anti-tigit neutralizing antibody  (AMS Biotechnology)

Journal: JHEP Reports

Article Title: The dual role of TIGIT in regulatory and effector T cells in chronic liver disease

doi: 10.1016/j.jhepr.2025.101405

Figure Lengend Snippet: Regulatory T cells, enriched with the co-inhibitory receptor TIGIT, highly express Treg suppressive functional markers in both the blood and liver of patients with CLD. Immune cells isolated from the blood and liver of patients with CLD and healthy controls were phenotyped using flow cytometry. Details of samples used are in . Each data point represents values for an individual patient. p values are displayed for statistically significant comparisons only ( p <0.05). (A) Flow cytometry gating strategy for TIGIT expression on CD25 + CD127 low Tregs, CD25 low CD127 + (Tconvs) and CD8 + T cells. (B) CD25 + CD127 low expression by CD3 + CD4 + T cells in healthy controls, AIH, and blood from patients with non-AIH liver disease (grouped as chronic disease liver) and donor and diseased liver tissue. (C) TIGIT expression on blood and liver Tregs across cohorts. Statistical analysis was performed using a one-way ANOVA. (D) Geometric mean of TIGIT on CD25 + CD127 low Tregs. (E) Expression of TIGIT on Tregs, Tconvs, and CD8 + T cells from both CLD and healthy control patients. Data represent cells isolated from the blood and liver individually. Statistical analysis was performed using one-way ANOVA. (F) Paired expression of TIGIT and CD226 on Tregs in each cohort. Statistical analysis was performed using mixed-effects analysis, with the Geisser–Greenhouse correction and Šídák’s multiple comparisons test, with individual variances for each comparison. (G) Comparison of TIGIT and CD226 expression by Tregs across all combined cohorts. Statistical analysis was performed using paired t test. (H) Geometric mean comparing expression of TIGIT to CD226 on CD25 + CD127 low Tregs. Statistical analysis was performed using the paired t test. AIH, autoimmune hepatitis; CDB, chronic liver disease blood; CLD, chronic liver disease; DiL, disease liver; DoL, donor liver; HCB, healthy control blood; Tconv, conventional T cells; TIGIT, T cell immunoreceptor with Ig and ITIM domains; Tregs, regulatory T cells.

Article Snippet: A TIGIT neutralising antibody (#71340, BPS Bioscience, San Diego, California, USA) was added (5 μg/ml) to select wells, blocking TIGIT interactions.

Techniques: Functional Assay, Isolation, Flow Cytometry, Expressing, Control, Comparison

Journal: JHEP Reports

Article Title: The dual role of TIGIT in regulatory and effector T cells in chronic liver disease

doi: 10.1016/j.jhepr.2025.101405

Figure Lengend Snippet: Hepatic recruitment and residency markers are upregulated on TIGIT + Tregs. TIGIT + FoxP3 + cells reside around hepatocytes in AIH livers and the TIGIT ligand-CD155 is expressed on inflamed hepatocytes. Ex vivo Tregs from patient peripheral blood and explant livers were phenotyped using flow cytometry. Details of samples used for flow cytometry experiments are in unless stated otherwise. Each data point represents values for an individual patient. (A) Representative histogram (CXCR3 and CD69) or contour plot (CD29/VLA-4) showing the expression of hepatic recruitment and residency markers by live CD3 + CD4 + CD25 + CD127 low TIGIT + Tregs as determined by flow cytometry. In the histograms, grey represents isotype, black represents positive stain. (B) Paired analysis comparing expression of the hepatic recruitment markers CXCR3, VLA-4, and tissue residency marker CD69 by blood and liver TIGIT + and TIGIT - Tregs using flow cytometry. Statistical analysis was performed using mixed-effects analysis, with the Geisser–Greenhouse correction and Šídák’s multiple comparisons test, with individual variances for each comparison. p values are displayed for statistically significant comparisons only ( p <0.05). (C) Immunohistochemistry staining of AIH explant liver sections and non-cirrhotic donor liver sections for TIGIT (brown). (C–E) The right panels represent a magnified view of the area shown on the left (red box). (D) Immunohistochemistry staining for CD155 (brown) on AIH and non-cirrhotic donor explant liver. (E) Representative confocal micrograph showing immunofluorescence staining for TIGIT (red) and FoxP3 (green) in an AIH explant section. Blue = DAPI. (F–G) Quantification of (F) CD155 intensity and (G) TIGIT frequency from immunohistochemistry staining of AIH explant liver and non-cirrhotic donor liver using eight randomly selected fields of view analysed using QuPath. Error bars represent standard error of mean (SEM). Statistical analysis was performed using an unpaired t test. p values are displayed for each statistical comparison made. AIH, autoimmune hepatitis; CDB, chronic liver disease blood; CLD, chronic liver diseases; DiL, disease liver; DoL, donor liver; HCB, healthy control blood; TIGIT, T cell immunoreceptor with Ig and ITIM domains; Tregs, regulatory T cells.

