Journal:

Article Title: An unusual Fc receptor-related protein expressed in human centroblasts

doi: 10.1073/pnas.022042699

Figure Lengend Snippet: Immunoglobulins do not bind FREB targeted to the cell surface of 293 cells. FREB was expressed on the cell surface of 293 cells as a FREB–CD4 fusion protein (Left). Control 293 cells were transfected with CD1a cDNA (Right). Cell surface expression of FREB and CD1a was confirmed by flow cytometry by using anti-FREB (a) and anti-CD1a (b) mAbs. Transfected cells were incubated with human IgA (c and d), IgE (e and f), IgM (g and h), and IgG (i and j), which had been previously aggregated by treatment at 65°C for 30 min. Immunoglobulins bound to transfected cells were detected by flow cytometry. Percentages of stained cells are indicated in the upper quadrants. Human IgA partially bound 293 cells, regardless of FREB expression (c and d). Cells stained only with the secondary antibodies fell within the lower quadrants. FSC, forward scatter.

Article Snippet: FREB was identified by using Texas red-conjugated anti-mouse IgG1 (Southern Biotechnology Associates), whereas CD20 and PCNA were detected with a biotinylated anti-mouse IgG2a antibody followed by FITC-conjugated streptavidin (Southern Biotechnology Associates).

Techniques: Transfection, Expressing, Flow Cytometry, Incubation, Staining

Journal:

Article Title: An unusual Fc receptor-related protein expressed in human centroblasts

doi: 10.1073/pnas.022042699

Figure Lengend Snippet: Preferential expression of FREB in germinal center centroblasts. Stainings of a secondary follicle with anti-CD20 (a, green) and anti-FREB (b, red) show preferential location of FREB in large cells of the germinal center corresponding to centroblasts. (c) FREB (red) shows a perinuclear pattern of expression around the nuclear PCNA (green). Coexpression of PCNA and FREB is particularly evident at the base of the germinal center (bottom), where centroblasts are predominant. In the upper part of the germinal center, where centrocytes predominate, PCNA-positive cells are mostly FREB negative. (d–g) Expressions of FREB (red) and IgD (d), IgA (e), IgM (f), or IgG (g; green) are mutually exclusive. In f, only rare cells coexpress FREB and IgM.

Article Snippet: FREB was identified by using Texas red-conjugated anti-mouse IgG1 (Southern Biotechnology Associates), whereas CD20 and PCNA were detected with a biotinylated anti-mouse IgG2a antibody followed by FITC-conjugated streptavidin (Southern Biotechnology Associates).

Techniques: Expressing

Journal: RSC Advances

Article Title: A quantitative ex vivo study of the interactions between reconstituted high-density lipoproteins and human leukocytes †

doi: 10.1039/c9ra08203d

Figure Lengend Snippet: Three-dimensional fluorescence microscopy for studying internalization of rHDL cargo. The anti-CD14 antibody is used to identify the surface of monocytes (red), the nucleus is stained with Hoechst (blue) and the DiO represents rHDL cargo (green). (A) Reconstructed 3D image of a monocyte with internalized DiO. (B) 2D image of a monocyte with orthogonal projection of the indicated slices, which clearly confirms the internalization of DiO.

Article Snippet: Anti-CD14 antibody (APC Mouse Anti-Human CD14, IgG1, κ, Clone 61D3) used for imaging flow cytometry was supplied by ThermoFisher Scientific (USA).

Techniques: Fluorescence, Microscopy, Staining

Journal: RSC Advances

Article Title: A quantitative ex vivo study of the interactions between reconstituted high-density lipoproteins and human leukocytes †

doi: 10.1039/c9ra08203d

Figure Lengend Snippet: Analysis of rHDL interaction with leukocytes using imaging flow cytometry for both DPPC and DPPG rHDL. (A) Selected DiO positive cells from two donors shown with brightfield (Ch01), CD14 (Ch11), DiO (Ch02), CD14/DiO and brightfield/DiO. The same contrast settings were used for monocytes and granulocytes for each of the rHDL formulations. (B) The imaging flow cytometry confirmed the preferred monocyte association of rHDL. The MFI was subtracted background from the specific cell type and adjusted by the integral of DiO absorbance from the SEC analysis of each rHDL formulation. (C) The brightfield images were used to define masks that defined the interior or the cells, as illustrated for a single monocyte in the inset, which also shows a mask of the cell surface. This allowed for the determination of the internalization of rHDL cargo, which showed more internalization positive monocytes than granulocytes. The data in B and C represent mean + SEM ( n = 2).

Article Snippet: Anti-CD14 antibody (APC Mouse Anti-Human CD14, IgG1, κ, Clone 61D3) used for imaging flow cytometry was supplied by ThermoFisher Scientific (USA).

