Journal: AIDS Research and Human Retroviruses

Article Title: Immunological Function Restoration with Lopinavir/Ritonavir Versus Efavirenz Containing Regimens in HIV-Infected Patients: A Randomized Clinical Trial

doi: 10.1089/aid.2013.0185

Figure Lengend Snippet: T Cell Subsets Analyzed in the Study, Grouped According to Eight Different Categories

Article Snippet: Numbers inside plots or histograms represent the percentage of positive events in each quadrant or region. table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 T cell subset category (antibody panel) T cell subset Differentiation stage of CD4 and CD8 cells (CD3-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD4 and CD8 cells (total) Naive cells (CD45RA + CD27 + ) Central memory cells (CD45RA − CD27 + ) Effector memory cells (CD45RA − CD27 − ) Effector cells (CD45RA + CD27 − ) Activation of CD4 and CD8 T cells (including naive, central memory, effector memory, and effector cells) (HLADR-FITC/CD27-PE/CD45RA-ECD/CD38-PECy5/CD4-PECy7 or CD8- PECy7) CD38 + DR − CD38 + DR + CD38 − DR + Thymic function (CD31) (CD31-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD31 expression on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) Expression of CD127 (CD127-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD127 expression on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) Senescence (CD57) (CD57-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD57 expression on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) Exhaustion (PD-1 and Tim-3) of CD3, CD4, and CD8 T cells (CD4-FITC/Tim-3-PE/CD3-ECD/CD8-PECy5/PD-1-biotin/streptavidin-PECy7) PD-1 + Tim-3 − PD-1 + Tim-3 + PD-1 − Tim-3 + Apoptosis (annexin binding) (annexin-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) Annexin + binding on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) T regulatory cells (FoxP3-FITC/CD127-PE/CD8-ECD/CD25-PECy5/CD4-PECy7) CD4 + CD25 + CD127 − FoxP3 + cells Open in a separate window T Cell Subsets Analyzed in the Study, Grouped According to Eight Different Categories Plasma interleukin (IL)-7 levels IL-7 was measured using a highly sensitive commercial immunoassay (Quantikine HS, R&D Systems, Barcelona, Spain), which has a lower limit of detection of 0.1 pg/ml following the manufacturer's instructions.

Techniques: Activation Assay, Expressing, Binding Assay

Journal: AIDS Research and Human Retroviruses

Article Title: Immunological Function Restoration with Lopinavir/Ritonavir Versus Efavirenz Containing Regimens in HIV-Infected Patients: A Randomized Clinical Trial

doi: 10.1089/aid.2013.0185

Figure Lengend Snippet: Representative example of flow cytometry data and gating strategy. A representative example for some of the antibody panels used is shown. (A) Expression of CD45RA and CD27 on CD4 and CD8 T cells to define the differentiation stage of these cells. (B, C) Expression of activation markers CD38 and HLA-DR on different subsets of CD4 (B) and CD8 (C) T cells as defined by CD45RA and CD27 expression. (D, E) Annexin binding on different subsets of CD4 (D) and CD8 (E) T cells as defined by CD45RA and CD27 expression. (F) Expression of exhaustion markers PD-1 and Tim-3 on CD4 and CD8 T cells. (G) Gating strategy used to define T regulatory (Treg) CD4 cells as CD127−CD25+FoxP3+. Numbers inside plots or histograms represent the percentage of positive events in each quadrant or region.