Article Snippet: A TIGIT neutralising antibody (#71340, BPS Bioscience, San Diego, California, USA) was added (5 μg/ml) to select wells, blocking TIGIT interactions.

Techniques: Ex Vivo, Flow Cytometry, Expressing, Staining, Marker, Comparison, Immunohistochemistry, Immunofluorescence, Control

Journal: JHEP Reports

Article Title: The dual role of TIGIT in regulatory and effector T cells in chronic liver disease

doi: 10.1016/j.jhepr.2025.101405

Figure Lengend Snippet: TIGIT + Tregs express significantly higher amounts of the suppressive functional markers CD39, CTLA-4, and FoxP3 and are more suppressive than TIGIT - Tregs, exerting suppression via IL-10 and soluble CTLA-4. Phenotyping of immune cells was performed using multiparameter flow cytometry from patient blood and explant liver samples. Details of samples used are in unless stated otherwise. Each data point represents values for an individual patient. (A) Representative histogram showing CTLA-4, FoxP3, and CD39 expression in live CD3 + CD4 + CD25 + CD127 - TIGIT + Tregs as determined by flow cytometry. Grey represents isotype, black represents positive stain. (B) Spider plot illustrating the co-expression of CTLA-4, CD39 and FoxP3 determined using flow cytometry analysis on blood samples of three patients with AIH. (C) tSNE plots of CD3 + CD4 + CD25 + CD127 - Tregs from blood from patients with AIH, highlighting the co-expression of TIGIT, CTLA-4, FoxP3, and CD39 on Tregs. (D) Paired comparisons of CTLA-4, FoxP3 and CD39 functional marker expression by TIGIT + and TIGIT - Tregs. Statistical analysis was performed using mixed-effects analysis, with the Geisser–Greenhouse correction and Šídák’s multiple comparisons test, with individual variances for each comparison. p values are displayed for statistically significant comparisons only ( p <0.05). (E) Representative flow cytometry contour plot demonstrating the result of magnetic sorting of expanded CD4 + CD25 + CD127 low/- Tregs for TIGIT + and TIGIT - Tregs from patient blood. (F) Expanded patient blood derived Tregs were cell-sorted for TIGIT + and TIGIT - Tregs and co-cultured with autologous matched responder CellTrace™ Violet-labelled CD4 + effector T cells at varying Treg:CD4 + T cell ratios (1:1 to 1:8) for 5 days. Responder cell suppression was assessed by measuring the reduction in proliferation, determined by flow cytometry, compared to cells cultured alone. Data were collected from three individual biological repeats and three experimental repeats per condition. Statistical analysis was performed using a two-way ANOVA, with p values shown for each statistical comparison made. Error bars represent standard error of mean (SEM). (G) IL-10 cytokine secretion levels in the cell co-culture supernatant of the suppression assay shown in F, measured using ELISA. Data were collected from two individual biological repeats and two experimental repeats per condition. Statistical analysis was performed using a two-way ANOVA, with p values shown for each statistical comparison made. Error bars represent standard error of mean (SEM). AIH, autoimmune hepatitis; CDB, chronic liver disease blood; CTLA-4, cytotoxic T lymphocyte-associated antigen-4; DiL, disease liver; DoL, donor liver; HCB, healthy control blood; TIGIT, T cell immunoreceptor with Ig and ITIM domains; Tregs, regulatory T cells; tSNE, t-distributed stochastic neighbour embedding.

Article Snippet: A TIGIT neutralising antibody (#71340, BPS Bioscience, San Diego, California, USA) was added (5 μg/ml) to select wells, blocking TIGIT interactions.

