Immune cell therapy for colorectal cancer liver metastasis

Abstract

The invention relates to the technical field of biological treatment, and provides application of adoptively transplanted beta-glucan induced monocytes in prevention and / or treatment of colorectal cancer liver metastasis. Training a donor, which can be a mammal, with beta-glucan, and obtaining bone marrow cells from the mammal donor; and further carrying out flow sorting to obtain mononuclear cells. By adoptively transplanting the monocytes trained by the beta-glcan (BG), the colorectal cancer can be remarkably inhibited from being transferred to the liver, the ratio of the liver to the body weight is reduced, and infiltration and proliferation of the colorectal cancer cells in the liver are inhibited. The invention provides a novel and efficient cellular immunotherapy for preventing and / or treating colorectal cancer liver metastasis.

Description

Technical Field

[0001] This invention relates to the field of biotherapy technology, and more particularly to an immunocellular therapy for treating liver metastases from colorectal cancer. Background Art

[0002] Colorectal cancer (CRC) is a common malignant tumor of the digestive system, and its incidence and mortality rates have been rising steadily in recent years, making it one of the most prevalent and deadliest malignant tumors worldwide. Despite significant advancements in systemic treatment and multidisciplinary collaboration in modern medicine, the vast majority of patients still die from tumor metastasis. The liver is the most common site of metastasis, with approximately 50% of patients developing liver metastases during the course of the disease. Once liver metastasis occurs, the disease has progressed to an advanced stage, resulting in a poor prognosis. Unlike colorectal carcinoma in situ, the pathogenesis of metastatic tumors is extremely complex, involving epithelial-mesenchymal transformation, infiltration of tumor cells into the bloodstream, survival in circulation, extravasation into target organs such as the liver, and finally, colonization and proliferation in the liver.

[0003] Currently, both international and domestic guidelines recommend the diagnosis and treatment of colorectal cancer metastases using a multidisciplinary team (MDT) model. However, traditional surgical treatments and systemic radiotherapy and chemotherapy cannot completely eliminate latent disseminated tumor cells, offering only limited survival benefits to patients. Therefore, effectively preventing and treating tumor metastasis in colorectal cancer patients is of great significance for improving patient survival rates.

[0004] Studies have shown that the tumor immunosuppressive microenvironment promotes tumor cell metastasis. Among these, bone marrow-derived suppressor cells (MDSCs), as key effector cells in immunosuppression, significantly proliferate in the peripheral blood of colorectal cancer patients, and their numbers are positively correlated with tumor stage and metastatic extent. MDSCs inhibit T cell activation and proliferation, aggregate in the liver interfering with local immune surveillance, and synergistically contribute to T cell apoptosis induced by hepatic stellate cells and the proliferation of regulatory T cells (Tregs), collectively constructing an immune tolerance niche and promoting tumor metastasis. Therefore, a key aspect of preventing and treating metastatic cancer lies in restoring the anti-tumor effects of bone marrow-derived immune cells.

[0005] Adoptive immunotherapy is mainly divided into two categories: non-genetically engineered and genetically engineered. The former includes tumor-infiltrating lymphocytes (TILs), cytokine-induced killer cells (CIKs), and dendritic cell / CIK (DC-CIK) therapy, while the latter encompasses chimeric antigen receptor T-cell (CAR-T) and T-cell receptor T-cell (TCR-T) therapy. Immunotherapy achieves precise tumor killing by modifying or activating immune cells in vitro, demonstrating broader targeting potential compared to immune checkpoint inhibitors, and showing significant advantages in precision and personalization. However, the widespread application of current immunotherapy is limited by technological complexity, high cost, and the unique biological barriers of solid tumors.

[0006] Traditionally, it was believed that innate immune cells lacked the function of immune memory. However, recent studies have shown that innate immune cells, such as monocytes and macrophages, can acquire immune memory similar to adaptive immunity when stimulated by certain vaccines or fungal cell wall components. This phenomenon is known as trained immunity. Beta-glucan (BG), an essential natural polysaccharide widely distributed in cereals and microorganisms, possesses a wide range of biological activities, including immunomodulatory, anti-inflammatory, antioxidant, antitumor, and microbiota-regulating functions, and is a potent inducer of trained immunity. Although BG has been used as an immunomodulator in cancer, the functional correlation between its induced trained immunity and malignant tumor metastasis remains unclear. Summary of the Invention

[0007] To address the shortcomings of existing technologies, this invention provides the application of β-glucan-induced trained immune cells in the preparation of drugs for the prevention and treatment of colorectal cancer liver metastases. The invention demonstrates the potential of treating colorectal cancer liver metastases through the injection of β-glucan-induced trained immune cells, providing a scientific basis for the development of novel and highly effective treatment regimens for colorectal cancer liver metastases.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is specifically described as follows: The first aspect of this invention discloses the application of mononuclear cells prepared by the following method in immunocellular therapy for the prevention and / or treatment of liver metastases from colorectal cancer, characterized in that the method comprises the following steps: a. Train donors using β-glucan to obtain bone marrow cells from donors; b. Flow cytometry sorts the bone marrow cells obtained in step (a) to obtain mononuclear cells.

