阅读说明：本技术 一种til临床改善胃癌的方法 (Method for clinically improving gastric cancer through TIL ) 是由 李军 于 2020-08-21 设计创作，主要内容包括：本发明公开了一种TIL临床改善胃癌的方法,TIL临床改善胃癌的方法是把肿瘤组织中的淋巴细胞分离出来并扩增,培养出成熟的TIL细胞,然后将再TIL细胞回输注射给胃癌患者,用于改善胃癌患者的生活质量。本发明中肿瘤浸润淋巴细胞的优势为：用机械处理和酶消化方法,从肿瘤局部分离出肿瘤浸润淋巴细胞,加入高剂量IL-2体外培养及改造,然后将其回输入患者体内,相较于其它免疫疗法,肿瘤浸润淋巴细胞具有识别肿瘤抗原靶点多、副作用少、可治疗实体瘤多,疗效更强等优点。本发明TIL免疫疗法具有高效的杀伤能力,能识别多个抗原靶点,使用自身免疫细胞,无副作用,而且TIL免疫疗法能够针对复发或转移的晚期胃癌患者使用。(The invention discloses a method for clinically improving gastric cancer by TIL, which comprises the steps of separating and amplifying lymphocytes in tumor tissues, culturing mature TIL cells, and then infusing the TIL cells back to a gastric cancer patient for improving the life quality of the gastric cancer patient. The advantages of the tumor infiltrating lymphocytes in the invention are as follows: compared with other immunotherapy methods, the tumor infiltrating lymphocytes have the advantages of more tumor antigen recognition targets, less side effects, more solid tumors which can be treated, stronger curative effect and the like. The TIL immunotherapy has high killing capacity, can identify a plurality of antigen targets, uses autoimmune cells, has no side effect, and can be used for patients with recurrent or metastatic late gastric cancer.)

1. A method for clinically improving gastric cancer by TIL is characterized in that: the method for clinically improving the gastric cancer by the TIL is to separate and amplify lymphocytes in tumor tissues, culture mature TIL cells and inject the TIL cells back to the gastric cancer patient for improving the life quality of the gastric cancer patient.

2. The method of claim 1, wherein the TIL is used for clinical improvement of gastric cancer, wherein the TIL is selected from the group consisting of: the TIThe method for clinically improving the gastric cancer specifically comprises the following steps: the gastric cancer patient is operated in hospital, the tumor tissue is taken out and transported to GMP laboratory for subsequent treatment and culture, after 21 days of culture, the TIL cells are cultured to be mature, and then transported back to hospital for transfusion to the gastric cancer patient, and the TIL cell injection amount of each patient is 7X 10⁹-7×10¹⁰And (4) respectively.

Technical Field

The invention belongs to the technical field of immune cell therapy, and particularly relates to a method for clinically improving gastric cancer by TIL.

Background

The causes of gastric cancer are: 1. regional environment and dietary life factors: the incidence of gastric cancer is obviously different regionally, and the incidence rate of gastric cancer is obviously higher in northwest and east coastal areas of China than in south areas. The incidence rate of far-end gastric cancer in people who eat smoked and salted foods for a long time is high, and is related to the high content of carcinogens or precancers such as nitrite, mycotoxin and polycyclic aromatic hydrocarbon compounds in the foods; smokers have a 50% higher risk of stomach cancer than non-smokers. 2. Helicobacter pylori (Hp) infection: the Hp infection rate of adults in high-incidence areas of gastric cancer in China is more than 60%. Helicobacter pylori can promote the conversion of nitrate into nitrite and nitrosamine to cause cancer; chronic inflammation of gastric mucosa caused by Hp infection and environmental pathogenic factors accelerate excessive proliferation of mucosal epithelial cells, so that distortion and carcinogenesis are caused; toxic products CagA and VacA of helicobacter pylori may have cancer promotion effect, and the detection rate of anti-CagA antibodies in gastric cancer patients is obviously higher than that of general people. 3. Precancerous lesions: gastric diseases include gastric polyps, chronic atrophic gastritis and gastric remnants after gastric resection, all of which may be associated with various degrees of chronic inflammatory processes, metaplasia or atypical hyperplasia of the gastric mucosa and intestinal epithelium, and may turn into cancer. Precancerous lesion refers to pathological and histological change of gastric mucosa which is easy to become cancerous, and is a boundary pathological change in the process of transforming benign epithelial tissue into cancer. The dysplasia of gastric mucosa epithelium belongs to precancerous lesions, which can be divided into light, medium and heavy according to the heterotypic degree of cells, and the severe dysplasia is sometimes difficult to distinguish from early gastric cancer with good differentiation. 4. Genetic and genetic: genetic and molecular biological researches show that the incidence of gastric cancer of relatives with blood relationship of gastric cancer patients is 4 times higher than that of the relatives in a control group. The canceration of gastric cancer is a process of multi-factor, multi-step, and multi-stage development, involving changes in oncogenes, cancer suppressor genes, apoptosis-related genes, metastasis-related genes, and the like, and the forms of gene changes are also diverse.

