanisms of extracellular adenine nucleotides-mediated inhibition of human Cd4+ T lymphocytes activation"

Article Title: Molecular mechanisms of extracellular adenine nucleotides-mediated inhibition of human Cd4+ T lymphocytes activation

Journal: Purinergic Signalling

doi: 10.1007/s11302-005-8077-9

ATPγS inhibits IFN-γ secretion in primary human CD4 +  T cells following activation mediated by anti-CD3 plus anti-CD28 but not by PMA plus anti-CD28. The cells were incubated for 24 h either with PMA (1 ng/ml) and two concentrations of soluble anti-CD28 (1 or 5 µg/ml) (A) or with a combination of pre-coated anti-CD3 and soluble anti-CD28 (1 µg/ml) (B), in the continuous presence or absence of ATPγS, that was added at the same time as PMA/CD28 or anti-CD3/anti-CD28. In the absence of stimulation, no IFN-γ was detectable in the supernatant (data not shown). Data represent the mean ± S.D. of triplicate experimental points obtained in one representative experiment of three.
Figure Legend Snippet: ATPγS inhibits IFN-γ secretion in primary human CD4 + T cells following activation mediated by anti-CD3 plus anti-CD28 but not by PMA plus anti-CD28. The cells were incubated for 24 h either with PMA (1 ng/ml) and two concentrations of soluble anti-CD28 (1 or 5 µg/ml) (A) or with a combination of pre-coated anti-CD3 and soluble anti-CD28 (1 µg/ml) (B), in the continuous presence or absence of ATPγS, that was added at the same time as PMA/CD28 or anti-CD3/anti-CD28. In the absence of stimulation, no IFN-γ was detectable in the supernatant (data not shown). Data represent the mean ± S.D. of triplicate experimental points obtained in one representative experiment of three.

Techniques Used: Activation Assay, Incubation

ATPγS has no effect on the calcium response induced by TCR stimulation with anti-CD3 and anti-CD28. Cells were loaded with FURA 2-AM, and intracellular calcium mobilization was followed on a spectrofluorometer (LS50B, Perkin Elmer). CD4 +  T lymphocytes were activated by cross-linking the anti-CD3/anti-CD28 mAb with goat anti-IgG (▾) without (A) or with 100 µM ATPγS added (▿) to the cells 2 min before cross-linking (B). Data are from one representative experiment out of three.
Figure Legend Snippet: ATPγS has no effect on the calcium response induced by TCR stimulation with anti-CD3 and anti-CD28. Cells were loaded with FURA 2-AM, and intracellular calcium mobilization was followed on a spectrofluorometer (LS50B, Perkin Elmer). CD4 + T lymphocytes were activated by cross-linking the anti-CD3/anti-CD28 mAb with goat anti-IgG (▾) without (A) or with 100 µM ATPγS added (▿) to the cells 2 min before cross-linking (B). Data are from one representative experiment out of three.

Techniques Used:

6) Product Images from "Proviral load and immune function in blood and lymph node during HIV-1 and HIV-2 infection"

Article Title: Proviral load and immune function in blood and lymph node during HIV-1 and HIV-2 infection

Journal: Clinical and Experimental Immunology

doi: 10.1046/j.1365-2249.1999.00914.x

CD4 % is lower in PBMC than in LNMC in HIV infection
Figure Legend Snippet: CD4 % is lower in PBMC than in LNMC in HIV infection

Techniques Used: Infection

7) Product Images from "Protection of Hepatocytes from Cytotoxic T Cell Mediated Killing by Interferon-Alpha"

Article Title: Protection of Hepatocytes from Cytotoxic T Cell Mediated Killing by Interferon-Alpha

Journal: PLoS ONE

doi: 10.1371/journal.pone.0000791

IFN-α stimulated hepatocytes are still able to act as APCs to CTL. HepG2 cells were stimulated for 16 hours with 1000 U/ml IFN-α, or left untreated. The cells were then pulsed with the specific cognate peptide and washed, prior to co-incubation with the CTL line 2. As controls non-peptide-pulsed cells were used, or all PBMC were peptide pulsed or stimulated with PMA/ionomycin. A) The raw FACS data showing the CD8+ expression on the y axis's and IFN-γ on the x axis's. B) The percentage secretion data from such experiment. HepG2p represents peptide pulsed cells with or without IFN-α stimulation as indicated. This data is representative of 4 such experiments.
Figure Legend Snippet: IFN-α stimulated hepatocytes are still able to act as APCs to CTL. HepG2 cells were stimulated for 16 hours with 1000 U/ml IFN-α, or left untreated. The cells were then pulsed with the specific cognate peptide and washed, prior to co-incubation with the CTL line 2. As controls non-peptide-pulsed cells were used, or all PBMC were peptide pulsed or stimulated with PMA/ionomycin. A) The raw FACS data showing the CD8+ expression on the y axis's and IFN-γ on the x axis's. B) The percentage secretion data from such experiment. HepG2p represents peptide pulsed cells with or without IFN-α stimulation as indicated. This data is representative of 4 such experiments.

Techniques Used: Activated Clotting Time Assay, CTL Assay, Incubation, FACS, Expressing

8) Product Images from "Evidence of a role for both anti-Hsp70 antibody and endothelial surface membrane Hsp70 in atherosclerosis"

Article Title: Evidence of a role for both anti-Hsp70 antibody and endothelial surface membrane Hsp70 in atherosclerosis

Journal: Cell Stress & Chaperones

doi: 10.1007/s12192-013-0404-4

Dose–response curves for ADCC. The procedures for OxLDL incubated and Ab treatment were the same as described in the legend for Fig. . As effectors normal rat PBMC were added to the culture instead of complement. Note increased 51Cr release
Figure Legend Snippet: Dose–response curves for ADCC. The procedures for OxLDL incubated and Ab treatment were the same as described in the legend for Fig. . As effectors normal rat PBMC were added to the culture instead of complement. Note increased 51Cr release

Techniques Used: Incubation

9) Product Images from "NKG2D expression in CD4+ T lymphocytes as a marker of senescence in the aged immune system"

Article Title: NKG2D expression in CD4+ T lymphocytes as a marker of senescence in the aged immune system

Journal: Age

doi: 10.1007/s11357-010-9200-6

CD69 expression in response to CMV antigens and to the influenza vaccine. PBMCs from elderly individuals were stimulated for 18 h and CD69 expression was evaluated by flow cytometry. a Representative dot plots of the frequency of CMV- and influenza vaccine-specific CD4 T cells with and without NKG2D expression. Percentages of positive cells in this representative experiment are expressed in dot plots. b Histograms summarize the percentage of CD69-cells positive for CMV and influenza vaccine in the CD4+ CD28 null NKG2D+ ( black bars ) and CD4+ CD28 null NKG2D- subsets ( white bars ) obtained from 11 elderly donors. Paired t test was used to compare paired frequencies
Figure Legend Snippet: CD69 expression in response to CMV antigens and to the influenza vaccine. PBMCs from elderly individuals were stimulated for 18 h and CD69 expression was evaluated by flow cytometry. a Representative dot plots of the frequency of CMV- and influenza vaccine-specific CD4 T cells with and without NKG2D expression. Percentages of positive cells in this representative experiment are expressed in dot plots. b Histograms summarize the percentage of CD69-cells positive for CMV and influenza vaccine in the CD4+ CD28 null NKG2D+ ( black bars ) and CD4+ CD28 null NKG2D- subsets ( white bars ) obtained from 11 elderly donors. Paired t test was used to compare paired frequencies

Techniques Used: Expressing, Flow Cytometry, Cytometry

CD4+ NKG2D+ T cells in peripheral blood from young and elderly individuals. ( a ) Representative dot plots showing the frequency of NKG2D expression in CD4+ T cells in elderly and young individuals. ( b ) Percentages of CD4+ T cells expressing NKG2D in a group of 100 elderly subjects compared with a group of 50 young subjects. Whole blood was stained with CD45-FITC/NKG2D-PE/CD3-PerCP/CD4-APC and 10 5  cells were acquired in each experiment. Frequencies of NKG2D+ cells in gated CD45+ CD3+ CD4+ lymphocytes were analyzed. Outlier values are represented by circles and extreme values by stars, and were calculated by adding 1.5 and 3 times the IR to the 75th percentile, respectively. The horizontal dotted line illustrates the 75th percentile in young donors (2.3%). The non-parametric Mann-Whitney U method was used to compare frequencies between groups
Figure Legend Snippet: CD4+ NKG2D+ T cells in peripheral blood from young and elderly individuals. ( a ) Representative dot plots showing the frequency of NKG2D expression in CD4+ T cells in elderly and young individuals. ( b ) Percentages of CD4+ T cells expressing NKG2D in a group of 100 elderly subjects compared with a group of 50 young subjects. Whole blood was stained with CD45-FITC/NKG2D-PE/CD3-PerCP/CD4-APC and 10 5 cells were acquired in each experiment. Frequencies of NKG2D+ cells in gated CD45+ CD3+ CD4+ lymphocytes were analyzed. Outlier values are represented by circles and extreme values by stars, and were calculated by adding 1.5 and 3 times the IR to the 75th percentile, respectively. The horizontal dotted line illustrates the 75th percentile in young donors (2.3%). The non-parametric Mann-Whitney U method was used to compare frequencies between groups