Article Snippet: Numbers inside plots or histograms represent the percentage of positive events in each quadrant or region. table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 T cell subset category (antibody panel) T cell subset Differentiation stage of CD4 and CD8 cells (CD3-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD4 and CD8 cells (total) Naive cells (CD45RA + CD27 + ) Central memory cells (CD45RA − CD27 + ) Effector memory cells (CD45RA − CD27 − ) Effector cells (CD45RA + CD27 − ) Activation of CD4 and CD8 T cells (including naive, central memory, effector memory, and effector cells) (HLADR-FITC/CD27-PE/CD45RA-ECD/CD38-PECy5/CD4-PECy7 or CD8- PECy7) CD38 + DR − CD38 + DR + CD38 − DR + Thymic function (CD31) (CD31-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD31 expression on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) Expression of CD127 (CD127-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD127 expression on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) Senescence (CD57) (CD57-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD57 expression on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) Exhaustion (PD-1 and Tim-3) of CD3, CD4, and CD8 T cells (CD4-FITC/Tim-3-PE/CD3-ECD/CD8-PECy5/PD-1-biotin/streptavidin-PECy7) PD-1 + Tim-3 − PD-1 + Tim-3 + PD-1 − Tim-3 + Apoptosis (annexin binding) (annexin-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) Annexin + binding on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) T regulatory cells (FoxP3-FITC/CD127-PE/CD8-ECD/CD25-PECy5/CD4-PECy7) CD4 + CD25 + CD127 − FoxP3 + cells Open in a separate window T Cell Subsets Analyzed in the Study, Grouped According to Eight Different Categories Plasma interleukin (IL)-7 levels IL-7 was measured using a highly sensitive commercial immunoassay (Quantikine HS, R&D Systems, Barcelona, Spain), which has a lower limit of detection of 0.1 pg/ml following the manufacturer's instructions.

Techniques: Flow Cytometry, Expressing, Activation Assay, Binding Assay

Journal: AIDS Research and Human Retroviruses

Article Title: Immunological Function Restoration with Lopinavir/Ritonavir Versus Efavirenz Containing Regimens in HIV-Infected Patients: A Randomized Clinical Trial

doi: 10.1089/aid.2013.0185

Figure Lengend Snippet: Box-plot graphs showing baseline (light gray boxes) and week 48 (dark gray boxes) levels of different T cell subsets in the whole population of patients. Vertical axes represent the proportion of cells and horizontal axes the different CD4 and CD8 T cells subsets. (A) Differentiation stage (coexpression of CD45RA and CD27) of CD4 and CD8 T cells. (B) Activation (coexpression of CD38 and HLA-DR) of different CD4 and CD8 T cells subsets. (C) Senescence (CD57 expression), exhaustion (PD1 expression), and apoptosis (Annexin binding) of different CD4 and CD8 T cell subsets. In this graph the right vertical axis applies only to senescence of CD4 T cells subsets. (D) CD127 expression, thymic function (CD31 expression), and Treg cells (defined as CD4+CD127−CD25+FoxP3+). In this graph the right vertical axis applies only to Treg cells. Levels of statistical significance for the pairwise comparison (Wilcoxon signed rank test) between baseline and week 48 values are as follows: p<0.05 (*); p<0.01 (**); p<0.001 (***). N, naive; CM, central memory; EM, effector memory; Ef, effector.

Article Snippet: Numbers inside plots or histograms represent the percentage of positive events in each quadrant or region. table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 T cell subset category (antibody panel) T cell subset Differentiation stage of CD4 and CD8 cells (CD3-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD4 and CD8 cells (total) Naive cells (CD45RA + CD27 + ) Central memory cells (CD45RA − CD27 + ) Effector memory cells (CD45RA − CD27 − ) Effector cells (CD45RA + CD27 − ) Activation of CD4 and CD8 T cells (including naive, central memory, effector memory, and effector cells) (HLADR-FITC/CD27-PE/CD45RA-ECD/CD38-PECy5/CD4-PECy7 or CD8- PECy7) CD38 + DR − CD38 + DR + CD38 − DR + Thymic function (CD31) (CD31-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD31 expression on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) Expression of CD127 (CD127-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD127 expression on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) Senescence (CD57) (CD57-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) CD57 expression on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) Exhaustion (PD-1 and Tim-3) of CD3, CD4, and CD8 T cells (CD4-FITC/Tim-3-PE/CD3-ECD/CD8-PECy5/PD-1-biotin/streptavidin-PECy7) PD-1 + Tim-3 − PD-1 + Tim-3 + PD-1 − Tim-3 + Apoptosis (annexin binding) (annexin-FITC/CD27-PE/CD45RA-ECD/CD8-PECy5/CD4-PECy7) Annexin + binding on CD4 and CD8 cells (including naive, central memory, effector memory, and effector cells) T regulatory cells (FoxP3-FITC/CD127-PE/CD8-ECD/CD25-PECy5/CD4-PECy7) CD4 + CD25 + CD127 − FoxP3 + cells Open in a separate window T Cell Subsets Analyzed in the Study, Grouped According to Eight Different Categories Plasma interleukin (IL)-7 levels IL-7 was measured using a highly sensitive commercial immunoassay (Quantikine HS, R&D Systems, Barcelona, Spain), which has a lower limit of detection of 0.1 pg/ml following the manufacturer's instructions.

