Journal: bioRxiv

Article Title: Adjuvant Discovery via a High Throughput Screen using Human Primary Mononuclear Cells

doi: 10.1101/2022.06.17.496630

Figure Lengend Snippet: A. Distribution of z scores, indicating induction of the cytokines (IFN, IL-10, and TNF). B. Distribution of z scores, representing the percentage of monocytes with activated HLA-DR and/or CD80/86 receptors. C. Distribution of z scores, representing the percentage of T-Cells with activated Ox40 receptors. D. Distribution of z scores, representing the percentage of B-Cells with activated HLA-DR and/or CD80/86 receptors. E. graphical representation of small molecule hit rate in varying number of donors. natural log of hit rate. F. Strong negative correlation between number of donors. Each dot represents one of the 2,296 compounds from the primary screen. The dashed line represents the donor threshold of a z score of 2 and representative z scores were chosen such that any compound above the threshold represents a hit in the screen.

Article Snippet: CD134 (OX40) Antibody, anti-human, APC, REAfinity TM (Clone- REA621) , Miltenyi Biotec , Order Number: 130-122-227.

Techniques:

Journal: bioRxiv

Article Title: Adjuvant Discovery via a High Throughput Screen using Human Primary Mononuclear Cells

doi: 10.1101/2022.06.17.496630

Figure Lengend Snippet: Key resource table

Article Snippet: CD134 (OX40) Antibody, anti-human, APC, REAfinity TM (Clone- REA621) , Miltenyi Biotec , Order Number: 130-122-227.

Techniques: Recombinant, Sterility, Injection, Modification, Staining, Saline, Enzyme-linked Immunosorbent Assay, Multiplex Assay, Software

Journal: iScience

Article Title: Naive and memory CD4 + T cell subsets can contribute to the generation of human Tfh cells

doi: 10.1016/j.isci.2021.103566

Figure Lengend Snippet:

Article Snippet: Anti-Human CD134/OX40 (ACT35)-158Gd , Fluidigm , Cat# 3158012B.

Techniques: Cell Analysis, Purification, Virus, Recombinant, Blocking Assay, Antibody Labeling, Transfection, Software

Journal: Frontiers in Immunology

Article Title: Enforced OX40 Stimulation Empowers Booster Vaccines to Induce Effective CD4 + and CD8 + T Cell Responses against Mouse Cytomegalovirus Infection

doi: 10.3389/fimmu.2017.00144

Figure Lengend Snippet: Activation of the OX40 axis during booster vaccination with a single MHC class II synthetic long peptide (SLP) vaccine propels increment of the vaccine-induced CD4 + T cell response . (A) Scheme of the experimental procedure and the vaccination timeline. Wild-type C57BL/6 mice were vaccinated (i) s.c. with M25 409–423 MHC class II SLP alone or (ii) with M25 409–423 MHC class II SLP (s.c.) along with anti-OX40 mAb (i.p.). Two weeks after prime vaccination mice from group (i) and (ii) were divided into two groups, respectively, and a booster immunization was administered. Half mice received only the M25 409–423 SLP and the other half were injected anti-OX40 mAb in addition to the M25 409–423 SLP. (B) The total size of the splenic M25 409–423 SLP vaccine-induced CD4 + T cells from each group was measured by intracellular cytokine staining. Representative plots depict percentages of IFN-γ versus TNF cytokine producing CD4 + T cell populations at day 8 post-booster vaccination. (C) Total numbers of splenic IFN-γ + producing M25 409–423 antigen-specific CD4 + T cells at day 8 post booster SLP vaccination and differential anti-OX40 mAb treatment are shown. (D) Total double (IFN-γ/TNF) and (E) triple (IFN-γ/TNF/IL-2) cytokine producers of M25 409–423 vaccine-specific CD4 + T cells measured in spleen at day 8 post-booster vaccination. Fold differences among each population are also depicted (F) . Pie charts show the percentages of the single (IFN-γ), double (IFN-γ/TNF), and triple (IFN-γ/TNF/IL-2) cytokine producers of each M25 409–423 -specific CD4 + T cell population upon vaccination with M25 409–423 SLP and anti-OX40 mAb. Data represent mean values and are representative of three independent experiments ( n = 5–6 per group). * P < 0.05; ** P < 0.01; ns, not significant.

Article Snippet: LB is a shareholder of Bioceros that holds a patent on anti-human OX40 mAbs.