Techniques Used: Expressing, Staining, MANN-WHITNEY

Distribution of CD4+ and CD8+ T cells into naïve, central memory, effector memory, and effector memory RA. CD45RA and CCR7 expression was analysed by flow cytometry in isolated CD4+ T cells from young people ( n  = 20) and elderly subjects with different frequencies of NKG2D (
Figure Legend Snippet: Distribution of CD4+ and CD8+ T cells into naïve, central memory, effector memory, and effector memory RA. CD45RA and CCR7 expression was analysed by flow cytometry in isolated CD4+ T cells from young people ( n  = 20) and elderly subjects with different frequencies of NKG2D (

Techniques Used: Expressing, Flow Cytometry, Cytometry, Isolation

Phenotypic characterization of CD4+ NKG2D+ T cells.  a  Expression of CD28 was analyzed in CD4+ NKG2D+ T cells from 20 elderly and 20 young individuals. Whole blood was stained with CD28-FITC/NKG2D-PE/CD3-PerCP/CD4-APC and 10 5  cells were acquired in each experiment. Histograms depict the percentage of CD28+ NKG2D+ ( black bar ) and CD28 null NKG2D+ ( white bar ) cells in gated CD3+ CD4+ lymphocytes. Percentages of CD28 null  and CD28+ cells within NKG2D+ subset in elderly and young individuals are summarized.  b  ( a ) Characterization of CD4+ NKG2D+ T cells related to expression of the CD28 marker. ( b ) CD4+ T cells from five aged donors were isolated and the expression of CD45RA-FITC, CD45RO-PercP, CD25-FITC, and HLA-DR-PerCP was analyzed in CD28 null NKG2D+ (Bb) and in CD28+ NKGD+ cells ( c ). Intracellular staining of granzyme B and perforin expression. The percentages of positive cells in the indicated cell populations in this representative experiment were expressed in each histogram plot. ( d ) Histograms summarize the percentage of positive cells for each marker in CD28 null NKG2D+ and in CD28+ NKG2D+ cells (mean ± SEM).  Asterisks  significant differences between the groups ( p
Figure Legend Snippet: Phenotypic characterization of CD4+ NKG2D+ T cells. a Expression of CD28 was analyzed in CD4+ NKG2D+ T cells from 20 elderly and 20 young individuals. Whole blood was stained with CD28-FITC/NKG2D-PE/CD3-PerCP/CD4-APC and 10 5 cells were acquired in each experiment. Histograms depict the percentage of CD28+ NKG2D+ ( black bar ) and CD28 null NKG2D+ ( white bar ) cells in gated CD3+ CD4+ lymphocytes. Percentages of CD28 null and CD28+ cells within NKG2D+ subset in elderly and young individuals are summarized. b ( a ) Characterization of CD4+ NKG2D+ T cells related to expression of the CD28 marker. ( b ) CD4+ T cells from five aged donors were isolated and the expression of CD45RA-FITC, CD45RO-PercP, CD25-FITC, and HLA-DR-PerCP was analyzed in CD28 null NKG2D+ (Bb) and in CD28+ NKGD+ cells ( c ). Intracellular staining of granzyme B and perforin expression. The percentages of positive cells in the indicated cell populations in this representative experiment were expressed in each histogram plot. ( d ) Histograms summarize the percentage of positive cells for each marker in CD28 null NKG2D+ and in CD28+ NKG2D+ cells (mean ± SEM). Asterisks significant differences between the groups ( p

Techniques Used: Expressing, Staining, Marker, Isolation

Distribution of EM and EMRA CD4+ T cells into subsets defined by CD28 and CD27 expression. a Schematic model of the EM and EMRA differentiation subsets according to CD28 and CD27 expression. b Representative dot plots of the subsets defined by CD27 and CD28 expression for individuals in each group. c Individual segments of the pie charts represent the proportions of cells with each combination of CD28 and CD27 in the EM and EMRA CD4 T cell subsets in young donors ( n = 20) and in the three NKG2D groups (
Figure Legend Snippet: Distribution of EM and EMRA CD4+ T cells into subsets defined by CD28 and CD27 expression. a Schematic model of the EM and EMRA differentiation subsets according to CD28 and CD27 expression. b Representative dot plots of the subsets defined by CD27 and CD28 expression for individuals in each group. c Individual segments of the pie charts represent the proportions of cells with each combination of CD28 and CD27 in the EM and EMRA CD4 T cell subsets in young donors ( n = 20) and in the three NKG2D groups (

Techniques Used: Expressing

10) Product Images from "Long-Term Exposure to Inflammation Induces Differential Cytokine Patterns and Apoptosis in Dendritic Cells"

Article Title: Long-Term Exposure to Inflammation Induces Differential Cytokine Patterns and Apoptosis in Dendritic Cells

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2019.02702

IFNγ release from co-cultures with long-term activated DCs is reduced. (A) DCs were activated with the DC1 maturation cocktail (LPS and IFNγ) for 1, 2, 3, or 4 days (DC1-1d, DC1-2d, DC1-3d, DC1-4d) prior to being set up in co-culture with allogeneic PBMCs at a 1:10 ratio. After 5 days, IFNγ release was measured in the MLR supernatants with a lower limit of sensitivity of 156 pg/mL. Co-cultures with iDCs and sDC were included for comparison. Cumulative data are shown from five independent experiments with nine unique donor pairs in total ( n = 9), consisting of seven DC donors and five PBMC donors. All experiments were performed in quadruplicates. Bars represent mean + standard deviation. Two-way ANOVA with Tukey's post-hoc test was performed on log10-transformed data to compare DC1 groups (**** p ≤ 0.0001, ** p ≤ 0.01, * p ≤ 0.05, and ns, not statistically significant). (B) Same as (A) , except isolated CD4 + and CD8 + T cells were used as responder cells instead of PBMCs. Data from three donors ( n = 3) are normalized to highest value of IFNγ release within each donor. Bars represent mean + standard deviation. Two-way ANOVA with Tukey's post-hoc test was performed on normalized data to compare DC1 groups (**** p ≤ 0.0001, ** p ≤ 0.01, * p ≤ 0.05). The lower limit of IFNγ detection was 39 pg/mL. (C) Co-cultures were set up as in (A) and supplemented with anti-IL-12p70 or recombinant human IL-12p70. Recombinant human IL-12p70 was also added to cultures of PBMCs without DCs (w/o DC). After 5 days, IFNγ release was measured. Cumulative data are shown from one experiment with three unique donors ( n = 3). Bars represent mean + standard deviation.
Figure Legend Snippet: IFNγ release from co-cultures with long-term activated DCs is reduced. (A) DCs were activated with the DC1 maturation cocktail (LPS and IFNγ) for 1, 2, 3, or 4 days (DC1-1d, DC1-2d, DC1-3d, DC1-4d) prior to being set up in co-culture with allogeneic PBMCs at a 1:10 ratio. After 5 days, IFNγ release was measured in the MLR supernatants with a lower limit of sensitivity of 156 pg/mL. Co-cultures with iDCs and sDC were included for comparison. Cumulative data are shown from five independent experiments with nine unique donor pairs in total ( n = 9), consisting of seven DC donors and five PBMC donors. All experiments were performed in quadruplicates. Bars represent mean + standard deviation. Two-way ANOVA with Tukey's post-hoc test was performed on log10-transformed data to compare DC1 groups (**** p ≤ 0.0001, ** p ≤ 0.01, * p ≤ 0.05, and ns, not statistically significant). (B) Same as (A) , except isolated CD4 + and CD8 + T cells were used as responder cells instead of PBMCs. Data from three donors ( n = 3) are normalized to highest value of IFNγ release within each donor. Bars represent mean + standard deviation. Two-way ANOVA with Tukey's post-hoc test was performed on normalized data to compare DC1 groups (**** p ≤ 0.0001, ** p ≤ 0.01, * p ≤ 0.05). The lower limit of IFNγ detection was 39 pg/mL. (C) Co-cultures were set up as in (A) and supplemented with anti-IL-12p70 or recombinant human IL-12p70. Recombinant human IL-12p70 was also added to cultures of PBMCs without DCs (w/o DC). After 5 days, IFNγ release was measured. Cumulative data are shown from one experiment with three unique donors ( n = 3). Bars represent mean + standard deviation.