Techniques: Activation Assay, Expressing, Binding Assay, Comparison

Journal: Frontiers in Immunology

Article Title: Evaluating the Genetics of Common Variable Immunodeficiency: Monogenetic Model and Beyond

doi: 10.3389/fimmu.2018.00636

Figure Lengend Snippet: (A) Chromatograms corresponding to the Sanger sequencing of the LRBA nonsense mutation region in L283 and five healthy relatives. (B) Western blot analysis of LRBA and GAPDH for L283 patient (P) and a healthy control C+. LRBA protein is not detectable in the LRBA-deficient patient. (C) CTLA4 expression is downregulated in LRBA- and CTLA4-deficient patients. CTLA4 expression was assessed in Treg cells (CD3+CD4+CD25hiFoxP3+ cells) in resting and in PHA-stimulated cells (24 h). Bars represent mean values and error bars represent SE of the mean values for adult healthy controls ( n = 5).

Article Snippet: Specifically, for Treg cell phenotyping and CTLA-4 expression PBMCs were left with medium (resting) or stimulated with PHA (5μg/ml, Sigma-Aldrich, St. Louis, MO, USA) for 24 h. Treg intracellular staining was performed with Treg Detection Kit (CD4/CD25/FoxP3) kit (Miltenyi Biotec, Germany) following manufacturer’s instructions.

Techniques: Sequencing, Mutagenesis, Western Blot, Control, Expressing

Journal: Nature Communications

Article Title: Increased SOAT2 expression in aged regulatory T cells is associated with altered cholesterol metabolism and reduced anti-tumor immunity

doi: 10.1038/s41467-025-56002-w

Figure Lengend Snippet: A Diagram of murine CD4 + CD25 + , CD4 + CD25 - T or CD4 - lymphocytes isolated by magnetic bead sorting from LSCC tumor tissue and spleen samples (Created in BioRender. Major, Z. (2024) https://BioRender.com/u18t594 ). B , C The proteins extracted from tumor ( B ) and spleen ( C ) Lymphocyte subsets in indicated groups were assessed by Western blotting analysis. The experiment was repeated 3 times independently with similar results. D , E The SOAT2 expression was assessed in tumor and spleen CD4 + CD25 + T lymphocytes by flow cytometry assay ( D ); quantitation of SOAT2 expression was shown ( n = 5) ( E ). F , G The SOAT2 expression was assessed in tumor and spleen CD4 + CD25 - &CD4 - T lymphocytes by flow cytometry assay ( F ); quantitation of SOAT2 expression was shown ( n = 5) ( G ). H Diagram of human CD4 + CD25 + T lymphocytes, CD4 + CD25 - T or CD4 - lymphocytes isolated by magnetic bead sorting from blood samples (Created in BioRender. Major, Z. (2024) https://BioRender.com/w00f660 ). I , J The mRNA and protein expression of SOAT2 in indicated groups were detected by quantitative RT-PCR ( I ) and Western blotting ( J , The experiment was repeated 3 times independently with similar results) analysis. Data represented means ± SD. Statistical difference was evaluated by unpaired two-tailed student’s t -test ( E , G ), and one-way ANOVA test ( I ). Source data are provided as a Source Data file. (* P < 0.05; *** P < 0.001; ns no significance).

Article Snippet: For mouse samples, CD4 + CD25 + FOXP3 + T lymphocyte cells (Treg cells), CD4 − T lymphocyte cells, CD8 + TILs (Teffs) were enriched and sorted from spleen and tumor tissues using a modified isolation protocol by Regulatory T Cell Isolation Kit and CD8 + TILs Isolation Kit (#130-091-041, #130-116-478, Miltenyi Biotec).