Techniques: Imaging, Flow Cytometry, Formulation

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

Article Title: First in Human Phase 1/2 ICONIC Trial of the ICOS agonist vopratelimab alone and with nivolumab: ICOS-hi CD4 T cell populations and predictors of response

doi: 10.1158/1078-0432.CCR-21-4256

Figure Lengend Snippet: Patient demographics (data cut: 07-22-2020)

Article Snippet: Patients were treated with vopratelimab monotherapy (JTX-2011; a humanized IgG1κ monoclonal antibody and ICOS agonist; Jounce Therapeutics, Inc., US), or vopratelimab in combination with nivolumab (OPDIVO®; a human IgG4κ anti-PD-1 monoclonal antibody; Bristol-Myers Squibb., US).

Techniques:

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

Article Title: First in Human Phase 1/2 ICONIC Trial of the ICOS agonist vopratelimab alone and with nivolumab: ICOS-hi CD4 T cell populations and predictors of response

doi: 10.1158/1078-0432.CCR-21-4256

Figure Lengend Snippet: Treatment-Related Adverse Events (TRAE) (data cut: 07-22-2020)

Article Snippet: Patients were treated with vopratelimab monotherapy (JTX-2011; a humanized IgG1κ monoclonal antibody and ICOS agonist; Jounce Therapeutics, Inc., US), or vopratelimab in combination with nivolumab (OPDIVO®; a human IgG4κ anti-PD-1 monoclonal antibody; Bristol-Myers Squibb., US).

Techniques:

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

Article Title: First in Human Phase 1/2 ICONIC Trial of the ICOS agonist vopratelimab alone and with nivolumab: ICOS-hi CD4 T cell populations and predictors of response

doi: 10.1158/1078-0432.CCR-21-4256

Figure Lengend Snippet: Preliminary efficacy of vopratelimab as monotherapy or in combination with nivolumab (Phase 1 and Phase 2) data cut: 07-22-2020

Article Snippet: Patients were treated with vopratelimab monotherapy (JTX-2011; a humanized IgG1κ monoclonal antibody and ICOS agonist; Jounce Therapeutics, Inc., US), or vopratelimab in combination with nivolumab (OPDIVO®; a human IgG4κ anti-PD-1 monoclonal antibody; Bristol-Myers Squibb., US).

Techniques:

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

Article Title: First in Human Phase 1/2 ICONIC Trial of the ICOS agonist vopratelimab alone and with nivolumab: ICOS-hi CD4 T cell populations and predictors of response

doi: 10.1158/1078-0432.CCR-21-4256

Figure Lengend Snippet: Various tumor biomarkers were evaluated for their ability to predict the emergence of a peripheral ICOS-hi CD4 T-cell population in response to vopratelimab monotherapy or with nivolumab. (A) The 18 TIS genes are associated with antigen presentation, lymphocyte abundance, interferon activity and T cell exhaustion, including activation of CD4 T-cells. Genes highlighted in blue are involved in antigen presentation to CD4 T cells (HLA-DRB1 and HLA-DQA1) and successful recruitment of T cells and antigen presenting cells (CCL5 and CXCL9). (B) ROC curve demonstrating the ability of TIS to predict ICOS-hi CD4 T-cell emergence (n=27, AUC=0.83) and the lack of prediction by PD-L1 IHC (n=24, AUC=0.55)

Article Snippet: Patients were treated with vopratelimab monotherapy (JTX-2011; a humanized IgG1κ monoclonal antibody and ICOS agonist; Jounce Therapeutics, Inc., US), or vopratelimab in combination with nivolumab (OPDIVO®; a human IgG4κ anti-PD-1 monoclonal antibody; Bristol-Myers Squibb., US).

Techniques: Immunopeptidomics, Activity Assay, Activation Assay

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

Article Title: First in Human Phase 1/2 ICONIC Trial of the ICOS agonist vopratelimab alone and with nivolumab: ICOS-hi CD4 T cell populations and predictors of response

doi: 10.1158/1078-0432.CCR-21-4256

Figure Lengend Snippet: ICONIC patients treated with vopratelimab monotherapy or in combination with nivolumab who had pre-treatment tumor samples that were TISvopra-positive showed better outcomes to treatment than those whose tumor samples were TISvopra-negative. In all panels, orange depicts TISvopra-positive patients and gray shows TISvopra-negative patients. (A) Waterfall plot showing maximum reduction in the sum of diameters of target tumors compared to baseline in patients with at least one post-baseline CT scan who were assessed for TISvopra status (n=67). (B) Swimmer plots showing time on treatment for patients who were evaluated for TISvopra (n=89). Arrows indicate patients for whom treatment is ongoing; Left panel: patients who are TISvopra positive; right panel: patients who are TISvopra negative. BOR is evaluated using RECIST 1.1 criteria; PR is depicted in green, SD is depicted in yellow, PD is depicted black, NE is depicted in gray. (C) Progression-free survival curves for TISvopra-positive vs. TISvopra-negative patients, as evaluated by investigator review. An event is defined as PD or death (see Methods). (D) Overall survival curves for TISvopra-positive vs. TISvopra-negative patients. An event is defined as death (see Methods). BOR, best overall response; CR, complete response; ICOS, inducible co-stimulator; PD, progressive disease; PR, partial response; RECIST, response evaluation criteria in solid tumors; SD, stable disease; TIS, tumor inflammation signature. Data cut for all panels is July 22, 2020.

