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imag human regulatory t lymphocyte separation kit  (Becton Dickinson)
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Becton Dickinson imag human regulatory t lymphocyte separation kit
HIV infection modulates <t>regulatory</t> <t>T</t> (Treg) cell frequencies and numbers. HIV‐infected participants were divided into three groups according to their plasmatic HIV viral loads [VL< 3 Log copies/ml (n = 8); 3–4 (n = 8); > 4 (n = 12)]. The total number of Treg cells decreased when the plasmatic HIV viral load increased (a), whereas an inverse tendency was observed when Treg cell frequencies were considered (b) although the differences were not significant using Kruskal–Wallis test. However, plasmatic HIV viral load correlated negatively with total Treg cell numbers (r = −0·51; P = 0·011; c) and positively with Treg cell frequencies (r = 0·55; P = 0·007; d) using Spearman test.
Imag Human Regulatory T Lymphocyte Separation Kit, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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HIV infection modulates regulatory T (Treg) cell frequencies and numbers. HIV‐infected participants were divided into three groups according to their plasmatic HIV viral loads [VL< 3 Log copies/ml (n = 8); 3–4 (n = 8); > 4 (n = 12)]. The total number of Treg cells decreased when the plasmatic HIV viral load increased (a), whereas an inverse tendency was observed when Treg cell frequencies were considered (b) although the differences were not significant using Kruskal–Wallis test. However, plasmatic HIV viral load correlated negatively with total Treg cell numbers (r = −0·51; P = 0·011; c) and positively with Treg cell frequencies (r = 0·55; P = 0·007; d) using Spearman test.

Journal: Immunology

Article Title: Phenotypic characterization of regulatory T cells from antiretroviral‐naive HIV ‐1‐infected people

doi: 10.1111/imm.12738

Figure Lengend Snippet: HIV infection modulates regulatory T (Treg) cell frequencies and numbers. HIV‐infected participants were divided into three groups according to their plasmatic HIV viral loads [VL< 3 Log copies/ml (n = 8); 3–4 (n = 8); > 4 (n = 12)]. The total number of Treg cells decreased when the plasmatic HIV viral load increased (a), whereas an inverse tendency was observed when Treg cell frequencies were considered (b) although the differences were not significant using Kruskal–Wallis test. However, plasmatic HIV viral load correlated negatively with total Treg cell numbers (r = −0·51; P = 0·011; c) and positively with Treg cell frequencies (r = 0·55; P = 0·007; d) using Spearman test.

Article Snippet: Partial purification of Treg cells CD4 + CD25 + Treg cells were isolated from PBMCs using the BD IMag human regulatory T lymphocyte separation kit (BD Biosciences) according to the manufacturer's instructions.

Techniques: Infection

Tracking FoxP3‐expressing cells using a combination of CD25+ CD127lo markers in either magnetically sorted regulatory T (Treg) cells or bulk peripheral blood mononuclear cells (PBMC). (a) Representative dot plots of the gating strategy for magnetically purified Treg cells. Following identification of lymphocytes based on forward and side scatter, CD3+ CD4+ T cells were selected from live lymphocytes and CD127lo T cells were gated from this population; 1000 events were collected from the CD127lo gate to detect cells that were positive for CD25 and FoxP3 expression. (b, c) Magnetic sorting of Treg cells resulted in a significant increase in the desired population in both HIV + (n = 16) and HIV – (n = 6) participants. (d–f) There was a positive correlation between CD127lo CD25+ and FoxP3+ irrespective of whether staining was done with bulk PBMCs (r = 0·94; P < 0·0001; d), partially purified Treg cells (r = 0·86; P < 0·0001; e) or fully purified Treg cells (r = 0·71; P = 0·0002; f) Horizontal bars represent the median. Values were compared using Kruskal–Wallis test followed by Dunn's multiple comparison test.(*P <0·05; **P <0·001; ***P <0·0001) Each dot represents a single individual and correlation was assessed using Spearman test.

