raltegravir  (MedChemExpress)

Journal: EMBO Molecular Medicine

Article Title: Combined dendritic cell and anti-TIGIT immunotherapy potentiates adaptive NK cells against HIV-1

doi: 10.1038/s44321-025-00255-x

Figure Lengend Snippet: ( A ) Schematic representation experiment of functional assays evaluating the elimination p24 + CD4 + T cells after culture with NK cells primed by Nano-PIC-MDDC. ( B ) Representative flow cytometry dot plots showing intracellular expression of HIV-1 p24 on CD4 + T cells from a PWH in the presence of Raltegravir alone or in combination with Romidepsin. A staining background control from an HIV negative donor is also shown. ( C ) Representative flow cytometry dot plot representing NKG2C and CD57 expression in gated CD56dim CD16 + NK cells. ( D ) Heatmaps representing Spearman correlation matrix between memory NK subsets within CD56dim or CD56lo/- CD16 + NK cells and proportions of p24+ cells after co-culture with NK primed by Nano-PIC-MDDC immunotherapy. Fold changes in p24+ frequencies are also included. Levels of positive and negative associations are highlighted in different intensities of red and blue, respectively. Significant associations have also been highlighted. * P < 0.05; ** P < 0.01; **** P < 0.0001. ( E ) ROC curve analysis, defining the cut-off based on NKG2C expression defining effective and non-responder PWH groups in our cohort. ( F ) Pie chart representing proportion of PWH from our cohort predicted as effective and non-responder PWH by ROC curve analysis and those PWH that were not classified by this model. ( G ) Fold change of intracellular expression of HIV-1 p24 on CD4 + T cells from aviremic ART PWH treated with Raltegravir and Romidepsin, in the absence or the presence of NK cells alone or stimulated with MDDCs treated with empty Nano or Nano-PIC. Data from three separate groups with effective-responders (Effect. R.; n = 13), unclassified (Unclas. R.; n = 7) and non-responders (Non-R.; n = 13) PWH to Nano-PIC-MDDC. ( H – J ) Proportions of total ( H ) NKG2C+ cells and different memory NK subsets based on combination of NKG2C and CD57 ( I , J ) on CD56dim or CD56lo/− CD16 + NK from effective responder ( n = 13, Effect. R; blue), unclassified ( n = 7, Unclas. R; gray) and non-responder ( n = 13, Non-R.; pink) PWH after activation with Nano-PIC-MDDCs. Data are represented in Box and Whiskers plots showing median and maximum and minimum values. ( K ) Proportions of CD107a+ IFNγ+ (left) and CD107a+ Granzyme B+ (right) cells included in gated NK subsets defined by differential NKG2C and CD57 expression from ( n = 10) selected effective responder PWH after PMA and Ionomycin stimulation (see methods). Data are represented in Box and Whiskers plots showing median and maximum and minimum values. Statistical significance was calculated using a Friedman or a Kruskal–Wallis tests for multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. .

Article Snippet: Raltegravir , Selleck Chemicals , S2005.

Techniques: Functional Assay, Flow Cytometry, Expressing, Staining, Control, Co-Culture Assay, Activation Assay

Journal: EMBO Molecular Medicine

Article Title: Combined dendritic cell and anti-TIGIT immunotherapy potentiates adaptive NK cells against HIV-1

