CN103823068A - Method for identifying anti-tumor lymphocytes - Google Patents
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- CN103823068A CN103823068A CN201310690699.4A CN201310690699A CN103823068A CN 103823068 A CN103823068 A CN 103823068A CN 201310690699 A CN201310690699 A CN 201310690699A CN 103823068 A CN103823068 A CN 103823068A
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Abstract
The invention discloses a method for identifying anti-tumor lymphocytes. The identification method comprises the following steps: stimulating lymphocytes of a small tumor tissue, which is taken from a cancer patient, in a specific culture medium and cultivating for 5 days; detecting the level of gamma interferon in the culture medium directly by adopting an enzyme-linked immunoassay method so as to identify the tumor killer lymphocytes. The method can directly measure the level of gamma interferon in the culture medium, is quick and low in time consumption.
Description
Technical field
The present invention relates to medical domain, in particular a kind of antitumor lymphocytic authentication method.
Background technology
Environmental pollution and inherent cause can cause the sudden change of DNA and gene, and cancer cell originates from human inner cell's growing without controlling due to DNA mutation and damage.Conventionally, growth of cancer cells forms tumour, and cancer cell also can be transferred to other places continued growth, thereby destroys normal histoorgan, causes patient's death.
Cancer has become common disease, frequently-occurring disease.According to the statistics of 2008, there was new cases of cancer 1,266 ten thousand in the whole world, dead 8,000,000, estimated will reach 1,500 ten thousand more than to new cases of cancer in 2015." 2012 Chinese tumour registration annual report " externally issued, and the report whole nation just has a people to be diagnosed as cancer in every 6 minutes, has every day 8550 people to become cancer patient, in every seven to eight people, just has a people to die from cancer.The situation is tense for whole nation pathogenesis of cancer, and incidence and mortality is lasting ascendant trend, and annual new cases of cancer approximately 3,500,000, because of cancer mortality approximately 2,500,000.Be accompanied by the uneasiness of the PM2.5 to thick air, the numeral of this succession of grey makes Chinese be stretched even tauter to the cognition of cancer.
At present cancer is the factor that second after cardiovascular causes mankind's death, and only about half of man and more than 1/3rd woman can produce cancer.It is 22% that China resident suffers from cancered probability all one's life, and cancer has become the mankind the first killer.At present, also do not have effectively way to tackle cancer, particularly the treatment of patients with advanced cancer.Since 20th century, take operative treatment, radiation therapy and the large conventional weapon of chemotherapy three as treatment and rehabilitation make huge contribution, but these traditional treatment means all have its limitation, fail to obtain overall promising result in treatment of cancer.Such as operative treatment, be only suitable for about 20% more early stage cancer patient, some operative treatment can obtain satisfied radical cure effect, and regrettably such situation is minority really, and most of case is not suitable for operative treatment.Chemotherapy is double-edged sword, systemic chemotherapy, and toxic and side effect is large, and the sick kind of minority can obtain radical cure effect, but most of chemotherapy just appeases or auxiliaring effect, and blindly the unhelpful quality of life of chemotherapy improves, and even increases health and financial burden because chemotherapy causes complication.In order to effect a radical cure cancer, in the urgent need to finding a kind of way of effective treatment cancer.
The immunization therapy of tumour is a kind of emerging, tumor treatment model with significant curative effect, is the anticancer new bio technology methods for the treatment of of a kind of autoimmunity.It is to use biotechnology and biopreparate to carry out feeding back to the method in patient body in vitro culture and amplification to the immunocyte gathering in patient body, excites, and strengthens body autoimmune function, thereby reaches the object for the treatment of tumour.The immunization therapy of tumour is the fourth-largest oncotherapy technology after operation, radiation and chemotherapy.