Techniques: Activation Assay, Injection, Staining

Journal: Frontiers in Immunology

Article Title: Enforced OX40 Stimulation Empowers Booster Vaccines to Induce Effective CD4 + and CD8 + T Cell Responses against Mouse Cytomegalovirus Infection

doi: 10.3389/fimmu.2017.00144

Figure Lengend Snippet: OX40 activation during booster vaccination with combinatorial MHC class II synthetic long peptide (SLP) vaccines leads to the induction of robust and polyfunctional CD4 + T cell responses . (A) Percentages (left) and total numbers (right) of the splenic epitope-specific IFN-γ + CD4 + T cell responses elicited at day 8 post-booster vaccination with a mixture of SLPs containing MHC class II epitopes alone (CD4 SLP mixture; white bars) or with SLPs containing MHC class II epitopes and anti-OX40 mAb (CD4 SLP mixture + anti-OX40 boost; black bars). (B) Pie charts show the percentages of the single (IFN-γ), double (IFN-γ/TNF), and triple (IFN-γ/TNF/IL-2) cytokine producers of each antigen-specific CD4 + T cell population at day 8 post-booster vaccination with a mixture of MHC class II SLPs alone or with MHC class II SLPs plus anti-OX40 mAb. (C) Total triple (IFN-γ/TNF/IL-2) cytokine producers of each antigen-specific CD4 + T cell population were measured in spleen at day 8 post-booster vaccination with a mixture of MHC class II SLPs and anti-OX40 mAb or with SLPs alone. Fold difference is indicated. Data represent mean values and are representative of three independent experiments ( n = 6 per group). * P < 0.05; ** P < 0.01.

Article Snippet: LB is a shareholder of Bioceros that holds a patent on anti-human OX40 mAbs.

Techniques: Activation Assay, Vaccines

Journal: Frontiers in Immunology

Article Title: Enforced OX40 Stimulation Empowers Booster Vaccines to Induce Effective CD4 + and CD8 + T Cell Responses against Mouse Cytomegalovirus Infection

doi: 10.3389/fimmu.2017.00144

Figure Lengend Snippet: OX40 ligation during booster vaccination with a mixture of MHC class I epitope-containing synthetic long peptides (SLPs) leads to the induction of strong and polyfunctional CD8 + T cell responses . (A) Longitudinal analysis of the epitope-specific CD8 + T cell responses in blood induced by combinatorial MHC class I epitope-containing SLP vaccines without (CD8 SLP mixture) and with anti-OX40 mAb administration (given only during booster vaccination) (CD8 SLP mixture + a-OX40 boost). Data represent mean values ± SEM ( n = 12 per group). (B) Total IFN-γ + cytokine-producing CD8 + T cells for each antigen-specific population detected in spleen at day 8 post-booster vaccination with CD8 SLP mixture (white bars) or with CD8 SLP mixture + anti-OX40 boost (black bars). (C) Pie charts show the percentages of the single (IFN-γ), double (IFN-γ/TNF), and triple (IFN-γ/TNF/IL-2)-specific cytokine producers of the antigen-specific CD8 + T cell populations at day 8 post-booster vaccination. (D) Total numbers of IFN-γ/TNF/L-2 cytokine producing CD8 + T cells for each antigen-specific population detected in spleen at day 8 post-booster vaccination. (E) At day 8 after booster vaccination with combined MHC class I SLPs and/or anti-OX40 mAb (during booster or during both prime/boost), the BCL-2 protein expression was measured within the antigen-specific CD8 + T cells in spleen by flow cytometry. Fold changes between groups are depicted. Data represent mean values + SEM ( n = 6 mice per group) and are representative of three independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: LB is a shareholder of Bioceros that holds a patent on anti-human OX40 mAbs.

Techniques: Ligation, Vaccines, Expressing, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Enforced OX40 Stimulation Empowers Booster Vaccines to Induce Effective CD4 + and CD8 + T Cell Responses against Mouse Cytomegalovirus Infection

doi: 10.3389/fimmu.2017.00144

Figure Lengend Snippet: Agonistic OX40 mAb administration during booster vaccination leads to improved memory synthetic long peptide (SLP)-elicited CD8 + T cell responses . (A) Total single IFN-γ + cytokine-producing CD8 + T cells for each antigen-specific population detected in spleen at day 60 post-booster vaccination with combinatorial MHC class I SLPs and anti-OX40 mAb (provided during booster vaccination) (CD8 SLP mixture + anti-OX40 boost) or with MHC class I SLPs alone (CD8 SLP mixture). Fold changes between groups are depicted. Data represent mean values + SEM ( n = 6 per group). (B) Pie charts show the percentages of the single (IFN-γ), double (IFN-γ/TNF), and triple (IFN-γ/TNF/IL-2) cytokine producers of each antigen-specific CD8 + T cell population in spleen at day 60 post-booster vaccination with a mixture of MHC class I SLPs and anti-OX40 mAb (administered only during booster vaccination) or with MHC class I SLPs alone. (C) Total triple IFN-γ + /TNF + /IL-2 + -producing CD8 + T cells for each antigen-specific population detected in spleen at day 60 post-booster vaccination. Fold changes between groups are depicted. Data represent mean values + SEM ( n = 6 per group). (D) Different groups of mice [unvaccinated (naive), mouse CMV (MCMV) infected (live-virus vaccine), vaccinated with a mixture of all 5 MHC class I SLPs, vaccinated with a mixture of all five MHC class I SLPs and anti-OX40 mAb (given only during booster vaccination), or treated with a-OX40 mAb only] were challenged at day 60 postvaccination/infection with 5 × 10 4 PFU salivary gland-derived MCMV Smith. At day 5, post challenge spleen, liver, and lungs were harvested and the viral genome copies were determined by qPCR. The viral titers of individual mice are depicted ( n = 5–8 mice per group). Mean ± SEM is indicated. The detection limit was below 1,000 genome copies as measured in naive mice. Experiments were performed twice with similar outcome. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant as compared to the unvaccinated group unless otherwise indicated.

