isotype control igg4  (Bio X Cell)

Journal: Journal of Translational Autoimmunity

Article Title: Anti-IgLON5 encephalitis is associated with anti-retinal immunological reactivity without retinal alteration

doi: 10.1016/j.jtauto.2026.100359

Figure Lengend Snippet: Indirect immunofluorescence on sections of monkey retina. The fluorescence observed is identified by a black arrow. (A) Patient 4 (IgG4); (B) Patient 1 (IgG4); (C) Patient 3 (IgG1); (D) Patient 5 (IgG4); (E) Patient 2 (IgG1); (F) Patient 1 (IgG4) after immunoadsorption of IgLON5 antibodies; (G) Control with macular edema (IgG1); (H) Control with anti-Hu encephalitis (IgG1); (I) Control with CAR syndrome (IgG4). The different layers of the retina are identified by their initials: pigment epithelium (pe), photoreceptor layer (pr), outer grain layer (og), outer plexiform layer (op), inner grain layer (ig), inner plexiform layer (ip), ganglion cell layer (gc), nerve fibre layer (nf).

Article Snippet: The reactivity of the patients’ sera and/or CSF against the retina was evaluated retrospectively with frozen samples (−80°, EXPLAINEUR biobank) through an indirect immunofluorescence technique using sections of monkey retina (Ref. FA1172-1005, Euroimmun), detected with an FITC-labelled secondary antibody anti-human IgAGM (Euroimmun conjugate) or directed against IgA (ref. F0204, DAKO), IgM (ref. F0203, DAKO), IgG1 (ref. 9052-02, Southern Biotech) and IgG4 (ref. 9200-02, Southern Biotech).

Techniques: Immunofluorescence, Fluorescence, Control

Journal: bioRxiv

Article Title: Combination antagonism of TNF superfamily signaling for T cell immunosuppression

doi: 10.64898/2026.04.27.721101

Figure Lengend Snippet: ( A ) Interleukin-2 (IL-2) and interferon gamma (IFN-γ) inhibition in CD3 + cells during allogeneic one-way MLR measured at day 2 and day 5, respectively, treated with 50 nM combination pharmacological inhibitors of TNF/TNFR targets of interest. Monotreatment groups included 50 nM of human IgG1. Cytokine inhibition was calculated as percent reduction from IgG1 control treatment. Ill denotes P <0.05 in student t-test for monotreatments vs. IgG1 control, # denotes P <0.05 in student t-tests for combination treatment groups vs. IgG1 controls and both constituent monotreatments. ( B ) Normalized IL-2 and IFN-γ expression in four promising combinations of TNF/TNFR inhibitors. ( C-D ) Proliferation in CD3 + cells during allogeneic one-way MLR measured at day 5 in the presence of combination pharmacological inhibition of TNF/TNFR targets of interest.* denotes P <0.05 in student t-test. Data from three unique stimulator-responder MLR pairs, mean ± SEM plotted. Large-molecule pharmacological TNF/TNFR inhibitors used: Adalimumab (abbr. Ada, α-TNF), Pateclizumab (α-LTA), Baminercept (abbr. Bam, LTβR decoy), Dapirolizumab (abbr. Dapiro, α-CD40L), Amlitelimab (abbr. Amlite, α-OX40L), CD30Li (α-CD30L), 4-1BB-Fc (4-1BB decoy), Quisovalimab (α-LIGHT), Duvakitug (α-TL1A), and GITRi (α-GITR).

Article Snippet: Adalimumab (Cat. HY-P9908), Pateclizumab (Cat. HY-P990034), Baminercept (Cat. HY-P99459), Amlitelimab (Cat. HY-P99434), Dapirolizumab (Cat. HY-P99842A), Quisovalimab (Cat. HY-P99810), Duvakitug (Cat. HY-P99842A), human IgG1 (Cat. HY-P99001) and human IgG4 (Cat. HY-P99003) were purchased from MedChemExpress.