Techniques: Isolation, Western Blot, Expressing, Flow Cytometry, Quantitation Assay, Quantitative RT-PCR, Two Tailed Test

Journal: Nature Communications

Article Title: Increased SOAT2 expression in aged regulatory T cells is associated with altered cholesterol metabolism and reduced anti-tumor immunity

doi: 10.1038/s41467-025-56002-w

Figure Lengend Snippet: A – D Ki-67 staining in indicated groups was analyzed by flow cytometry to measure lymphocyte proliferation ( A , C ); quantitation of Ki67 of Max (%) was shown ( n = 5) ( B , D ). E , F The apoptotic Treg cells in indicated groups were analyzed by flow cytometry with 7-AAD and PE-Annexin V staining; the percentage of apoptotic Treg cells in indicated groups were shown ( n = 3). G , H The migration of Treg cells in indicated groups were detected by transwell assays ( n = 5). I , J FOXP3 staining in indicated groups was analyzed by flow cytometry to measure Treg cells stability. K The differentiation ability of naive CD4 + T cells to CD4 + CD25 + FOXP3 + T lymphocytes sorted in indicated groups under Treg polarizing conditions was detected by flow cytometry assay. Data represented means ± SD. Statistical difference was evaluated by one-way ANOVA test ( B , D , E , F , G , H ). Source data are provided as a Source Data file. (* P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: For mouse samples, CD4 + CD25 + FOXP3 + T lymphocyte cells (Treg cells), CD4 − T lymphocyte cells, CD8 + TILs (Teffs) were enriched and sorted from spleen and tumor tissues using a modified isolation protocol by Regulatory T Cell Isolation Kit and CD8 + TILs Isolation Kit (#130-091-041, #130-116-478, Miltenyi Biotec).

Techniques: Staining, Flow Cytometry, Quantitation Assay, Migration

Journal: Frontiers in Immunology

Article Title: New Paradigm in the Role of Regulatory T Cells During Pregnancy

doi: 10.3389/fimmu.2019.00573

Figure Lengend Snippet: Immunological balance at the feto-maternal interface during early pregnancy. EVTs did not express polymorphic HLA-A, B whereas HLA-C and non-polymorphic HLA-E, G, and F were expressed. Maternal CD8 + T cells and NK cells can directly recognize paternal HLA-C and CD4 + T cells can indirectly recognize it. HLA- E and G protect EVTs from NK-cell mediated cytotoxicity. Treg cells can recognize fetal antigens via maternal antigen presenting cells (APCs) and induce tolerance in an antigen-specific manner. EVT, Extravillous trophoblast; NK, natural killer cell; Treg; regulatory T cell; APC, antigen-presenting cell.

Article Snippet: Semi-allogenic fetuses are not rejected by the maternal immune system because feto-maternal tolerance induced by CD4 + CD25 + FoxP3 + regulatory T (Treg) cells is established during pregnancy.

Techniques:

Journal: Frontiers in Immunology

Article Title: New Paradigm in the Role of Regulatory T Cells During Pregnancy

doi: 10.3389/fimmu.2019.00573

Figure Lengend Snippet: Paternal antigen-specific Treg cells in mouse models.

Article Snippet: Semi-allogenic fetuses are not rejected by the maternal immune system because feto-maternal tolerance induced by CD4 + CD25 + FoxP3 + regulatory T (Treg) cells is established during pregnancy.

Techniques: Derivative Assay

Journal: Frontiers in Immunology

Article Title: New Paradigm in the Role of Regulatory T Cells During Pregnancy

doi: 10.3389/fimmu.2019.00573

Figure Lengend Snippet: Single-cell based TCR repertoire analysis method. To study the clonality of effector Treg cells, a single-cell based T cell receptor (TCR) repertoire analysis method was used. Paired samples of maternal peripheral blood mononuclear cells and decidual lymphocytes were obtained. CD4+CD25+CD45RA-CD127low/- effector Treg cells were single-cell sorted. The cDNAs of complementarity determining lesion 3 (CDR3) in TCRβ chain and FoxP3 were amplified by RT-PCR. The nucleotides and amino acid sequences of CDR3 were analyzed.

Article Snippet: Semi-allogenic fetuses are not rejected by the maternal immune system because feto-maternal tolerance induced by CD4 + CD25 + FoxP3 + regulatory T (Treg) cells is established during pregnancy.

Techniques: Amplification, Reverse Transcription Polymerase Chain Reaction