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Chimeric Antigen T cell Therapy CART

CAR T cell Therapy: Over the course of time immunological research has been improved by the study of different receptor. The receptors ar...

CAR T cell Therapy:
Over the course of time immunological research has been improved by the study of different receptor. The receptors are studied on the basis of their structure and their interaction with different other antigenic components like the anti-immune components of different cancerous cells and some viruses and microbes. These studies provided lot of help in the treatment of different immunological diseases. One of the major approaches is CAR T Cell Therapy which uses specific receptors for treatment of cancerous cells. These receptors are modified in vitro and then utilized for treatment. This is process is a categorized part of immunotherapy.

What is CAR?

CAR refers to “Chimeric Antigen Receptor” which is a protein made in laboratory used to attach at T cell surface so that it act as receptor for antigens present on cancerous cells. These CARs consist of Extracellular Chains which single chained and are variable domains consisting of trans-membrane regions. The intracellular region is also containing tyrosine based activation motifs which functions for activation of its effector cells against antigens that it recognizes. The single chain variable region functions for attachment to tumor antigens present on tumor cell surfaces. The intracellular domain function for activation of T cells, its proliferation.
Figure 1. Structure of CAR (Source: Wikipedia )
The Car structure is flexible in a sense that its single chain variable fragment region can recognize a lot of different forms of antigens and is independent of major histocompatibility complex (on which the receptors of T cells are dependent).The CARs can recognize antigens with different types of functional groups such as proteins, carbohydrates and gangliosides.

How are CARs generated?

The chimeric antigen receptors are generated in laboratory by using the clones of specific gene responsible for single chain variable fragment specific for carcino-embryonic antigen of a human carcino cell. The domain of gene was cloned by its insertion into Jurkat cells which are immortalized lines human T cells, by using pBi vectors (bidirectional vectors). The cDNA regions that are cloned were obtained from hybridoma and modified by PCR processing to remove the specified region coding for anti carcino-embryonic antigens.
Different screening techniques were used to confirm the insertion in the clones and for T cell activation confirmation, different T cell check assays were used like cytokine production assay.

How They Work?

When the CAR T cells recognize the tumor antigens and bind to it, it causes the activation of T cells by phosphorylation of the tyrosine based intracellular motifs. This results in the secretions of different chemical signals like cytokines which is followed by cytotoxic T cell activation and proliferation to maintain a balanced attack against the specific tumor cells.
The CAR T cells after antigen binding also causes the secretion of granzyme granules and perforins and also interferon gamma IFN-γ. These also activate the cell death receptor indulging apoptosis by using Fas-L and TNF-R. Thus this action of multi-immune agents causes the destruction of tumor cells.
The actions of CAR T cells are thought to be improved by using multiple signaling receptors to activate the multiple components of immune system at a time to efficiently destroy the tumor cells. TRUCK: T cell redirected for universal cytokine killing, is a mechanism which was introduced to redirect CAR-T cells so that it becomes able to release protein signals like IL-20 (Interleukin 20) and others to activate innate immune system against tumor antigen alongside adaptive immune activity to potentiate the process by co-activation of other immune components like NK cells.

How the therapy is performed?

During this therapy the T cells of patient are taken from patients through apheresis and then these T cells are modified by biotechnological modification using techniques of PCR amplification and the product it then cloned and multiplied. After genetic modification and cloning CAR T cells are produced which are then injected back to the patient. Before the injection of CAR T cells the patients are pre-tested by a short course of chemotherapy to check the effectiveness of the patient for the process. Too weak immune patients are first aided to get rid of some level of weakness and then treated.

Side Effects of CAR T cell Therapy:

The CAR T cell Therapy can also have some side effects and it require proper handling as some patients have been died at its pre stages of treatments.

1.      Cytokine Release Syndrome CRS:
This is the excessive release of cytokines into the blood stream of the patients. This causes due to excessive activation and proliferation of CAR T cells soon after the injection of CAR-Ts into the patient blood. This can result into lot of health problems like high fever, nausea, muscle aches etc. Different drugs are used along side CAR infusion into patient to prevent the CRS.

2.      Effects on Nervous system:

The improper infusion can also result in the causing of defects in the brain the nervous system, which if not handled properly can be severely dangerous. Researchers are working to still find the main reason of these effects of CAR T cell therapy.
3.      B cell loss, B cell Aplasia:
This includes the destruction of normal B cells by CAR t cells during its action of cancerous cells. This can then causes other immune disabilities leading to slowness and even stopping of certain immune functions against antigens.

4.      Tumor Lysis Syndrome TLS:

This is due to toxic effects caused by breakdown of the dying cells of tumors which are destructed by immune components. This mainly is due to metabolic mistakes durig action of CAR T cells on the cancerous cells and usually starts after certain weeks of treatments. To handle this certain medications and drugs are used after the therapy. 

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