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Anti-Nuclear Antibody Screening Using HEp-2 Cells
Published on: June 23, 2014
Generation and characterization of novel anti-DR4 and anti-DR5 antibodies developed by genetic immunization
Agathe Dubuisson1,2,3, Cécile Favreau1,3, Eric Fourmaux2
1Université Bourgogne Franche-Comté, INSERM, LNC UMR1231, F-21079, Dijon, France.
Abstract:
Development of therapeutic antibodies in oncology has attracted much interest in the past decades. More than 30 of them have been approved and are being used to treat patients suffering from cancer. Despite encouraging results, and albeit most clinical trials aiming at evaluating monoclonal antibodies directed against TRAIL agonist receptors have been discontinued, DR4 or DR5 remain interesting targets, since these receptors are overexpressed by tumour cells and are able to trigger their death. In an effort to develop novel and specific anti-DR4 and anti-DR5 antibodies with improved properties, we used genetic immunization to express native proteins in vivo. Injection of DR4 and DR5 cDNA into the tail veins of mice elicited significant humoral anti-DR4 and anti-DR5 responses and fusions of the corresponding spleens resulted in numerous hybridomas secreting antibodies that could specifically recognize DR4 or DR5 in their native forms. All antibodies bound specifically to their targets with a very high affinity, from picomolar to nanomolar range. Among the 21 anti-DR4 and anti-DR5 monoclonal antibodies that we have produced and purified, two displayed proapoptotic properties alone, five induced apoptosis after cross-linking, four were found to potentiate TRAIL-induced apoptosis and three displayed antiapoptotic potential. The most potent anti-DR4 antibody, C#16, was assessed in vivo and was found, alone, to inhibit tumour growth in animal models. This is the first demonstration that DNA-based immunization method can be used to generate novel monoclonal antibodies targeting receptors of the TNF superfamily that may constitute new therapeutic agents.
Insights
Researchers developed novel monoclonal antibodies targeting DR4 and DR5 receptors using DNA-based immunization. One antibody, C#16, demonstrated significant tumor growth inhibition in vivo, offering a promising new cancer therapy approach.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
Background:
- Therapeutic antibodies are crucial in oncology, with over 30 approved for cancer treatment.
- Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) agonist receptors DR4 and DR5 are overexpressed in tumors and can trigger cancer cell death.
- Despite challenges with previous TRAIL-targeting antibodies, DR4 and DR5 remain promising therapeutic targets.
Purpose of the Study:
- To develop novel, specific monoclonal antibodies targeting DR4 and DR5 with enhanced properties.
- To utilize DNA-based immunization for generating antibodies against native DR4 and DR5 proteins.
- To evaluate the therapeutic potential of these newly developed antibodies in preclinical models.
Main Methods:
- Genetic immunization by injecting DR4 and DR5 cDNA into mice tail veins.
- Generation and screening of hybridomas to identify antibodies specific for native DR4 and DR5.
- Affinity and functional characterization of anti-DR4 and anti-DR5 monoclonal antibodies, including apoptosis induction and TRAIL potentiation assays.
- In vivo assessment of the most potent anti-DR4 antibody (C#16) for tumor growth inhibition.
Main Results:
- Successful generation of 21 anti-DR4 and anti-DR5 monoclonal antibodies with high affinity (picomolar to nanomolar range).
- Antibodies exhibited diverse functional properties: 2 induced apoptosis directly, 5 after cross-linking, 4 potentiated TRAIL-induced apoptosis, and 3 showed anti-apoptotic potential.
- The lead anti-DR4 antibody, C#16, demonstrated significant tumor growth inhibition as a monotherapy in vivo.
Conclusions:
- DNA-based immunization is an effective method for generating therapeutic antibodies against TNF superfamily receptors.
- Novel anti-DR4 and anti-DR5 monoclonal antibodies, particularly C#16, show promise as new therapeutic agents for cancer treatment.
- Targeting DR4 and DR5 with specifically engineered antibodies represents a viable strategy for enhancing anti-cancer therapies.
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