Techniques Used: Co-Culture Assay, Standard Deviation, Transformation Assay, Isolation, Recombinant

11) Product Images from "Decreased CD95 expression on naive T cells from HIV-infected persons undergoing highly active anti-retroviral therapy (HAART) and the influence of IL-2 low dose administration"

Article Title: Decreased CD95 expression on naive T cells from HIV-infected persons undergoing highly active anti-retroviral therapy (HAART) and the influence of IL-2 low dose administration

Journal: Clinical and Experimental Immunology

doi: 10.1046/j.1365-2249.2000.01223.x

rIL-2 administration during HAART affects the expression of CD25 (IL-2Rα) in HIV-infected individuals undergoing therapy. (a) Changes in the percentage of CD25-expressing total peripheral blood mononuclear cells (PBMC) in all groups, investigated by single-cell analysis at t = 0 (□) and at t = 12 weeks (▪). Mean percentages of CD25 + cell increase, relative to each group following therapy, are reported in the text. Analysis of different subsets of PBMC expressing CD25 was carried out by flow cytometry on the three groups of patients, and percentages reported in (b,c,d). A significant rIL-2 treatment-induced increase in the percentages of CD3 + , CD8 + , CD16 + and CD19 + lymphocytes was observed after therapy in groups 2 and 3, when compared with baseline values at starting treatment. Data represent the arithmetic mean percentage; error bars indicate s.e.m. P values are indicated: * P
Figure Legend Snippet: rIL-2 administration during HAART affects the expression of CD25 (IL-2Rα) in HIV-infected individuals undergoing therapy. (a) Changes in the percentage of CD25-expressing total peripheral blood mononuclear cells (PBMC) in all groups, investigated by single-cell analysis at t = 0 (□) and at t = 12 weeks (▪). Mean percentages of CD25 + cell increase, relative to each group following therapy, are reported in the text. Analysis of different subsets of PBMC expressing CD25 was carried out by flow cytometry on the three groups of patients, and percentages reported in (b,c,d). A significant rIL-2 treatment-induced increase in the percentages of CD3 + , CD8 + , CD16 + and CD19 + lymphocytes was observed after therapy in groups 2 and 3, when compared with baseline values at starting treatment. Data represent the arithmetic mean percentage; error bars indicate s.e.m. P values are indicated: * P

Techniques Used: Expressing, Infection, Single-cell Analysis, Flow Cytometry, Cytometry

rIL-2 administration during HAART does not affect the expression of CD122 (IL-2Rβ) in HIV-infected individuals undergoing therapy. (a) Mean percentages of CD122-expressing total peripheral blood mononuclear cells (PBMC) in all groups, investigated by single-cell analysis at t = 0 (□) and at t = 12 weeks (▪). Mean percentages of CD122 + cells did not change significantly after therapy in all groups. Different subsets of PBMC expressing CD122 were then analysed and are reported in (b,c,d). No significant treatment-induced changes in the percentage of CD122 + lymphocytes were observed after therapy, when compared with baseline values at starting treatment. Data represent the arithmetic mean percentage; error bars indicate s.e.m. P values are indicated: * P
Figure Legend Snippet: rIL-2 administration during HAART does not affect the expression of CD122 (IL-2Rβ) in HIV-infected individuals undergoing therapy. (a) Mean percentages of CD122-expressing total peripheral blood mononuclear cells (PBMC) in all groups, investigated by single-cell analysis at t = 0 (□) and at t = 12 weeks (▪). Mean percentages of CD122 + cells did not change significantly after therapy in all groups. Different subsets of PBMC expressing CD122 were then analysed and are reported in (b,c,d). No significant treatment-induced changes in the percentage of CD122 + lymphocytes were observed after therapy, when compared with baseline values at starting treatment. Data represent the arithmetic mean percentage; error bars indicate s.e.m. P values are indicated: * P

Techniques Used: Expressing, Infection, Single-cell Analysis

Decrease of CD95 death receptor expression on naive subset of total peripheral blood mononuclear cells (PBMC) after differentiated therapy in HIV-infected individuals. (a) Changes in the percentage of naive (CD45RA + CD62L + ) cells expressing the CD95 receptor, before (□) and after 12 weeks of therapy (▪) in each group of patients. Mean percentages of decrease, relative to each group following therapy, are reported in the text. Data represent the arithmetic mean percentages; error bars indicate s.e.m. P values are indicated: * P
Figure Legend Snippet: Decrease of CD95 death receptor expression on naive subset of total peripheral blood mononuclear cells (PBMC) after differentiated therapy in HIV-infected individuals. (a) Changes in the percentage of naive (CD45RA + CD62L + ) cells expressing the CD95 receptor, before (□) and after 12 weeks of therapy (▪) in each group of patients. Mean percentages of decrease, relative to each group following therapy, are reported in the text. Data represent the arithmetic mean percentages; error bars indicate s.e.m. P values are indicated: * P

Techniques Used: Expressing, Infection

Frequencies of naive cells in HIV-infected individuals undergoing anti-retroviral therapy. Detection of peripheral blood mononuclear cells (PBMC) with a naive phenotype (identified as expression of CD45RA + and CD62L + receptors) was performed by single-cell analysis using a 4-fluorescence cytometric protocol at t = 0 (before therapy; □) and at t = 12 weeks (after 12 weeks of therapy; ▪). The percentage of naive cells analysed in each patient is shown in (a) (group 1), (b) (group 2), and (c) (group 3). In parallel, CD4 + T cells with naive phenotype were also analysed and relative percentages are shown in (d) (group 1), (e) (group 2), and (f) (group 3).
Figure Legend Snippet: Frequencies of naive cells in HIV-infected individuals undergoing anti-retroviral therapy. Detection of peripheral blood mononuclear cells (PBMC) with a naive phenotype (identified as expression of CD45RA + and CD62L + receptors) was performed by single-cell analysis using a 4-fluorescence cytometric protocol at t = 0 (before therapy; □) and at t = 12 weeks (after 12 weeks of therapy; ▪). The percentage of naive cells analysed in each patient is shown in (a) (group 1), (b) (group 2), and (c) (group 3). In parallel, CD4 + T cells with naive phenotype were also analysed and relative percentages are shown in (d) (group 1), (e) (group 2), and (f) (group 3).

Techniques Used: Infection, Expressing, Single-cell Analysis, Fluorescence

Analysis of CD95 expression on peripheral blood mononuclear cells (PBMC) obtained from HIV-infected individuals undergoing therapy. The expression of the CD95 receptor on total PBMC was investigated by single-cell analysis at t = 0 (□) and at t = 12 weeks (▪). (a) Changes in the percentage of total PBMC expressing the CD95 receptor in each group, before and after differentiated therapies. Percentages of CD95 + cell decrease, relative to each group following therapy, are shown. Results of phenotypical single-cell analysis of CD95 + PBMC subsets are shown in (b) (percentage of CD4 + CD95 + T cells), (c) (percentage of CD8 + CD95 + T cells), and (d) (percentage of natural killer (NK) CD16 + CD95 + cells). Data represent the arithmetic mean percentage; error bars indicate s.e.m. P values are indicated: * P
Figure Legend Snippet: Analysis of CD95 expression on peripheral blood mononuclear cells (PBMC) obtained from HIV-infected individuals undergoing therapy. The expression of the CD95 receptor on total PBMC was investigated by single-cell analysis at t = 0 (□) and at t = 12 weeks (▪). (a) Changes in the percentage of total PBMC expressing the CD95 receptor in each group, before and after differentiated therapies. Percentages of CD95 + cell decrease, relative to each group following therapy, are shown. Results of phenotypical single-cell analysis of CD95 + PBMC subsets are shown in (b) (percentage of CD4 + CD95 + T cells), (c) (percentage of CD8 + CD95 + T cells), and (d) (percentage of natural killer (NK) CD16 + CD95 + cells). Data represent the arithmetic mean percentage; error bars indicate s.e.m. P values are indicated: * P

Techniques Used: Expressing, Infection, Single-cell Analysis

12) Product Images from "Relationship between functional ability in older people, immune system status, and intensity of response to CMV"

Article Title: Relationship between functional ability in older people, immune system status, and intensity of response to CMV

Journal: Age

doi: 10.1007/s11357-011-9240-6

CD69 expression and proliferative capacity of CD4+ and CD8+ depending on the functional capacity. Whole blood from the BI groups of elders (group 0 + 1, n = 12 and group 2 + 3, n = 14) was stimulated for 18 h and expression of CD69 in CD4+ and CD8+ T cells was evaluated by flow cytometry. Proliferative capacity of CD4+ and CD8+ T cells subsets was also evaluated in the two groups (group 0 + 1, n = 19 and group 2 + 3, n = 16) by labeling the PBMC with CFSE. Cells were stained and 1 × 10 5 cells were acquired per experiment. a Representative dot plots showing the frequency of CD69 expression in CD4+ and CD8+ subset from elderly with different functional status. Cells were stimulated using anti-CD3 (10 ng/mL). Percentage of positive cells in each subpopulation in this representative experiment is expressed in the upper right corner and summarized results from all donors (median and IR) were also expressed in dot plots. b Proliferative capacity of CD4+ and CD8+ T cells subsets in response to anti-CD3. PBMC were labeled with CFSE (1.5 μM) and cultured in presence of anti-CD3 (10 ng/mL) for 5 days. Percentage of dividing CD4+ and CD8+ T cells is represented. Bars represent results from the grouped elders (mean ± SEM). c Expression of CD69 into de CD4+ and CD8+ T cell subset was analyzed in the same way as in Fig. 5a in response to a CMV supernatant (10 4 PFU/mL). d Proliferative capacity of CD4+ and CD8+ T-cell subsets in response to the CMV supernatant. Bars represent resulted from the grouped elders (mean ± SEM). The Student’s t test (when data were normally distributed) and Mann–Whitney non-parametric (when data were not normally distributed) methods were used to compare frequencies between groups. p values are depicted in the panels
Figure Legend Snippet: CD69 expression and proliferative capacity of CD4+ and CD8+ depending on the functional capacity. Whole blood from the BI groups of elders (group 0 + 1, n = 12 and group 2 + 3, n = 14) was stimulated for 18 h and expression of CD69 in CD4+ and CD8+ T cells was evaluated by flow cytometry. Proliferative capacity of CD4+ and CD8+ T cells subsets was also evaluated in the two groups (group 0 + 1, n = 19 and group 2 + 3, n = 16) by labeling the PBMC with CFSE. Cells were stained and 1 × 10 5 cells were acquired per experiment. a Representative dot plots showing the frequency of CD69 expression in CD4+ and CD8+ subset from elderly with different functional status. Cells were stimulated using anti-CD3 (10 ng/mL). Percentage of positive cells in each subpopulation in this representative experiment is expressed in the upper right corner and summarized results from all donors (median and IR) were also expressed in dot plots. b Proliferative capacity of CD4+ and CD8+ T cells subsets in response to anti-CD3. PBMC were labeled with CFSE (1.5 μM) and cultured in presence of anti-CD3 (10 ng/mL) for 5 days. Percentage of dividing CD4+ and CD8+ T cells is represented. Bars represent results from the grouped elders (mean ± SEM). c Expression of CD69 into de CD4+ and CD8+ T cell subset was analyzed in the same way as in Fig. 5a in response to a CMV supernatant (10 4 PFU/mL). d Proliferative capacity of CD4+ and CD8+ T-cell subsets in response to the CMV supernatant. Bars represent resulted from the grouped elders (mean ± SEM). The Student’s t test (when data were normally distributed) and Mann–Whitney non-parametric (when data were not normally distributed) methods were used to compare frequencies between groups. p values are depicted in the panels

