Immune effects and uses of farnesyl diphosphate, a cholesterol synthesis intermediate metabolite

Abstract

The present invention relates to the field of biotechnology, specifically to the immune effects and uses of farnesyl diphosphate, an intermediate metabolite of cholesterol synthesis. The present invention relates to the use of farnesyl diphosphate or its analogs, derivatives, potentiators, or inhibitors in regulating dendritic cell chemotaxis and / or lymphocyte activation, or in the preparation of a drug or kit for regulating dendritic cell chemotaxis and / or lymphocyte activation, as well as corresponding drugs or kits and methods of regulation. The present invention can be applied to regulating dendritic cell chemotaxis and / or lymphocyte activation, and / or further used to regulate the body's immune response and homeostasis, prevent and treat allergic diseases such as allergic rhinitis, autoimmune diseases such as systemic lupus erythematosus, and select tumor immunotherapy regimens and / or prognosis assessment, and has broad application prospects.

Description

Technical Field

[0001] The present invention relates to the fields of biotechnology and medicine, and specifically to the use of farnesyl diphosphate, an intermediate metabolite of cholesterol synthesis, in regulating the chemotactic ability of dendritic cells and / or lymphocyte activation, and further in regulating the body's immune response and immune-related diseases. Background Art

[0002] Dendritic cells are a key bridge connecting innate immunity and adaptive immunity, playing a key regulatory role in activating the body's anti-pathogen immune response and maintaining autoimmune tolerance. The functional regulation of dendritic cells determines the overall balance of the immune response. Dendritic cells are widely distributed in non-lymphoid tissues and lymphoid tissues. The migration of dendritic cells in the body is crucial for their maturation, activation and functional regulation. However, an increasing number of studies have shown that disorders in dendritic cell migration may lead to excessive aggregation and activation of dendritic cells at sites of inflammation, causing excessive tissue inflammation and even triggering the occurrence of autoimmune diseases. Dendritic cell-related chemokines and chemokine receptors have become potential diagnostic markers and therapeutic targets for autoimmune diseases.

[0003] DC migration is regulated by chemokines, cytokines, and other inflammatory mediators. Immature DCs distributed in the periphery sense danger signals through pattern recognition receptors on their surface. After ingesting pathogens, they mature and activate, upregulating expression of the chemokine receptor CCR7. CCL19 / CCL21, secreted by lymph node stromal cells, interacts with CCR7 expressed on DCs, promoting their migration to the T cell compartments of secondary lymphoid organs, initiating and regulating T cell-mediated adaptive immune responses. During DC chemotaxis, glycolysis levels are significantly elevated, as evidenced by enhanced activation of the transcription factor hypoxia-inducible factor 1alpha (HIF1α), increased expression of the glycolytic enzyme lactate dehydrogenase (LDHA), and increased lactate production. Expression of CCL19 / CCL21 has been shown to be closely associated with the development and progression of various autoimmune diseases, such as multiple sclerosis (MS), rheumatoid arthritis (RA), and inflammatory bowel disease (IBD). Mice deficient in CCL19 / CCL21 or its receptor CCR7 exhibit significantly reduced incidence of experimental autoimmune encephalomyelitis and antigen-induced arthritis, and significantly decreased DC-mediated Th1 and Th17 cell activation. Therefore, CCL19 / CCL21-dependent DC migration and functional activation play a key role in maintaining the homeostasis of immune responses and immune regulation.

[0004] Cholesterol is a key lipid component of cell membranes and organelle membranes. Cholesterol synthesis and degradation are regulated by a series of cholesterol metabolic enzymes, forming a complex cholesterol metabolic pathway ( Figure 1 The cholesterol biosynthesis pathway begins with acetyl-CoA. Acetyl-CoA is converted to mevalonate under the catalysis of hydroxymethylglutaryl-CoA reductase (Hmgcr). Mevalonate is further converted to farnesyl diphosphate (FPP). FPP is then converted to cholesterol through a series of pathways.

[0005] Studies have found that the cholesterol pathway plays an important role in T cell anti-tumor immunity and is closely related to the occurrence and development of tumors. Statins are commonly used cholesterol-lowering drugs. Simvastatin can significantly inhibit the rate-limiting enzyme Hmgcr in cholesterol synthesis and is widely used in the clinical treatment of hypercholesterolemia. Statins have also been reported to regulate T cell exhaustion in HIV-infected patients and reduce tumor-related mortality. The above shows that the cholesterol pathway is a key regulatory component of T cell anti-tumor and antiviral immune responses, and blocking the cholesterol pathway may become an effective adjuvant for tumor immunotherapy.

