Immunoadjuvant of diffuse large B-cell lymphoma and application of immunoadjuvant

An immune adjuvant, B cell technology, applied in the field of biomedicine, can solve the problems of lack of specificity, long-term toxicity of drugs, lack of drugs, etc., and achieve the effect of reducing the rate of cell proliferation

Active Publication Date: 2020-02-21
FUJIAN MEDICAL UNIV UNION HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the lack of specificity of chemokines and their receptor antagonists currently under development in controlling tumor growth and metastasis, and the long-term drug toxicity of currently approved chemokines and their receptor antagonists, Side effects such as inflammation and cytokineemia make the development of immune preparations for chemokines and their receptors unsatisfactory. So far, there are no reports on CCL20-related inhibitors and immune adjuvants

Method used

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  • Immunoadjuvant of diffuse large B-cell lymphoma and application of immunoadjuvant
  • Immunoadjuvant of diffuse large B-cell lymphoma and application of immunoadjuvant
  • Immunoadjuvant of diffuse large B-cell lymphoma and application of immunoadjuvant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] An immune adjuvant for diffuse large B-cell lymphoma, including a nucleic acid adjuvant composed of targeted ODN, the phosphate backbone of the targeted ODN is fully thio-modified, and the sequence of the targeted ODN is as shown in SEQ ID NO: 1 shown.

[0041] After dissolving the three targeted ODNs shown in Table 3 (wherein the third one is the targeted ODN shown in SEQ ID NO: 1), they were respectively dissolved in phosphate buffered saline solution and adjusted to concentrations of 5 μg / ml and 10 μg respectively. / ml, 15μg / ml and 20μg / ml for use. Take the human-derived DLBCL cell lines SU-DHL-2 and SU-DHL-6 in the logarithmic growth phase, and adjust the cell concentration to 5x10 with 1640 medium containing 10% fetal bovine serum 6 / ml, inoculate in 24-well plate, 1ml / well, add three targeted ODNs of different concentrations to the cells respectively, at 37°C, 5% CO 2 Cultivate in the cell incubator for 0, 12, 24, 48, and 72 hours, collect the cells incubated wi...

Embodiment 2

[0044] The human DLBCL cell lines SU-DHL-2 and SU-DHL-6 in logarithmic growth phase were inoculated in 96-well plates, divided into experimental group and control group, with 5000 cells per well, and SEQ ID NO: 1 The indicated targeted ODN was added to the cells of the experimental group at a concentration of 10 μg / ml, co-cultured at 37°C in a 5% CO2 cell incubator, and the cell viability was detected once every 24 hours using the CCK8 kit ( Figure 8-9 ). Add 10 μl CCK8 detection reagent to each well 2 hours before detection, 37°C, 5% CO 2 After incubation for 2 h, the OD value of each well at 450 nm was measured with a microplate reader. Relative proliferation activity = OD value of treatment group / OD value of blank control group. The results showed that, compared with the control group, the cell proliferation rate in the experimental group decreased significantly, indicating that co-incubation of the targeted ODN shown in SEQ ID NO: 1 with DLBCL cells can significantly re...

Embodiment 3

[0046] Harvest mouse B lymphoma cells A20 in the logarithmic growth phase, adjust the concentration to 2x10 6 / 200 μl, add doxorubicin (final concentration: 100 μg / ml), bathe in 37°C water for 30 minutes, wash with PBS three times, add an equal amount of normal saline for later use. Female DBA / 2 mice weighing about 25 g were randomly divided into three groups: control group, 1x10 5 tumor cell group and 1x10 6 Tumor cell group. The above-mentioned inactivated tumor cells were inoculated under the right ribs of the mice in the latter two groups, and the same amount of normal saline was inoculated in the control group. After 14 days, the three groups of mice were inoculated with 1×10 6 , 1×10 5 , 1×10 6 , 1×10 6 , 1×10 5 , 1×10 6 tumor cells. The tumor formation rate and tumor regression phenomenon of mice in each group were observed.

[0047] The results are shown in Table 4. It can be seen from Table 4 that all the mice in the control group and the immunized mice form...

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Abstract

The invention discloses an immunoadjuvant based on diffuse large B-cell lymphoma of a chemotactic factor CCL20 and an application of the immunoadjuvant. Target ODN of a total-phosphorothioate chemotactic factor CCL20mRNA 3'UTR is used as the immunoadjuvant to be applied to the diffuse large B-cell lymphoma. The immunoadjuvant exerts favorable antitumor effects on DLBCL in vivo and in vitro research, and through co-hatching of the immunoadjuvant and DLBCL cells, the cell proliferation rate of the DLBCL can be obviously reduced; after the immunoadjuvant and inactivated tumor cells are mixed forlocal injection, the general information of mice is favorable, and the mice do not have abnormal conditions of convulsions, disturbance, pilo-erection, twitch and the like. The immunoadjuvant does nothave abnormal phenomena of red and swollen, inflammation, ulceration, scleroma and the like in local injection parts. The mice without bearing cancer do not have the death phenomenon, the immunoadjuvant is promoted to be a DLBCL immunoadjuvant, can be used for clinical treatment, and has wide prospects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an immune adjuvant for diffuse large B-cell lymphoma based on chemokine CCL20 and its application. Background technique [0002] As the fourth cancer therapy besides surgery, chemotherapy, and radiotherapy, immunotherapy has attracted widespread attention because it uses tumor-specific antigens to attack cancer cells, and has the characteristics of invasiveness and few side effects. The current immunotherapy is expensive and has a weak immune effect, which limits its clinical application. Although the safety of new tumor vaccines has been significantly improved compared with traditional vaccines, their immune effects and immunogenicity are weak. Therefore, it is particularly important to use adjuvants to enhance and / or shape antigen-specific immune responses in tumor immunotherapy. [0003] Chemokines are cytokines that induce the directed migration of inflammato...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K39/00A61P35/00
CPCA61K39/39A61K39/0011A61P35/00A61K2039/55561A61K2039/804
Inventor 沈建箴黄倩付海英张凤陈荣
Owner FUJIAN MEDICAL UNIV UNION HOSPITAL
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