Journal: The Journal of Clinical Investigation

Article Title: Lymphoid tissue fibrosis is associated with impaired vaccine responses

doi: 10.1172/JCI97377

Figure Lengend Snippet: This figure shows LN analysis from 3 different participants before vaccination and again at week 2 after vaccination. Participant 1996 has recognizable follicles before vaccination and formation of follicles after vaccination. The accumulation of PD1 staining cells within the secondary follicle (D) with green and blue staining cells in the secondary follicle (staining yellow) show an expected reaction to vaccination. Panel B shows participant 1682 with fewer, more poorly formed follicles at baseline and a lack of recognizable secondary follicles with vaccination. PD1 staining cells are not inside of the follicle structure. Participant 1688 has no recognizable follicles prior to vaccination and no response to vaccination. Scale bar indicates 20 μm.

Article Snippet: Slides were then washed 3 times in PBS for a total of 60 minutes and stained for 2 hours at RT with titrated amounts of the following conjugated antibodies: PD1-Alx488 (catalog FAB7115G; R&D Systems), CD20-efluor615 (clone L26; eBioscience), Ki67-Alx647 (clone B56; BD Biosciences), CD57-Alx680 (clone NK-1; in-house conjugated).

Techniques: Staining

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Clinical significance of PD-L1 + exosomes in plasma of Head and Neck Cancer patients

doi: 10.1158/1078-0432.CCR-17-2664

Figure Lengend Snippet: Protein levels of exosomes and RFVs of PD-1+ or PD-L1+ exosomes in plasma of patients with HNSCC. A. protein concentrations of exosomes (μg/ml plasma) are significantly elevated in patients with active disease (AD) relative to those with no evidence of disease after therapy (NED); B. RFVs for PD-L1+ exosomes is significantly higher in AD patients, patients with positive lymph nodes and patients with the high (III/IV) UICC stage tumors at diagnosis. * p< 0.05; *** p< 0.0008. C. RFVs for PD-1 exosomes in plasma are not statistically different in HNSCC patients stratified by disease activity, lymph node or UICC status. The data presented as box plots are normalized to the frequency of exosomes/1mL of patients’ plasma in this and the subsequent figures.

Article Snippet: Next, anti-PD-1 Ab, #AF1086 lot #ICA02 (purchased from R&D Systems) was used to block effects of exosome-mediated suppression.

Techniques: Activity Assay

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Clinical significance of PD-L1 + exosomes in plasma of Head and Neck Cancer patients

doi: 10.1158/1078-0432.CCR-17-2664

Figure Lengend Snippet: RFVs for PD-L1+ and PD-1+ exosomes/mL plasma and levels of sPD-L1 in plasma of HNSCC patients. A. No correlation between RFVs for PD-L1+ and PD-1+ exosomes in plasma of HNSCC patients was observed: Spearman’s correlation at p=0.7, r=0.05. B. No correlation between RFVs for PD-L1+ exosomes in patients’ plasma and soluble PD-L1; Spearman’s correlation at p=0.85, r=0.03.

Article Snippet: Next, anti-PD-1 Ab, #AF1086 lot #ICA02 (purchased from R&D Systems) was used to block effects of exosome-mediated suppression.

Techniques:

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Clinical significance of PD-L1 + exosomes in plasma of Head and Neck Cancer patients

doi: 10.1158/1078-0432.CCR-17-2664

Figure Lengend Snippet: CD69 and PD-1 expression on activated CD8+ T cells and down-regulation of CD69 expression by PD-L1high but not by PD-L1low exosomes. A. Representative flow cytometry showing expression levels of CD69 or PD-1 on the surface of activated CD8+ T cells. Decreasing levels of CD8+CD69+ on the surface of T cells co-incubated with PD-L1high exosomes. B, Downregulation of CD69 expression (% and MFI) on CD8+ T-cells after co-incubation with PD-L1high exosomes. In C, blocking of exosome-mediated down-regulation of CD69 expression on CD8+ T cells by anti-PD-1 Ab. Note that inhibition mediated by PD-L1high exosomes was almost completely reversed (% and MFI) by adding the PD-1 inhibitor. * p< 0.05; ** p<0.005

Article Snippet: Next, anti-PD-1 Ab, #AF1086 lot #ICA02 (purchased from R&D Systems) was used to block effects of exosome-mediated suppression.

Techniques: Expressing, Flow Cytometry, Incubation, Blocking Assay, Inhibition