[0006] The cholesterol pathway is also receiving increasing attention in innate immunity and anti-pathogen inflammatory responses. Cholesterol accumulation in the cell membrane can promote the formation of TLR4-MD2 and TLR4-CD14 complexes, thereby enhancing LPS-induced innate immune responses. Correspondingly, Abca1 and Abcg1 can promote cholesterol efflux from macrophages, disrupting the formation of lipid valve structures in the cell membrane and endosomes, thereby inhibiting macrophage inflammatory responses. Furthermore, cholesterol crystals can induce inflammasome activation, thereby promoting the formation of atherosclerotic plaques.

[0007] However, the specific roles and links of the cholesterol metabolite FPP and the metabolic enzyme Hmgcr in regulating dendritic cell migration and subsequent lymphocyte activation, as well as autoimmune diseases, remain unclear. To facilitate clinical and scientific applications, there is an urgent need to research and develop specific metabolic molecules that can regulate dendritic cell chemotaxis and / or lymphocyte activation and / or autoimmune diseases. Summary of the Invention

[0008] The present invention aims to provide a substance for regulating dendritic cell chemotaxis and / or lymphocyte activation: farnesyl diphosphate or its analogs, derivatives, potentiators, or inhibitors. Another object of the present invention is to provide the use of the above substances in the preparation of a drug or kit for regulating dendritic cell chemotaxis and / or lymphocyte activation, as well as the corresponding drug or kit.

[0009] The present invention has found that simvastatin treatment or interference with Hmgcr expression can inhibit the chemotactic ability of dendritic cells under the stimulation of chemokines CCL19 and / or CCL21, mitochondrial activation and fusion, T lymphocyte proliferation, differentiation, germinal center B cell production, and pathological damage and inflammatory response of systemic lupus erythematosus. Supplementation of FPP can enhance the chemotactic ability of dendritic cells under the stimulation of chemokines CCL19 and / or CCL21, mitochondrial activation and fusion, T lymphocyte proliferation, differentiation, and germinal center B cell production. By achieving positive or negative regulation of the immune response, the effect of suppressing inflammatory damage in inflammatory diseases, blocking the progression of autoimmune diseases, or enhancing dendritic cell tumor vaccines is exerted, thereby achieving the purpose of treating the disease. On this basis, the present invention has been completed.

[0010] The first aspect of the present invention provides the use of farnesyl diphosphate or its analogs, derivatives, potentiators or inhibitors in regulating the chemotactic ability of dendritic cells and / or lymphocyte activation.

[0011] The second aspect of the present invention provides the use of farnesyl diphosphate or its analogs, derivatives, potentiators or inhibitors in the preparation of drugs or kits for regulating dendritic cell chemotaxis and / or lymphocyte activation.

[0012] Furthermore, the farnesyl diphosphate is a metabolic molecule generated during the synthesis of cholesterol.

[0013] Furthermore, the dendritic cells are derived from mammals, preferably mice, humans, rats, dogs, monkeys, gorillas, pigs, horses, cattle or sheep, more preferably mice.

[0014] Furthermore, the chemotactic ability of dendritic cells is selected from: chemotaxis of dendritic cells under stimulation of chemokine CCR7 ligands (CCL19+CCL21, the same below) (in vitro), chemotactic migration of dendritic cells from peripheral skin tissue to stimulated lymph nodes (in vivo, which can be assisted by immune complex induction), and mitochondrial activation and fusion of dendritic cells under stimulation of CCR7L ligands; lymphocyte activation is selected from: T cell proliferation, differentiation into effector T cells (including Th1, Th17 and Tfh cells), germinal center B cell formation, and mediation of inflammatory immune response.

[0015] Furthermore, the farnesyl diphosphate or its analogs, derivatives, or potentiators promotes dendritic cell chemotaxis and / or lymphocyte activation. Furthermore, compared to control dendritic cells or lymphocytes not exposed to farnesyl diphosphate or its analogs, derivatives, or potentiators, farnesyl diphosphate or its analogs, derivatives, or potentiators promotes dendritic cell chemotaxis, mitochondrial activation and fusion, and lymphocyte activation.