Techniques Used: Expressing, Functional Assay, Flow Cytometry, Cytometry, Labeling, Staining, Cell Culture, MANN-WHITNEY

13) Product Images from "NKG2D expression in CD4+ T lymphocytes as a marker of senescence in the aged immune system"

Article Title: NKG2D expression in CD4+ T lymphocytes as a marker of senescence in the aged immune system

Journal: Age

doi: 10.1007/s11357-010-9200-6

CD69 expression in response to CMV antigens and to the influenza vaccine. PBMCs from elderly individuals were stimulated for 18 h and CD69 expression was evaluated by flow cytometry. a Representative dot plots of the frequency of CMV- and influenza vaccine-specific CD4 T cells with and without NKG2D expression. Percentages of positive cells in this representative experiment are expressed in dot plots. b Histograms summarize the percentage of CD69-cells positive for CMV and influenza vaccine in the CD4+ CD28 null NKG2D+ ( black bars ) and CD4+ CD28 null NKG2D- subsets ( white bars ) obtained from 11 elderly donors. Paired t test was used to compare paired frequencies
Figure Legend Snippet: CD69 expression in response to CMV antigens and to the influenza vaccine. PBMCs from elderly individuals were stimulated for 18 h and CD69 expression was evaluated by flow cytometry. a Representative dot plots of the frequency of CMV- and influenza vaccine-specific CD4 T cells with and without NKG2D expression. Percentages of positive cells in this representative experiment are expressed in dot plots. b Histograms summarize the percentage of CD69-cells positive for CMV and influenza vaccine in the CD4+ CD28 null NKG2D+ ( black bars ) and CD4+ CD28 null NKG2D- subsets ( white bars ) obtained from 11 elderly donors. Paired t test was used to compare paired frequencies

Techniques Used: Expressing, Flow Cytometry, Cytometry

Differentiated status of CD4+ subsets defined by IL-2/IFN-γ production and TREC content. a PBMCs were stimulated for 6 h with soluble anti-CD3 (1 μg/mL). The responder cells were analyzed for intracellular IL-2-APC staining in the indicated populations. b Cells were treated as described previously and IFN-γ expression was analyzed by intracellular staining. Percentages of positive cells in the indicated populations in this representative experiment are expressed on the histogram-plots. Histograms summarize the percentage of IFN-γ-positive cells in CD4+ CD28 null and in CD4+ CD28+ lymphocytes NKG2D− and NKG2D + (mean ± SEM) from the six elderly donors tested. A paired t test was used to compare paired means. c Dose-response curves of the IFN-γ production by CD4 subsets defined by CD28 and NKG2D expression in response to anti-CD3. The cells were cultured for 6 h in medium alone or with increasing concentrations of anti-CD3 (1–1,000 ng/mL). A fluorescence analysis was carried out as previously described. One representative experiment of three is shown. d Quantification of TREC copy number. CD28+, CD28 null NKG2D − , and CD28 null NKG2D+ populations were isolated by sorting (magnetic bead separation) and the TREC copy number was determined by real-time PCR. Experiments were conducted in duplicate and bars represented results from three aging donors (mean ± SEM)
Figure Legend Snippet: Differentiated status of CD4+ subsets defined by IL-2/IFN-γ production and TREC content. a PBMCs were stimulated for 6 h with soluble anti-CD3 (1 μg/mL). The responder cells were analyzed for intracellular IL-2-APC staining in the indicated populations. b Cells were treated as described previously and IFN-γ expression was analyzed by intracellular staining. Percentages of positive cells in the indicated populations in this representative experiment are expressed on the histogram-plots. Histograms summarize the percentage of IFN-γ-positive cells in CD4+ CD28 null and in CD4+ CD28+ lymphocytes NKG2D− and NKG2D + (mean ± SEM) from the six elderly donors tested. A paired t test was used to compare paired means. c Dose-response curves of the IFN-γ production by CD4 subsets defined by CD28 and NKG2D expression in response to anti-CD3. The cells were cultured for 6 h in medium alone or with increasing concentrations of anti-CD3 (1–1,000 ng/mL). A fluorescence analysis was carried out as previously described. One representative experiment of three is shown. d Quantification of TREC copy number. CD28+, CD28 null NKG2D − , and CD28 null NKG2D+ populations were isolated by sorting (magnetic bead separation) and the TREC copy number was determined by real-time PCR. Experiments were conducted in duplicate and bars represented results from three aging donors (mean ± SEM)

Techniques Used: Staining, Expressing, Cell Culture, Fluorescence, Isolation, Real-time Polymerase Chain Reaction

14) Product Images from "5-Azacytidine treatment sensitizes tumor cells to T-cell mediated cytotoxicity and modulates NK cells in patients with myeloid malignancies"

Article Title: 5-Azacytidine treatment sensitizes tumor cells to T-cell mediated cytotoxicity and modulates NK cells in patients with myeloid malignancies

Journal: Blood Cancer Journal

doi: 10.1038/bcj.2014.14

Cancer testis antigen (CTA)-specific T cells in the peripheral blood of patients. Detection of CTA- or viral-specific T cells in PBMCs by MHC multimers, expressed in percentage of CD8 T cells. ( a ) The sum of CTA-specific T cells as measured in peripheral blood at different time points during treatment. Eight patients were tested, results from the six patients with detectable responses are shown. First cycle represents a sample obtained before treatment. The following responses were found for each patient: AZA 1 (SART-3 WLE , SART-3 QIR , Sp17 ILD ), AZA 2 (MAGE-A2 LVH , MAGE-A2 KMV , TAG-1 SLG ), AZA 4 (MAGE-A2 LVH , MAGE-A2 LVQ , NY-ESO-1 QLS ), AZA 5 (MAGE-A1 EAD ), AZA 12 (MAGE-A2 LVH , MAGE-A2 KMV , CDCA1 KLA , TAG-1 SLG , NY-ESO-1 SLL , MAGE-A1 EAD ) and AZA 16 (MAGE-A2 LVH , MAGE-A2 KMV , GnTV VLP , TAG-1 SLG ). ( b ) The frequency of individual CTA-specific T cells detected after an in vitro peptide pre-stimulation was performed at different time points during treatment. ( c ) The sum of virus-specific T cells detected over the course of treatment. The following responses were detected: AZA 1 (EBV RLR , EBV RLR , FLU ILR ), AZA 2 (EBV GLC , FLU ILR ), AZA 4 (EBV GLC , EBV YVL , FLU GIL ), AZA 5 (FLU BP-VSD ), AZA 12 (CMV VTE , CMV YSE , CMV NLV , FLU GIL ), AZA 14 (CMV YSE , CMV VTE , FLU BP-VSD ), AZA 16 (CMV NLV , EBV GLC ) and AZA 17 (CMV YSE , CMV VTE , FLU BP-VSD ). MHC-multimer-specific T cells are given in percentage of CD8 cells. Significance is indicated by * P
Figure Legend Snippet: Cancer testis antigen (CTA)-specific T cells in the peripheral blood of patients. Detection of CTA- or viral-specific T cells in PBMCs by MHC multimers, expressed in percentage of CD8 T cells. ( a ) The sum of CTA-specific T cells as measured in peripheral blood at different time points during treatment. Eight patients were tested, results from the six patients with detectable responses are shown. First cycle represents a sample obtained before treatment. The following responses were found for each patient: AZA 1 (SART-3 WLE , SART-3 QIR , Sp17 ILD ), AZA 2 (MAGE-A2 LVH , MAGE-A2 KMV , TAG-1 SLG ), AZA 4 (MAGE-A2 LVH , MAGE-A2 LVQ , NY-ESO-1 QLS ), AZA 5 (MAGE-A1 EAD ), AZA 12 (MAGE-A2 LVH , MAGE-A2 KMV , CDCA1 KLA , TAG-1 SLG , NY-ESO-1 SLL , MAGE-A1 EAD ) and AZA 16 (MAGE-A2 LVH , MAGE-A2 KMV , GnTV VLP , TAG-1 SLG ). ( b ) The frequency of individual CTA-specific T cells detected after an in vitro peptide pre-stimulation was performed at different time points during treatment. ( c ) The sum of virus-specific T cells detected over the course of treatment. The following responses were detected: AZA 1 (EBV RLR , EBV RLR , FLU ILR ), AZA 2 (EBV GLC , FLU ILR ), AZA 4 (EBV GLC , EBV YVL , FLU GIL ), AZA 5 (FLU BP-VSD ), AZA 12 (CMV VTE , CMV YSE , CMV NLV , FLU GIL ), AZA 14 (CMV YSE , CMV VTE , FLU BP-VSD ), AZA 16 (CMV NLV , EBV GLC ) and AZA 17 (CMV YSE , CMV VTE , FLU BP-VSD ). MHC-multimer-specific T cells are given in percentage of CD8 cells. Significance is indicated by * P

