theralizumab (TAB08) / TheraMab - LARVOL DELTA

Investigation of immunotherapy for cutaneous squamous cell carcinoma using a tissue slice culture system (ESDR 2024) - "Conversely, ciclosporin suppressed immune responses in these tissue slices, reducing T cell proliferation, and granzyme B and CD25 expression (48.2%, 12.4% and 58.9% reduction respectively, p < 0.01). Using this system, parallel screening of multiple potential immunotherapies identified that TGN1412, an anti-CD28 agonist, can enhance cSCC T cell function, both in isolation and in combination with PD-L1 blockade...We have demonstrated that the immune responses in the slices can be manipulated with antibody and small-molecule therapies. Furthermore, this platform offers an opportunity to test and compare novel immunotherapies alone and in combination to identify new potential immunotherapeutic strategies for cSCC."

IO biomarker • Genetic Disorders • Non-melanoma Skin Cancer • Oncology • Skin Cancer • Squamous Cell Carcinoma • Squamous Cell Skin Cancer • CD69 • GZMB • IFNG • IL2RA

Conditionally active CD28xVISTA bispecific antibodies induce myeloid-driven tumor-specific T-cell co-stimulation for improved cancer immunotherapy (AACR 2024) - "Finally, cytokine release from human PBMCs, co-cultured with human umbilical vein endothelial cells (HUVECs) and treated with the pH-selective BS2 (or TGN1412 as a positive control), was measured using a bead-based multiplex immune assay. Our data show that the combination of anti-mPD-1 and our prototype CD28xVISTA bsAb (BS2) with pH-selective VISTA binding significantly inhibited tumor growth in vivo... Our study demonstrates the feasibility of cis and trans CD28 co-stimulation using a CD28xVISTA bsAb (BS2). This approach, which bypasses the need for tumor-associated antigens (TAAs) required by other CD28xTAA bispecifics in development, suggests a potentially safer alternative for T-cell engagement and stimulation. Moreover, our novel myeloid-directed TME-selective co-stimulation strategy with a CD28xVISTA bsAb may broaden the potential for T-cell engagement approaches in solid tumors by enabling rational combinations with existing CD3xTAA T-cell engagers without..."

IO biomarker • Genito-urinary Cancer • Oncology • Prostate Cancer • Solid Tumor • CD4 • CD8 • IL2 • IL2RA • VSIR

CD28 humanized mouse model for efficacy and safety assessment of CD28-targeting therapies (AACR 2024) - "Furthermore, TGN1412 administration in CD28 mice induces cytokine release, and a slightly body weight loss, but no major change in body temperature...A non-conserved amino acid variant regulates differential signalling between human and mouse CD28. Nat Commun 2018; 9:1-16"

IO biomarker • Preclinical • Colorectal Cancer • Gastrointestinal Cancer • Oncology • Solid Tumor • CD28

Discovery of agonist CD28-specific single domain antibodies from alpaca (AACR 2024) - "Development of superagonistic CD28-targeting antibody has been paused after TeGenero's TGN1412 Phase 1 trial in 2006 due to life-threatening cytokine release syndrome...All purified VHH-Fc showed sub-nanomolar affinity by ELISA and bind to primary T cells by flow cytometry. Three VHH-Fc induced IL-2 production in a primary T cells activation assay only when cross-linked and in the presence of CD3 co-stimulation and are suitable for further bispecific T cell engager development."

Oncology • CD28 • CD3E • CD80 • CD86 • IL2

CD19-CD28: An affinity-optimized CD28 agonist for combination with glofitamab (CD20-TCB) as off-the-shelf immunotherapy. (PubMed, Blood) - "We developed a bispecific CD19-targeted CD28 agonist (RG6333, CD19-CD28) to enhance the efficacy of glofitamab and similar TCBs by delivering signal 2 to tumor-infiltrating T cells. CD19-CD28 distinguishes itself from the superagonistic antibody TGN1412, as its activity requires the simultaneous presence of a TCR signal and CD19 target binding...Our findings highlight CD19-CD28 as a safe and highly efficacious off-the-shelf combination partner for glofitamab, similar TCBs, and other costimulatory agonists. CD19-CD28 is currently in a Phase 1 clinical trial in combination with glofitamab."

IO biomarker • Journal • Hematological Malignancies • Lymphoma • Oncology • CD20

CAR T CELLS TARGETING CD28 SHOW PRECLINICAL ACTIVITY AGAINST T-ALl (EBMT 2024) - " Next, we designed 20 CAR molecules against CD28 based on five publicly available scFvs and identified two lead molecules by in vitro testing (FIG_1I&J) termed 28CAR_2 (scFv: TGN1412) and 28CAR_14 (scFv: CD28.3)... Taken together, our data highlight CD28 CAR T-cells as a novel synthetic immunotherapy against T-ALL and beyond."