[0016] Furthermore, the inhibitor of farnesyl diphosphate inhibits dendritic cell chemotaxis and / or lymphocyte activation. Still further, the inhibitor of farnesyl diphosphate inhibits dendritic cell chemotaxis, mitochondrial activation and fusion, and lymphocyte activation compared to control dendritic cells or lymphocytes not exposed to the inhibitor of farnesyl diphosphate.

[0017] Furthermore, the farnesyl diphosphate or its analogs, derivatives, and synergists are selected from: farnesyl diphosphate and its chemical structure analogs, derivatives, reagents or drugs that can increase the expression of farnesyl diphosphate; the inhibitor of farnesyl diphosphate is selected from: RNAi, antisense oligonucleotides, interfering viruses, and specific inhibitors (including but not limited to simvastatin) and / or molecular compounds targeting farnesyl diphosphate and its upstream metabolic enzyme Hmgcr (hydroxymethylglutaryl coenzyme A reductase) for hindering or reducing the expression and / or function of farnesyl diphosphate and its upstream metabolic enzyme Hmgcr.

[0018] Furthermore, the farnesyl diphosphate or its analogs, derivatives, enhancers or inhibitors are further used to regulate the body's immune response and homeostasis, prevent and treat allergic diseases such as contact dermatitis, autoimmune diseases such as systemic lupus erythematosus, tumor immunotherapy regimen selection and / or prognosis assessment.

[0019] In a preferred embodiment of the present invention, the autoimmune disease is systemic lupus erythematosus.

[0020] A third aspect of the present invention provides a drug or kit for regulating dendritic cell chemotaxis and / or lymphocyte activation, comprising:

[0021] i) farnesyl diphosphate or its analogs, derivatives, potentiators or inhibitors;

[0022] ii) pharmaceutically or immunologically acceptable carriers or excipients.

[0023] Furthermore, the medicine or kit further comprises: immature or mature dendritic cells, chemotactic or non-chemotactic dendritic cells, chemokines CCL19 and / or CCL21, T lymphocytes, and B lymphocytes.

[0024] In a fourth aspect, the present invention provides a method for regulating the chemotactic ability of dendritic cells and / or lymphocyte activation, comprising the step of contacting dendritic cells and / or lymphocytes and / or mice with farnesyl diphosphate or its analogs, derivatives, enhancers or inhibitors.

[0025] Furthermore, the method further comprises:

[0026] Before, during or after the contacting step, the dendritic cells are contacted with a chemokine CCR7 ligand, the lymphocytes are contacted with dendritic cells, and the mice are contacted with a corresponding agent.

[0027] All numerical ranges provided herein are intended to expressly include all values ​​between the endpoints of the ranges and ranges therebetween. Features described herein or in the embodiments may be combined. All features disclosed herein may be used in any combination, and each feature disclosed herein may be replaced by any alternative feature that serves the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the features disclosed are intended only to be general examples of equivalent or similar features.

[0028] As used in the present invention, "containing", "having" or "including" include "comprising", "mainly consisting of", "substantially consisting of", and "consisting of"; "mainly consisting of", "substantially consisting of" and "consisting of" are subordinate concepts of "containing", "having" or "including".

[0029] Drugs or test kits

[0030] The present invention also provides a drug or kit for regulating dendritic cell chemotaxis and / or lymphocyte activation, and / or for regulating the body's immune response and homeostasis, preventing and treating allergic diseases such as contact dermatitis, autoimmune diseases such as systemic lupus erythematosus, selecting tumor immunotherapy regimens and / or assessing prognosis. The drug or kit comprises an effective amount of a sequence selected from the following group, or its expression product, or an inhibitor or agonist thereof, and a pharmaceutically or immunologically acceptable carrier or excipient.

[0031] The "pharmaceutically or immunologically acceptable" ingredients are suitable for use in humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), that is, substances with a reasonable benefit / risk ratio. The "effective amount" refers to an amount that can produce a function or activity in humans and / or animals and is acceptable to humans and / or animals.

[0032] The term "pharmaceutically or immunologically acceptable carrier" refers to a carrier used for the administration of a therapeutic agent or vaccine, including various excipients, diluents, and adjuvants. The term refers to pharmaceutical or vaccine carriers that are not themselves essential active ingredients and are not unduly toxic upon administration. Suitable carriers are well known to those of ordinary skill in the art. A comprehensive discussion of pharmaceutically acceptable excipients can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., NJ 1991).

