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cytomegalovirus cmv pp65 peptide pools  (Miltenyi Biotec)
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(A) Representative flow plots and (B) summary data for the frequency of IFN-γ-producing CD56 dim NK cells following 6-hour of stimulation with antibody-coated Raji cells (Target cells) in individuals with subtype A and non-A subtype infection. Control = Uncoated cells. (C) Representative flow plots and (D) pie charts showing IFN-γ production in FcεRγ + CD57 + CD56 dim and FcεRγ − CD57 + CD56 dim NK cells in participants with subtype A and non-A subtype infection. (E) The frequencies of IFN-γ + FcεRγ − CD57 + CD56 dim NK cells (dashed line) and log 10 viral loads (grey solid line) at all time points. (F) Representative plots for the identification of antigen-reactive NK cells based on double expression (IFN-γ and CD107a) following 6-h stimulation with media alone (control) or with HIV-1 (Gag and Env) or CMV <t>(pp65)</t> peptide pools. (G) a Percentage of responders (subtype A) to HIV-1 peptide pools at each time point or CMV pools at 1-month time point. (H) Frequency of HIV-reactive NK cells at all time points. Pie charts representing the proportion of antigen-reactive NK cells within adaptive (NKG2C + ) or canonical (NKG2C − ) NK cells. Significance determined by two-tailed Mann– Whitney U test or Wilcoxon matched-paired signed rank test. *p < 0.05, **p < 0.01. The non-parametric Spearman test was used for correlation analysis (two-tailed).
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(A) Representative flow plots and (B) summary data for the frequency of IFN-γ-producing CD56 dim NK cells following 6-hour of stimulation with antibody-coated Raji cells (Target cells) in individuals with subtype A and non-A subtype infection. Control = Uncoated cells. (C) Representative flow plots and (D) pie charts showing IFN-γ production in FcεRγ + CD57 + CD56 dim and FcεRγ − CD57 + CD56 dim NK cells in participants with subtype A and non-A subtype infection. (E) The frequencies of IFN-γ + FcεRγ − CD57 + CD56 dim NK cells (dashed line) and log 10 viral loads (grey solid line) at all time points. (F) Representative plots for the identification of antigen-reactive NK cells based on double expression (IFN-γ and CD107a) following 6-h stimulation with media alone (control) or with HIV-1 (Gag and Env) or CMV <t>(pp65)</t> peptide pools. (G) a Percentage of responders (subtype A) to HIV-1 peptide pools at each time point or CMV pools at 1-month time point. (H) Frequency of HIV-reactive NK cells at all time points. Pie charts representing the proportion of antigen-reactive NK cells within adaptive (NKG2C + ) or canonical (NKG2C − ) NK cells. Significance determined by two-tailed Mann– Whitney U test or Wilcoxon matched-paired signed rank test. *p < 0.05, **p < 0.01. The non-parametric Spearman test was used for correlation analysis (two-tailed).
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(A) Representative flow plots and (B) summary data for the frequency of IFN-γ-producing CD56 dim NK cells following 6-hour of stimulation with antibody-coated Raji cells (Target cells) in individuals with subtype A and non-A subtype infection. Control = Uncoated cells. (C) Representative flow plots and (D) pie charts showing IFN-γ production in FcεRγ + CD57 + CD56 dim and FcεRγ − CD57 + CD56 dim NK cells in participants with subtype A and non-A subtype infection. (E) The frequencies of IFN-γ + FcεRγ − CD57 + CD56 dim NK cells (dashed line) and log 10 viral loads (grey solid line) at all time points. (F) Representative plots for the identification of antigen-reactive NK cells based on double expression (IFN-γ and CD107a) following 6-h stimulation with media alone (control) or with HIV-1 (Gag and Env) or CMV <t>(pp65)</t> peptide pools. (G) a Percentage of responders (subtype A) to HIV-1 peptide pools at each time point or CMV pools at 1-month time point. (H) Frequency of HIV-reactive NK cells at all time points. Pie charts representing the proportion of antigen-reactive NK cells within adaptive (NKG2C + ) or canonical (NKG2C − ) NK cells. Significance determined by two-tailed Mann– Whitney U test or Wilcoxon matched-paired signed rank test. *p < 0.05, **p < 0.01. The non-parametric Spearman test was used for correlation analysis (two-tailed).
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(A) Representative flow plots and (B) summary data for the frequency of IFN-γ-producing CD56 dim NK cells following 6-hour of stimulation with antibody-coated Raji cells (Target cells) in individuals with subtype A and non-A subtype infection. Control = Uncoated cells. (C) Representative flow plots and (D) pie charts showing IFN-γ production in FcεRγ + CD57 + CD56 dim and FcεRγ − CD57 + CD56 dim NK cells in participants with subtype A and non-A subtype infection. (E) The frequencies of IFN-γ + FcεRγ − CD57 + CD56 dim NK cells (dashed line) and log 10 viral loads (grey solid line) at all time points. (F) Representative plots for the identification of antigen-reactive NK cells based on double expression (IFN-γ and CD107a) following 6-h stimulation with media alone (control) or with HIV-1 (Gag and Env) or CMV <t>(pp65)</t> peptide pools. (G) a Percentage of responders (subtype A) to HIV-1 peptide pools at each time point or CMV pools at 1-month time point. (H) Frequency of HIV-reactive NK cells at all time points. Pie charts representing the proportion of antigen-reactive NK cells within adaptive (NKG2C + ) or canonical (NKG2C − ) NK cells. Significance determined by two-tailed Mann– Whitney U test or Wilcoxon matched-paired signed rank test. *p < 0.05, **p < 0.01. The non-parametric Spearman test was used for correlation analysis (two-tailed).
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(A) Representative flow plots and (B) summary data for the frequency of IFN-γ-producing CD56 dim NK cells following 6-hour of stimulation with antibody-coated Raji cells (Target cells) in individuals with subtype A and non-A subtype infection. Control = Uncoated cells. (C) Representative flow plots and (D) pie charts showing IFN-γ production in FcεRγ + CD57 + CD56 dim and FcεRγ − CD57 + CD56 dim NK cells in participants with subtype A and non-A subtype infection. (E) The frequencies of IFN-γ + FcεRγ − CD57 + CD56 dim NK cells (dashed line) and log 10 viral loads (grey solid line) at all time points. (F) Representative plots for the identification of antigen-reactive NK cells based on double expression (IFN-γ and CD107a) following 6-h stimulation with media alone (control) or with HIV-1 (Gag and Env) or CMV <t>(pp65)</t> peptide pools. (G) a Percentage of responders (subtype A) to HIV-1 peptide pools at each time point or CMV pools at 1-month time point. (H) Frequency of HIV-reactive NK cells at all time points. Pie charts representing the proportion of antigen-reactive NK cells within adaptive (NKG2C + ) or canonical (NKG2C − ) NK cells. Significance determined by two-tailed Mann– Whitney U test or Wilcoxon matched-paired signed rank test. *p < 0.05, **p < 0.01. The non-parametric Spearman test was used for correlation analysis (two-tailed).
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(A) Representative flow plots and (B) summary data for the frequency of IFN-γ-producing CD56 dim NK cells following 6-hour of stimulation with antibody-coated Raji cells (Target cells) in individuals with subtype A and non-A subtype infection. Control = Uncoated cells. (C) Representative flow plots and (D) pie charts showing IFN-γ production in FcεRγ + CD57 + CD56 dim and FcεRγ − CD57 + CD56 dim NK cells in participants with subtype A and non-A subtype infection. (E) The frequencies of IFN-γ + FcεRγ − CD57 + CD56 dim NK cells (dashed line) and log 10 viral loads (grey solid line) at all time points. (F) Representative plots for the identification of antigen-reactive NK cells based on double expression (IFN-γ and CD107a) following 6-h stimulation with media alone (control) or with HIV-1 (Gag and Env) or CMV (pp65) peptide pools. (G) a Percentage of responders (subtype A) to HIV-1 peptide pools at each time point or CMV pools at 1-month time point. (H) Frequency of HIV-reactive NK cells at all time points. Pie charts representing the proportion of antigen-reactive NK cells within adaptive (NKG2C + ) or canonical (NKG2C − ) NK cells. Significance determined by two-tailed Mann– Whitney U test or Wilcoxon matched-paired signed rank test. *p < 0.05, **p < 0.01. The non-parametric Spearman test was used for correlation analysis (two-tailed).

