{"id":994,"date":"2017-08-22T03:37:41","date_gmt":"2017-08-22T08:37:41","guid":{"rendered":"http:\/\/www.creative-biolabs.com\/blog\/?p=994"},"modified":"2018-04-18T01:04:57","modified_gmt":"2018-04-18T06:04:57","slug":"the-development-of-car-gene-construction","status":"publish","type":"post","link":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/the-development-of-car-gene-construction\/","title":{"rendered":"The Development of CAR Gene Construction"},"content":{"rendered":"

CAR-T is chimeric antigen receptor T cell immunotherapy, which modifies T cell receptor by CAR and transforms it into the scFV segment in the binding domain of monoclonal antibodies. A classic CAR-T molecule generally consists of a single chain antibody (scFv) domain, cytoplasmic signaling domains and transmembrane domains. The modified CAR-T cells can specifically recognize tumor-associated antigens without restriction of MHC and optimize the targeting ability, killing activity and persistence of effector T cells.<\/p>\n

Universal CAR T cells are generally engineered to express on their surfaces receptors that recognize a specific antigen. These cells have been used up to now to recognize and kill tumor cells – for example, B cell leukemias carrying the pan-B cell marker CD19. The CAR T cells and their progeny, including memory T cells, remain in the body and continue to carry out their functions, grate provides immune surveillance in case cancer cells exceptions again. But they uniquely recognize just CD19 – a problem, in that they kill even Normal B cells, so-called off-target.<\/p>\n

In 1989, the first generation of CAR was published in the Proceedings of the American Academy of Sciences (PNAS). The basic design includes a tumor-associated antigen-binding region, an extracellular hinge region, a transmembrane region, and a TCR complex \u03b6\u00a0chain as an intracellular signal region.The first generation of CAR can only cause transient T cell proliferation and lower cytokine secretion, but can not provide long-term T cell expansion signal and sustained in vivo anti-tumor effect with the lack of co-stimulatory signal. CAR-T cells have been apoptotic before contacting with a large number of tumor cells. Julie combined CE7R single-chain antibody with CD3\u03b6 for the cell adhesion molecule-L1 and\u00a0constructed CAR vector by\u00a0DNA\u00a0of\u00a0CAR <\/u>plasmid<\/u><\/a>\u00a0with the introduction of “suicide gene” HyTK, treating recurrent neuroblastoma. Although there were no adverse effects associated with cell dose, each infusion of CAR-T cells did not survive for more than 1 week in vivo, and only one patient with the smallest tumor was partially relieved. In a word, the treatment effect was not satisfactory.<\/p>\n

As for the second-generation of\u00a0CAR gene construction<\/u><\/a>, the second signal molecules (usually CD28, CD134 or CD137 ), the necessary signal for T cell survival, were assembled into T cells to increase the CAR-T immune memory effects and cytotoxicity on tumor cells.<\/p>\n

\"Three<\/a><\/p>\n

The third-generation CAR-T structure not only includes “signal 1” and “signal 2”, but also contains additional co-stimulatory signals while CD28, CD134, CD137 molecules are assembled into CAR-T cells. Till constructed third-generation CAR-T cells (scFv CD20-CD28-CD137-CD3\u03b6) with retrovirus as a vector for the treatment of 3 patients with non-Hodgkin’s lymphoma. By PCR, the effector cells survived in vivo for more than 12 months.<\/p>\n

CAR-T cells were introduced into cytokines (such as IL-12) in the 4th<\/sup>\u00a0generation of CAR technology. When CAR-T cell receptor was used to recognize the target tumor, IL-12 was expressed at the tumor site, and all kinds of innate immune cells kill the tumor. The key point of CAR gene construction is to make a CAR T cell with a surface receptor binding to the dye fluorescein which is connected through a short linker to a molecule that binds specifically to a research expression of tumor body.<\/p>\n","protected":false},"excerpt":{"rendered":"

CAR-T is chimeric antigen receptor T cell immunotherapy, which modifies T cell receptor by CAR and transforms it into the scFV segment in the binding domain of monoclonal antibodies. A classic CAR-TRead More…<\/a><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[109],"tags":[79,81,82,52,80],"_links":{"self":[{"href":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/wp-json\/wp\/v2\/posts\/994"}],"collection":[{"href":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/wp-json\/wp\/v2\/comments?post=994"}],"version-history":[{"count":4,"href":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/wp-json\/wp\/v2\/posts\/994\/revisions"}],"predecessor-version":[{"id":1123,"href":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/wp-json\/wp\/v2\/posts\/994\/revisions\/1123"}],"wp:attachment":[{"href":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/wp-json\/wp\/v2\/media?parent=994"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/wp-json\/wp\/v2\/categories?post=994"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.creative-biolabs.com\/blog\/index.php\/wp-json\/wp\/v2\/tags?post=994"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}