Journal: OncoImmunology

Article Title: MICA/B-targeted antibody promotes NK cell–driven tumor immunity in patients with intrahepatic cholangiocarcinoma

doi: 10.1080/2162402x.2022.2035919

Figure Lengend Snippet: Figure 6. Anti-MICA/B 7C6 mAb boosts anti-tumor cytotoxic function of peripheral NK cells from iCCA patients against the HuCCT-1 cell line. A): peripheral NK cell degranulation, evaluated as frequency of CD107a+ NK cells, in iCCA patients (n = 13) and HC (n = 16) in the presence of 7C6 mAb or IgG1-Fc. Parametric paired and unpaired t tests were used to compare data. B): dot plots showing the frequency of CD3-CD56+ CD107a+ NK cells in a HC and a patient (iCCA) in the presence of 7C6 mAb or IgG1-Fc. C): the proportion of circulating IFNγ+ NK cells in patients (n = 10) and HC (n = 11) in the presence of 7C6 mAb compared with IgG1-Fc. To compare paired data, the parametric t test and the non-parametric Wilcoxon t test were used. To compare unpaired data, the parametric t test and the non-parametric Mann-Whitney U test were used. D): representative dot plots showing the frequency of CD3-CD56+ IFNγ+ NK cells in a HC and in a patient (iCCA) in the presence of 7C6 mAb or IgG1-Fc.

Article Snippet: The recombinant human IgG1 Fc (BioXcell, Lebanon, NH, USA) and the humanized anti-MICA/B 7C6-IgG1 mAb were added at a final concentration of 10 μg/ml.

Techniques: MANN-WHITNEY

Journal: OncoImmunology

Article Title: MICA/B-targeted antibody promotes NK cell–driven tumor immunity in patients with intrahepatic cholangiocarcinoma

doi: 10.1080/2162402x.2022.2035919

Figure Lengend Snippet: Figure 7. Anti-MICA/B 7C6 mAb boosts anti-tumor cytotoxic function of peripheral NK cells from iCCA patients against patient-derived iCCA cell lines. A): peripheral NK cell degranulation, evaluated as CD107a+NK frequency, in iCCA patients (n = 12) and HC (n = 8) in the presence of 7C6 mAb or IgG1-Fc using patient- derived primary tumor cell lines as targets. Parametric paired and unpaired t tests were used to compare data. B): dot plots showing the frequency of CD3-CD56 + CD107a+ NK cells in a HC and in a patient (iCCA) in the presence of 7C6 mAb or IgG1-Fc. C): proportion of circulating IFNγ+NK cells in patients (n = 10) and in HC (n = 8) in the presence of 7C6 mAb compared with IgG1-Fc. To compare paired data, we used the parametric t test and the non-parametric Wilcoxon t test. To compare unpaired data, the parametric t test and the non-parametric Mann-Whitney U test were used. D): representative dot plots showing the frequency of CD3-CD56+ IFNγ +NK cells in a HC and in a patient (iCCA) in the presence of 7C6 mAb or IgG1-Fc.

Article Snippet: The recombinant human IgG1 Fc (BioXcell, Lebanon, NH, USA) and the humanized anti-MICA/B 7C6-IgG1 mAb were added at a final concentration of 10 μg/ml.

Techniques: Derivative Assay, MANN-WHITNEY

Journal: OncoImmunology

Article Title: MICA/B-targeted antibody promotes NK cell–driven tumor immunity in patients with intrahepatic cholangiocarcinoma

doi: 10.1080/2162402x.2022.2035919

Figure Lengend Snippet: Figure 8. 7C6 mAb enhances the anti-tumor effect of liver- and tumor-infiltrating NK cells in iCCA patients. A): Frequency of degranulating CD107a+NK cells in LIL (n = 13) and TIL (n = 10) of iCCA patients in the presence of anti-MICA/B 7C6 mAb or IgG1-Fc using autologous tumor-derived cell lines as targets. Parametric paired and unpaired t tests were used to compare data. B): representative dot plots showing the frequency of CD3-CD56+ CD107a+ LIL- and TIL-NK cells in the presence of 7C6 mAb or IgG1-Fc. C): proportion of IFNγ+ NK cells in LIL (n = 10) and TIL (n = 8) of iCCA patients in the presence of 7C6 mAb compared with IgG1-Fc using autologous tumor-derived cell lines as targets. The parametric t test and non-parametric Wilcoxon t test were used to compare paired data. The parametric t test was used to compare unpaired data.

Article Snippet: The recombinant human IgG1 Fc (BioXcell, Lebanon, NH, USA) and the humanized anti-MICA/B 7C6-IgG1 mAb were added at a final concentration of 10 μg/ml.

Techniques: Derivative Assay

Journal: OncoImmunology

Article Title: MICA/B-targeted antibody promotes NK cell–driven tumor immunity in patients with intrahepatic cholangiocarcinoma

doi: 10.1080/2162402x.2022.2035919

Figure Lengend Snippet: Figure 9. Cytotoxicity assay of HC PBMC, patient PBMC, LIL and TIL cells. A, B): Frequency of CFSE+LIVE/DEAD (LD)+ HuCCT-1 cell line targets when HC PBMC (n = 5), patient PBMC (n = 10), LIL (n = 8) and TIL (n = 5) were used as effector cells in the presence of 7C6 mAb and isotype control (IgG1). The parametric paired t tests were used to compare data. C, D): Frequency of CFSE+LD+ patient-derived cell line targets when HC PBMC (n = 5), patient PBMC (n = 8), LIL (n = 8) and TIL (n = 4) were used as effectors in the presence of 7C6 and isotype control. The parametric paired t test was used to compare data in panel C. The non-parametric Wilcoxon t test was used to compare paired data in panel D. Target cell death was determined as frequency of CFSE+LD+ cells.