doi: 10.1038/s44321-025-00255-x

Figure Lengend Snippet: ( A ) Raw data and fold change in proportions of HIV-1 p24+ CD4+ T cells from all n = 33 PWH recruited for the study cultured with Romidepsin and Raltegavir in the absence or the presence of autologous NK cells alone or stimulated with Nano-empty or Nano-PIC-MDDC. ( B ) Proportions of HIV-1 p24+ CD4+ T cells in the presence of NK cells stimulated with Nano-PIC-MDDC from three separate effective responder (Effect. R; n = 13), unclassified (Unclas. R. n = 7) and non-responder (Non-R.; n = 13) PWH groups. ( C ) Representative flow cytometry dot plots showing intracellular expression of p24 in Nano-PIC-MDDC (upper plot) or co-cultured autologous CD4 + T cells (lower plot) from a tested PWH. ( D ) Proportions of HIV-1 p24+ cells analyzed by FACS (left plots, light blue) and IPDA analysis of intact (dark blue) or defective (gray) HIV-DNA in CD4+ T cells from n = 4 PWH cultured with Romidepsin and Raltegravir in the absence or the presence of autologous NK cells alone or stimulated with Nano-PIC-MDDC. ( E ) Proportions of NKG2C- CD57+ subset on CD56dim or CD56lo/- CD16 + NK from effective responder ( n = 13, Effect. R; blue), unclassified ( n = 7, Unclas. R; gray) and non-responder ( n = 13, Non-R.; pink) PWH after activation with Nano-PIC-MDDC. ( F ) Proportions of total CD107a+ (left), total FNγ+ (right) and total TNFα+ (below) included in gated memory NK precursors NKG2C + CD57−, memory differentiated NKG2C + CD57+ and effector NKG2C− CD57+ subsets from n = 10 selected effective responder PWH after Nano-PIC-MDDCs in presence of PMA and Ionomycin stimulation for 4 h and Brefeldin A and Monensin. Data from ( A , B , E , F ) are presented in Box and Whiskers plots showing median values and maximum and minimum error bars. The data in ( D ) are represented with bar plots. Statistical Significance was calculated using a Friedman or a Kruskal–Wallis test for multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Raltegravir , Selleck Chemicals , S2005.

Techniques: Cell Culture, Flow Cytometry, Expressing, Activation Assay

Journal: EMBO Molecular Medicine

Article Title: Combined dendritic cell and anti-TIGIT immunotherapy potentiates adaptive NK cells against HIV-1

doi: 10.1038/s44321-025-00255-x

Figure Lengend Snippet: ( A ) Representative flow cytometry dot plots for the NK receptor ligands MIC/ab (NKG2D), HLA-E (NKG2C/A) and DR4 (TRAIL), in gated p24+ or p24- CD4 + T cells from PWH after 16h culture with PHA + IL-2. A FMO control is included to each NK ligand. ( B ) Representative Flow cytometry dot plot showing expression of TRAIL on total CD56dim CD16+ NK, total NKG2C-, NKG2C+ CD57- and NKG2C+ CD57+ NK from a representative effective responder PWH. ( C ) Fold change in proportions of TRAIL+ cells in NKG2C- (left) and adaptive NKG2C+ CD57- (middle) and NKG2C+ CD57+ (right) subsets after Nano-PIC-MDDC from effective responder ( n = 10; Effect. R; blue), unclassified ( n = 6; Unclas. R; gray) and non-responder ( n = 8; Non-R.; pink) PWH included on CD56lo/- CD16 + NK. ( D ) Raw proportions of TRAIL+ cells within adaptive NKG2C+ CD57+ and NKG2C+ CD57− cell subsets in the same responders PWH groups defined in ( C ) included in CD56dim CD16+ NK subpopulation after stimulation with Nano-PIC-MDDC. Statistical significance was calculated using a Mann–Whitney test and Bonferroni correction. ( E , F ) Ratio of TIGIT+ versus TRAIL+ cells in precursor NKG2C+ CD57- adaptive ( E ) and mature NKG2C + CD57+ ( F ) cells in CD56dim (left) and CD56lo/− (right) CD16 + NK subsets from the same PWH responder groups. Statistical significance was calculated using a one tale Mann–Whitney test and Bonferroni correction. ( G , H ) Proportions of HIV-p24 + CD4 + T cells from n = 6 ( G ) and n = 7 ( H ) PWH previously identified as effective responders treated with Raltegravir and Romidepsin and cultured in the absence or the presence of autologous unstimulated NK cells or NK treated with Nano-PIC-MDDCs in the presence of either IgG Isotypic control or with anti-TRAIL blocking mAb ( G ) or in the presence of anti-NKG2C mAb ( H ). Statistical significance was calculated using a one tale Wilcoxon matched pairs test and Bonferroni correction or a Friedman test for multiple comparisons. ( I ) Spearman correlations between proportions of TIM3 and TRAIL after Nano-PIC-MDDC within CD56dim (upper) and CD56lo/− (bottom) CD16 + NK. Statistical P and R values considering all data (black), without unclassified group (gray). ( J ) Analysis of proportions of TRAIL+ cells within adaptive NKG2C + CD57 + NK from n = 7 PWH stimulated with Nano-PIC-MDDC in the presence of isotype or anti-TIM3 mAb. Statistical significance was calculated were calculated using a two-tailed Wilcoxon matched pairs. Data from ( C – H , J ) are presented in Box and Whiskers plots showing median values and maximum and minimum error bars. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Raltegravir , Selleck Chemicals , S2005.

