Application of memory lymphocyte population in liver cancer treatment

A technology of lymphocytes and memory, applied in the direction of animal cells, vertebrate cells, cell culture active agents, etc., can solve the problems of short survival time of T cells in vivo, lack of universality, and low success rate of cure

Active Publication Date: 2019-04-16
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although TIL, TCR-T and CAR-T have certain tumor therapeutic effects, their successful cure rate is still not high, and they are not universal. survival time in vivo

Method used

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  • Application of memory lymphocyte population in liver cancer treatment
  • Application of memory lymphocyte population in liver cancer treatment
  • Application of memory lymphocyte population in liver cancer treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] In this example, memory lymphocytes were obtained according to the following method:

[0107] 1. Medium

[0108] GT-T551 medium, 5×10 5 U / L Interleukin-2, 5ng / mL Interleukin-7, 5ng / mL Interleukin-15, 3μg / ml Anti-CD3 antibody, 5% by volume autologous plasma, pH 7.2 ~7.4.

[0109] The culture flask needs to be coated with Anti-CD3 antibody in advance, and the volume of the coating solution is 5ml / 75cm 2 Culture flasks need to be placed overnight at 4°C.

[0110] Table 1 Raw material list

[0111] Reagent name

brand

GT-T551

Takara

interleukin-2

Jiangsu Kingsley

interleukin-7

R&D

Interleukin-15

R&D

Anti-CD3 antibody

Baipsies

[0112] 2. Training method

[0113] (1) Use heparin as an anticoagulant to extract fresh peripheral blood donated by liver cancer patients, centrifuge at 1600rpm / min for 5-10min, extract autologous plasma, and the lower layer is blood cells. Autologous plasma was inactivate...

Embodiment 2

[0118] Example 2 Evaluation Test of Curative Effect of Memory Lymphocytes on Transplanted Liver Cancer in Nude Mice

[0119] In this example, tumor antigens derived from SMMC-7721 cells were selected, and the differentiation, proliferation, and effector abilities of memory lymphocytes under the stimulation of DC cells loaded with tumor antigens were observed, so as to provide references for relevant mechanisms of clinical application.

[0120] 1. Materials and methods

[0121] The DC cells used in the experimental example were obtained from the adherent part of healthy human peripheral blood PBMC under the culture conditions containing GM-CSF, IL-4 and TNF-α, and were identified by flow cytometry as CD45 + CD11b + CD11c + .

[0122] The tumor antigens derived from SMMC-7721 used in the experimental examples were obtained by dissolving the SMMC-7721 cell line repeatedly frozen and thawed in liquid nitrogen in 0.1% HCLO (physiological saline as a solvent).

[0123] 2. Experi...

Embodiment 3

[0134] Example 3 Evaluation Test of Curative Effect of Memory Lymphocytes on Transplanted Liver Cancer in Nude Mice

[0135] In this example, SMMC-7721 cells were selected to establish a xenograft tumor model in nude mice, and the growth inhibitory effect of memory lymphocytes on xenograft tumors in nude mice was observed, so as to provide reference for clinical application.

[0136] 1. Materials and methods

[0137] Used test product (memory lymphocyte liquid) in the embodiment is colorless transparent liquid, and cell content / specification is 5 * 10 7 / mL, where CD3 + CD45RA - CD45RO + CD62L + The cell ratio is not less than 70%.

[0138] The experimental animals used in the examples were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., and were SPF grade BALB / c nude mice, each weighing 16-18g, 6-7 weeks old, a total of 20 females, divided into 2 groups experimenting.

[0139] The grouping method is as follows: the animals were weighed on...

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Abstract

The invention provides a memory lymphocyte population, a culture medium,a method for obtaining the memory lymphocyte population and application. The memory lymphocyte population contains at least oneof the following marker molecules of leukocyte differentiation antigens CD3, CD4, CD8, CD16, CD56, CD62L and CD45RO. The memory lymphocyte population can rapidly activate, proliferate and differentiate under specific stimulation of the hepatocellular carcinoma antigens, and IFN-gamma can be secreted, and then the immune response of the body is assisted to be strengthened; IFN-gammacan be retainedin the body, thus a better tumor killing effect is achieved;in addition, the risk of metastasis and recurrencecan be effectively reduced, and the memory lymphocyte population is especially applicableto hepatocellular carcinoma with microvascular invasion.

Description

technical field [0001] The present invention relates to the field of biology. Specifically, the present invention relates to the application of memory lymphocyte population in the treatment of liver cancer. More specifically, the present invention relates to a memory lymphocyte population, a culture medium, and a method for obtaining and using the memory lymphocyte population. Background technique [0002] Cancer is a kind of disease in which cells proliferate malignantly and invade normal tissues of the body. Its incidence rate is high, and more than 14.6% of the death cases are caused by cancer every year. The battle against cancer has never stopped, and new progress has been made, but there is still no perfect cure for cancer. [0003] The most common cancer treatment method is surgical resection, followed by radiation therapy and chemotherapy, but all three have certain defects, such as the inability to remove all cancer cells, resulting in the failure of residual canc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783A61K35/17A61P35/00
CPCA61K35/17A61P35/00C12N5/0636C12N2501/2302C12N2501/2307C12N2501/2315C12N2501/515C12N2506/11
Inventor 陈立功孙忠杰赵宏蔡建强马骏凡郭潇齐海龙薛庆磊张叔人
Owner TSINGHUA UNIV
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