Fig 2: Expression of CXCL16 on monocytes is associated with enhanced clinical disease activity and cardiovascular risk. (A) Flow cytometry analysis of CXCL16 expression on monocyte subpopulations defined by gating for CD14 and CD16 expression in PBMC: classical monocytes (CD14++ CD16-, R1 green), intermediate monocytes (CD14++ CD16+, R2 blue) and non-classical monocytes (CD14low CD16++, R3, red), mean and SD of healthy controls (n= 14; unfilled dots) and patients with psoriasis vulgaris (n= 39; filled dots), Welch’s t test. (B) Correlation of CXCL16 expression on all monocytes of psoriatic patients (n=39) and clinical disease activity (PASI), Pearson’s correlation. (C) Correlation for each monocyte subset defined by color indicated in (A) and PASI score, Pearson’s correlation. (D) Mean and SD of PASI (unpaired t test) and PROCAM (Mann-Whitney test) of normal-weight (unfilled triangles; n=19) and obese psoriatic patients (filled triangles; n=20). (E) Correlation of chemokine expression with cardiovascular risk (PROCAM score); and body-mass-index (BMI). Each dot represents an individual patient, Pearson correlation. (F) CXCL16 expression on monocytes in psoriatic patients without atherosclerosis (n= 6; unfilled trapezoid) or with atherosclerosis (n= 9; filled trapezoid), mean and SD, unpaired t test. *p < 0.05; **p < 0.01; ***< 0.001. ns, not significant.

Fig 3: Monkey CCR4+ Treg depletion for animals M11016 and 11216 in the peripheral blood using the CCR4 immunotoxin. (A) Representative flow cytometry analysis of the CCR4+ cell depletion in the peripheral blood using the antibodies against human CD4 and CCR4. (B) CCR4+ cell depletion in the peripheral blood was monitored by flow cytometry using the antibodies against human CD4 and CCR4 (CD4+ CCR4+). (C) Representative flow cytometry analysis of the CCR4+Foxp3+ Treg depletion in the peripheral blood using the antibodies against human CCR4 and Foxp3 (CCR4+Foxp3+ among the gated CD4+ cells). (D) CCR4+ Treg depletion in the peripheral blood was monitored by flow cytometry using the antibodies against human CCR4 and Foxp3 (CCR4+Foxp3+ among the gated CD4+ cells). (E) The other CD4+ cells in the peripheral blood were monitored by flow cytometry using antibodies against human CD4 and CCR4 (CD4+ CCR4−). (F) The CD8+ T cells in the peripheral blood were monitored by flow cytometry using the antibodies against human CD3, CD4, and CD8 (CD3+ CD4− CD8+). (G) The B cells in the peripheral blood were monitored by flow cytometry using antibodies against human CD20, CD3, CD16, and HLA‐DR (CD20+ CD3− CD16− HLA‐DR +). (H) The NK cells in the peripheral blood were monitored using antibodies against human CD3, CD16, and NKp80 (CD3− CD16+ NKp80+). (I) Monocytes in the peripheral blood were monitored by flow cytometry using antibodies against human CD14, CD16, and CD11b (CD14+ CD11b+ or CD14+ CD16+, PBMC gating). (J) The effector Tregs in the peripheral blood were monitored by flow cytometry using antibodies against CD45RA and Foxp3 (CD45RA ‐Foxp3+ among the gated CD4+ cells).

Fig 4: Monkey CCR4+ Treg depletion for animals M11016 and 11216 in the lymph node using the CCR4 immunotoxin. The lymph node biopsies were performed before and after each course of the immunotoxin treatment as well as in the end of the study. (A) Flow cytometry analysis of the lymph node biopsy samples using antibodies against human CD4 and CCR4. (B) Flow cytometry analysis of the lymph node biopsy samples using antibodies against human CCR4 and Foxp3 (CCR4+Foxp3+ among the gated CD4+ cells). (C) Lymph node CCR4+ Treg depletion was monitored by flow cytometry (CCR4+ cells: CD4+ CCR4+, CCR4+ Tregs: CCR4+Foxp3+ among the gated CD4+ cells, effector Tregs: CD45RA ‐Foxp3+ among the gated CD4+ cells). Other cell populations in the lymph node were also monitored by flow cytometry (other CD4+ cells: CD4+ CCR4−; CD8+ cells: CD8+ CD3+ CD4−; B cells: CD20+ CD3− CD16− HLA‐DR +).

Fig 5: More Pronounced NK Cell Subset Redistribution in bnAb versus Control Subjects(A) Representative pseudocolor flow cytometry plots from bnAb, control, and HIV-seronegative individuals showing NK cell subsets distinguished on the basis of CD56 and CD16 expression. Gated on live, CD19-CD14-CD4-CD3- lymphocytes.(B–D) Boxplots of the percentage of NK cells that were CD56bright (B), CD56dim (C), and CD56-CD16+ (CD56neg) (D) in individuals in the bnAb (red; n = 22), control (blue; n = 19), and HIV-seronegative (black; n = 19) groups. Each symbol represents data from an individual subject and the box-and-whisker plots show the median, quartiles, and range. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. p values are corrected for multiple comparisons analysis and viral load.(E and F) Spearman correlations of (E) RAB11FIP5 mRNA levels in total PBMC and (F) plasma HIV-1 neutralization breadth (PC1) with the percentage of CD56dim (black) and CD56-CD16+ (blue) NK subsets within total NK cells.See also Figure S3 and Table S2.