Article Snippet: The recombinant human IgG1 Fc (BioXcell, Lebanon, NH, USA) and the humanized anti-MICA/B 7C6-IgG1 mAb were added at a final concentration of 10 μg/ml.

Techniques: Cytotoxicity Assay, Control, Derivative Assay

Journal: Journal of Immunology Research

Article Title: Constitutive Changes in Circulating Follicular Helper T Cells and Their Subsets in Patients with Graves' Disease

doi: 10.1155/2018/8972572

Figure Lengend Snippet: Flow analysis of circulating Tfh cells in GD patients. Human PBMCs from GD patients (BT: 36; AT: 21) and 20 HC were stained with anti-CD4, anti-CXCR5, anti-ICOS, anti-CD45RA, and anti-PD-1. (a) The cells were gated initially on lymphocytes and then circulating Tfh cells were analyzed by flow cytometry; (b) the numbers of circulating CD4 + CXCR5 + CD45RA − Tfh (cTfh) cells; (c) the numbers of CD4 + CXCR5 + CD45RA − ICOS + T cells; (d) CD4 + CXCR5 + CD45RA − PD-1 + Tfh cells; (e) the correlation between PD-1 + Tfh cell proportions and TPO-Ab levels in GD patients. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; ns: no significant difference.

Article Snippet: The peripheral blood mononuclear cells (PBMCs) were separated immediately by density gradient separation with Ficoll-Hypaque solution (CL5020, CEDARLANE, Canada).

Techniques: Staining, Flow Cytometry

Journal: Journal of Immunology Research

Article Title: Constitutive Changes in Circulating Follicular Helper T Cells and Their Subsets in Patients with Graves' Disease

doi: 10.1155/2018/8972572

Figure Lengend Snippet: Expanded frequency of circulating plasma cells in GD patients. Human PBMCs from GD patients (BT: 36; AT: 21) and 20 HC were stained with anti-CD19, anti-CD38, and anti-CD27. (a) The cells were gated initially on lymphocytes and then on CD19 + B cells; afterwards, the numbers of CD27 high CD38 high PCs were analyzed by flow cytometry. (b) Changes in PCs in GD patients. (c) Relation of PC proportions and serum TPO-Ab levels in GD patients. (d) Relation of Tfh2 subset proportions with PCs in GD patients. (e) The correlation between the proportions of PCs and ICOS + Tfh cells in GD patients. (f) The correlation between the proportions of PCs and PD-1 + Tfh cells in GD patients. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; ns: no significant difference.

Article Snippet: The peripheral blood mononuclear cells (PBMCs) were separated immediately by density gradient separation with Ficoll-Hypaque solution (CL5020, CEDARLANE, Canada).

Techniques: Clinical Proteomics, Staining, Flow Cytometry

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Obstructive Sleep Apnea–Induced Hypertension Is Associated With Increased Gut and Neuroinflammation

doi: 10.1161/JAHA.122.029218

Figure Lengend Snippet: Plasma levels of interleukin‐17a ( A ) and interleukin‐10 ( B ). Systolic blood pressure ( C ) in sham and OSA rats treated with a nonimmune IgG or interleukin‐17a neutralizing antibody. n=5 in ( A and B ), n=5–6 in ( C ); * P <0.05 for Sham+IgG vs OSA+IgG, # P <0.05 of OSA+nIL‐17a vs OSA+IgG, *** P <0.001 using two‐way ANOVA ( A , B ) or 2‐way repeated measures ANOVA and multiple comparisons with Tukey's multiple comparisons test ( C ). Error bars represent ±SEM. BP indicates blood pressure; IgG, immunoglobulin G; and OSA, obstructive sleep apnea.

Article Snippet: Biotinylated anti‐rat interleukin‐17a (Rockland, No. 212‐406‐b32) was added and incubated for 2 hours at room temperature.

Techniques: Clinical Proteomics

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Obstructive Sleep Apnea–Induced Hypertension Is Associated With Increased Gut and Neuroinflammation

doi: 10.1161/JAHA.122.029218

Figure Lengend Snippet: Effects of interleukin‐17a neutralization on Tregs, T H 1, T H 17, TNFα+, and macrophages in the ileum ( A ), cecum ( B ), and brain ( C ) of sham and OSA rats. n=4 in ileum, n=6 in cecum and brain; * P <0.05, ** P <0.005, *** P <0.0005, **** P <0.0001 using 2‐way ANOVA with Tukey's multiple comparisons test. Error bars represent ±SEM. OSA indicates obstructive sleep apnea; T H , T helper cell; and Tregs, T regulatory cells.

Article Snippet: Biotinylated anti‐rat interleukin‐17a (Rockland, No. 212‐406‐b32) was added and incubated for 2 hours at room temperature.

Techniques: Neutralization