Techniques Used: In Vitro, Gas Chromatography

15) Product Images from "Distinct and Overlapping Effector Functions of Expanded Human CD4+, CD8?+ and CD4-CD8?- Invariant Natural Killer T Cells"

Article Title: Distinct and Overlapping Effector Functions of Expanded Human CD4+, CD8?+ and CD4-CD8?- Invariant Natural Killer T Cells

Journal: PLoS ONE

doi: 10.1371/journal.pone.0028648

Isolation and expansion of human peripheral blood iNKT cells and analysis of CD4 and CD8α expression. A, Representative flow cytometry dot plot showing Vα24 and Vβ11 TCR chain expression by freshly-isolated human PBMC. B, Vα24 and Vβ11 TCR chain expression by PBMC after enrichment using anti-iNKT cell magnetic beads but without expansion (left panel) and CD4 and CD8α expression by gated Vα24 + Vβ11 + cells (right panel). Plots are representative of PBMCs from 8 healthy donors. C, Vα24 and Vβ11 TCR chain expression by an expanded sorted iNKT cell line. Numbers in plots show percentages of cells in each quadrant. D and E, Distribution of CD4 + , CD8α + and DN cells among freshly-isolated iNKT cell lines from 8 randomly obtained healthy donors (D) and among expanded iNKT cells from 20 donors (E). Horizontal lines show means; p values in shaded boxes show comparisons of CD4 + , CD8α + and DN iNKT cell frequencies using the Kruskal-Wallis test; asterisks indicate significant differences between individual groups (indicated by bars) using post hoc Dunn's multiple comparison tests; ** p
Figure Legend Snippet: Isolation and expansion of human peripheral blood iNKT cells and analysis of CD4 and CD8α expression. A, Representative flow cytometry dot plot showing Vα24 and Vβ11 TCR chain expression by freshly-isolated human PBMC. B, Vα24 and Vβ11 TCR chain expression by PBMC after enrichment using anti-iNKT cell magnetic beads but without expansion (left panel) and CD4 and CD8α expression by gated Vα24 + Vβ11 + cells (right panel). Plots are representative of PBMCs from 8 healthy donors. C, Vα24 and Vβ11 TCR chain expression by an expanded sorted iNKT cell line. Numbers in plots show percentages of cells in each quadrant. D and E, Distribution of CD4 + , CD8α + and DN cells among freshly-isolated iNKT cell lines from 8 randomly obtained healthy donors (D) and among expanded iNKT cells from 20 donors (E). Horizontal lines show means; p values in shaded boxes show comparisons of CD4 + , CD8α + and DN iNKT cell frequencies using the Kruskal-Wallis test; asterisks indicate significant differences between individual groups (indicated by bars) using post hoc Dunn's multiple comparison tests; ** p

Techniques Used: Isolation, Expressing, Flow Cytometry, Cytometry, Magnetic Beads

16) Product Images from "Low level laser (LLL) attenuate LPS-induced inflammatory responses in mesenchymal stem cells via the suppression of NF-κB signaling pathway in vitro"

Article Title: Low level laser (LLL) attenuate LPS-induced inflammatory responses in mesenchymal stem cells via the suppression of NF-κB signaling pathway in vitro

Journal: PLoS ONE

doi: 10.1371/journal.pone.0179175

Percentage of positive PBMC after stimulation with PHA in the absence or presence of 3 types of treated MSCs. After reduction of LLL treated MSCs, CD25+ cells decreased (**P
Figure Legend Snippet: Percentage of positive PBMC after stimulation with PHA in the absence or presence of 3 types of treated MSCs. After reduction of LLL treated MSCs, CD25+ cells decreased (**P

Techniques Used:

17) Product Images from "Utility of Major Leukocyte Subpopulations for Monitoring Secondary Cytomegalovirus Infections in Renal-Allograft Recipients by PCR"

Article Title: Utility of Major Leukocyte Subpopulations for Monitoring Secondary Cytomegalovirus Infections in Renal-Allograft Recipients by PCR

Journal: Journal of Clinical Microbiology

doi:

Correlation of pp65 antigen-positive cells with CMV DNA levels in mixed PBL (○), PMNL (x) and PBMC (•) of patients with active CMV infection determined by Spearman regression analysis. (A) Forty-three samples from 23 patients were analyzed before onset of ganciclovir therapy. Significant correlation was observed for mixed PBL ( r S = 0.979; P
Figure Legend Snippet: Correlation of pp65 antigen-positive cells with CMV DNA levels in mixed PBL (○), PMNL (x) and PBMC (•) of patients with active CMV infection determined by Spearman regression analysis. (A) Forty-three samples from 23 patients were analyzed before onset of ganciclovir therapy. Significant correlation was observed for mixed PBL ( r S = 0.979; P

Techniques Used: Infection

18) Product Images from "Assessment of thymic output in common variable immunodeficiency patients by evaluation of T cell receptor excision circles"

Article Title: Assessment of thymic output in common variable immunodeficiency patients by evaluation of T cell receptor excision circles

Journal: Clinical and Experimental Immunology

doi: 10.1046/j.1365-2249.2002.01893.x

Analysis of TRECs (log 10 ) in CD4 + and CD8 + T cells in CVID (i) and in healthy individuals (ii). PBMC were isolated from heparinized venus blood of CVID patients and normal donors by Ficoll-Isopaque (Lymphoprep-Nycomed, Oslo, Norway) gradient centrifugation. CD4 + and CD8 + T cells were separated using magnetic beads coated with anti-CD4 and anti-CD8 MoAbs. Real time PCR analysis was performed on CD4 + and CD8 + T cells with TREC specific primers to detect recent thymic emigrants, and on the GAPDH to standardize for DNA content. TREC levels in CD4 + (a) and in CD8 + (b) T cells were significantly reduced in CVID patients when compared with age-matched healthy individuals.
Figure Legend Snippet: Analysis of TRECs (log 10 ) in CD4 + and CD8 + T cells in CVID (i) and in healthy individuals (ii). PBMC were isolated from heparinized venus blood of CVID patients and normal donors by Ficoll-Isopaque (Lymphoprep-Nycomed, Oslo, Norway) gradient centrifugation. CD4 + and CD8 + T cells were separated using magnetic beads coated with anti-CD4 and anti-CD8 MoAbs. Real time PCR analysis was performed on CD4 + and CD8 + T cells with TREC specific primers to detect recent thymic emigrants, and on the GAPDH to standardize for DNA content. TREC levels in CD4 + (a) and in CD8 + (b) T cells were significantly reduced in CVID patients when compared with age-matched healthy individuals.

Techniques Used: Isolation, Gradient Centrifugation, Magnetic Beads, Real-time Polymerase Chain Reaction

19) Product Images from "Oral supplementation with Lactobacillus delbrueckii subsp. bulgaricus 8481 enhances systemic immunity in elderly subjects"

Article Title: Oral supplementation with Lactobacillus delbrueckii subsp. bulgaricus 8481 enhances systemic immunity in elderly subjects

Journal: Age

doi: 10.1007/s11357-012-9434-6

Changes in percentage in NK cells, TREC content, T cell subsets distribution, and percentage of CD45RA+CD31+ into CD4+ and CD8+ T cells 6 months after stopping the probiotic intake. Measurements were made in nine elderly people at baseline, 6 months
Figure Legend Snippet: Changes in percentage in NK cells, TREC content, T cell subsets distribution, and percentage of CD45RA+CD31+ into CD4+ and CD8+ T cells 6 months after stopping the probiotic intake. Measurements were made in nine elderly people at baseline, 6 months

Techniques Used:

Proximity to the thymus of the T cell subsets and TREC content. Whole blood from the elderly of the probiotic group was stained with anti-CD45RA-FITC, anti-CD31-PE, and these CD4+ and CD8+ T cell subsets were evaluated by flow cytometry.  a  Representative
Figure Legend Snippet: Proximity to the thymus of the T cell subsets and TREC content. Whole blood from the elderly of the probiotic group was stained with anti-CD45RA-FITC, anti-CD31-PE, and these CD4+ and CD8+ T cell subsets were evaluated by flow cytometry. a Representative

Techniques Used: Staining, Flow Cytometry, Cytometry

Distribution of T cell subsets in the probiotics group. CD4+ and CD8+ T cells were categorized into NAÏVE, CM, EM, and EMRA cells. Distribution of EMRA in CD4+ and CD8+ T cells into subsets defined by CD28 and CD27 expression. Expression of CD45RA,
Figure Legend Snippet: Distribution of T cell subsets in the probiotics group. CD4+ and CD8+ T cells were categorized into NAÏVE, CM, EM, and EMRA cells. Distribution of EMRA in CD4+ and CD8+ T cells into subsets defined by CD28 and CD27 expression. Expression of CD45RA,

Techniques Used: Expressing

20) Product Images from "Evidence of a role for both anti-Hsp70 antibody and endothelial surface membrane Hsp70 in atherosclerosis"

Article Title: Evidence of a role for both anti-Hsp70 antibody and endothelial surface membrane Hsp70 in atherosclerosis