[0033] Such carriers include (but are not limited to): saline, buffer, glucose, water, glycerol, ethanol, and combinations thereof. Generally, drug / vaccine formulations should be compatible with the mode of administration. For example, the drug of the present invention can be prepared using physiological saline or an aqueous solution containing glucose and other adjuvants by conventional methods to produce an injectable form. The drug is preferably manufactured under sterile conditions. The preparations of the present invention can also be formulated as sustained-release preparations.

[0034] The medicaments and kits of the present invention may be equipped with reagents or reagent sets as needed, based on the principles and methods of prevention, treatment, and prognosis. For example, the medicaments or kits of the present invention may further include: immature or mature dendritic cells, chemotactic or non-chemotactic dendritic cells, chemokines CCL19 and / or CCL21, T lymphocytes, and B lymphocytes.

[0035] In addition, the kit of the present invention may further include: a container, a control substance (including a positive or negative control), instructions for use, a buffer, etc. as needed, and those skilled in the art may select them according to specific circumstances.

[0036] The advantages of the present invention are:

[0037] 1. They demonstrated that farnesyl diphosphate can regulate the chemotactic ability of dendritic cells and further regulate the adaptive immune and inflammatory responses that dendritic cells rely on;

[0038] 2. The present invention can be used to regulate the chemotactic ability of dendritic cells and / or lymphocyte activation, and / or further be used to regulate the body's immune response and inflammatory response, prevent and treat allergic diseases such as contact dermatitis, autoimmune diseases such as systemic lupus erythematosus, and select tumor immunotherapy regimens and / or prognosis assessment, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 : Intracellular cholesterol metabolic pathway.

[0040] Figure 2 :Effects of simvastatin and farnesyl diphosphate treatment on the chemotactic ability of dendritic cells in vitro;

[0041] Flow cytometry analysis of the effects of simvastatin (SIM, the same below) and farnesyl diphosphate (FPP) treatment on the chemotactic ability of dendritic cells in vitro. The results shown in the figure are mean ± standard deviation (n = 4).

[0042] Figure 3 :The effect of interfering Hmgcr and FPP treatment on the chemotactic ability of dendritic cells in vivo;

[0043] (A) Flow cytometric analysis of the effects of Hmgcr and FPP treatment on the chemotactic ability of dendritic cells in vivo.

[0044] (B) Figure 3 A. Corresponding statistical results. The results shown in the figure are mean ± standard deviation (n=4).

[0045] Figure 4 :Effects of simvastatin and FPP treatment on the chemotactic ability of dendritic cells induced by antigen complexes in vivo;

[0046] (A) Immunofluorescence detection of the effects of simvastatin and FPP treatment on the chemotaxis of dendritic cells induced by antigen complexes in vivo.

[0047] (B) Figure 4 A. Corresponding statistical results. The results shown in the figure are mean ± standard deviation (n=6).

[0048] Figure 5 :Effects of simvastatin and FPP treatment on lymphocyte activation induced by antigen complex;

[0049] (A) Flow cytometric analysis of the effects of simvastatin and FPP treatment on Tfh cell and germinal center activation induced by antigen complexes.

[0050] (B) Figure 5 A. Corresponding statistical results. The results shown in the figure are mean ± standard deviation (n = 6-9).

[0051] Figure 6 :Effects of simvastatin and FPP treatment on mitochondrial activation in chemotactic dendritic cells;

[0052] (A) Absolute numerical detection of the effects of simvastatin and FPP treatment on the respiratory capacity of mitochondria in dendritic cells stimulated by chemokines. The results shown in the figure are mean ± standard deviation (n = 3-4).

[0053] (B) Simvastatin and FPP treatment affected the percentage of mitochondrial respiratory capacity of chemokine-stimulated dendritic cells. The results shown in the figure are mean ± standard deviation (n = 3-4).

[0054] Figure 7 :The effects of interfering with Hmgcr and FPP treatment on mitochondrial fusion in chemotactic dendritic cells;

[0055] (A) Electron microscopy examination of the effects of Hmgcr and FPP interference on mitochondrial fusion in chemokine-stimulated dendritic cells.

[0056] (B) Figure 7 A. Corresponding statistical results. The results shown in the figure are mean ± standard deviation (n = 25-30).

