In March 2016, Mark J Osborn et al published in Molecular Therapy a major article for genome editing (doi:10.1038/mt.2015.197), about knock-out of CD3 in human T-cells. The goal is to improve T-cell-based immunotherapies to fight tumours using engineered allogenic T-cells from healthy donors. It is a very good example of how CRISPR-CAS9 can help medecine. And even if you are not very comfortable with CAR T-cells and the treatments of malignancies, I would recommend you read it and especially take a look at figure 2. Indeed, dear friends of genome editing, the authors made a clear and fair comparison of several KO strategies, covering all the main options. Thus, it is not only a major step for anti-tumour treatments but it is also an excellent overview that reveals the best approaches. So, before reading this post any further, you might like to read the article mentioned above. [Read more…]
IL-17A pathway – a key target for immunotherapies
Fig 1: Function of IL-17 in diverse processes
Interleukin-17 (IL-17) is a family of 6 closely related cytokines, designated IL-17A-F, that play a central role in mediating inflammation, autoimmunity, and host defense. IL-17 is mainly secreted by a specific subset of T helper cells known as TH17 cells (for an overview see Fig 1). The IL-17 cytokines mediate their biological functions via surface receptors on target cells. IL-17A binds to IL-17 receptor A (IL-17RA), which stimulates the production of other pro-inflammatory cytokines including IL-6 and IL-8. IL-17A and its receptor play a pathogenic role in many inflammatory and autoimmune diseases such as rheumatoid arthritis. IL-17 signaling is also involved in mucosal immunity and host defense against extracellular bacterial and fungal infections (Staph, Candida, Pneumonia, etc.). [Read more…]
Conformational antibody array for mAb process & formulation development
A recent publication by ArrayBridge, Inc., in cooperation with Wayne State University, focuses on monoclonal antibody (mAb) Higher Order Structure Analysis.
The clinical and biological properties of protein-based therapeutics, or biologics, are closely related to their Higher Order Structures (HOS), which in turn can be altered by many physical and chemical conditions. The paper describes a novel technology to monitor changes in mAb HOS – the Protein Conformational Array (PCA) ELISA – which is based on a unique bank of more than 30 well-characterized antibodies enabling the measurement of protein epitope change on the surface of the mAb. [Read more…]
Cryopreserved Leukopaks: a solution for Cell Therapy Logistics
Human cells are critical raw materials for research and manufacturing of cell therapy products. However, access to freshly procured cells can be limited, creating a crucial need for a suitable alternative to fresh cells that are viable and functional — especially when transporting materials globally.
At tebu-bio, we chose HemaCare to investigate the viability and functionality of lymphocytes, both fresh and cryopreserved, from leukopaks (leukapheresis collections) procured within their FDA registered cGMP donor collection facility. [Read more…]
Immunotherapy Screening – OX40:OX40L pathway
The treatment of diseases by inducing, enhancing, or surpressing an immune response is referred to as Immunotherapy. T-cell activation and inactivation requires the coordination of various co-inhibitory and co-stimulatory signals and most immunotherapies modulate these signals.
Therapeutic manipulation of immunopathways has lead to promising clinical results for the treatment of a number of diseases such as cancer, autoimmune diseases and inflammatory diseases. Research in this field is rapidly evolving as scientists seek to identify the next generation of therapies.
Over the past 12 months I have introduced a number of pathways and proteins involved, which represent potential targets for drug discovery campaigns and I’ve presented assays to measure inhibitor effects on these pathways (B7-1 / CD28 and B7-1 / CTLA4; PD-1/PD-L1/PD-L2; BLTA:HVEM, CD47:SIRPα; GITR:GITRL; CD40:CD40L; CD137:CD137L; IDO).
IDO and TDO – Cell based & biochemical assays
Which roles do IDO and TDO play in Immunotherapy? Immune evasion is one of the identifying hallmarks of cancer and researchers are investigating the complex mechanisms that enable cancer cells to evade the host’s immune system. In the context of a tumor, Tryptophan (Trp or L-Trp) catabolizing enzymes have been shown to assist cancer cells in immune evasion. [Read more…]
9 pathway-specific screening assays in Immunotherapy
The immune system is a system of cells and organs whose function is to defend an organism from foreign pathogens. With the ability to mount a response against virtually any foreign material and return to a quiescent state following neutralization of the threat, this fascinating organ system displays remarkable specificity and plasticity. To achieve this, there is a multifaceted balancing act between the many activators and suppressors which maintains homeostasis of the body’s perhaps most complex organ system.
Immunotherapy Screening – IDO pathway
In previous blogs, I invited you to join me in exploring the relevance of the following pathways:
Today I’d like to focus on the IDO pathway. [Read more…]
Cancer immunotherapy and PBMCs
Patients who receive cancer immunotherapy treatments in clinical trials must have their peripheral blood mononuclear cell (PBMC) samples collected at baseline and at later time points. Immune monitoring facilities provide laboratory testing of these PBMC samples in order to gauge the response of the patient’s immune system to the test treatment.
A primary focus of immune monitoring facilities is therefore to develop cutting-edge technologies whilst also standardizing and validating immune assays with rigorous quality-control standards to ensure data reliability. After all, the weight of clinical trial results depends on the accuracy, precision, and reliability of data generated from these PBMC samples.
The fragile nature of biological samples makes standardization of laboratory procedures an especially important focus. Parameters that can affect data include the anticoagulant used for preserving PBMC samples, the time frame between sampling and processing, storage/shipping temperature en route to the central processing lab, the cryopreservation and thawing process, as well as the cell culture media.[1]
Immunotherapy Screening – CD137:CD137L pathway
In previous blogs, I invited you to read about the relevance of the B7-1 : CD28, B7-1 : CTLA4, the BLTA:HVEM, CD47:SIRPα , the GITR:GITRL, the CD40:CD40L and the PD-1/PD-L1/PD-L2 pathway for immunotherapy screenings and discussed the products available to work on these pathways. Today, I will focus on the CD137:CD137L pathway.
CD137 is another co-stimulatory protein that is expressed on activated T cells. Unlike CD40:CD40L signaling, which primarily involves helper T-cells, CD137 has a crucial role in the development of cytotoxic T-cells and anti-tumor immunity. Its ligand, CD137L, is mainly expressed on antigen-presenting cells, such as activated B cells, macrophages, and dendritic cells, as well as on human tumor cells.
Co-stimulation through CD137:CD137L enhances T-cell activation, promotes the rejection of cardiac allografts and skin transplants, and eradicates experimentally induced tumors in animal models. Several clinical studies are on-going that use agonistic anti-CD137 antibodies to induce an anti-cancer response to solid tumors. [Read more…]