Utilize special killer lymphocyte to carry out immunotherapy of tumors and originate from the eighties, utilizing antineoplastic lymphocyte to carry out immunotherapy of tumors is a kind of special, effective and advanced treatment of cancer means.In recent years, the application that this technology is succeeded in cutaneum carcinoma end-stage patients' treatment.Clinical testing proves, 3/4ths cutaneum carcinoma end-stage patients cancerous tumors after treatment obviously disappears, and the cutaneum carcinoma end-stage patients of half return to one's perfect health.Therefore, utilizing antineoplastic lymphocyte to carry out tumour cell immunization therapy is a kind of very effective treatment of cancer new technology, has broad application prospects.
The process of utilizing special killer lymphocyte to carry out immunotherapy of tumors comprises obtaining of neoplasmic tissue sample, from tumor sample, separate tumor-killing lymphocyte, the lymphocytic a large amount of amplifications of tumor-killing and activation, the observation of tumor-killing lymphocyte input cancer patient and clinical effectiveness.The lymphocytic separation of tumor-killing is crucial.Carry out a large amount of research to separate tumor-killing lymphocyte from skin cancer patient, and obtained good effect, can separate and obtain tumor-killing lymphocyte from more than 80 percent skin cancer patient.Studies show that, also can separate and obtain tumor-killing lymphocyte from other cancer patients patient.Therefore, the lymphocytic immunotherapy of tumors of tumor-killing also can be applied to other cancer patients.
The key of immunotherapy of tumors is in cancer human body, to separate, identify tumor-killing lymphocyte.Conventionally, from cancer patient, excise a fritter tumor tissues, through stimulation and the cultivation of 7-10 days, obtain millions of tumor infiltrating lymphocytes.Then, carry out the common cultivation of tumor infiltrating lymphocyte and tumour cell.After twenty four hours, utilize enzymoimmunoassay to detect lymphocyte and whether discharge IFN-γ, thereby determine whether lymphocyte has tumor-killing.The lymphocytic method of evaluation tumor-killing of this classics, consuming time longer, and need tumour cell as target cell.Often from cancer patient tumor tissues, primary tumor cell is cultivated in induction, very difficult.So, identify the tumor-killing lymphocyte from cancer patient in the urgent need to the method for simple and fast, so that effectively for the immunization therapy of tumour.
Summary of the invention
Technical matters to be solved by this invention is for the deficiencies in the prior art, and a kind of antitumor lymphocytic authentication method is provided.
Technical scheme of the present invention is as follows:
A kind of antitumor lymphocytic authentication method, comprises the following steps:
(1) get the tumor tissue of 0.5 centimetre of big or small cancer patient, in Biohazard Safety Equipment, be cut into 1 millimeter of square tumor tissues fritter with scalpel;
(2) tumor tissues is put in to 24 well culture plates, adds RPMI-1640 nutrient culture media and interleukin 2, then 24 well culture plates are placed in to 37 ℃ and cultivate with 5%CO2 incubator;
(3) cultivate two days later, then add RPMI-1640 nutrient culture media and interleukin 2,24 well culture plates are placed in to 37 ℃ and 5%CO2 incubator continuation cultivation;
(4) the 5th days, draw 0.5 milliliter of nutrient culture media from every hole nutrient culture media, be placed in-20 ℃ of Refrigerator stores; Measure the content of gamma interferon in nutrient culture media with gamma interferon enzyme linked immunological kit; In nutrient culture media, containing the lymphocyte in the corresponding holes of gamma interferon more than 100 pg/ml, is the lymphocyte with antitumor activity.
Described authentication method, in step (2), one (1) described tumor tissues fritter is put in the 24 every holes of well culture plate; 1 milliliter of RPMI-1640 nutrient culture media and 1000IU interleukin 2 are added in every hole.
Described authentication method, in step (3), 1 milliliter of RPMI-1640 nutrient culture media and 200IU interleukin 2 are added in every hole.
Feature of the present invention is directly to measure IFN-γ level in nutrient culture media.So, fast, consuming time shorter, need to not cultivate primary tumor cell as target cell from the induction of cancer patient tumor tissues.Principle of the present invention is that the cancer cell in tumor tissues itself also can stimulate lymphocyte in the time that the tumor infiltrating lymphocyte in tumor tissues is subject to defined medium stimulation.That is to say, the cancer cell itself in tumor tissues, as target cell, stimulates lymphocyte to discharge IFN-γ, identifies the antitumor lymphocyte from cancer patient with this.