Journal: bioRxiv

Article Title: An early functional adaptive NK cell signature drives optimal CD8 + T-cell activation and predicts sustained HIV-1 viral control

doi: 10.1101/2025.03.17.643703

Figure Lengend Snippet: (A) Representative flow plots and (B) summary data for the frequency of IFN-γ-producing CD56 dim NK cells following 6-hour of stimulation with antibody-coated Raji cells (Target cells) in individuals with subtype A and non-A subtype infection. Control = Uncoated cells. (C) Representative flow plots and (D) pie charts showing IFN-γ production in FcεRγ + CD57 + CD56 dim and FcεRγ − CD57 + CD56 dim NK cells in participants with subtype A and non-A subtype infection. (E) The frequencies of IFN-γ + FcεRγ − CD57 + CD56 dim NK cells (dashed line) and log 10 viral loads (grey solid line) at all time points. (F) Representative plots for the identification of antigen-reactive NK cells based on double expression (IFN-γ and CD107a) following 6-h stimulation with media alone (control) or with HIV-1 (Gag and Env) or CMV (pp65) peptide pools. (G) a Percentage of responders (subtype A) to HIV-1 peptide pools at each time point or CMV pools at 1-month time point. (H) Frequency of HIV-reactive NK cells at all time points. Pie charts representing the proportion of antigen-reactive NK cells within adaptive (NKG2C + ) or canonical (NKG2C − ) NK cells. Significance determined by two-tailed Mann– Whitney U test or Wilcoxon matched-paired signed rank test. *p < 0.05, **p < 0.01. The non-parametric Spearman test was used for correlation analysis (two-tailed).

Article Snippet: After overnight rest, PBMCs were stimulated for 6 hours with 2 μg/ml of cohort-specific HIV-1 Gag or ENV peptide pools (provided by IAVI) or cytomegalovirus (CMV)-pp65 peptide pools (Miltenyibiotec) in the presence of αCD28/αCD49d co-Stim antibodies (1 μg/ml), GolgiStop (containing Monensin, 2 μmol/l), GolgiPlug (containing brefeldin A, 10 μg/ml) (BD Biosciences) and anti-CD107α BV421 antibody (BD Biosciences, Catalog # 562623, dilution 1 in 200).

Techniques: Infection, Control, Expressing, Two Tailed Test, MANN-WHITNEY