Journal: Cell Stress & Chaperones

doi: 10.1007/s12192-013-0404-4

Dose–response curves for ADCC. The procedures for OxLDL incubated and Ab treatment were the same as described in the legend for Fig. . As effectors normal rat PBMC were added to the culture instead of complement. Note increased 51Cr release
Figure Legend Snippet: Dose–response curves for ADCC. The procedures for OxLDL incubated and Ab treatment were the same as described in the legend for Fig. . As effectors normal rat PBMC were added to the culture instead of complement. Note increased 51Cr release

Techniques Used: Incubation

21) Product Images from "Blockade of Toll-like receptor 2 prevents spontaneous cytokine release from rheumatoid arthritis ex vivo synovial explant cultures"

Article Title: Blockade of Toll-like receptor 2 prevents spontaneous cytokine release from rheumatoid arthritis ex vivo synovial explant cultures

Journal: Arthritis Research & Therapy

doi: 10.1186/ar3261

Pam3CSK4 induced IL-6 and IL-8 in mononuclear cells from RA patients . SFMCs ( A ; n = 6) and PBMCs ( B ; n = 11) were stimulated with TLR2 agonist Pam3CSK4 at 200 ng/ml. Levels of IL-6 and IL-8 in the culture supernatants were determined and compared to unstimulated cells (Basal) at six hours. Values are expressed as the mean ± SEM. * P
Figure Legend Snippet: Pam3CSK4 induced IL-6 and IL-8 in mononuclear cells from RA patients . SFMCs ( A ; n = 6) and PBMCs ( B ; n = 11) were stimulated with TLR2 agonist Pam3CSK4 at 200 ng/ml. Levels of IL-6 and IL-8 in the culture supernatants were determined and compared to unstimulated cells (Basal) at six hours. Values are expressed as the mean ± SEM. * P

Techniques Used:

22) Product Images from "Natural regulatory T cells: number and function are normal in the majority of patients with lupus nephritis"

Article Title: Natural regulatory T cells: number and function are normal in the majority of patients with lupus nephritis

Journal: Clinical and Experimental Immunology

doi: 10.1111/j.1365-2249.2008.03665.x

A higher proportion of CD4 + T cells express CD25 in patients with active systemic lupus erythematosus (SLE), although the proportion of CD25 hi remains similar to controls. Peripheral blood mononuclear cells were stained with anti-CD4-APC and an IgG1-PE isotype control antibody to establish levels of background staining. Lymphocytes were gated using forward-scatter/side-scatter characteristics. (a) Representative example of a PE-labelled isotype control and anti-CD25 staining from a control individual. (b) Staining pattern of an individual with active SLE with a particularly marked increase in the proportion of CD4 + T cells expressing CD25 (1·4%). Note also the marked reduction in the proportion of lymphocytes that are CD4 + (8.88% versus 37% in a). (c) Comparing 18 controls, 12 patients with active SLE and six with inactive disease, there was a significant increase in the percentage of CD25 + in those with active disease. P
Figure Legend Snippet: A higher proportion of CD4 + T cells express CD25 in patients with active systemic lupus erythematosus (SLE), although the proportion of CD25 hi remains similar to controls. Peripheral blood mononuclear cells were stained with anti-CD4-APC and an IgG1-PE isotype control antibody to establish levels of background staining. Lymphocytes were gated using forward-scatter/side-scatter characteristics. (a) Representative example of a PE-labelled isotype control and anti-CD25 staining from a control individual. (b) Staining pattern of an individual with active SLE with a particularly marked increase in the proportion of CD4 + T cells expressing CD25 (1·4%). Note also the marked reduction in the proportion of lymphocytes that are CD4 + (8.88% versus 37% in a). (c) Comparing 18 controls, 12 patients with active SLE and six with inactive disease, there was a significant increase in the percentage of CD25 + in those with active disease. P

Techniques Used: Staining, Expressing

Phenotypic comparison of CD25 subsets. Active lupus is associated with phenotypic changes in the CD25 – subset. Peripheral blood mononuclear cells were triple-stained for CD4, CD25 and expression of markers associated with CD4 + CD25 hi T regs or T cell activation. Subsets were gated using CD25 expression level and the proportion of each subset expressing the marker determined by comparison with staining with the isotype control antibody. Values shown are median percentage positive, with figures in brackets the 25th and 75th percentiles. Eleven controls, 11 with active systemic lupus erythematosus, five with inactive disease. (a) CD4 + T cells expressing CD25 at levels above the upper limit of CD25 expression by CD4 – lymphocytes. (b) The 2% CD4 + T cells expressing the highest CD25 levels (definition of CD25 hi used for FACS separation). (c) CD25 int . (d) CD25 – . CCR7 from 10 controls, nine patients with active and five with inactive disease. ×: P -value comparing controls and patients with inactive lupus; ♦, P -value comparing controls and patients with active lupus, †: P -value comparing patients with active and inactive lupus. Statistical comparison was made using the Kruskal–Wallis test with Dunn's post-test. FACS, fluorescence activated cell sorter; HLA-DR, human leucocyte antigen D-related.
Figure Legend Snippet: Phenotypic comparison of CD25 subsets. Active lupus is associated with phenotypic changes in the CD25 – subset. Peripheral blood mononuclear cells were triple-stained for CD4, CD25 and expression of markers associated with CD4 + CD25 hi T regs or T cell activation. Subsets were gated using CD25 expression level and the proportion of each subset expressing the marker determined by comparison with staining with the isotype control antibody. Values shown are median percentage positive, with figures in brackets the 25th and 75th percentiles. Eleven controls, 11 with active systemic lupus erythematosus, five with inactive disease. (a) CD4 + T cells expressing CD25 at levels above the upper limit of CD25 expression by CD4 – lymphocytes. (b) The 2% CD4 + T cells expressing the highest CD25 levels (definition of CD25 hi used for FACS separation). (c) CD25 int . (d) CD25 – . CCR7 from 10 controls, nine patients with active and five with inactive disease. ×: P -value comparing controls and patients with inactive lupus; ♦, P -value comparing controls and patients with active lupus, †: P -value comparing patients with active and inactive lupus. Statistical comparison was made using the Kruskal–Wallis test with Dunn's post-test. FACS, fluorescence activated cell sorter; HLA-DR, human leucocyte antigen D-related.

Techniques Used: Staining, Expressing, Activation Assay, Marker, FACS, Fluorescence

23) Product Images from "Long-Term Exposure to Inflammation Induces Differential Cytokine Patterns and Apoptosis in Dendritic Cells"

Article Title: Long-Term Exposure to Inflammation Induces Differential Cytokine Patterns and Apoptosis in Dendritic Cells

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2019.02702

IFNγ release from co-cultures with long-term activated DCs is reduced. (A) DCs were activated with the DC1 maturation cocktail (LPS and IFNγ) for 1, 2, 3, or 4 days (DC1-1d, DC1-2d, DC1-3d, DC1-4d) prior to being set up in co-culture with allogeneic PBMCs at a 1:10 ratio. After 5 days, IFNγ release was measured in the MLR supernatants with a lower limit of sensitivity of 156 pg/mL. Co-cultures with iDCs and sDC were included for comparison. Cumulative data are shown from five independent experiments with nine unique donor pairs in total ( n = 9), consisting of seven DC donors and five PBMC donors. All experiments were performed in quadruplicates. Bars represent mean + standard deviation. Two-way ANOVA with Tukey's post-hoc test was performed on log10-transformed data to compare DC1 groups (**** p ≤ 0.0001, ** p ≤ 0.01, * p ≤ 0.05, and ns, not statistically significant). (B) Same as (A) , except isolated CD4 + and CD8 + T cells were used as responder cells instead of PBMCs. Data from three donors ( n = 3) are normalized to highest value of IFNγ release within each donor. Bars represent mean + standard deviation. Two-way ANOVA with Tukey's post-hoc test was performed on normalized data to compare DC1 groups (**** p ≤ 0.0001, ** p ≤ 0.01, * p ≤ 0.05). The lower limit of IFNγ detection was 39 pg/mL. (C) Co-cultures were set up as in (A) and supplemented with anti-IL-12p70 or recombinant human IL-12p70. Recombinant human IL-12p70 was also added to cultures of PBMCs without DCs (w/o DC). After 5 days, IFNγ release was measured. Cumulative data are shown from one experiment with three unique donors ( n = 3). Bars represent mean + standard deviation.
Figure Legend Snippet: IFNγ release from co-cultures with long-term activated DCs is reduced. (A) DCs were activated with the DC1 maturation cocktail (LPS and IFNγ) for 1, 2, 3, or 4 days (DC1-1d, DC1-2d, DC1-3d, DC1-4d) prior to being set up in co-culture with allogeneic PBMCs at a 1:10 ratio. After 5 days, IFNγ release was measured in the MLR supernatants with a lower limit of sensitivity of 156 pg/mL. Co-cultures with iDCs and sDC were included for comparison. Cumulative data are shown from five independent experiments with nine unique donor pairs in total ( n = 9), consisting of seven DC donors and five PBMC donors. All experiments were performed in quadruplicates. Bars represent mean + standard deviation. Two-way ANOVA with Tukey's post-hoc test was performed on log10-transformed data to compare DC1 groups (**** p ≤ 0.0001, ** p ≤ 0.01, * p ≤ 0.05, and ns, not statistically significant). (B) Same as (A) , except isolated CD4 + and CD8 + T cells were used as responder cells instead of PBMCs. Data from three donors ( n = 3) are normalized to highest value of IFNγ release within each donor. Bars represent mean + standard deviation. Two-way ANOVA with Tukey's post-hoc test was performed on normalized data to compare DC1 groups (**** p ≤ 0.0001, ** p ≤ 0.01, * p ≤ 0.05). The lower limit of IFNγ detection was 39 pg/mL. (C) Co-cultures were set up as in (A) and supplemented with anti-IL-12p70 or recombinant human IL-12p70. Recombinant human IL-12p70 was also added to cultures of PBMCs without DCs (w/o DC). After 5 days, IFNγ release was measured. Cumulative data are shown from one experiment with three unique donors ( n = 3). Bars represent mean + standard deviation.

