Killing cell for high-efficiency and stable expression of antibodies and use thereof

Abstract

The invention relates to a killing cell for high-efficiency and stable expression of antibodies and a use thereof. Specifically, the invention provides the transgenic killing cell; a genome of the killing cell is stably integrated with an expression cassette containing an encoding sequence of human Fc segment-containing antibodies, or containing an encoding sequence of chimeric antigen receptors and inhibitory antibodies or activated type antibodies, and both ends of the expression cassette contain inverted terminal repeated sequences of a transposon. The killing cell can stably express the human Fc segment-containing antibodies or the chimeric antigen receptors and antigen-binding fragments derived from interested antibodies with high level while maintaining cell killing toxic effect. In addition, for preventing systemic toxicity and autoimmune diseases caused by over expression of antibodies due to in-vivo continuous proliferation of immune cells for stable expression of the antibodies, a molecular braking system is also introduced. With use of monoclonal antibody agents appearing on the market, the killing cell integrated with the antibody expression cassette can be quickly eliminated, and the safety of treatment is effectively improved.

Description

technical field [0001] The invention belongs to the field of cell biology and oncology, and in particular relates to a killer cell expressing an antibody efficiently and stably and its use. Background technique [0002] Cancer has now become the number one killer of human health. The fast pace of life, huge work pressure, unhealthy eating habits, and poor environment are all accomplices to the occurrence of cancer, making the high incidence and younger trend of cancer more and more obvious. At present, the efficacy of traditional treatment methods has reached the bottleneck, and it is urgent to explore a more effective treatment method to improve the survival rate and quality of life of cancer patients. Immunotherapy against malignant tumors has developed rapidly in recent years and has achieved remarkable clinical efficacy. In 2011, Nature and JCO, the top journal of clinical oncology, respectively published review articles with the same title "The era of tumor immunothera...

AI Technical Summary

Problems solved by technology

[0006] Although there have been reports of transfection of exogenous genes into, for example, NK cells or T cells, the commonly used gene transfection vector systems have a low transfection rate for effector immune cells with cell killing toxicity, or it is difficult to make exogenous genes in their cells express at a high level

The use of adenoviral vectors (non-integrated) can mediate the efficient short-term expression of foreign genes in NK cells or T cells, but the proliferation of activated NK cells or T cells is very fast, and the expression cassettes of foreign genes carried are in It will be lost rapidly during cell passage, and the expression is difficult to last; the integration of foreign genes in the genome of NK cells or T cells can be mediated by retrovirus or lentivirus, and stable expression can be achieved in theory, but the antibody contains light chain and heavy chain , the coding sequence is long and the molecular weight is large, which leads to great difficulties in the packaging and preparation of retroviruses or lentiviruses carrying full-length antibody expression cassettes, and it is difficult to express antibodies efficiently. It can only be used to express single-chain antibodies with simple structures (lack of Fc segment fragment, incomplete function and short half-life)

Therefore, before this, there are no reports of transgenic cells that have cytotoxicity and can stably express antibodies containing human Fc fragments at high levels

Images