Techniques: Flow Cytometry, Control, Expressing, MANN-WHITNEY, Cell Culture, Blocking Assay, Two Tailed Test

Journal: EMBO Molecular Medicine

Article Title: Combined dendritic cell and anti-TIGIT immunotherapy potentiates adaptive NK cells against HIV-1

doi: 10.1038/s44321-025-00255-x

Figure Lengend Snippet: ( A ) Proportions of CD4+ T cells from PWH expressing the NK receptor ligands MICa/b (left), HLA-E (middle) and DR4 (right) within HIV-1 p24+ (purple) and p24- (yellow) populations in n = 7 PWH. Data are represented in Box and Whiskers plots showing median and maximum and minimum values in error bars. ( B ) Fold change proportions of TRAIL+ cells within adaptive NKG2C+ CD57− precursors (left) and mature NKG2C+ CD57+ (middle) and NKG2C- (right) NK subsets after Nano-PIC-MDDC from effective responder ( n = 10; Effect. R; blue), unclassified responder ( n = 6; Unclas. R; gray) and non-responder ( n = 8; Non-R.; pink) PWH included in CD56dim CD16 + . Data are represented in Box and Whiskers plots showing median and maximum and minimum values in error bars. ( C ) Proportions of TRAIL+ cells within adaptive NKG2C + CD57+ and NKG2C+ CD57- cell subsets from effective responder ( n = 10; Effect. R.; blue), unclassified responder ( n = 6; Unclas. R; gray) and non-responder ( n = 8; Non-R; pink) PWH included in CD56lo/- CD16 + NK subpopulation. Statistical significance was calculated using a two-tailed Wilcoxon matched pairs or a Mann–Whitney tests Statistical and Bonferroni correction was applied for multiple comparisons. Significance after removed outliers recognized by a Grubbs´test were highlighted in green. * P < 0.05; ** P < 0.01. Data are represented in Box and Whiskers plots showing median and maximum and minimum values in error bars. ( D ) Spearman correlation between proportions of cells expressing TIGIT versus TRAIL in CD56dim CD16+ NK cells from PWH. Data from effective and non or unclassified responders PWH were highlighted in blue, pink and gray, respectively. Statistical P and R values considering all data (black), without unclassified group (gray) or removing outlier data recognized by a Grubb´s test (green) are shown. ( E ) Proportions of TRAIL in NKG2C+ CD57 + NK cells after Nano-PIC-MDDC from n = 8 PWH, in the presence of either IgG Isotypic control or anti-TIGIT antibodies. Statistical significance was calculated using a two-tailed Wilcoxon matched pairs test. ( F ) Ratio of TIGIT+ /TRAIL+ cells in NKG2C- CD57+ effector NK expressing from effective ( n = 10; Effect. R; blue), unclassified ( n = 6; Unclas.R; gray) and non ( n = 8; Non-R.; pink) responders PWH in CD56dim (left) and CD56lo/− (right) CD16 + NK subsets. Statistical significance was calculated using a two-tailed Mann–Whitney test and Bonferroni correction for multiple comparisons. Data are represented in Box and Whiskers plots showing median and maximum and minimum values in error bars. ( G ) Fold change of p24+ CD4+ T cells from n = 13 PWH previously identified as effective responders treated with Raltegravir and Romidepsin and cultured in the absence or the presence of autologous NK cells treated with Nano-PIC-MDDCs and either IgG Isotypic control or with anti-TRAIL antibodies. Data are represented in Box and Whiskers plots showing median and maximum and minimum values in error bars. Statistical significance was calculated using a Friedman anova and Dunn´s post-hoc test for multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001. .

Article Snippet: Raltegravir , Selleck Chemicals , S2005.

Techniques: Expressing, Two Tailed Test, MANN-WHITNEY, Control, Cell Culture