Accompanying drawing explanation
Fig. 1 is the mensuration of the lymphocyte active anticancer of breast cancer patients LMC5130.
Fig. 2 is the mensuration of the lymphocyte active anticancer of lung cancer patient LMC576.
Fig. 3 is the mensuration of the lymphocyte active anticancer of kidney patient LMC584.
Fig. 4 is the mensuration of the lymphocyte active anticancer of patients with colorectal cancer LMC586.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
0.5 centimetre of big or small tumor mass of excision and normal galactophore tissue's piece of closing on thereof from breast cancer patients (patient identification code is LMC5130).In Biohazard Safety Equipment, with scalpel, tumor tissues is cut into 1 millimeter of square tumor tissues fritter, mammary gland normal structure is also cut into 1 millimeter of square normal structure fritter.Tumor tissues and mammary gland normal structure are put in to 24 well culture plates, and fritter tumor tissues or a normal structure placed in each hole, and every kind of sample is placed in 4 holes and cultivates.Mammary gland normal structure is labeled as 5130N-1,5130N-2,5130N-3 and 5130N-4; Breast tumor tissues is labeled as 5130Tu-1,5130Tu-2,5130Tu-3 and 5130Tu-4.1 milliliter of RPMI-1640 nutrient culture media and 1000IU interleukin 2 are added in each hole.24 well culture plates are placed in to 37 ℃ and 5%CO
2in incubator, cultivate.Cultivate two days later, 1 milliliter of RPMI-1640 nutrient culture media and 200IU interleukin 2 are added in every hole.24 well culture plates are placed in to 37 ℃ and 5%CO
2in incubator, continue to cultivate.The 5th day, draw 0.5 milliliter from every hole nutrient culture media, be placed in-20 ℃ of Refrigerator stores.Measure the content of gamma interferon in nutrient culture media with gamma interferon enzyme linked immunological kit.
Measurement result as shown in Figure 1, as can be seen from Figure 1, from breast cancer patients LMC5130 tumor tissues, separate, detect antineoplastic lymphocyte strain-5130Tu-1 and antineoplastic lymphocyte strain-5130Tu-3, in the nutrient culture media of this two strains lymphocyte sample, detect that the gamma interferon that exceedes 100 pg/ml discharges.
0.5 centimetre of big or small tumor mass of excision and normal lung tissue's piece of closing on thereof from lung cancer patient (patient identification code is LMC576).In Biohazard Safety Equipment, with scalpel, tumor tissues is cut into 1 millimeter of square tumor tissues fritter, lung normal structure is also cut into 1 millimeter of square normal structure fritter.Tumor tissues and lung normal structure are put in to 24 well culture plates, and fritter tumor tissues or a normal structure placed in each hole, and every kind of sample is placed in 4 holes and cultivates.Lung normal structure is labeled as 576N-1,576N-2,576N-3 and 576N-4; Lung cancer tumor tissues is labeled as 576Tu-1,576Tu-2,576Tu-3 and 576Tu-4.1 milliliter of RPMI-1640 nutrient culture media and 1000IU interleukin 2 are added in each hole.24 well culture plates are placed in to 37 ℃ and 5%CO
2in incubator, cultivate.Cultivate two days later, 1 milliliter of RPMI-1640 nutrient culture media and 200IU interleukin 2 are added in every hole.24 well culture plates are placed in to 37 ℃ and 5%CO
2in incubator, continue to cultivate.The 5th day, draw 0.5 milliliter from every hole nutrient culture media, be placed in-20 ℃ of Refrigerator stores.Measure the content of gamma interferon in nutrient culture media with gamma interferon enzyme linked immunological kit.
Measurement result as described in Figure 2, as can be seen from Figure 2, separates, detects antineoplastic lymphocyte strain-576T-1,576T-2,576T-3 and 576T-4 from lung cancer patient LMC576 tumor tissues.From lung normal structure, also detect antineoplastic lymphocyte strain-576N-1 and 576N-2.In the nutrient culture media of these lymphocyte samples, detect that the gamma interferon that exceedes 100 pg/ml discharges.