[0057] Figure 8:The effect of FPP treatment on the ability of chemotactic dendritic cells to stimulate T cell proliferation and activation;

[0058] Flow cytometry analysis of the effects of FPP treatment on the ability of chemotactic dendritic cells to stimulate T cell differentiation toward Th1, Th17, and Tfh, as well as their proliferation.

[0059] Figure 9 :Effects of simvastatin treatment on pathological damage and inflammatory response in systemic lupus erythematosus;

[0060] (A) Detection of the effects of simvastatin treatment on the appearance of SLE mice.

[0061] (B) HE staining analysis of the effects of simvastatin treatment on the skin pathology of SLE mice.

[0062] (C) HE staining and immunofluorescence analysis of the effects of simvastatin treatment on renal pathology in SLE mice.

[0063] (D) Flow cytometric analysis of the effect of simvastatin treatment on dendritic cell migration in SLE mice.

[0064] (E) Flow cytometric analysis of the effects of simvastatin treatment on the activation of lymph node Tfh cells and germinal center B cells in SLE mice. DETAILED DESCRIPTION

[0065] The specific implementation methods provided by the present invention are described in detail below with reference to the examples.

[0066] It should be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention. Those skilled in the art may make appropriate modifications and changes to the present invention, and these modifications and changes are all within the scope of the present invention.

[0067] For experimental procedures in the following examples where specific conditions are not specified, conventional methods in the art may be employed, for example, as described in Molecular Cloning: A Laboratory Manual (3rd ed., Cold Spring Harbor Laboratory Press, New York, 1989) or according to the conditions recommended by the supplier. DNA sequencing methods are conventional in the art and are also available from commercial companies.

[0068] Unless otherwise indicated, percentages and parts are by weight. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the present invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0069] Example 1: Dendritic cell culture process

[0070] Mouse bone marrow cells were cultured in RPMI-1640 (PAA) cell culture medium containing 10% (v / v) FCS (PAA) at 37°C for six days supplemented with 100 ng / mL mouse GM-CSF and 10 ng / mL mouse IL-4 cytokines (R&D Systems, Minneapolis, MN). After stimulation with LPS (100 ng / mL, Sigma) for 24 hours, mature dendritic cells were induced and sorted using anti-CD11c magnetic beads (Miltenyi Biotech). The purified dendritic cells were then isolated.

[0071] Example 2: Effects of simvastatin and FPP treatment on the chemotactic ability of dendritic cells in vitro.

[0072] The dendritic cells obtained in Example 1 were stimulated with simvastatin (50 μM) alone or in combination with farnesyl diphosphate (50 μM) for 12 h, and then cultured on the upper layer of a 24-well transwell culture plate (1×10 5 Cells were plated in 100 μl of culture medium per well (8 μm pore size, Corning, Life Science). 600 μl of culture medium containing chemokines CCL19 (50 ng / ml) and CCL21 (50 ng / ml) (R&D) or the same volume of culture medium without chemokines was added to the lower layer. After 6 hours, cells in the lower layer were counted by flow cytometry. The number of cells attracted by the chemokines was calculated by subtracting the number of cells in the lower layer without chemokines from the number in the lower layer with chemokines. The test results are shown in Table 1. Figure 2 .

[0073] The data showed that simvastatin could inhibit dendritic cell chemotaxis, while FPP supplementation could enhance the chemotaxis of dendritic cells.

[0074] Example 3: Effects of interfering with Hmgcr and FPP treatment on the chemotactic ability of dendritic cells in vivo.

[0075] The dendritic cells obtained in Example 1 were treated with Hmgcr interfering RNA for 36 hours and then treated with FPP for 12 hours. Then, they were labeled with CFSE (0.5 mM) for 10 minutes and 2×10 6 The cells were injected subcutaneously into the unilateral footpad of mice. Lymph nodes on the same side were harvested 48 hours later and CFSE was detected. + CD11c + The ratio of cells represents the ability of CFSE-labeled DCs to chemotaxis toward lymph nodes in vivo. Figure 3 A shows flow cytometry data, Figure 3 B is Figure 3 A. Corresponding statistical results.

[0076] Therefore, interfering with Hmgcr can inhibit the chemotaxis of dendritic cells, while replenishing FPP can enhance the chemotaxis of dendritic cells.

[0077] The Hmgcr interfering RNA sequence is as follows:

[0078] Hmgcr SiRNA sense(5'-3'):GGGAGUUCAAACUGUAUUATT

[0079] Hmgcr SiRNA antisense(5'-3'):UAAUACAGUUUGAACUCCCTT.