Techniques: Inhibition, Control, Expressing

Journal: bioRxiv

Article Title: Combination antagonism of TNF superfamily signaling for T cell immunosuppression

doi: 10.64898/2026.04.27.721101

Figure Lengend Snippet: ( A-B ) Interleukin-2 (IL-2) and interferon gamma (IFN-γ) inhibition in CD3 + cells during allogeneic one-way MLR measured at day 2 and day 5, respectively, treated with 50 nM combination pharmacological inhibitors of TNF/TNFR targets of interest. Monotreatment groups included 50 nM of human IgG1. Cytokine expression normalized to IgG1 control treatment. * denotes P <0.05 in student t-test for monotreatments vs. IgG1 control. Data from three individual stimulator-responder MLR pairs, mean ± SEM plotted.

Article Snippet: Adalimumab (Cat. HY-P9908), Pateclizumab (Cat. HY-P990034), Baminercept (Cat. HY-P99459), Amlitelimab (Cat. HY-P99434), Dapirolizumab (Cat. HY-P99842A), Quisovalimab (Cat. HY-P99810), Duvakitug (Cat. HY-P99842A), human IgG1 (Cat. HY-P99001) and human IgG4 (Cat. HY-P99003) were purchased from MedChemExpress.

Techniques: Inhibition, Expressing, Control

Journal: NPJ Breast Cancer

Article Title: TGF-α/EGFR-mediated lymphatic metastasis reveals a repositionable therapeutic target in breast cancer

doi: 10.1038/s41523-026-00941-0

Figure Lengend Snippet: Studies were performed to analyze whether blockade of TGF-α could inhibit chemotactic invasion and lymphatic metastasis of 4T1-EGFR cells. a Results from transwell invasion assays showing that Fepixnebart, a neutralizing antibody against TGFα (aTGFα), significantly inhibited chemotactic invasion of 4T1-EGFR cells toward svLEC-conditioned media. An isotype matched IgG antibody was used as control. For each condition, three fields per well were imaged and quantified across three independent experiments based on crystal violet-stained areas. Statistical analysis was performed using an unpaired two-tailed Student’s T-test. b Schematic overview of the in vivo experimental setup. 4T1 cells were orthotopically implanted into mice, followed by tail vein injection of Fepixnebart. Tumor-draining lymph nodes were collected for analysis 10 days later. c Bar graphs showing quantification of CK⁺ tumor cells in inguinal (ILN) and axillary (ALN) lymph nodes 10 days after orthotopic injection of 4T1-EGFR cells into mammary fat pads. Mice received a single intravenous injection of Fepixnebart (10 mg/kg; n = 9) or isotype IgG control ( n = 8) at the time of tumor cell implantation. Data are shown as mean ± SEM; statistical analysis by unpaired two-tailed Student’s t-test.

Article Snippet: Simultaneously, mice received a single intravenous dose of Fepixnebart (10 mg/kg) or an isotype-matched human IgG4 control antibody (HY-P99003, MedChemExpress).

Techniques: Control, Staining, Two Tailed Test, In Vivo, Injection

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A ) Expression of CD27 and IgD on CD19 + B cells. Right: Percentage of IgD + CD27 − naïve B cells and IgD − CD27 − DN B cells. ( B ) Expression of CD27 and CD38 on CD19 + B cells. Right: Percentages of CD27 hi CD38 hi ASCs and CD27 + CD38 − memory B cells. ( C ) Expression of CD138 on CD27 hi CD38 hi ASCs. Right: Percentage of CD138 + ASCs and CD138 − ASCs. ( D ) Expression of CD11c and CD21 on CD19 + B cells. Right: Percentage of CD11c + CD21 − CD19 + B cells. [(A) to (D)] HC ( n = 63), irAE ( n = 33), RAC ( n = 46), and ICI ( n = 20). ( E to G ) GSEA was performed on the B cells between irAE and ICI. (E) Significantly enriched pathways in B cells from irAE and ICI. GSEA plots of IFN-α and IFN-γ response (F), and oxidative phosphorylation (G). ( H ) The volcano plots of the citrullinated or noncitrullinated relative IgG or IgM isotype autoantigen levels comparing RA versus HC, irAE versus ICI, irAE versus HC, or irAE versus RA. The autoantigens were labeled when P < 0.01. TNF-α reactivities resulted from the administration of anti–TNF-α therapy for the treatment of RA. ( I ) Immunoglobulin isotype levels in the plasma were measured by multiplex assay. HC ( n = 22), irAE ( n = 34), RAC ( n = 46), and ICI ( n = 26). ( J ) CXCL13 levels in the plasma were measured by enzyme-linked immunosorbent assay (ELISA). HC ( n = 20), irAE ( n = 34) RAC ( n = 47), and ICI ( n = 23). ( K ) B-cell activating factor (BAFF) levels in the plasma were measured by multiplex assay. HC ( n = 21), irAE ( n = 34), RAC ( n = 47), and ICI ( n = 18). Data in graphs represent mean ± SEM. Significance was tested by one-way ANOVA [(A) to (D) and (I) to (K)] and logistic regression (H). [(A) to (D) and (F) to (K)] ICI, ICI control.