Techniques Used: Co-Culture Assay, Standard Deviation, Transformation Assay, Isolation, Recombinant

Related Articles

Isolation:

Article Title: Interleukin-33 promoting Th1 lymphocyte differentiation dependents on IL-12
Article Snippet: .. 2.5 CD4+ T cell purification and culture Human cord blood was obtained from informed consented mothers and peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation through Lymphoprep (Nycomed). .. CD4+ T cells from human PBMC and murine spleen were purified by negative selection (AutoMACS; MiltenyiBiotec).

Article Title: Natural regulatory T cells: number and function are normal in the majority of patients with lupus nephritis
Article Snippet: .. Peripheral blood mononuclear cells (PBMC) and CD4+ T cells were isolated by density gradient centrifugation on Lymphoprep (Nycomed, Marlow, UK). .. Isolated cells were stained with anti-CD4-fluorescein isothiocyanate (FITC) (clone Q1420, Sigma, Gillingham, UK) and anti-CD25-phycoerythrin (PE) (clone 2A3, BD PharmingenTM , Oxford, UK) in phosphate-buffered saline (PBS)/2% HS.

Article Title: 5-Azacytidine treatment sensitizes tumor cells to T-cell mediated cytotoxicity and modulates NK cells in patients with myeloid malignancies
Article Snippet: .. PBMC collection and processing Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood by gradient centrifugation (Lymphoprep, 1.077 g/ml, Nycomed Pharma AS, Oslo, Norway) at room temperature within a few hours after the blood was obtained from the patient. .. PBMCs were cryopreserved in 90% heat-inactivated fetal calf serum (FCS, Gibco, Life Technologies, Naerum, Denmark) and 10% dimethyl sulfoxide (Sigma-Aldrich, Broendby, Denmark) with 5–30 × 106 cells/ampoule and stored at −150 °C.

Article Title: Relationship between functional ability in older people, immune system status, and intensity of response to CMV
Article Snippet: .. To analyze the proliferation status of CD4+ and CD8+ T cells, peripheral blood mononuclear cells (PBMC) were isolated by centrifugation on Ficoll-Hypaque gradients (Lymphoprep; Nycomed, Oslo, Norway). .. CD4+ and CD8+ T cells from elderly were isolated with magnetic beads (Myltenyi Biotec GmbH, Bergisch Gladbach, Germany).

Article Title: Protection of Hepatocytes from Cytotoxic T Cell Mediated Killing by Interferon-Alpha
Article Snippet: .. Isolation of Peripheral Blood Mononuclear Cells (PBMC): PBMCs were isolated from whole blood collected and mixed with either heparin or EDTA, and separated by density gradient centrifugation over lymphoprep (Nycomed, UK). .. CTL lines Lines were generated from the PBMC of healthy donors.

Article Title: Molecular mechanisms of extracellular adenine nucleotides-mediated inhibition of human Cd4+ T lymphocytes activation
Article Snippet: .. Isolation of resting CD4+ T cells from peripheral blood Peripheral blood mononuclear cells (PBMC) were isolated from buffy coats of healthy blood donors by density gradient centrifugation on Lymphoprep (Nycomed, Oslo, Norway). .. After five washes in Hank's balanced salt solution (HBSS) (Gibco Life Technologies, Paisley, UK) CD4+ T cells were isolated from PBMC using the MACS negative depletion system (Miltenyi Biotec, Auburn, CA).

Article Title: Evidence of a role for both anti-Hsp70 antibody and endothelial surface membrane Hsp70 in atherosclerosis
Article Snippet: .. To determine antibody-dependent cellular cytotoxicity (ADCC), peripheral blood mononuclear cells (PBMC) were isolated from healthy rats by density centrifugation (Lymphoprep, density 1,083; Nycomed Pharmaceuticals Oslo, Norway) as described previously (Jurgens et al. ). ..

Centrifugation:

Article Title: Relationship between functional ability in older people, immune system status, and intensity of response to CMV
Article Snippet: .. To analyze the proliferation status of CD4+ and CD8+ T cells, peripheral blood mononuclear cells (PBMC) were isolated by centrifugation on Ficoll-Hypaque gradients (Lymphoprep; Nycomed, Oslo, Norway). .. CD4+ and CD8+ T cells from elderly were isolated with magnetic beads (Myltenyi Biotec GmbH, Bergisch Gladbach, Germany).

Article Title: Evidence of a role for both anti-Hsp70 antibody and endothelial surface membrane Hsp70 in atherosclerosis
Article Snippet: .. To determine antibody-dependent cellular cytotoxicity (ADCC), peripheral blood mononuclear cells (PBMC) were isolated from healthy rats by density centrifugation (Lymphoprep, density 1,083; Nycomed Pharmaceuticals Oslo, Norway) as described previously (Jurgens et al. ). ..

Article Title: Proviral load and immune function in blood and lymph node during HIV-1 and HIV-2 infection
Article Snippet: .. PBMC and LNMC were purified by density centrifugation on Lymphoprep (Nycomed, Oslo, Norway). ..

Gradient Centrifugation:

Article Title: Interleukin-33 promoting Th1 lymphocyte differentiation dependents on IL-12
Article Snippet: .. 2.5 CD4+ T cell purification and culture Human cord blood was obtained from informed consented mothers and peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation through Lymphoprep (Nycomed). .. CD4+ T cells from human PBMC and murine spleen were purified by negative selection (AutoMACS; MiltenyiBiotec).

Article Title: Natural regulatory T cells: number and function are normal in the majority of patients with lupus nephritis
Article Snippet: .. Peripheral blood mononuclear cells (PBMC) and CD4+ T cells were isolated by density gradient centrifugation on Lymphoprep (Nycomed, Marlow, UK). .. Isolated cells were stained with anti-CD4-fluorescein isothiocyanate (FITC) (clone Q1420, Sigma, Gillingham, UK) and anti-CD25-phycoerythrin (PE) (clone 2A3, BD PharmingenTM , Oxford, UK) in phosphate-buffered saline (PBS)/2% HS.

Article Title: 5-Azacytidine treatment sensitizes tumor cells to T-cell mediated cytotoxicity and modulates NK cells in patients with myeloid malignancies
Article Snippet: .. PBMC collection and processing Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood by gradient centrifugation (Lymphoprep, 1.077 g/ml, Nycomed Pharma AS, Oslo, Norway) at room temperature within a few hours after the blood was obtained from the patient. .. PBMCs were cryopreserved in 90% heat-inactivated fetal calf serum (FCS, Gibco, Life Technologies, Naerum, Denmark) and 10% dimethyl sulfoxide (Sigma-Aldrich, Broendby, Denmark) with 5–30 × 106 cells/ampoule and stored at −150 °C.

Article Title: Protection of Hepatocytes from Cytotoxic T Cell Mediated Killing by Interferon-Alpha
Article Snippet: .. Isolation of Peripheral Blood Mononuclear Cells (PBMC): PBMCs were isolated from whole blood collected and mixed with either heparin or EDTA, and separated by density gradient centrifugation over lymphoprep (Nycomed, UK). .. CTL lines Lines were generated from the PBMC of healthy donors.

Article Title: Molecular mechanisms of extracellular adenine nucleotides-mediated inhibition of human Cd4+ T lymphocytes activation
Article Snippet: .. Isolation of resting CD4+ T cells from peripheral blood Peripheral blood mononuclear cells (PBMC) were isolated from buffy coats of healthy blood donors by density gradient centrifugation on Lymphoprep (Nycomed, Oslo, Norway). .. After five washes in Hank's balanced salt solution (HBSS) (Gibco Life Technologies, Paisley, UK) CD4+ T cells were isolated from PBMC using the MACS negative depletion system (Miltenyi Biotec, Auburn, CA).

Purification:

Article Title: Interleukin-33 promoting Th1 lymphocyte differentiation dependents on IL-12
Article Snippet: .. 2.5 CD4+ T cell purification and culture Human cord blood was obtained from informed consented mothers and peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation through Lymphoprep (Nycomed). .. CD4+ T cells from human PBMC and murine spleen were purified by negative selection (AutoMACS; MiltenyiBiotec).

Article Title: Proviral load and immune function in blood and lymph node during HIV-1 and HIV-2 infection
Article Snippet: .. PBMC and LNMC were purified by density centrifugation on Lymphoprep (Nycomed, Oslo, Norway). ..

