Preparation method and application of tumor tissue complete antigen

Abstract

The invention discloses a preparation method and application of a tumor tissue complete antigen. The method comprises the following steps: (1) placing the lobulated fresh bioptic tumor tissue into water steam to inactivate the protease in the cytoplasm for 10-30min to obtain the tumor tissue which inactivates the protease in the cytoplasm; (2) homogenizing the tumor tissue obtained in the step (1) to obtain chyliform liquid; and (3) mixing the chyliform liquid and normal saline to obtain the tumor tissue complete antigen. The invention also discloses a method for obtaining tumor vaccines by using the tumor tissue complete antigen to pulse the dendritic cell.

Description

technical field [0001] The invention relates to the field of tumor immunotherapy, in particular to a preparation method and use of tumor tissue whole antigens. Background technique [0002] Dendritic cells (DC) are antigen-presenting cells with the most powerful function of activating tumor-specific CTL in vivo. At present, a sufficient amount of patient's own DC can be obtained by in vitro isolation or culture, and can be prepared into a tumor vaccine after being loaded with tumor antigens in vitro, and can induce strong anti-tumor immune function in vivo after reinfusion into the patient. [0003] DC vaccine was first used in the clinical treatment of malignant lymphoma. Hue et al. reported for the first time that DC vaccine was used to treat 4 patients with follicular B-cell lymphoma, and achieved good curative effect; in 2002, Timmerman et al. reported that DC was used in clinical treatment of 35 patients with B-cell lymphoma, and the results also achieved certain curat...

AI Technical Summary

Problems solved by technology

[0018] Vaccines prepared by shocking DC with tumor freeze-thawed antigens can produce better anti-tumor effects. The advantage is that there is no need to identify tumor-specific antigens or purify specific antigenic proteins. It is easy to prepare and can be applied to most patients. The disadvantage is that freeze-thawed antigens Using tumor tissue or cell freeze-thawed supernatant, most of the membrane antigens cannot be broken down and removed, so the freeze-thawed antigens only contain intracytoplasmic antigens, and the antigenicity is reduced

If all freeze-thaw products are used, the freeze-thaw products are not uniform, most of the membrane antigens are too large to be processed by endocytosis, and the prepared DC vaccine contains too much cell debris and cannot be used

[0019] The method of freezing and thawing antigens cannot inactivate proteases, and proteases degrade tumor antigens in large quantities when they are stored and added to DC impact culture; at the same time, they cannot effectively break up cell membranes, resulting in too large fragments to be effectively taken up by DCs. Usually, centrifugation and filtration methods are used in preparation Discarded, resulting in a large loss of tumor antigens

Ultrasound has poor ability to disrupt tissue, and generally can only be used to disrupt cells, and it cannot inactivate proteases

Tissues can be lysed with reagents such as detergents, but they cause protein denaturation, making it difficult to refold and affect subsequent use (detergents can also affect DC activity)

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