Journal: Immunology

Article Title: Phenotypic characterization of regulatory T cells from antiretroviral‐naive HIV ‐1‐infected people

doi: 10.1111/imm.12738

Figure Lengend Snippet: Tracking FoxP3‐expressing cells using a combination of CD25+ CD127lo markers in either magnetically sorted regulatory T (Treg) cells or bulk peripheral blood mononuclear cells (PBMC). (a) Representative dot plots of the gating strategy for magnetically purified Treg cells. Following identification of lymphocytes based on forward and side scatter, CD3+ CD4+ T cells were selected from live lymphocytes and CD127lo T cells were gated from this population; 1000 events were collected from the CD127lo gate to detect cells that were positive for CD25 and FoxP3 expression. (b, c) Magnetic sorting of Treg cells resulted in a significant increase in the desired population in both HIV + (n = 16) and HIV – (n = 6) participants. (d–f) There was a positive correlation between CD127lo CD25+ and FoxP3+ irrespective of whether staining was done with bulk PBMCs (r = 0·94; P < 0·0001; d), partially purified Treg cells (r = 0·86; P < 0·0001; e) or fully purified Treg cells (r = 0·71; P = 0·0002; f) Horizontal bars represent the median. Values were compared using Kruskal–Wallis test followed by Dunn's multiple comparison test.(*P <0·05; **P <0·001; ***P <0·0001) Each dot represents a single individual and correlation was assessed using Spearman test.

Article Snippet: Partial purification of Treg cells CD4 + CD25 + Treg cells were isolated from PBMCs using the BD IMag human regulatory T lymphocyte separation kit (BD Biosciences) according to the manufacturer's instructions.

Techniques: Expressing, Purification, Staining

Identification of regulatory T (Treg) cell subsets in antiretroviral‐naive HIV infection using CD45RA, CD27, CD62L and CCR7 cell surface markers. (a) Representative dot plots of the gating strategy of different Treg cell subsets. Purified Treg cells obtained from peripheral blood mononuclear cells (PBMCs) of HIV + (n = 31) and HIV – (n = 17) participants were stained using monoclonal antibodies specific for CD3, CD4, CD25, CD127, CD45RA, CD27, CD62L and CCR7 and then analysed by multiparametric flow cytometry. Dot plots showing co‐expression of both CD27 and CD45RA on CD4+ CD25+ CD127lo Treg cells are represented in the middle panel. The data on CCR7 and CD62L are shown in each CD27 CD45RA subset. Distribution of Treg cell subsets analysed on CD45RA + or CD45RA – Treg cells are shown in (b) and (c), respectively. Whereas seronegative people displayed predominantly naive (P < 0·05) and central memory (P < 0·001) phenotypes, antiretroviral therapy‐naive HIV‐infected participants showed significantly elevated numbers of effector (P < 0·001) and effector memory (P < 0·05) Treg cell subsets. Bars represent the median and Mann–Whitney U‐test was used to compare each subset between HIV‐infected and uninfected controls. *P<0·05; **P<0·001.

Journal: Immunology

Article Title: Phenotypic characterization of regulatory T cells from antiretroviral‐naive HIV ‐1‐infected people

doi: 10.1111/imm.12738

Figure Lengend Snippet: Identification of regulatory T (Treg) cell subsets in antiretroviral‐naive HIV infection using CD45RA, CD27, CD62L and CCR7 cell surface markers. (a) Representative dot plots of the gating strategy of different Treg cell subsets. Purified Treg cells obtained from peripheral blood mononuclear cells (PBMCs) of HIV + (n = 31) and HIV – (n = 17) participants were stained using monoclonal antibodies specific for CD3, CD4, CD25, CD127, CD45RA, CD27, CD62L and CCR7 and then analysed by multiparametric flow cytometry. Dot plots showing co‐expression of both CD27 and CD45RA on CD4+ CD25+ CD127lo Treg cells are represented in the middle panel. The data on CCR7 and CD62L are shown in each CD27 CD45RA subset. Distribution of Treg cell subsets analysed on CD45RA + or CD45RA – Treg cells are shown in (b) and (c), respectively. Whereas seronegative people displayed predominantly naive (P < 0·05) and central memory (P < 0·001) phenotypes, antiretroviral therapy‐naive HIV‐infected participants showed significantly elevated numbers of effector (P < 0·001) and effector memory (P < 0·05) Treg cell subsets. Bars represent the median and Mann–Whitney U‐test was used to compare each subset between HIV‐infected and uninfected controls. *P<0·05; **P<0·001.