Article Snippet: Patients were treated with vopratelimab monotherapy (JTX-2011; a humanized IgG1κ monoclonal antibody and ICOS agonist; Jounce Therapeutics, Inc., US), or vopratelimab in combination with nivolumab (OPDIVO®; a human IgG4κ anti-PD-1 monoclonal antibody; Bristol-Myers Squibb., US).

Techniques: Computed Tomography

Journal: BMC Oral Health

Article Title: Different concentrations of C5a affect human dental pulp mesenchymal stem cells differentiation

doi: 10.1186/s12903-021-01833-4

Figure Lengend Snippet: Monoclonal antibodies used in this study

Article Snippet: MSCA-1 , PE , Mouse IgG1κ , Miltenyi Biotec , 130-099-198.

Techniques: Bioprocessing

Journal: Frontiers in Immunology

Article Title: HEV-associated dendritic cells are observed in metastatic tumor-draining lymph nodes of cutaneous melanoma patients with longer distant metastasis-free survival after adjuvant immunotherapy

doi: 10.3389/fimmu.2023.1231734

Figure Lengend Snippet: Favoring Antigen-Presenting Structures (FAPS) are present in mTDLN of GO patients. Biomarkers were determined by IHC or multiplex IF, as described under Methods; representative images are shown. (A) Patient#17 is enriched in peri- and intra-tumoral CD11c + cells. (B) Patient#17 CD11c + cells (green) contain melanin and tyrosinase (orange, white arrow). (C) A FAPS example is shown (Patient#13). CD11c + cells (white arrow) are shown surrounding PNAd + HEV (black arrow). (D) CD11c + cells (green) surrounding HEV (white) contained melanin and tyrosinase (orange) (Patient#17). (E) DC-LAMP + cells (white arrow) surrounding HEV with cuboidal epithelium (black arrow) are shown (Patient#13). (F) FAPS structure showing PNAd + (white) HEV interactions between CD62L + cells (red) and CD11c + cells (green) are indicated by arrows (Patient#13). Original magnifications: (A) 400X; (B, D, F) 200X; (C, E) 1000X. Scale bars: (A) 50µm; (B, D, F) 10µm; (C, E) 20µm.

Article Snippet: Primary mAbs: CD11c (clone EP 1347Y, 1:200, Abcam); MART-1 conjugated to AF647 [IgG1, κ, clone 2A9 ( )]; TYR conjugated to AF647 (clone T311, Santa Cruz Biotechnology); CD62L mAb conjugated to AlexaFluor-647 (clone DREG56, 1:50, IgG1 κ, Santa Cruz Biotechnology).

Techniques: Multiplex Assay

Journal: Frontiers in Immunology

Article Title: HEV-associated dendritic cells are observed in metastatic tumor-draining lymph nodes of cutaneous melanoma patients with longer distant metastasis-free survival after adjuvant immunotherapy

doi: 10.3389/fimmu.2023.1231734

Figure Lengend Snippet: Multiplex immunofluorescence analysis of FAPS. Multiplex staining of a mTDLN peritumoral area of one GO patient (#13) with anti-CD3 (cyan), anti-CD20 (blue), anti-CD62L (pink), anti-PNAd (red), anti-CD11c (yellow), and anti-DC-LAMP (white) antibodies was performed as described under Methods. DC-LAMP + and CD11c + cells surround PNAd + HEV. In the lower part area of the figure, marked with orange arrows, sprouts of PNAd + HEV (red), and CD62L + naïve lymphocytes (pink) making contacts with HEV and DC-LAMP + dendritic cells (white) may be observed.

Article Snippet: Primary mAbs: CD11c (clone EP 1347Y, 1:200, Abcam); MART-1 conjugated to AF647 [IgG1, κ, clone 2A9 ( )]; TYR conjugated to AF647 (clone T311, Santa Cruz Biotechnology); CD62L mAb conjugated to AlexaFluor-647 (clone DREG56, 1:50, IgG1 κ, Santa Cruz Biotechnology).

Techniques: Multiplex Assay, Immunofluorescence, Staining