Article Snippet: Pembrolizumab (100 μg/ml; Keytruda) or isotype control IgG4 (Bio X Cell, catalog no. CP147) was added into the culture media 2 days after stimulation, and B cells were further cultured for another 5 days.

Techniques: Expressing, Phospho-proteomics, Immunopeptidomics, Labeling, Clinical Proteomics, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to C ) Human naïve B cells were isolated from PBMCs and cultured under indicated conditions for 7 days; pembrolizumab or isotype control IgG4 (IgG4 Iso) was added on day 2; n = 8. (A) Expression of CD38 and CD27 on B cells. Right: Percentages of CD27 + CD38 − and CD27 + CD38 + B cells. (B) Expression of CD138 on CD27 + CD38 + B cells. Right: Percentage of CD138 + CD27 + CD38 + B cells. (C) Different immunoglobulin isotypes were measured in the culture supernatant of (A) and (B) by multiplex assay. ( D to F ) B cells from Humanized PD-1 (HuPD-1) mice were isolated and cultured with LPS, IL-4, BAFF, or ODN 2006, anti-IgM, IL-21, IL-4 or R848, anti-IgM, anti-CD40, IL-21, and IFN-γ for 3 days; pembrolizumab or isotype control was added on day 1. (D) Expression of IgG2c on activated B cells. FSC-H, Forward scatter height. Right: Percentage of IgG2c + B cells; n = 5. (E) Expression of IgG1 on activated B cells. Right: Percentage of IgG1 + B cells; n = 5. (F) Different immunoglobulin isotypes in the supernatant were measured by multiplex assay; n = 5. Data in graphs represent mean ± SEM. Significance was tested by two-way ANOVA.

Article Snippet: Pembrolizumab (100 μg/ml; Keytruda) or isotype control IgG4 (Bio X Cell, catalog no. CP147) was added into the culture media 2 days after stimulation, and B cells were further cultured for another 5 days.

Techniques: Isolation, Cell Culture, Control, Expressing, Multiplex Assay

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to G ) PBMCs from the patients with irAE were cultured in the plate coated with anti-CD3 and anti-CD28 (10 μg/ml) in the presence of IgG1 isotype control or anti–human IL-6R (50 μg/ml), anti–human IL-12p40 (50 μg/ml), and anti–human IFNAR1 (50 μg/ml) for 3 days; n = 9. (A) Expression of CD38 and CD127 on CD8 + T cells. Right: Percentage of CD38 + CD127 − CD8 + T cells. (B and C) CD8 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (B) Expression of granzyme B and IFN-γ. Right: Percentage of granzyme B + IFN-γ + CD8 + T cells. (C) Expression of perforin and IFN-γ. Right: Percentage of perforin + IFN-γ + CD8 + T cells. [(D) and (E)] MFIs of GluCy5 (D), TMRM, and MTDR (E) in CD8 + T cells were presented. [(F) and (G)] CD4 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (F) Expression of IL-21 and IL-2. Right: Percentage of IL-21 + IL-2 + CD4 + T cells. (G) Expression of CD38 and CXCR5 on CD4 + T cells. Right: Percentage of CD38 + CXCR5 − CD4 + T cells. Data in graphs represent mean ± SEM. Significance was tested paired Student’s t test [(A) to (G)].

Article Snippet: Pembrolizumab (100 μg/ml; Keytruda) or isotype control IgG4 (Bio X Cell, catalog no. CP147) was added into the culture media 2 days after stimulation, and B cells were further cultured for another 5 days.

Techniques: Cell Culture, Control, Expressing