CAR T-Cell Therapy • IO biomarker • Preclinical • Acute Lymphocytic Leukemia • Hematological Malignancies • Leukemia • Multiple Myeloma • Oncology • Pediatrics • T Acute Lymphoblastic Leukemia • CD28 • CD7 • CD80 • CD86 • LAG3

CAR T CELLS TARGETING CD28 SHOW PRECLINICAL ACTIVITY AGAINST T-ALl (EBMT 2024) - " Next, we designed 20 CAR molecules against CD28 based on five publicly available scFvs and identified two lead molecules by in vitro testing (FIG_1I&J) termed 28CAR_2 (scFv: TGN1412) and 28CAR_14 (scFv: CD28.3)... Taken together, our data highlight CD28 CAR T-cells as a novel synthetic immunotherapy against T-ALL and beyond."

CAR T-Cell Therapy • IO biomarker • Preclinical • Acute Lymphocytic Leukemia • Hematological Malignancies • Leukemia • Multiple Myeloma • Oncology • Pediatrics • T Acute Lymphoblastic Leukemia • CD28 • CD7 • CD80 • CD86 • LAG3

CD28 CAR T Cells for T-ALL (EBMT-EHA 2024) - " Next, we designed 20 CAR molecules against CD28 based on five publicly available scFvs and identified two lead molecules by in vitro testing (FIG_1I-J) termed 28CAR_2 (scFv: TGN1412) and 28CAR_14 (scFv: CD28.3)... Taken together, our data highlight CD28 CAR T-cells as a novel synthetic immunotherapy against T-ALL and beyond."

CAR T-Cell Therapy • IO biomarker • Acute Lymphocytic Leukemia • Hematological Malignancies • Leukemia • Multiple Myeloma • Oncology • Pediatrics • T Acute Lymphoblastic Leukemia • CD28 • CD7 • CD80 • CD86 • LAG3

Using Reference Reagents to Confirm Robustness of Cytokine Release Assays for the Prediction of Monoclonal Antibody Safety. (PubMed, J Vis Exp) - "However, different labs using different control antibodies can threaten the harmonization of CRAs, and clinically relevant controls (such as TGN1412) can be difficult to source, which can lead to less accurate or reliable results or data which are difficult to compare between laboratories...Each antibody is provided with an isotype-matched negative control antibody. This panel of reference reagents has previously been shown to have good inter-lab reproducibility and are suitable controls to increase the confidence and robustness of safety data from a variety of CRA platforms."

Journal • Inflammation

Generation of a novel fully human non-superagonistic anti-CD28 antibody with efficient and safe T-cell co-stimulation properties. (PubMed, MAbs) - "However, the development of CD28-targeting therapies ceased after TeGenero's Phase 1 trial in 2006 evaluating a superagonistic anti-CD28 antibody (TGN1412) resulted in severe life-threatening side effects...In an in vitro activity assay using human PBMCs, the combination of E1P2 with CD3 bispecific antibodies enhanced tumor cell killing and T-cell proliferation. Collectively, these data demonstrate the therapeutic potential of E1P2 to improve the activity of T-cell receptor/CD3 activating constructs in targeted immunotherapeutic approaches against cancer or infectious diseases."

Journal • Hematological Disorders • Hematological Malignancies • Immune Modulation • Infectious Disease • Oncology

Mutation of the TGN1412 anti-CD28 Monoclonal Antibody Lower Hinge Confers Specific FcγRIIb Binding and Retention of Super-Agonist Activity. (PubMed, Immunol Cell Biol) - "In addition to having FcγRIIb specificity, these engineered TGN1412 antibodies also retained their super-agonistic ability, demonstrating that CD28 and FcγRIIb-specific binding are together sufficient for agonistic function. The FcγRIIb-specific nature of IgG4-L E has utility for mAb-mediated immune agonism therapies that are dependent on FcγRIIb interaction and of anti-inflammatory mAbs in allergy and autoimmunity that harness FcγRIIb inhibitory signalling."

Journal • Allergy • Immune Modulation • Immunology • Inflammation

RG6333 (CD19-CD28), a CD19-Targeted Affinity-Optimized CD28 Bispecific Antibody, Enhances and Prolongs the Anti-Tumor Activity of Glofitamab (CD20-TCB) in Preclinical Models (ASH 2022) - P1 | "In contrast, TGN1412, a CD28 superagonist antibody, induced strong T cell activation, proliferation and cytokine secretion in vitro and in huNSG. Scheduling studies with glofitamab and RG6333 in huNSG suggest a safe and potent treatment regimen by using Gazyva pre-treatment followed by a staggered infusion of glofitmab and RG6333 applying an interval of three days at the first treatment cycle...Interestingly, the alternation of RG6333 with an alternative 4-1BB costimulatory agent (RG6076; CD19-4-1BBL) completely prevented tumor relapse during glofitamab treatment for more than 120 days when RG6333 was given for the first treatment cycles followed by RG6076 at later cycles...Optimal scheduling including alternation of costimulatory bispecific antibodies suggest a powerful off-the-shelf T cell redirection approach as an alternative to CAR-T cell therapies. RG6333 is currently in a phase I, open-label, dose-escalation study in combination with glofitamab (NCT05219513)."