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 88
    Nycomed pbmc collection
    Cancer testis antigen (CTA)-specific T cells in the peripheral blood of patients. Detection of CTA- or viral-specific T cells in <t>PBMCs</t> by MHC multimers, expressed in percentage of CD8 T cells. ( a ) The sum of CTA-specific T cells as measured in peripheral blood at different time points during treatment. Eight patients were tested, results from the six patients with detectable responses are shown. First cycle represents a sample obtained before treatment. The following responses were found for each patient: AZA 1 (SART-3 WLE , SART-3 QIR , Sp17 ILD ), AZA 2 (MAGE-A2 LVH , MAGE-A2 KMV , TAG-1 SLG ), AZA 4 (MAGE-A2 LVH , MAGE-A2 LVQ , NY-ESO-1 QLS ), AZA 5 (MAGE-A1 EAD ), AZA 12 (MAGE-A2 LVH , MAGE-A2 KMV , CDCA1 KLA , TAG-1 SLG , NY-ESO-1 SLL , MAGE-A1 EAD ) and AZA 16 (MAGE-A2 LVH , MAGE-A2 KMV , GnTV VLP , TAG-1 SLG ). ( b ) The frequency of individual CTA-specific T cells detected after an in vitro peptide pre-stimulation was performed at different time points during treatment. ( c ) The sum of virus-specific T cells detected over the course of treatment. The following responses were detected: AZA 1 (EBV RLR , EBV RLR , FLU ILR ), AZA 2 (EBV GLC , FLU ILR ), AZA 4 (EBV GLC , EBV YVL , FLU GIL ), AZA 5 (FLU BP-VSD ), AZA 12 (CMV VTE , CMV YSE , CMV NLV , FLU GIL ), AZA 14 (CMV YSE , CMV VTE , FLU BP-VSD ), AZA 16 (CMV NLV , EBV GLC ) and AZA 17 (CMV YSE , CMV VTE , FLU BP-VSD ). MHC-multimer-specific T cells are given in percentage of CD8 cells. Significance is indicated by * P
    Pbmc Collection, supplied by Nycomed, used in various techniques. Bioz Stars score: 88/100, based on 224 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmc collection/product/Nycomed
    Average 88 stars, based on 224 article reviews
    Price from $9.99 to $1999.99
    pbmc collection - by Bioz Stars, 2020-09
    88/100 stars
      Buy from Supplier

    Image Search Results


    Cancer testis antigen (CTA)-specific T cells in the peripheral blood of patients. Detection of CTA- or viral-specific T cells in PBMCs by MHC multimers, expressed in percentage of CD8 T cells. ( a ) The sum of CTA-specific T cells as measured in peripheral blood at different time points during treatment. Eight patients were tested, results from the six patients with detectable responses are shown. First cycle represents a sample obtained before treatment. The following responses were found for each patient: AZA 1 (SART-3 WLE , SART-3 QIR , Sp17 ILD ), AZA 2 (MAGE-A2 LVH , MAGE-A2 KMV , TAG-1 SLG ), AZA 4 (MAGE-A2 LVH , MAGE-A2 LVQ , NY-ESO-1 QLS ), AZA 5 (MAGE-A1 EAD ), AZA 12 (MAGE-A2 LVH , MAGE-A2 KMV , CDCA1 KLA , TAG-1 SLG , NY-ESO-1 SLL , MAGE-A1 EAD ) and AZA 16 (MAGE-A2 LVH , MAGE-A2 KMV , GnTV VLP , TAG-1 SLG ). ( b ) The frequency of individual CTA-specific T cells detected after an in vitro peptide pre-stimulation was performed at different time points during treatment. ( c ) The sum of virus-specific T cells detected over the course of treatment. The following responses were detected: AZA 1 (EBV RLR , EBV RLR , FLU ILR ), AZA 2 (EBV GLC , FLU ILR ), AZA 4 (EBV GLC , EBV YVL , FLU GIL ), AZA 5 (FLU BP-VSD ), AZA 12 (CMV VTE , CMV YSE , CMV NLV , FLU GIL ), AZA 14 (CMV YSE , CMV VTE , FLU BP-VSD ), AZA 16 (CMV NLV , EBV GLC ) and AZA 17 (CMV YSE , CMV VTE , FLU BP-VSD ). MHC-multimer-specific T cells are given in percentage of CD8 cells. Significance is indicated by * P

    Journal: Blood Cancer Journal

    Article Title: 5-Azacytidine treatment sensitizes tumor cells to T-cell mediated cytotoxicity and modulates NK cells in patients with myeloid malignancies

    doi: 10.1038/bcj.2014.14

    Figure Lengend Snippet: Cancer testis antigen (CTA)-specific T cells in the peripheral blood of patients. Detection of CTA- or viral-specific T cells in PBMCs by MHC multimers, expressed in percentage of CD8 T cells. ( a ) The sum of CTA-specific T cells as measured in peripheral blood at different time points during treatment. Eight patients were tested, results from the six patients with detectable responses are shown. First cycle represents a sample obtained before treatment. The following responses were found for each patient: AZA 1 (SART-3 WLE , SART-3 QIR , Sp17 ILD ), AZA 2 (MAGE-A2 LVH , MAGE-A2 KMV , TAG-1 SLG ), AZA 4 (MAGE-A2 LVH , MAGE-A2 LVQ , NY-ESO-1 QLS ), AZA 5 (MAGE-A1 EAD ), AZA 12 (MAGE-A2 LVH , MAGE-A2 KMV , CDCA1 KLA , TAG-1 SLG , NY-ESO-1 SLL , MAGE-A1 EAD ) and AZA 16 (MAGE-A2 LVH , MAGE-A2 KMV , GnTV VLP , TAG-1 SLG ). ( b ) The frequency of individual CTA-specific T cells detected after an in vitro peptide pre-stimulation was performed at different time points during treatment. ( c ) The sum of virus-specific T cells detected over the course of treatment. The following responses were detected: AZA 1 (EBV RLR , EBV RLR , FLU ILR ), AZA 2 (EBV GLC , FLU ILR ), AZA 4 (EBV GLC , EBV YVL , FLU GIL ), AZA 5 (FLU BP-VSD ), AZA 12 (CMV VTE , CMV YSE , CMV NLV , FLU GIL ), AZA 14 (CMV YSE , CMV VTE , FLU BP-VSD ), AZA 16 (CMV NLV , EBV GLC ) and AZA 17 (CMV YSE , CMV VTE , FLU BP-VSD ). MHC-multimer-specific T cells are given in percentage of CD8 cells. Significance is indicated by * P

    Article Snippet: PBMC collection and processing Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood by gradient centrifugation (Lymphoprep, 1.077 g/ml, Nycomed Pharma AS, Oslo, Norway) at room temperature within a few hours after the blood was obtained from the patient.

    Techniques: In Vitro, Gas Chromatography

    CD4 % is lower in PBMC than in LNMC in HIV infection

    Journal: Clinical and Experimental Immunology

    Article Title: Proviral load and immune function in blood and lymph node during HIV-1 and HIV-2 infection

    doi: 10.1046/j.1365-2249.1999.00914.x

    Figure Lengend Snippet: CD4 % is lower in PBMC than in LNMC in HIV infection

    Article Snippet: PBMC and LNMC were purified by density centrifugation on Lymphoprep (Nycomed, Oslo, Norway).

    Techniques: Infection

    IFN-α stimulated hepatocytes are still able to act as APCs to CTL. HepG2 cells were stimulated for 16 hours with 1000 U/ml IFN-α, or left untreated. The cells were then pulsed with the specific cognate peptide and washed, prior to co-incubation with the CTL line 2. As controls non-peptide-pulsed cells were used, or all PBMC were peptide pulsed or stimulated with PMA/ionomycin. A) The raw FACS data showing the CD8+ expression on the y axis's and IFN-γ on the x axis's. B) The percentage secretion data from such experiment. HepG2p represents peptide pulsed cells with or without IFN-α stimulation as indicated. This data is representative of 4 such experiments.

    Journal: PLoS ONE

    Article Title: Protection of Hepatocytes from Cytotoxic T Cell Mediated Killing by Interferon-Alpha

    doi: 10.1371/journal.pone.0000791

    Figure Lengend Snippet: IFN-α stimulated hepatocytes are still able to act as APCs to CTL. HepG2 cells were stimulated for 16 hours with 1000 U/ml IFN-α, or left untreated. The cells were then pulsed with the specific cognate peptide and washed, prior to co-incubation with the CTL line 2. As controls non-peptide-pulsed cells were used, or all PBMC were peptide pulsed or stimulated with PMA/ionomycin. A) The raw FACS data showing the CD8+ expression on the y axis's and IFN-γ on the x axis's. B) The percentage secretion data from such experiment. HepG2p represents peptide pulsed cells with or without IFN-α stimulation as indicated. This data is representative of 4 such experiments.

    Article Snippet: Isolation of Peripheral Blood Mononuclear Cells (PBMC): PBMCs were isolated from whole blood collected and mixed with either heparin or EDTA, and separated by density gradient centrifugation over lymphoprep (Nycomed, UK).

    Techniques: Activated Clotting Time Assay, CTL Assay, Incubation, FACS, Expressing

    Dose–response curves for ADCC. The procedures for OxLDL incubated and Ab treatment were the same as described in the legend for Fig. . As effectors normal rat PBMC were added to the culture instead of complement. Note increased 51Cr release

    Journal: Cell Stress & Chaperones

    Article Title: Evidence of a role for both anti-Hsp70 antibody and endothelial surface membrane Hsp70 in atherosclerosis

    doi: 10.1007/s12192-013-0404-4

    Figure Lengend Snippet: Dose–response curves for ADCC. The procedures for OxLDL incubated and Ab treatment were the same as described in the legend for Fig. . As effectors normal rat PBMC were added to the culture instead of complement. Note increased 51Cr release

    Article Snippet: To determine antibody-dependent cellular cytotoxicity (ADCC), peripheral blood mononuclear cells (PBMC) were isolated from healthy rats by density centrifugation (Lymphoprep, density 1,083; Nycomed Pharmaceuticals Oslo, Norway) as described previously (Jurgens et al. ).

    Techniques: Incubation