[0080] Example 4: Effects of Simvastatin and FPP Treatment on Antigen Complex-Induced Dendritic Cell Chemotaxis and Lymphocyte Activation in Vivo

[0081] Two hours after oral administration of SIM (20 mg / kg) or combined oral administration of FPP (20 mg / kg), mice were subcutaneously injected with immune complexes formed by OVA-OVA antibodies conjugated to A647. 48 hours later, immunofluorescence was used to detect the expression of CD3 (orange fluorescence), B220 (blue fluorescence), CD11c (red fluorescence), and OVA (green fluorescence) in the lymph nodes to indicate the ability of dendritic cells carrying OVA antigens to migrate to the lymph nodes. Ten days later, the expression of follicular helper T cells (Tfh cells: CD4 + CXCR5 hi PD-1 + ) and germinal center B cells (B220 + GL7 + ) ratio.

[0082] Figure 4 A shows that oral administration of simvastatin can inhibit the proportion of cells co-expressing CD11c / OVA, that is, it can inhibit the migration of dendritic cells in vivo induced by antigen complexes, while supplementation of FPP can enhance the chemotactic ability of dendritic cells in vivo; Figure 4 B is Figure 4 A. Corresponding statistical results.

[0083] Figure 5 A shows that oral administration of simvastatin can inhibit the activation of Tfh cells and germinal center B cells, while supplementation of FPP can enhance the activation of Tfh cells and germinal center B cells; Figure 5 B is Figure 5 Statistical results of A.

[0084] Therefore, simvastatin can inhibit the chemotaxis of dendritic cells and lymphocyte activation in vivo, while FPP supplementation can enhance the chemotaxis of dendritic cells and lymphocyte activation in vivo.

[0085] Example 5: Effects of Simvastatin and FPP Treatment on Mitochondrial Activation in Chemotactic Dendritic Cells

[0086] Dendritic cells obtained in Example 1 were stimulated with simvastatin (50 μM) alone or in combination with farnesyl diphosphate (50 μM) for 12 hours, followed by administration of the CCR7 ligands CCL19 and CCL21 (50 ng / ml) for 3 hours. Cellular oxygen consumption (OCR) was measured using an XF-96 Extracellular Flux Analyzer (Seahorse Bioscience), and mitochondrial spare respiratory capacity (SRC) was calculated to indicate mitochondrial activity. Figure 6 A and 6B show the absolute value and percentage of spare respiratory capacity, respectively. The results showed that oral administration of simvastatin could inhibit chemokine-stimulated mitochondrial activation of dendritic cells, while supplementation of FPP could enhance chemokine-stimulated mitochondrial activation of dendritic cells.

[0087] Therefore, simvastatin can inhibit the mitochondrial activity of chemotactic dendritic cells, while FPP replenishment can enhance the mitochondrial activity of chemotactic dendritic cells.

[0088] Example 6: Effects of Interference with Hmgcr and FPP Treatment on Mitochondrial Fusion in Chemotactic Dendritic Cells

[0089] Dendritic cells obtained in Example 1 were treated with Hmgcr interfering RNA for 36 h, and then treated with CCR7 ligands CCL19 and CCL21 (50 ng / ml) or farnesyl diphosphate (FPP, 50 μM) for 6 h. Mitochondrial length was then measured using cryo-electron microscopy to indicate mitochondrial fusion. Figure 7 A shows that interfering with Hmgcr can reduce the mitochondrial fusion of dendritic cells stimulated by chemokines, while replenishing FPP can enhance the mitochondrial fusion of dendritic cells stimulated by chemokines. Figure 7 B is Figure 7 Statistical results of A.

[0090] Therefore, interfering with Hmgcr can inhibit the mitochondrial fusion of chemotactic dendritic cells, while replenishing FPP can enhance the mitochondrial fusion of chemotactic dendritic cells.

[0091] The Hmgcr interfering RNA sequence is the same as that in Example 3.

[0092] Example 7: Effects of FPP treatment on the ability of chemotactic dendritic cells to stimulate T cell proliferation and activation.