Preclinical • Diffuse Large B Cell Lymphoma • Hematological Malignancies • Inflammation • Lymphoma • Non-Hodgkin’s Lymphoma • Oncology • CD20 • CD4 • CD8

Novel GPC3xCD3 (NILK-2501) and GPC3xCD28 (NILK-3801) κλ Bispecific Antibodies for Next Generation Immunotherapy of GPC3-Expressing Solid Cancers (APASL 2023) - "The superagonistic monoclonal anti-human CD28 antibody (IgG4κ) TGN1412 was used as comparator. Activity is maintained while it allows well tailorable dose response with reduced cytokine release. Compounds are currently in extensive pre-clinical assessment 999"

IO biomarker • Gastrointestinal Cancer • Hepatocellular Cancer • Hepatology • Liver Cancer • Oncology • Solid Tumor • CD4 • CD8 • GPC3 • IL6 • TNFA

A Bifunctional Tumor Activated Immunomodulator (TRACIr) Targeting PD-L1 And CD28 Is a Potent Enhancer of T Cell-Mediated Anti-Tumor Activity (SITC 2022) - "However, the first phase 1 clinical trial of the CD28 agonistic antibody TGN1412 failed due to an unexpected and rapid systemic proinflammatory cytokine response...PD-L1 blocking activity was comparable with atezolizumab, avelumab, nivolumab, and pembrolizumab...Finally, TRACIr was well tolerated in NHPs at high doses and exhibited half-life extended pharmacokinetics. Conclusions Preclinical activity and safety profiles of PDL1xCD28 TRACIr support its further development as an attractive bifunctional T cell modulator."

Breast Cancer • Lung Cancer • Non Small Cell Lung Cancer • Oncology • Renal Cell Carcinoma • Solid Tumor • Triple Negative Breast Cancer

A novel fully human CD28 antibody that cross-reacts with CTLA-4 and mouse CD28 for potential applications in cancer immunotherapy (SITC 2022) - "Conclusions VE19ZH is a promising module for cancer immunotherapy with unique properties: (i) Fully human mAb for minimal immunogenicity (ii) Potent co-stimulator for full T cell activation (iii) Conventional agonist of CD28 and not super-agonistic like TGN1412 (iv) cross reacts with mouse CD28 for better assessment in immunocompetent mouse models (v) Binds to human CTLA-4 for potential checkpoint inhibition. The potential of VE19ZH to boost T cell response via CD28 activation and CTLA-4 blockade is currently being investigated in vitro and in vivo."

Preclinical • Oncology • CD80 • CD86 • CTLA4 • IL2

NOVEL MULTIFUNCTIONAL TETRAVALENT CD38 NKP46 FLEX-NK ENGAGERS ACTIVELY TARGET AND KILL MULTIPLE MYELOMA CELLS (EHA 2022) - "Cytokine release assessment of CYT-338 in the human PBMC assay showed no evidence of cytokine release while high levels of cytokine release was observed with daratumumab, anti-CD28 (TGN1412) and CD3 antibody controls. Conclusion These results suggest that the CYT-338 engager has a favorable NK cell engager profile for targeting CD38 expressing multiple myeloma distinct from daratumumab."

Hematological Malignancies • Multiple Myeloma • Oncology

A humanized mouse model to evaluate the systemic adverse effects of cytokine release syndrome induced by immunoactivating drugs (CIMT 2022) - "The lack of predictive preclinical models to evaluate antibodymediated adverse effects can lead to clinical failures, such as TGN1412, a humanized anti-CD28 monoclonal antibody that caused life-threatening cytokine release syndrome (CRS) during the firstin-man trial...We have demonstrated that the novel mouse model using PBMC-humanized NSG™ or derivative strains can successfully detect CRS induced by immunoactivating antibodies in a dose- and timedependent manner. Our data support that the PBMC humanized mouse model for CRS assessment is a valuable tool to assess in vivo safety of experimental human immunoregulatory therapies, including immune checkpoint inhibitors, CAR T, and bispecific antibodies."

