Application of PD-L1/CTLA-4 in preparation of immunosuppressant

A technology of CTLA-4 and PD-L1, which is applied in the field of biomedicine to achieve the effect of suppressing immune rejection, great application prospects, and good immunosuppressive effect

Pending Publication Date: 2020-06-02
中山大学深圳
View PDF9 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of PD-L1 and CTLA-4 in immunosuppression, especially

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of PD-L1/CTLA-4 in preparation of immunosuppressant
  • Application of PD-L1/CTLA-4 in preparation of immunosuppressant
  • Application of PD-L1/CTLA-4 in preparation of immunosuppressant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Construction of PD-L1 / CTLA-4 membrane nanovesicles

[0039] Construction of cell lines: HEK 293T cells were transfected with packaging plasmids VSV-G, pHIV-gag-pol and targeting plasmids Lenti-PD-L1-OFP or Lenti-CTLA-4-GFP using Lipofectamine 2000 (Life Technologies). 12 and 24 hours after transfection (p.t.), the supernatant containing lentivirus was collected and filtered (0.45 μm). Then the collected virus was infected with HEK293T cells, or ASC cells and original 264.7 cells were infected with lentivirus, and screened with puromycin (2 μg / ml) to obtain stably expressed target cells. Firstly, a cell line stably expressing PD-L1-OFP was constructed, and then the cells were re-infected with CTLA-4-GFP lentivirus, and the co-infection cell line was obtained through puromycin selection.

[0040] The localization results of PD-L1 / CTLA-4 expression on the 293T cell membrane were detected by confocal laser microscopy. The result is as figure 1 As shown in a, la...

Embodiment 2

[0044] Example 2 In vitro biological behavior of PD-L1 / CTLA-4 membrane nanovesicles

[0045] (1) Nanovesicle cell binding assay: Jurkat cells were incubated with PD-L1-OFP NVs (50 μg / ml, protein weight) for 30 minutes, and centrifuged with a Ceoporep-4 centrifuge (Intech, China) to make slides. Then, wheat germ agglutinin (WGA) and Alexa-Fluor 488 conjugate were added to stain the cell membrane for 10 min, bmdc were seeded in a confocal culture dish, and DC cells were stimulated with 10 U-TNF-α. CTLA-4-OFP NVs (50 μg / ml, protein weight) were added and incubated for 30 min as previously described. Then, wheat germ agglutinin (WGA) and Alexa-Fluor 350 conjugate were added, incubated at 37° C. for 10 min, and the membrane was stained. 293T-CD80-OFP cells or 293T-PD-1-GFP cells were seeded in confocal culture dishes, respectively. CTLA-4-OFP NVs (50 μg / ml, protein weight) or PD-L1-OFP NVs (50 μg / ml, protein weight) were added to the medium and incubated for 30 minutes. Images w...

Embodiment 3

[0050] Example 3 The use of PD-L1 / CTLA-4 membrane nanovesicles in skin transplantation models

[0051] The skin graft models were established respectively. The use of experimental animals and all experiments were reviewed and approved by the Animal Ethics Committee of Sun Yat-sen Medical College, Sun Yat-sen University, with approval number 2018000577.

[0052] For skin grafting, mice were anesthetized, shaved and sterilized with 75% ethanol. The skin of male C57BL / 6 mice (0.8cm 2) were transplanted into the dorsal side of 8-week-old BALB / c mice. Fifteen recipients were randomly divided into 5 groups: PC group (normal saline, n=3), group 2 (PD-L1 NVs, 25mg / kg, n=3), group 3 (CTLA-4NVs, 25mg / kg kg, n=3), group 4 (PD-L1 / CTLA-4NVs, 25 mg / kg, n=3), NC group (autologous transplantation, n=3). Transplanted mice were dosed daily by tail vein injection for the first 3 days. After 3 days, NVs were injected every other day, and the survival curve of the mice was observed until 14 d...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Particle sizeaaaaaaaaaa
Electric potentialaaaaaaaaaa
Login to view more

Abstract

The invention discloses use of PD-L1 and/or CTLA-4 in immunosuppression. According to the use, PD-L1/CTLA-4 cytomembrane nano-vesicles expressing the PD-L1 and the CTLA-4 are prepared, actions of thePD-L1/CTLA-4 cytomembrane nano-vesicles are researched from a cellular level and an animal experiment level, and it is discovered that the PD-L1/CTLA-4 cytomembrane nano-vesicles can be bonded to a Tcell surface suppressant acceptor PD-1, present a cell surface acceptor B7 with a competitive antigen of an activated acceptor CD28 to inhibit activation of T cells and then regulate and control immune response of an organism and inhibit immunologic rejections of the organism to transplanted organs. It is shown that the PD-L1/CTLA-4 nano-vesicle suppressant can be used for preparing preparations for inhibiting the activation of the T cells and has an application prospect.

Description

technical field [0001] The invention relates to the technical field of biomedicine. More specifically, it relates to the application of PD-L1 / CTLA-4 in the preparation of immunosuppressants. Background technique [0002] The immune system is an important system for the body to perform immune responses and immune functions. It has the functions of identifying and eliminating antigenic foreign bodies and maintaining the stability and physiological balance of the body's internal environment. However, in human organ transplantation, the transplanted organ often causes the body's allogeneic immune rejection, which makes the transplanted organ unable to survive normally in the body, and even causes serious adverse reactions in the body due to immune rejection. [0003] Allogeneic immune rejection refers to the process of specific immune response of the organ recipient to the graft after organ transplantation between different individuals of the same species. Activation of T cell...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61K38/17A61K9/51A61K47/46A61P37/06C12N15/85
CPCA61K38/1774A61K9/5176A61P37/06C12N15/85Y02A50/30
Inventor 陈红波查华联徐占雪程芳贺超吴彦萍
Owner 中山大学深圳
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap