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Immunoadjuvant and its application in diffuse large b-cell lymphoma

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

Active Publication Date: 2022-07-26
FUJIAN MEDICAL UNIV UNION HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 and its application in diffuse large b-cell lymphoma
  • Immunoadjuvant and its application in diffuse large b-cell lymphoma
  • Immunoadjuvant and its application in diffuse large b-cell lymphoma

Examples

Experimental program
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Effect test

Embodiment 1

[0040] An immune adjuvant for diffuse large B-cell lymphoma, including a nucleic acid adjuvant composed of a targeted ODN, the phosphate backbone of the targeted ODN is fully sulfur-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 is the targeted ODN shown in SEQ ID NO: 1) respectively with phosphate buffered saline solution, the concentrations were adjusted to 5 μg / ml and 10 μg respectively. / ml, 15μg / ml and 20μg / ml for use. Take the human DLBCL cell lines SU-DHL-2 and SU-DHL-6 in logarithmic growth phase, and adjust the cell concentration to 5x10 with 1640 medium containing 10% fetal bovine serum 6 / ml, seeded in a 24-well plate, 1ml / well, three targeted ODNs with different concentrations were added to the cells, at 37°C, 5% CO 2 Culture in a cell incubator for 0, 12, 24, 48, 72 h, collect cells incubated with ODN, place them in a 1.5 ml small centrifuge t...

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, and divided into experimental group and control group, with 5000 cells per well, and the SEQ ID NO: 1 The indicated targeting ODN was added to the cells of the experimental group at a concentration of 10 μg / ml, and co-cultured at 37 °C in a 5% CO2 cell incubator, and the cell viability was detected by the CCK8 kit every 24 h ( Figure 8-9 ). 2h before detection, add 10 μl CCK8 detection reagent to each well, 37°C, 5% CO 2 After incubation for 2 h, the OD value of each well at 450 nm was measured by 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 was significantly decreased, indicating that the co-incubation of the targeted ODN shown in SEQ ID NO: 1 with DLBCL cells could significantly...

Embodiment 3

[0046] Harvest mouse B lymphoma cells A20 in logarithmic growth phase at an adjusted concentration of 2x10 6 / 200 μl, add doxorubicin (final concentration is 100 μg / ml), water bath at 37° C. for 30 min, wash 3 times with PBS, and add an equal amount of normal saline for use. Female DBA / 2 mice weighing about 25g were randomly divided into three groups: control group, 1x10 5 Tumor cell group and 1x10 6 tumor cell group. The above inactivated tumor cells were inoculated under the right flank of the latter two groups of mice, and the control group was inoculated with an equal volume of normal saline. After 14 days, 1 × 10 1 × 10 were inoculated into the skin of the contralateral lower limbs of the three groups of mice. 6 , 1×10 5 , 1×10 6 , 1×10 6 , 1×10 5 , 1×10 6 tumor cells. The tumor formation rate and the presence or absence of tumor regression were observed in each group of mice.

[0047] The results are shown in Table 4. It can be seen from Table 4 that all the mi...

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Abstract

The invention discloses an immune adjuvant for diffuse large B-cell lymphoma based on chemokine CCL20 and its application. It is used in large B-cell lymphoma. The immune adjuvant has a good anti-tumor effect in DLBCL in vitro and in vivo studies. Co-incubation with DLBCL cells can significantly reduce the cell proliferation rate of DLBCL. It acts as an immune adjuvant and inactivates tumors. After the mixed local injection of cells, the mice were generally in good condition, with no abnormal conditions such as convulsions, agitation, piloerection, and convulsions. There were no abnormalities such as redness, inflammation, ulceration, and induration at the local injection site, and non-tumor-bearing mice did not die. , suggesting that it can be used as an immune adjuvant for DLBCL, which can be used in clinical practice and has broad prospects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular 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 in addition to surgery, chemotherapy, and radiotherapy, immunotherapy has attracted widespread attention because it uses tumor-specific antigens to attack cancer cells, with the characteristics of invasiveness and few side effects. The current immunotherapy has high cost and weak immune effect, which limits its clinical application. Although the safety of new tumor vaccines has been significantly improved compared with traditional vaccines, the immune effects and immunogenicity are weaker. Therefore, the application of adjuvants to enhance and / or shape antigen-specific immune responses is particularly important in tumor immunotherapy. [0003] Chemokines are a class of cytokines that induce directional migration of infl...

Claims

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Application Information

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Patent Type & Authority Patents(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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