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Sphingolipids for generating regulatory cd4+ t cells

A technology of cells and uses, applied in the field of sphingolipids used to generate regulatory CD4+ T cells, can solve the problems of inactivation of immune cells, harmful side effects of drugs, etc.

Pending Publication Date: 2022-02-15
DEUTES KREBSFORSCHUNGSZENT STIFTUNG DES OFFENTLICHEN RECHTS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This need stems from the fact that current treatments utilizing immunosuppressive drugs also inactivate functioning immune cells, with the drugs carrying the risk of harmful side effects

Method used

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  • Sphingolipids for generating regulatory cd4+ t cells
  • Sphingolipids for generating regulatory cd4+ t cells
  • Sphingolipids for generating regulatory cd4+ t cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] Sptlc2 deficiency of T cells increases tumor growth but reduces Treg cell formation. Sptlc2 Flox / Flox Cd4-Cre (Fl / Fl, 8 mice) and Sptlc2 + / + Cd4-Cre (+ / +, 11 mice) mice implanted with 2×10 5 melanoma B16 cells. The results are shown in figure 1 middle. We observed that genetic deficiency of Sptlc2 in T cells attenuates antitumor immunity ( figure 1 A), and we further observed that a subset of T cells, called regulatory T cells (Treg cells), is reduced by SPTLC2 deficiency ( figure 1 B).

Embodiment 2

[0118] Sptlc2 deficiency in Treg cells affects the immunosuppressive function of Treg cells. We used a flow cytometry sorter from Sptlc2 Flox / Flox Foxp3Cre-YFP mice or Sptlc2 + / + Purification of Treg and non-Treg cells from Foxp3Cre-YFP mice. Foxp3 is expressed in the nucleus and FACS staining of FOXP3 protein requires cell fixation and permeabilization, which kills the cells and is not suitable for subsequent cell culture. In this Foxp3Cre-YFP mouse strain, FOXP3 protein expression is reported by YFP protein expression. We were unable to directly FACS sort cells expressing FOXP3 without fixing and permeabilizing the cells, and thus preserving cell viability. YFP positive CD4 + Treg cells and YFP negative CD4 + Non-Treg cells were sorted by FACS. Non-Treg cells were labeled with the fluorescent dye Celltrace Violet (CTV, to determine cell proliferation) and co-cultured with or without Sptlc2-deficient or sufficient Treg cells for three days in the presence of the T cell...

Embodiment 3

[0120] Sptlc2 deficiency in Treg cells enhances autoimmunity in a mouse model of EAE. in Sptlc2 Flox / Flox Foxp3Cre-YFP mice or Sptlc2 + / + EAE was induced in Foxp3Cre-YFP mice. Briefly, each mouse was treated with 200 μg of MOG emulsified in Freund's complete adjuvant 35-55 Peptide subcutaneous immunization. Pertussis toxin (400 ng per mouse) was injected intraperitoneally. EAE symptoms were scored daily. Compared with wild-type control mice, Sptlc2 Flox / Flox Foxp3Cre-YFP mice developed more severe EAE. The results are shown in image 3 middle.

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Abstract

The present invention relates to a substance of formula (I), whereby R1 is an alkyl or alkenyl group having 6 to 20 carbon atoms; R2 is H or missing, whereby O is bound via a double bond, R3 is H or an acyl group -C(O)R5, whereby R5 is an alkyl or alkylene group having 1 to 10 carbon atoms, and R4 is H or a phosphate group for use as a medicament and for use in a method of preventing or treating a subject suffering from an autoimmune disease. The present invention further relates to a method for generating regulatory T cells (Treg cells) in vitro comprising the steps of providing precursor CD4 + T cells, cultivating the precursor CD4 + T cells provided in step 1) in the presence of the substance as defined herein, and, optionally, isolating the generated regulatory T cells (Treg cells).

Description

Background technique [0001] Regulatory T cells (Treg cells or Treg) were originally described as CD4 + CD25 + T cells. Treg cells are mainly related to the regulation of the immune system. Due to their mechanism of formation, Treg cells are divided into natural Treg cells (nTreg), which differentiate in the thymus and then translocate to the periphery of the body, and induced / adaptive Treg cells (iTreg), which arise in the periphery of the body. Treg cells are characterized by the expression of CD4, as well as with activated effector T cells (CD25 low , also known as CD25 - Compared with CD25 expression in T cells), the expression of the alpha chain of the interleukin-2 receptor is increased (CD25 high , also known as CD25 + ). Therefore, Treg cells can be distinguished from effector T cells by the expression level of CD25. About 2% to 10% CD4 + T cells express high levels of CD25 (CD25 high ) and Treg cells. In addition to the high expression of CD25, Treg cells ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/685A61K31/661A61K31/133A61P37/06C12N5/0783
CPCA61K31/685A61K31/133A61K31/661A61P37/06C12N5/0636C12N2501/999C12N2501/515C12N2501/51C12N2501/15C12N2501/2302C12N2500/30C12N2501/90C12N2500/36C12N2500/38C12N5/0637A61K39/4621A61K39/46433A61K39/4611A61K35/17A61P37/00C07F9/10C12N2500/42C12N2501/505C07C215/10C07K14/495C07K14/55C07K16/2809C07K16/2818C12N2501/04
Inventor 崔国梁马思聪
Owner DEUTES KREBSFORSCHUNGSZENT STIFTUNG DES OFFENTLICHEN RECHTS
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