The gastric cancer is a malignant tumor originated from gastric mucosa epithelium, the incidence rate of the gastric cancer is the first in various malignant tumors in China, the incidence rate of the gastric cancer is obviously different regionally, and the incidence rate of the gastric cancer is obviously higher in northwest and east coastal areas of China than in south areas. The good hair age is more than 50 years old, and the ratio of the incidence rates of men and women is 2: 1. gastric cancer tends to be younger due to changes in dietary structure, increased working pressure, infection with helicobacter pylori, and the like. Gastric cancer can occur in any part of the stomach, more than half of which occur in antrum, greater curvature, lesser curvature, and anterior and posterior walls. Most of gastric cancers belong to adenocarcinoma, have no obvious symptoms in the early stage, or have nonspecific symptoms such as epigastric discomfort, eructation and the like, are often similar to the symptoms of chronic stomach diseases such as gastritis, gastric ulcer and the like, and are easy to ignore, so the early diagnosis rate of the gastric cancers in China is still low at present. The prognosis of gastric cancer is related to the pathological stage, location, tissue type, biological behavior, and therapeutic measures of gastric cancer.

Gastric cancer is classified by gross morphology: 1. early gastric cancer: it refers to a gastric cancer in which cancer tissues are localized in the gastric mucosa and submucosa, and can be classified into a bulge type, a flat type and a recess type according to the morphology of naked eyes. 2. Advanced gastric cancer: it refers to the gastric cancer whose cancer tissue infiltration depth exceeds the submucosal layer, and can be classified into polyp type, localized ulcer type, infiltration ulcer type and diffuse infiltration type. Gastric cancer is classified by histopathology: can be divided into adenocarcinoma, adenosquamous carcinoma, squamous carcinoma, carcinoid, etc., and the majority is gastric adenocarcinoma. Adenocarcinoma can be classified into papillary carcinoma, tubular adenocarcinoma, poorly differentiated adenocarcinoma, mucinous adenocarcinoma, and signet ring cell carcinoma according to the tissue structure. According to the differentiation degree of cells, the cells can be divided into 3 types of high differentiation, medium differentiation and low differentiation. They can be classified into intestinal type and gastric type according to their tissue origin. Stomach cancer is classified by the diseased site: it can be classified into gastric fundus and cardia cancer, gastric antrum cancer, etc., and the gastric cancer at different sites determines the difference of the operation types.

In the early stage of gastric cancer, most patients have no obvious symptoms, and few patients have nausea, vomiting or upper gastrointestinal symptoms similar to ulcer diseases, so that sufficient attention is hardly paid. As the tumor grows, the stomach function is affected, but the symptoms are obvious, but the specificity is lacked. Pain and weight loss are the most common clinical symptoms of advanced gastric cancer. Patients often have more definite upper gastrointestinal symptoms, such as upper abdominal discomfort, fullness after eating, aggravation of upper abdominal pain along with the progress of the disease, appetite reduction and hypodynamia. It is also characterized by different tumor sites. Carcinoma of the gastric fundus of the cardia may have poststernal pain and progressive dysphagia; gastric cancer near the pylorus shows pyloric obstruction. When the tumor destroys blood vessels, there may be symptoms of digestive tract hemorrhage such as hematemesis and dark stool; if the tumor invades the pancreatic capsule, persistent pain radiating to the back of the waist can appear; perforation of tumor ulcers can cause severe pain and even signs of peritoneal irritation; jaundice can occur when hepatic lymph node metastasis occurs in the tumor or when the common bile duct is pressed; when distant lymph nodes metastasize, the swollen lymph node can be reached on the left clavicle. Patients with advanced gastric cancer often have the symptoms of anemia, emaciation, malnutrition, even cachexia and the like.

The following pathways exist for the spread and metastasis of gastric cancer: 1. direct infiltration: carcinoma of the gastric fundus of the cardia and stomach, which is apt to invade the lower part of the esophagus, may infiltrate into the duodenum. After the gastric cancer with poor infiltrative growth of differentiation breaks through serosa, the gastric cancer is easy to diffuse to adjacent organs such as omentum, colon, liver, pancreas and the like. 2. Blood circulation transfer: in the advanced stage, cancer cells enter the portal vein or systemic circulation and spread to other parts of the body, forming metastases. The common organs with metastasis include liver, lung, pancreas, and bones, and liver metastasis is usually considered. 3. And (3) peritoneal planting and transferring: when the stomach cancer tissue infiltrates outside the serosa, tumor cells fall off and are planted on the peritoneum and the visceral serosa to form metastatic nodules. Peritoneal seeding most easily occurs in the upper abdomen, mesentery. The implantation in the rectum and bladder is the sign of late gastric cancer. Metastatic carcinoma of the anterior fovea rectum can be found by digital rectal examination; ovarian metastatic tumors can develop in female patients with gastric cancer. 4. Lymphatic metastasis: lymph metastasis is the main metastasis way of gastric cancer, the lymph metastasis rate of the gastric cancer in the advanced stage is as high as about 70%, and lymph metastasis can also occur in early stage gastric cancer. The lymph node metastasis rate of gastric cancer is positively correlated with the depth of invasion of the cancer foci. Lymph node metastasis from gastric carcinoma is usually gradual, but also abrupt lymph metastasis can occur, i.e., no metastasis at the first station and metastasis at the second station. Terminal stage gastric cancer may metastasize to the supraclavicular lymph nodes via the chest catheter, or to the umbilicus via the ligamentum teres hepaticum.