[0093] The dendritic cells obtained in Example 1 were given OVA 323-339 Pre-sensitization for 2 hours, followed by CFSE-labeled CD4 + T cells were subjected to mixed lymphocyte reactions and treated with the CCR7 ligands CCL19 and CCL21 (50 ng / ml) or in combination with farnesyl diphosphate (FPP, 50 μM). Four days later, IFN-γ, IL-17, and IL-21 were labeled to indicate Th1, Th17, and Tfh cell differentiation, respectively, and CFSE dilution ratio was used to indicate T cell proliferation and activation. Figure 8 The results showed that FPP could enhance the ability of chemotactic dendritic cells to stimulate T cells to differentiate into Th1, Th17 and Tfh, as well as their proliferation.

[0094] Therefore, FPP can enhance the ability of chemotactic dendritic cells to stimulate T cell proliferation and activation.

[0095] Example 8: Effect of simvastatin treatment on pathological damage and inflammatory response in systemic lupus erythematosus

[0096] Mice were pretreated with simvastatin (20 mg / kg) for 3 weeks, and then 500 μl of pristane (Sigma) was injected intraperitoneally to induce a systemic lupus erythematosus model. Mice were sacrificed 20 weeks after induction for analysis. Figure 9 A shows that simvastatin can reduce skin damage in systemic lupus erythematosus. Figure 9 B shows that simvastatin can reduce inflammatory cell infiltration and tissue destruction in the skin of systemic lupus erythematosus. Figure 9 C shows that simvastatin can reduce glomerular swelling in systemic lupus erythematosus, namely, renal complement C3c deposition. Figure 9 D showed that simvastatin can reduce the migration of dendritic cells in systemic lupus erythematosus, Figure 9 E shows that simvastatin can reduce the activation of Tfh cells and germinal center B cells in systemic lupus erythematosus lymph nodes.

[0097] Therefore, simvastatin treatment inhibits the pathological damage and inflammatory response of systemic lupus erythematosus.

[0098] In summary, simvastatin treatment or interference with Hmgcr expression can inhibit the chemotactic ability of dendritic cells stimulated by the chemokines CCL19 and / or CCL21, mitochondrial activation and fusion, T lymphocyte proliferation and differentiation, germinal center B cell generation, and pathological damage in systemic lupus erythematosus. Supplementation with farnesyl diphosphate can enhance the chemotactic ability of dendritic cells stimulated by the chemokines CCL19 and / or CCL21, mitochondrial activation and fusion, T lymphocyte proliferation and differentiation, and germinal center B cell generation. Regulation of farnesyl diphosphate can positively or negatively modulate immune responses, inhibiting inflammatory damage in inflammatory diseases, blocking the progression of autoimmune diseases, or enhancing the efficacy of dendritic cell tumor vaccines, thereby achieving therapeutic goals.

[0099] The preferred embodiments of the present invention have been specifically described above, but the present invention is not limited to the described embodiments. Those skilled in the art may make various equivalent modifications or substitutions without departing from the spirit of the present invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.

Claims

1. Use of farnesyl diphosphate in the preparation of a kit for regulating dendritic cell chemotaxis and / or lymphocyte activation; the kit further comprises chemokines CCL19 and / or CCL21.

2. The use according to claim 1, characterized in that The dendritic cells are derived from mammals.

3. The use according to claim 1, characterized in that The chemotactic ability of dendritic cells is selected from: chemotaxis of dendritic cells in vitro under stimulation of chemokine CCR7 ligands CCL19+CCL21, chemotactic migration of dendritic cells from peripheral skin tissue to stimulated lymph nodes, and mitochondrial activation and fusion of dendritic cells under stimulation of CCR7 ligands; lymphocyte activation is selected from: T cell proliferation, differentiation into effector T cells, including Th1, Th17 and Tfh cells, germinal center B cell formation / activation, and mediation of inflammatory immune responses.

4. The use according to claim 1, characterized in that The farnesyl diphosphate promotes dendritic cell chemotaxis and / or lymphocyte activation.

5. The use according to claim 1, characterized in that The farnesyl diphosphate is used to regulate the body's immune response and homeostasis, and prevent and treat allergic diseases and autoimmune diseases.

6. A kit for regulating dendritic cell chemotaxis and / or lymphocyte activation, characterized in that: Include: i) farnesyl diphosphate; ii) chemokines CCL19 and / or CCL21; ii) pharmaceutically or immunologically acceptable carriers or excipients.

7. The kit according to claim 6, characterized in that The kit further comprises: immature or mature dendritic cells, chemotactic or non-chemotactic dendritic cells, T lymphocytes, and B lymphocytes.

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