0.5 centimetre of big or small tumor mass of excision and the normal kidney tissue piece that closes on thereof from kidney patient (patient identification code is LMC584).In Biohazard Safety Equipment, with scalpel, tumor tissues is cut into 1 millimeter of square tumor tissues fritter, kidney normal structure is also cut into 1 millimeter of square normal structure fritter.Tumor tissues and kidney normal structure are put in to 24 well culture plates, and fritter tumor tissues or a normal structure placed in each hole, and every kind of sample is placed in 4 holes and cultivates.Kidney normal structure is labeled as 584N-1,584N-2,584N-3 and 584N-4; Kidney tumor tissues is labeled as 584Tu-1,584Tu-2,584Tu-3 and 584Tu-4.1 milliliter of RPMI-1640 nutrient culture media and 1000IU interleukin 2 are added in each hole.24 well culture plates are placed in to 37 ℃ and 5%CO
2in incubator, cultivate.Cultivate two days later, 1 milliliter of RPMI-1640 nutrient culture media and 200IU interleukin 2 are added in every hole.24 well culture plates are placed in to 37 ℃ and 5%CO
2in incubator, continue to cultivate.The 5th day, draw 0.5 milliliter from every hole nutrient culture media, be placed in-20 ℃ of Refrigerator stores.Measure the content of gamma interferon in nutrient culture media with gamma interferon enzyme linked immunological kit.
Measurement result is shown in Fig. 3, as can be seen from Figure 3, from kidney patient LMC584 tumor tissues, separate, detect antineoplastic lymphocyte strain-584Tu-2 and 584Tu-3, in the nutrient culture media of this two strains lymphocyte sample, detect that the gamma interferon that exceedes 100 pg/ml discharges.
0.5 centimetre of big or small tumor mass of excision and the normal mucosa piece that closes on thereof from patients with colorectal cancer (patient identification code is LMC586).In Biohazard Safety Equipment, with scalpel, tumor tissues is cut into 1 millimeter of square tumor tissues fritter, large intestine normal structure is also cut into 1 millimeter of square normal structure fritter.Tumor tissues and large intestine normal structure are put in to 24 well culture plates, and fritter tumor tissues or a normal structure placed in each hole, and every kind of sample is placed in 4 holes and cultivates.Large intestine normal structure is labeled as 586N-1,586N-2,586N-3 and 586N-4; Colorectal cancer tumor tissues is labeled as 586Tu-1,586Tu-2,586Tu-3 and 586Tu-4.1 milliliter of RPMI-1640 nutrient culture media and 1000IU interleukin 2 are added in each hole.24 well culture plates are placed in to 37 ℃ and 5%CO
2in incubator, cultivate.Cultivate two days later, 1 milliliter of RPMI-1640 nutrient culture media and 200IU interleukin 2 are added in every hole.24 well culture plates are placed in to 37 ℃ and 5%CO
2in incubator, continue to cultivate.The 5th day, draw 0.5 milliliter from every hole nutrient culture media, be placed in-20 ℃ of Refrigerator stores.Measure the content of gamma interferon in nutrient culture media with gamma interferon enzyme linked immunological kit.
Measurement result as shown in Figure 4, as can be seen from Figure 4, in patients with colorectal cancer LMC586 tumor tissues, separate, detect antineoplastic lymphocyte strain-586Tu-3, in the nutrient culture media of this lymphocyte sample, detect that the gamma interferon that exceedes 100 pg/ml discharges.
Should be understood that, for those of ordinary skills, can be improved according to the above description or convert, and all these improvement and conversion all should belong to the protection domain of claims of the present invention.