Adverse events • Cytokine release syndrome • Preclinical • Immune Modulation • Inflammation • Oncology • CASP3 • TNFA

Tumor-targeted CD28 bispecific POWERbodyTM for safe and synergistic T cell-mediated immunotherapy (AACR 2022) - "Targeting CD28 for systemic T cell activation caused severe cytokine storm and multiorgan failure in an anti-CD28 antibody TGN1412 phase 1 trial...When combined with TAA×CD3 TCEs and/or checkpoint inhibitors, they show strong synergistic T cell-mediated antitumor responses with robust safety profiles. Taken together, our tumor specific CD28 bispecific POWERbody TCEs exhibit enormous potential to fulfill the promises of safe and durable T cell-mediated immunotherapies when combined with CD3 bispecific POWERbody TCEs and/or checkpoint inhibitors."

Oncology • CD276 • HER-2 • TROP2

A novel, rapid, sensitive, and reproducible in vivo platform to assess efficacy and toxicity of bispecific antibodies (AACR 2022) - "The platform was validated using an anti-CD28 superagonist (TGN1412), which none of the pre-clinical in vitro assays and in vivo studies, including non-human primates, executed before clinical trials were able to predict the observed clinical toxicities...This model will also help to optimize the therapeutic drug dose range to improve the safety profile of BiTE and other immune-oncology drugs. We believe this platform will greatly benefit not only the scientific community but potentially cancer patients as well."

Preclinical • Hematological Malignancies • Lymphoma • Non-Hodgkin’s Lymphoma • Oncology

"The CD28 superagonist antibody TGN1412 disaster is a prime example of this. Worked fine in mice but nearly killed 6 human volunteers at 1/500th equivalent dose." (@BFraggles)

Dose Escalation Study of TAB08 in Patients With Advanced Solid Neoplasms (clinicaltrials.gov) - P1b; N=12; Completed; Sponsor: Theramab LLC; Recruiting ➔ Completed; N=38 ➔ 12

Clinical • Enrollment change • Trial completion • Oncology • Solid Tumor

Regulatory and strategic considerations for addressing immunogenicity and related responses in biopharmaceutical development programs. (PubMed, J Clin Transl Sci) - "Anti-drug antibodies can impact the safety and efficacy of drug products, and related immune responses, like the cytokine release syndrome that occurred in the infamous TGN-1412 clinical trial, can be challenging to predict with nonclinical models...This process begins at the discovery stage with the application of "quality by design," continues into the clinic with the development of quality assays and management strategies, and culminates in the effective presentation of this information in regulatory documents. This review provides an overview of some of the key strategic and regulatory considerations for biotherapeutics as they pertain to immunogenicity and related responses."

Journal • Review • Gene Therapies • Inflammation

Myelopoiesis of acute inflammation: lessons from TGN1412-induced cytokine storm. (PubMed, Cancer Immunol Immunother) - "Granulocytic dysplasia continued for 20 days in association with increased expression of CD69 and IL-4, but reduced IL-10; with resolution, this profile reversed to higher IL-10 expression and counter-balanced circannual cycling of IL-4 and IL-10 thereafter over 7 months. Along with immune cell subset and cytokine correlates monitored over 2 years, these observations offer unique insights into the expected changes in myelopoiesis and natural resolution in otherwise healthy young individuals in response to acute inflammation and cytokine storm in the absence of concomitant infection or comorbidity."

Cytokine storm • Journal • Hematological Disorders • Immunology • Inflammation • IL10 • IL4

Immune reconstitution and clinical recovery following anti-CD28 antibody (TGN1412)-induced cytokine storm. (PubMed, Cancer Immunol Immunother) - "Some also had signs or symptoms of psychological, mucosal or immune dysregulation. These observations may discern immunopathology, treatment targets, and long-term monitoring strategies for other patients undergoing immunotherapy or with cytokine storm."

Clinical • Cytokine storm • Journal • CNS Disorders • Cognitive Disorders • Depression • Dermatology • Gastrointestinal Disorder • Hematological Disorders • Infectious Disease • Neutropenia • Novel Coronavirus Disease • Oncology • Pruritus • Psychiatry • CD4 • CD8

Functional differences and similarities in activated peripheral blood mononuclear cells by lipopolysaccharide or phytohemagglutinin stimulation between human and cynomolgus monkeys. (PubMed, Ann Transl Med) - "However, the failures of preclinical studies to predict a life-threatening "cytokine storm", which, for instance, rapidly occurred in six healthy volunteers with the CD28 superagonist monoclonal antibody (mAb) TGN1412 in the first-in-human phase I clinical trial, have emphasized a need to clarify the differences between human and monkey immune systems...Moreover, there were some differences in antigen processing and presentation, and the interferon gamma (INF-γ)-mediated signaling pathway in these species detected by gene expression profile study. In conclusion, this is the first study to compare data on the responses of PBMCs to PHA and LPS in humans versus cynomolgus monkeys, and these findings may provide crucial insights into translating non-human primate (NHP) studies into human trials."

Journal • Inflammation • CEP55 • IFNG • TOP2A • TYMS