Article Snippet: Partial purification of Treg cells CD4 + CD25 + Treg cells were isolated from PBMCs using the BD IMag human regulatory T lymphocyte separation kit (BD Biosciences) according to the manufacturer's instructions.

Techniques: Infection, Purification, Staining, Flow Cytometry, Expressing, MANN-WHITNEY

Relationship between helper CD4+ T‐cell count and regulatory T (Treg) cells. HIV‐infected participants were grouped into three categories according to their helper CD4+ T‐cell counts [CD4 < 350 cells/mm3 (n = 5); 350–499 cells/mm3 (n = 9); >500 cells/mm3 (n = 17)]. When compared with uninfected participants (n = 17), the median number of Treg cells diminished proportionately with the helper CD4+ T‐cell count (a). The lowest values were observed in HIV‐infected participants with CD4 < 350 cells/mm3 (P < 0·05). However, this group showed significant increase in Treg cell percentages within total CD4+ T cells compared with other groups (b) with helper CD4+ T‐cell count between 350 and 499 cells/mm3 (P < 0·05) or > 500 cells/mm3 (P < 0·001). (c, d) Helper CD4+ T‐cell count of HIV‐infected participants correlated positively with the total number of Treg cells and negatively with the percentages of Treg cells. Bars represent the median values, which were compared using Kruskal–Wallis test followed by Dunn's multiple comparison test. (*P <0·05; **P <0·001) Each dot represents a single individual and the correlation was calculated using Spearman test.

Journal: Immunology

Article Title: Phenotypic characterization of regulatory T cells from antiretroviral‐naive HIV ‐1‐infected people

doi: 10.1111/imm.12738

Figure Lengend Snippet: Relationship between helper CD4+ T‐cell count and regulatory T (Treg) cells. HIV‐infected participants were grouped into three categories according to their helper CD4+ T‐cell counts [CD4 < 350 cells/mm3 (n = 5); 350–499 cells/mm3 (n = 9); >500 cells/mm3 (n = 17)]. When compared with uninfected participants (n = 17), the median number of Treg cells diminished proportionately with the helper CD4+ T‐cell count (a). The lowest values were observed in HIV‐infected participants with CD4 < 350 cells/mm3 (P < 0·05). However, this group showed significant increase in Treg cell percentages within total CD4+ T cells compared with other groups (b) with helper CD4+ T‐cell count between 350 and 499 cells/mm3 (P < 0·05) or > 500 cells/mm3 (P < 0·001). (c, d) Helper CD4+ T‐cell count of HIV‐infected participants correlated positively with the total number of Treg cells and negatively with the percentages of Treg cells. Bars represent the median values, which were compared using Kruskal–Wallis test followed by Dunn's multiple comparison test. (*P <0·05; **P <0·001) Each dot represents a single individual and the correlation was calculated using Spearman test.

Article Snippet: Partial purification of Treg cells CD4 + CD25 + Treg cells were isolated from PBMCs using the BD IMag human regulatory T lymphocyte separation kit (BD Biosciences) according to the manufacturer's instructions.