Disclosure of Invention

The invention provides a method for clinically improving gastric cancer by TIL (total insulin secretion) in order to overcome the defects in the prior art.

The invention is realized by the following technical method: the invention discloses a method for clinically improving gastric cancer by TIL, which comprises the steps of separating and amplifying lymphocytes in tumor tissues, culturing mature TIL cells, and infusing the TIL cells back to a gastric cancer patient for improving the life quality of the gastric cancer patient.

The method for clinically improving the gastric cancer by the TIL comprises the following steps: the gastric cancer patient is operated in hospital, the tumor tissue is taken out and transported to GMP laboratory for subsequent treatment and culture, after 21 days of culture, the TIL cells are cultured to be mature, and then transported back to hospital for transfusion to the gastric cancer patient, and the TIL cell injection amount of each patient is 7X 10⁹-7×10¹⁰And (4) respectively.

The TIL tumor infiltration lymphocyte is a T cell which is infiltrated in tumor tissues and can effectively identify tumor cell neoantigen, and the T cell which can identify the tumor cell neoantigen has the capacity of efficiently killing tumor cells, including tumor passage cells transferred from the focus to other parts.

The GMP laboratory is a biological laboratory which meets the international cell preparation quality management system standard, and can provide the technical applications of stem cell separation and purification, culture and amplification, collection and cryopreservation, detection and quality control, cell resuscitation, product transportation, cell return and the like for meeting the requirements of patients on treating diseases.

The invention has the beneficial effects that: the advantages of tumor infiltrating lymphocytes are: compared with other immunotherapy methods, the tumor infiltrating lymphocytes have the advantages of more tumor antigen recognition targets, less side effects, more solid tumors which can be treated, stronger curative effect and the like.

Doctor of the national cancer institute surgical department, maine roleb (Steve Rosenberg), pioneered the use of tumor infiltrating lymphocytes. TIL tumor-infiltrating lymphocytes are cells derived from the infiltration of tumor tissue, and are known immune cells that are capable of specifically recognizing and attacking cancer. The application of the tumor infiltrating lymphocytes in the invention in improving the gastric cancer is to separate and amplify the lymphocytes in the tumor tissues and then return the lymphocytes to the patient, thus achieving the purpose of improving the life quality of the patient with the gastric cancer.

The objective remission rate of the TIL immune cells for treating solid tumors reaches 44 percent, and the disease control rate reaches 89 percent. Advantages of the TIL immunotherapy of the invention over other treatment methods: 1. surgical resection method: the advantages are that: is capable of directly excising the focus; disadvantages are that: has the risk of bleeding or infection, has long healing time, and can not be operated if the tumor spreads or metastasizes, thereby injuring the primordial qi of the patient. 2. Radiotherapy: the advantages are that: accurate, high-efficiency, small side effect and the like; disadvantages are that: only aiming at specific parts, some parts can not be used, and the traditional Chinese medicine composition has harm to normal tissue cells and low side effect. 3. Chemotherapy: the advantages are that: the tumor cells can be effectively killed and reduced when the tumor spreads or metastasizes; disadvantages are that: the side effect is large, the immune system loss is serious, the infection risk is increased, and the drug resistance is easy to generate. 4. Targeted drug therapy: the advantages are that: the method is accurate and efficient; disadvantages are that: only aims at a single antigen target, and is easy to generate drug resistance and side effects. The TIL immunotherapy disclosed by the invention has high killing capacity, can identify a plurality of antigen targets, uses autoimmune cells, has no side effect, and can be used for patients with recurrent or metastatic late gastric cancer.

Detailed Description

The present invention will be described in detail with reference to specific embodiments.

The invention discloses a method for clinically improving gastric cancer by TIL, which comprises the steps of separating and amplifying lymphocytes in tumor tissues, culturing mature TIL cells, and infusing the TIL cells back to a gastric cancer patient for improving the life quality of the gastric cancer patient. The method for clinically improving the gastric cancer by the TIL comprises the following steps: the gastric cancer patient is operated in hospital, the tumor tissue is taken out and transported to GMP laboratory for subsequent treatment and culture, after 21 days of culture, the TIL cells are cultured to be mature, and then transported back to hospital for transfusion to the gastric cancer patient, and the TIL cell injection amount of each patient is 7X 10⁹-7×10¹²And (4) respectively.

The improvement case is as follows: 1 patient with 46-year-old brain metastasis and gastric cancer who had undergone resection and chemotherapy for gastric cancer and had an average injection of TIL cells at 7X 10¹⁰After 15 months, the tumor disappeared from the medical image, and the patient was clinically free.

Finally, it should be noted that the above contents are only used to illustrate the technical method of the present invention, and not to limit the protection scope of the present invention. Simple modifications and equivalents of the technical method of the present invention may be made by those skilled in the art without departing from the spirit and scope of the technical method of the present invention.