Claims (3)
1. an antitumor lymphocytic authentication method, is characterized in that, comprises the following steps:
(1) get the tumor tissue of 0.5 centimetre of big or small cancer patient, in Biohazard Safety Equipment, be cut into 1 millimeter of square tumor tissues fritter with scalpel;
(2) tumor tissues is put in to 24 well culture plates, adds RPMI-1640 nutrient culture media and interleukin 2, then 24 well culture plates are placed in to 37 ℃ and 5%CO
2in incubator, cultivate;
(3) cultivate two days later, then add RPMI-1640 nutrient culture media and interleukin 2,24 well culture plates are placed in to 37 ℃ and 5%CO
2in incubator, continue to cultivate;
(4) the 5th days, draw 0.5 milliliter of nutrient culture media from every hole nutrient culture media, be placed in-20 ℃ of Refrigerator stores; Measure the content of gamma interferon in nutrient culture media with gamma interferon enzyme linked immunological kit; In nutrient culture media, containing the lymphocyte in the corresponding holes of gamma interferon more than 100 pg/ml, is the lymphocyte with antitumor activity.
2. authentication method according to claim 1, is characterized in that, in step (2), one (1) described tumor tissues fritter is put in the 24 every holes of well culture plate; 1 milliliter of RPMI-1640 nutrient culture media and 1000IU interleukin 2 are added in every hole.
3. authentication method according to claim 1, is characterized in that, in step (3), 1 milliliter of RPMI-1640 nutrient culture media and 200IU interleukin 2 are added in every hole.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1560234A (en) * | 2004-02-18 | 2005-01-05 | 南京大学医学院附属鼓楼医院 | Process of caltivating TIL cell by self hydrothorax supernatant |
| US20070264275A1 (en) * | 2003-06-25 | 2007-11-15 | Inserm | Peptides Derived from the Protein Mmp-2, and the Use Thereof in Antitumoral Immunotherapy |
| WO2011013129A1 (en) * | 2009-07-30 | 2011-02-03 | Tel Hashomer Medical Research Infrastructure And Services Ltd. | Selection of lymphocytes for the treatment of cancer |
| CN102174469A (en) * | 2011-01-26 | 2011-09-07 | 宋鑫 | Method for effectively culturing tumor infiltrating lymphocytes (TILs) |
| CN103374548A (en) * | 2012-04-27 | 2013-10-30 | 上海诺昊亚医学诊断技术有限公司 | High-efficiency amplification culture solution for tumor infiltration lymphocytes |
| CN103396992A (en) * | 2013-08-15 | 2013-11-20 | 中国人民解放军第二军医大学 | Culture and application of oligoclonal tumor-infiltrating lymphocytes for liver cancer |
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070264275A1 (en) * | 2003-06-25 | 2007-11-15 | Inserm | Peptides Derived from the Protein Mmp-2, and the Use Thereof in Antitumoral Immunotherapy |
| CN1560234A (en) * | 2004-02-18 | 2005-01-05 | 南京大学医学院附属鼓楼医院 | Process of caltivating TIL cell by self hydrothorax supernatant |
| WO2011013129A1 (en) * | 2009-07-30 | 2011-02-03 | Tel Hashomer Medical Research Infrastructure And Services Ltd. | Selection of lymphocytes for the treatment of cancer |
| CN102174469A (en) * | 2011-01-26 | 2011-09-07 | 宋鑫 | Method for effectively culturing tumor infiltrating lymphocytes (TILs) |
| CN103374548A (en) * | 2012-04-27 | 2013-10-30 | 上海诺昊亚医学诊断技术有限公司 | High-efficiency amplification culture solution for tumor infiltration lymphocytes |
| CN103396992A (en) * | 2013-08-15 | 2013-11-20 | 中国人民解放军第二军医大学 | Culture and application of oligoclonal tumor-infiltrating lymphocytes for liver cancer |
Non-Patent Citations (7)
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Effective date of registration: 20161226 Address after: The city of Hangzhou in West Zhejiang province 311121 No. 998 Zhejiang future science and Technology City Building No. 5 room 410B Patentee after: Hangzhou he Siti Biotechnology Co. Ltd. Address before: Room 1, unit 3, building 11, Dongshan, Xihu District, Zhejiang, Hangzhou, 310013, China Patentee before: Zhou Juhua |