Techniques: Functional Assay, Flow Cytometry, Expressing, Staining, Marker, Comparison, Derivative Assay, Cell Culture, Co-Culture Assay, Suppression Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: JHEP Reports

Article Title: The dual role of TIGIT in regulatory and effector T cells in chronic liver disease

doi: 10.1016/j.jhepr.2025.101405

Figure Lengend Snippet: TIGIT agonism enhances the suppressive activity of TIGIT + Tregs but reduces their proliferative capacity. (A) Expanded patient blood derived Tregs were cell-sorted for TIGIT + and TIGIT - Tregs and co-cultured with autologous matched responder CellTrace™ Violet-labelled CD4 + effector T cells at varying Treg:CD4 + T cell ratios (1:1 to 1:8) for 5 days in the presence or absence of TIGIT agonist. Responder cell suppression was assessed by measuring the reduction in proliferation, determined by flow cytometry, compared with cells cultured alone. Data were collected from three individual biological repeats and three experimental repeats per condition. Statistical analysis was performed using a two-way ANOVA, with p values shown for each statistical comparison made. Error bars represent standard error of mean (SEM). (B) TIGIT + and TIGIT - Tregs were cultured individually in the presence or absence of a TIGIT agonist (solid line and dotted line respectively) for 3 days. Expression of CD39, CTLA-4, FoxP3, IFN-γ, TNF-α and Ki67 was determined by flow cytometry. Data were collected from three individual biological repeats and three experimental repeats per condition. Statistical analysis was performed using a two-way ANOVA, with p values shown for statistically significant comparisons only ( p <0.05). Error bars represent standard error of mean (SEM). (C) Representative contour plots showing the frequency of FoxP3 expression on TIGIT + Tregs on day 2 of co-culture with and without a TIGIT agonist, analysed by flow cytometry. (D) Magnetically isolated TIGIT + and TIGIT - expanded Tregs were cultured with the addition of α-CD3, α-CD28 and α-TIGIT antibodies in various combinations ( i.e. α-CD3, α-CD3 and α-TIGIT or α-CD3, α-CD28 and α-TIGIT) for 3 days. Proliferation was determined by the reduction in fluorescence in CFSE labelling between day 0 and day 3. Data were collected from four individual biological repeats and two experimental repeats per condition. Statistical analysis was performed using a two-way ANOVA, with p values displayed for each statistical comparison made. IFN-γ, interferon-gamma; TIGIT, T cell immunoreceptor with Ig and ITIM domains; TNF-α, tumour necrosis factor-alpha; Tregs, regulatory T cells.

Article Snippet: A TIGIT neutralising antibody (#71340, BPS Bioscience, San Diego, California, USA) was added (5 μg/ml) to select wells, blocking TIGIT interactions.

Techniques: Activity Assay, Derivative Assay, Cell Culture, Flow Cytometry, Comparison, Expressing, Co-Culture Assay, Isolation, Fluorescence

Journal: JHEP Reports

Article Title: The dual role of TIGIT in regulatory and effector T cells in chronic liver disease

doi: 10.1016/j.jhepr.2025.101405

Figure Lengend Snippet: The intrahepatic environment is deprived of IL-2, and low-dose IL-2 enhances the expression of the suppressive markers CTLA-4 and FoxP3 on TIGIT + Tregs. (A–C) Cell-sorted ex vivo CD4 + T cells isolated from blood were co-cultured with IL-2 at varying concentrations: no IL-2, low IL-2 (0.3 ng/ml), high IL-2 (6 ng/ml), extremely high IL-2 (12,000 ng/ml). Data were collected from seven individual biological repeats and three experimental repeats per condition. (A) Flow cytometry gating strategy demonstrating the expression of CTLA-4 and FoxP3 on TIGIT + Tregs from magnetically sorted CD4 + T cells. (B) TIGIT expression on Tregs determined by flow cytometry after 24 h of CD4 + T cells cultured with varying concentrations of IL-2. Error bars represent the standard error of the mean (SEM). (C) Expression of CTLA-4 and FoxP3 on Tregs, as determined by flow cytometry, after 24 h of co-culture with varying IL-2 concentrations (n = 7). Statistical analysis was performed using a mixed-effects model with Geisser–Greenhouse correction and Tukey’s multiple comparisons test with individual variances computed for each comparison. Error bars represent the standard error of the mean (SEM). (D) Serum levels of IL-6 and IL-2 measured by ELISA in patients with inflamed chronic immune-mediated liver disease and healthy controls. Each data point represents values for an individual patient. Statistical analysis was performed using a one-way ANOVA, p values are displayed for statistically significant comparisons only ( p <0.05). Error bars represent the standard error of the mean (SEM). CTLA-4, cytotoxic T lymphocyte-associated antigen-4; TIGIT, T cell immunoreceptor with Ig and ITIM domains; Tregs, regulatory T cells.

Article Snippet: A TIGIT neutralising antibody (#71340, BPS Bioscience, San Diego, California, USA) was added (5 μg/ml) to select wells, blocking TIGIT interactions.