[0009] In some applications of the mononuclear cells described in the first aspect in immunocellular therapy for the prevention and / or treatment of liver metastases from colorectal cancer, the mononuclear cells are administered by injection.

[0010] In some applications of the mononuclear cells described in the first aspect in immunocellular therapy for the prevention and / or treatment of liver metastases from colorectal cancer, the donor is a mammal, such as a mouse, rabbit, dog, or human.

[0011] In some applications of the mononuclear cells described in the first aspect in immunocellular therapy for the prevention and / or treatment of liver metastases from colorectal cancer, the mononuclear cells are Ly6C-expressing cells. hi CD45 + CD11b + Bone marrow mononuclear cells.

[0012] In some applications of mononuclear cells described in the first aspect in immunocellular therapy for the prevention and / or treatment of liver metastases from colorectal cancer, the β-glucan is β-glucan or a composition containing β-glucan.

[0013] In some applications of the mononuclear cells described in the first aspect in immunocellular therapy for the prevention and / or treatment of liver metastases from colorectal cancer, the β-glucan is derived from yeast, fungi, algae, or grains.

[0014] In some applications of the mononuclear cells described in the first aspect in immunocellular therapy for the prevention and / or treatment of colorectal cancer liver metastases, the adoptive transplantation of β-glucan-trained mononuclear cells is used to reduce the ratio of metastatic colorectal cancer liver to body weight.

[0015] In some applications of the mononuclear cells described in the first aspect in immunocellular therapy for the prevention and / or treatment of liver metastases of colorectal cancer, the adoptive transplantation of β-glucan-trained mononuclear cells is used to alleviate the infiltration and proliferation of colorectal cancer cells in the liver.

[0016] A second aspect of the present invention provides a therapeutic composition comprising β-glucan-induced isolated mononuclear cells, specifically, the composition comprising the mononuclear cells obtained in steps a) and b) of the first aspect.

[0017] In some of the therapeutic compositions described in the second aspect, the composition further comprises a pharmaceutically acceptable carrier, diluent, excipient and / or additive, and preferably, the composition further comprises a chemotherapeutic drug, immune checkpoint inhibitor or targeted drug that works synergistically with the monocytes.

[0018] In some of the therapeutic compositions described in the second aspect, the monocytes, after induction, may exhibit enhanced chemotaxis, phagocytic activity, and expression of pro-inflammatory factors.

[0019] A third aspect of the present invention provides the use of a second aspect therapeutic composition in the prevention and / or treatment of liver metastases from colorectal cancer, said therapeutic composition being administered by intravenous injection, portal vein injection, hepatic artery perfusion, or intraperitoneal injection.

[0020] A third aspect of the present invention provides the use of a second aspect therapeutic composition in the prevention and / or treatment of liver metastases of colorectal cancer, said therapeutic composition being used as an adjunct to postoperative treatment, inhibition of advanced colorectal cancer liver metastases, or prevention of liver metastasis recurrence.

[0021] The reagents used in this invention are all purchased from the open and legal market and have not undergone further purification.

[0022] Compared with the prior art, the beneficial technical effects of the present invention are as follows: Compared to oral or injectable β-glucan preparations, adoptive transplantation of β-glucan-trained monocytes can reduce or avoid discomfort such as diarrhea, chills, fever, injection site pain, headache, back pain, joint pain, nausea, vomiting, swollen lymph nodes, dizziness, abnormal blood pressure, rash, and fatigue. Furthermore, some colorectal cancer patients have damaged hematopoietic stem cells; these patients benefit even more from transplanting β-glucan-trained monocytes.

[0023] Terminology Definition To facilitate understanding of the technical terms used in this patent document, the following definitions and explanations are provided for key terms: Treatment (or treating) is a method of achieving a beneficial or desired clinical outcome, including but not limited to one or more of the following: alleviating one or more symptoms caused by the disease, reducing the severity of the disease, stabilizing the disease (e.g., preventing or delaying disease exacerbation), preventing or delaying the spread of the disease (e.g., metastasis), preventing or delaying disease recurrence, delaying or slowing disease progression, improving disease status, providing remission of the disease (partial or complete), reducing the dosage of one or more other medications required to treat the disease, delaying disease progression, improving quality of life, and / or prolonging survival. Therefore, the term "treatment" also includes a reduction in HCC pathological outcomes. The methods disclosed herein cover any one or more of these treatment aspects.

[0024] β-glucan (BG): a natural polysaccharide widely found in yeast, fungi, algae, and grains, possessing biological activities such as immunomodulation, anti-inflammation, antioxidant, and antitumor activity. In this patent, β-glucan is used to train donors to induce trained immune properties in monocytes.

[0025] Trained immunity refers to the enhanced immune response exhibited by innate immune cells (such as monocytes or macrophages) upon exposure to specific stimuli (such as β-glucan), including increased chemotaxis, phagocytic activity, and expression of pro-inflammatory factors. This phenomenon is similar to the memory function of adaptive immunity.