Article Snippet: LB is a shareholder of Bioceros that holds a patent on anti-human OX40 mAbs.

Techniques: Infection, Virus, Derivative Assay

Journal: Frontiers in Immunology

Article Title: Enforced OX40 Stimulation Empowers Booster Vaccines to Induce Effective CD4 + and CD8 + T Cell Responses against Mouse Cytomegalovirus Infection

doi: 10.3389/fimmu.2017.00144

Figure Lengend Snippet: Combination of MHC class I and II epitope-containing synthetic long peptide (SLP) vaccines and OX40 agonistic mAb during booster vaccination leads to robust induction of both CD4 + and CD8 + T cell responses . C57BL/6 mice were vaccinated s.c. with a combination of all MHC class I epitope-containing SLPs (CD8 SLP) or with a combination of all MHC class I and II epitope-containing SLPs (CD4 + CD8 SLP). In both groups agonistic OX40 mAb was administered i.p. during booster vaccination. (A) Kinetics of the antigen-specific CD8 + T cells measured by MHC class I tetramer staining in blood. Data shown are mean values ± SEM ( n = 18 mice/group) (B) Total MHC class I tetramer-specific CD8 + T cells induced by combinatorial MHC class I or MHC class I plus MHC class II SLP vaccination compared to MCMV infection at day 8 post-booster vaccination in spleen. Data represent mean values ( n = 5 mice/group). (C) The cytokine production capacity of the splenic SLP vaccine-induced CD8 + T cells was examined by intracellular cytokine staining at day 8 after booster vaccination. Pie charts depict the percentages of the single (IFN-γ), double (IFN-γ/TNF), and triple (IFN-γ/TNF/IL-2) cytokine producers of each antigen-specific CD8 + T cell populations at day 8 post-booster vaccination. (D) Phenotypic analysis of the combinatorial SLP vaccine-induced CD8 + T cells in spleen at day 8 post-booster vaccination. Data represent mean values ( n = 5 mice/group) and are representative of three independent experiments.

Article Snippet: LB is a shareholder of Bioceros that holds a patent on anti-human OX40 mAbs.

Techniques: Vaccines, Staining, Infection

Journal: Frontiers in Immunology

Article Title: Enforced OX40 Stimulation Empowers Booster Vaccines to Induce Effective CD4 + and CD8 + T Cell Responses against Mouse Cytomegalovirus Infection

doi: 10.3389/fimmu.2017.00144

Figure Lengend Snippet: Prime-boost vaccination with a combination of MHC class I and II epitope-containing synthetic long peptides (SLPs) exhibits increased potency to protect against lytic mouse CMV (MCMV) infection . Unvaccinated (naive), MCMV (live-virus vaccine), combined MHC class II SLPs (CD4 mixture), combined MHC class I SLPs (CD8 mixture), combined MHC class I and II SLPs (CD4 + CD8 mixture), and anti-OX40 mAb only treated C57BL/6 mice were challenged 60 days post-booster vaccination/infection with 5 × 10 4 PFU salivary gland-derived MCMV Smith. At day 5 post challenge, liver, lungs, and spleen were harvested and the viral genome copies were quantified by qPCR. To further evaluate the efficacy of the vaccines, the viral titers of chronically infected (MCMV chronic/day 60) mice were also measured. The viral titers of individual mice are depicted ( n = 4–8 per group). Statistical significance between the unvaccinated group and the rest of the groups is indicated with asterisks above each group. A statistical comparison between the CD4 + CD8 mixture group and the MCMV (live-virus vaccine) and MCMV chronic group was also performed (vertical statistical bar). For both comparisons the statistical difference was found the same (** P < 0.01). The detection limit was below 1,000 genome copies as measured in naive mice. Experiment was performed twice with similar outcome. Statistical difference is indicated ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant as compared to the unvaccinated group unless otherwise indicated.

Article Snippet: LB is a shareholder of Bioceros that holds a patent on anti-human OX40 mAbs.

Techniques: Infection, Virus, Derivative Assay, Vaccines, Comparison