Techniques: Expressing, Ex Vivo, Isolation, Cell Culture, Flow Cytometry, Co-Culture Assay, Comparison, Enzyme-linked Immunosorbent Assay

Journal: JHEP Reports

Article Title: The dual role of TIGIT in regulatory and effector T cells in chronic liver disease

doi: 10.1016/j.jhepr.2025.101405

Figure Lengend Snippet: TIGIT-expressing CD8 + T cells and Tconvs are equipped with liver homing and residency markers. Immune cells isolated from the blood and liver of patients with CLD and healthy controls were phenotyped using flow cytometry. Details of samples used are in . Each data point represents values for an individual patient. p values are displayed for statistically significant comparisons only ( p <0.05). Error bars represent the standard error of the mean (SEM). (A) The expression level of TIGIT on Tconvs and CD8 + T cells in blood and liver. Statistical analysis was performed using a one-way ANOVA. (B) Expression of TIGIT on Tconvs and CD8 + T cells from both CLD and healthy control patients. Data represent cells isolated from the blood and liver individually. Statistical analysis was performed using one-way ANOVA. (C–D) Paired analysis comparing expression of the hepatic recruitment markers CXCR3, VLA-4, and tissue residency marker CD69 by blood and liver TIGIT + and TIGIT - (C) CD8 + T cells and (D) Tconv T cells using flow cytometry. Statistical analysis was performed using a mixed-effects analysis, with the Geisser–Greenhouse correction and Šídák’s multiple comparisons test, with individual variances for each comparison. (E) Paired expression of TIGIT and CD226 by Tconvs and CD8 + T cells from the blood and liver in disease and healthy cohorts. Statistical tests were conducted using mixed-effects analysis, with the Geisser–Greenhouse correction and Tukey’s multiple comparisons test, with individual variances computed for each comparison. CDB, chronic liver disease blood; CLD, chronic liver diseases; DiL, disease liver; DoL, donor liver; HCB, healthy control blood; Tconv, conventional T cells; TIGIT, T cell immunoreceptor with Ig and ITIM domains.

Article Snippet: A TIGIT neutralising antibody (#71340, BPS Bioscience, San Diego, California, USA) was added (5 μg/ml) to select wells, blocking TIGIT interactions.

Techniques: Expressing, Isolation, Flow Cytometry, Control, Marker, Comparison

Journal: JHEP Reports

Article Title: The dual role of TIGIT in regulatory and effector T cells in chronic liver disease

doi: 10.1016/j.jhepr.2025.101405

Figure Lengend Snippet: TIGIT interactions on TIGIT-expressing CD8 + T cells inhibit hepatocyte apoptosis. (A) Representative histogram of granzyme B and perforin expression, gated on live CD3 + CD8 + TIGIT + T cells. Grey represents isotype, black represents positive stain. (B) Paired comparison of granzyme B and perforin expression by TIGIT + and TIGIT - CD8 + T cells. Statistical tests were conducted using mixed-effects analysis, with the Geisser–Greenhouse correction and Tukey’s multiple comparisons test, with individual variances computed for each comparison. Details of samples used for flow cytometry are in . p values are displayed for statistically significant comparisons only ( p <0.05). (C) PBMCs from AIH patient blood were used for flow cytometry staining for naive T cell markers, CD45RA and CCR7, gated on live CD3 + CD8 + TIGIT + or TIGIT - T cells. (D) Paired comparison of antigen-experienced marker CD40L expression on TIGIT + and TIGIT - CD8 + T cells. Statistical analysis was performed using a mixed-effects analysis, with the Geisser–Greenhouse correction and Šídák’s multiple comparisons test, with individual variances for each comparison. (E) ICC staining of PHHs in co-culture with either TIGIT - or TIGIT + (magenta) labelled CD8 + T cells (CellTracker™ Red/CTR, orange). T cells were isolated from blood derived from patients with AIH using FACS and rested overnight before labelling and co-culture with PHHs. (F) Hepatocyte cell death after 24 h in co-culture with TIGIT - or TIGIT + CD8 + T cells or TIGIT - or TIGIT + Tconv cells in the presence or absence of a TIGIT neutralising antibody (monoclonal; 5 μg/ml) Data were collected from two individual biological repeats. Statistical analysis was performed using one-way ANOVA, with p values shown for statistically significant comparisons only ( p <0.05). Error bars represent the standard error of the mean (SEM). (G) Time-lapse images showing hepatocyte cell death. Images were taken every 30 min of PHH and TIGIT - CD8 + T cells labelled with CellTrace Red (CTR) in culture. The red arrow points towards an immune cell, and the white arrow points towards an apoptosing hepatocyte. The grey image shows the acquisition of phase gradient. (H) Representative images of ICC staining showing the expression of granzyme B (white) on either TIGIT - CD8 + T cells or TIGIT + CD8 T cells (CTR, magenta) and a magnified image of the interaction between a TIGIT - CD8 + T cell (CTR, magenta) and hepatocytes (CellTracker™ Green/CTG, yellow) or the lack of interaction between a TIGIT + CD8 + T cell and hepatocytes. AIH, autoimmune hepatitis; AIHB, AIH patient blood; CDB, chronic liver disease blood; DiL, disease liver; DoL, donor liver; HCB, healthy control blood; ICC, immunocytochemistry; PBMCs, peripheral blood mononuclear cells; PHHs, primary human hepatocytes; Tconv, conventional T cells; TIGIT, T cell immunoreceptor with Ig and ITIM domains.