Techniques: Cell Counting, Infection

Antiretroviral therapy‐naive HIV‐infection reduces CD39 and CD73 expression in naive and central memory regulatory T (Treg) cells. (a, c, e) Representative dot plots illustrating the gating strategy of CD39 and CD73 expression. The median numbers of each Treg subset expressing CD39, CD73 or both markers are shown in (b), (d) and (f), respectively. The differences between HIV + and HIV – participants were calculated using Mann–Whitney U‐test. Effector and effector memory Treg cells from antiretroviral therapy‐naive HIV‐infected participants showed significant increase in CD39 (P < 0·05) and CD73 (P < 0·001) expression. In contrast, these markers were highly expressed instead on naive (P < 0·05 and P < 0·0001 respectively for CD39 and CD73) and central memory (P < 0·05 for CD73) Treg cells from uninfected individuals. A similar trend was observed for the combined expression of both CD39 and CD73. *P <0·05, **P <0·001, ****P <0·0001.

Journal: Immunology

Article Title: Phenotypic characterization of regulatory T cells from antiretroviral‐naive HIV ‐1‐infected people

doi: 10.1111/imm.12738

Figure Lengend Snippet: Antiretroviral therapy‐naive HIV‐infection reduces CD39 and CD73 expression in naive and central memory regulatory T (Treg) cells. (a, c, e) Representative dot plots illustrating the gating strategy of CD39 and CD73 expression. The median numbers of each Treg subset expressing CD39, CD73 or both markers are shown in (b), (d) and (f), respectively. The differences between HIV + and HIV – participants were calculated using Mann–Whitney U‐test. Effector and effector memory Treg cells from antiretroviral therapy‐naive HIV‐infected participants showed significant increase in CD39 (P < 0·05) and CD73 (P < 0·001) expression. In contrast, these markers were highly expressed instead on naive (P < 0·05 and P < 0·0001 respectively for CD39 and CD73) and central memory (P < 0·05 for CD73) Treg cells from uninfected individuals. A similar trend was observed for the combined expression of both CD39 and CD73. *P <0·05, **P <0·001, ****P <0·0001.

Article Snippet: Partial purification of Treg cells CD4 + CD25 + Treg cells were isolated from PBMCs using the BD IMag human regulatory T lymphocyte separation kit (BD Biosciences) according to the manufacturer's instructions.

Techniques: Infection, Expressing, MANN-WHITNEY

Expression of HLA‐DR and CD38 on regulatory T (Treg) cell subsets. In (a) the gating strategy for HLA‐DR Treg cell surface expression is shown. The median values of HLA‐DR and CD38 are represented as bars with range (b and c). The differences in HLA‐DR/CD38 expression for each Treg subset was assessed between HIV + and HIV – participants using Mann–Whitney U‐test (*P <0·05; **P <0·001). While naive Treg cells from uninfected people displayed a significant increase in the expression of HLA‐DR/CD38 (P < 0·05), as shown in (b) and (c), these markers were highly expressed by effector and effector memory Treg cells from antiretroviral therapy‐naive HIV‐infected participants (P < 0·001).

Journal: Immunology

Article Title: Phenotypic characterization of regulatory T cells from antiretroviral‐naive HIV ‐1‐infected people

doi: 10.1111/imm.12738

Figure Lengend Snippet: Expression of HLA‐DR and CD38 on regulatory T (Treg) cell subsets. In (a) the gating strategy for HLA‐DR Treg cell surface expression is shown. The median values of HLA‐DR and CD38 are represented as bars with range (b and c). The differences in HLA‐DR/CD38 expression for each Treg subset was assessed between HIV + and HIV – participants using Mann–Whitney U‐test (*P <0·05; **P <0·001). While naive Treg cells from uninfected people displayed a significant increase in the expression of HLA‐DR/CD38 (P < 0·05), as shown in (b) and (c), these markers were highly expressed by effector and effector memory Treg cells from antiretroviral therapy‐naive HIV‐infected participants (P < 0·001).

Article Snippet: Partial purification of Treg cells CD4 + CD25 + Treg cells were isolated from PBMCs using the BD IMag human regulatory T lymphocyte separation kit (BD Biosciences) according to the manufacturer's instructions.

Techniques: Expressing, MANN-WHITNEY, Infection