Article Snippet: A TIGIT neutralising antibody (#71340, BPS Bioscience, San Diego, California, USA) was added (5 μg/ml) to select wells, blocking TIGIT interactions.

Techniques: Expressing, Staining, Comparison, Flow Cytometry, Marker, Co-Culture Assay, Isolation, Derivative Assay, Control, Immunocytochemistry

Journal: Biomarker Research

Article Title: Targeting TIGIT for cancer immunotherapy: recent advances and future directions

doi: 10.1186/s40364-023-00543-z

Figure Lengend Snippet: Selected Preclinical studies of anti-TIGIT agents in solid and hematological tumors

Article Snippet: Anti-TIGIT (#71340, 5 μg/mL, BPS Biosciences) [ ] , Not mentioned , CD8 + T cells from human gastric cancer tissues , Treatment of anti-TIGIT or combination of anti-TIGIT and SOX enhanced the proliferation and IFN-γ production ability of CD8+ T cells. , S-1 plus oxaliplatin (SOX) , Gastric Cancer.

Techniques: Knock-In, Cell Function Assay, Injection, Activation Assay, Amplification, Incubation, Lysis

Journal: Biomarker Research

Article Title: Targeting TIGIT for cancer immunotherapy: recent advances and future directions

doi: 10.1186/s40364-023-00543-z

Figure Lengend Snippet: Mechanisms of immunosuppression of TIGIT in TME. A. CD155 binds with TIGIT with higher affinity compared to CD226 leading to deactivation of T or NK cells. In addition, CD155 is associated with drug resistance, and tumor cell migration and metastasis. Fap2, secreted from F. nucleatum, triggers inhibition by binding to TIGIT. B. The interaction of CD112 and CD112R, or TIGIT, inhibits proliferation of T-cells and NK cells by reducing IFN-γ production. CD155 increases IL-10 production by binding to TIGIT. C. TIGIT on Treg activates its immunosuppression on Th1, Th17, and effector T cells. D. The binding of TIGIT and CD155 on memory B cells exhibits deactivated DCs and inhibits Th1/17/2. E. The production of IL-10 from DCs inhibits TILs

Article Snippet: Anti-TIGIT (#71340, 5 μg/mL, BPS Biosciences) [ ] , Not mentioned , CD8 + T cells from human gastric cancer tissues , Treatment of anti-TIGIT or combination of anti-TIGIT and SOX enhanced the proliferation and IFN-γ production ability of CD8+ T cells. , S-1 plus oxaliplatin (SOX) , Gastric Cancer.

Techniques: Migration, Inhibition, Binding Assay

Journal: Biomarker Research

Article Title: Targeting TIGIT for cancer immunotherapy: recent advances and future directions

doi: 10.1186/s40364-023-00543-z

Figure Lengend Snippet: The expression of TIGIT can be influenced by a variety of factors. Notably, Interferon-I (IFN-I), chemotherapy agents, GITR antibodies, and 1α,25-Dihydroxyvitamin D3 (1α,25(OH)(2)D(3)) have been observed to downregulate its expression. In contrast, microwave ablation, glucose deprivation, hypoxic conditions, as well as the presence of CCL23, IL-10, inhaled IL-15, and anti-PD-1/PD-L1 therapies have been found to enhance the expression of TIGIT, thereby contributing to an immunosuppressive TME

Article Snippet: Anti-TIGIT (#71340, 5 μg/mL, BPS Biosciences) [ ] , Not mentioned , CD8 + T cells from human gastric cancer tissues , Treatment of anti-TIGIT or combination of anti-TIGIT and SOX enhanced the proliferation and IFN-γ production ability of CD8+ T cells. , S-1 plus oxaliplatin (SOX) , Gastric Cancer.

Techniques: Expressing