[0026] Adoptive transfer: Cellular immunotherapy, which involves transferring in vitro isolated, processed, or modified immune cells (such as monocytes) into a recipient to enhance the immune response or treat a disease. This patent refers to the transplantation of β-glucan-trained monocytes into a colorectal cancer liver metastasis model.

[0027] Monocytes: Immune cells derived from bone marrow, possessing functions of phagocytosis, chemotaxis, and secretion of pro-inflammatory factors. This patent specifically refers to CD45+CD11b+ bone marrow monocytes expressing Ly6Chi, trained with β-glucan for the treatment of colorectal cancer liver metastases.

[0028] Colorectal cancer liver metastasis: The process by which colorectal cancer cells spread to the liver via the bloodstream or other routes, forming metastatic tumors. In this patent, liver metastasis is one of the leading causes of death in colorectal cancer patients, and the treatment goal is to inhibit the infiltration and proliferation of tumor cells in the liver.

[0029] Donor: In this patent, "donor" refers to the biological individual that provides bone marrow cells for the preparation of β-glucan-trained mononuclear cells. Donors are typically mammals, including but not limited to mice, rabbits, dogs, or humans.

[0030] Myeloid-derived suppressor cells (MDSCs) are a heterogeneous class of immunosuppressive cells that can inhibit T cell activation and proliferation, promoting tumor immune escape. This patent mentions that MDSCs significantly proliferate in the peripheral blood of colorectal cancer patients, and this proliferation is correlated with the degree of tumor metastasis.

[0031] Flow cytometry sorting: a technique for separating specific cell populations using flow cytometry. In this patent, bone marrow mononuclear cells expressing specific markers (such as CD45+CD11b+Ly6Chi) are obtained through flow cytometry sorting.

[0032] Chemotaxis: The ability of cells to migrate in a directed manner guided by a chemical gradient. In this patent, β-glucan-trained monocytes exhibit enhanced chemotaxis, which helps them localize in the liver and exert anti-tumor effects.

[0033] Phagocytic activity: The ability of immune cells (such as monocytes) to engulf and clear pathogens, tumor cells, or other foreign substances. In this patent, β-glucan-trained monocytes exhibit enhanced phagocytic activity.

[0034] Pro-inflammatory cytokines are signaling molecules secreted by immune cells, such as IL-1β and TNF-α, which can promote inflammatory responses and immune activation. In this patent, β-glucan-trained monocytes express higher levels of pro-inflammatory cytokines.

[0035] Liver-to-Body Weight Ratio: The ratio of liver weight to the total weight of an animal, commonly used to assess liver disease or tumor burden. In this patent, adoptive transplantation of β-glucan-trained monocytes significantly reduces this ratio, reflecting the inhibition of liver metastasis.

[0036] Immune checkpoint inhibitors: a class of drugs that enhance anti-tumor immune responses by blocking immunosuppressive signals (such as the PD-1 / PD-L1 pathway). This patent mentions their potential use in combination with β-glucan-trained monocytes to improve efficacy.

[0037] HE staining (Hematoxylin and Eosin Staining): A staining method commonly used in histological analysis. Hematoxylin stains the cell nucleus, and eosin stains the cytoplasm, used to observe tissue structure and tumor cell infiltration.

[0038] Ki67 staining: An immunohistochemical staining method used to detect Ki67 protein expression, reflecting cell proliferation activity. In this patent, it is used to assess the proliferation level of liver metastatic tumor cells.

[0039] In-Vivo Imaging: Non-invasive imaging in living animals using fluorophores (such as D-Luciferin) to monitor tumor growth or metastasis. This patent uses the IVIS Lumina II system to assess the fluorescence intensity of liver metastases.

[0040] Tail vein injection: A method of injecting drugs or cells into the body through the tail vein of a mouse. In this patent, it is used to inject trained mononuclear cells into mice for treatment.

[0041] DMEM medium: A commonly used liquid culture medium in laboratories containing nutrients necessary for cell growth. In this patent, it is used to culture cancer cells and treat bone marrow cells.

[0042] D-Luciferin: A luminescent substance that, when injected into an animal, emits fluorescence in cancer cells, allowing for tumor observation using imaging instruments. In this patent, it is used for in vivo imaging in small animals. Attached Figure Description

[0043] Figure 1 Technical flowchart; Figure 2 Representative in vivo fluorescence imaging and fluorescence intensity statistics of PBS-trained and β-glucan-trained mononuclear cells in mice on day 28 after colorectal cancer liver metastasis. Figure 3 Statistical graphs of liver appearance and liver-to-body weight ratio on day 28 after transplantation of PBS-trained monocytes and β-glucan-trained monocytes following colorectal cancer liver metastasis. Figure 4 HE and Ki67 staining images of mouse liver tissue fixed and embedded sections on day 28 after transplantation of PBS-trained monocytes and β-glucan-trained monocytes following colorectal cancer liver metastasis. Detailed Implementation

[0044] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention.

[0045] Example 1: Training mouse monocytes with β-glucan.

[0046] (1) Male C57BL / 6 mice, 6-8 weeks old, weighing about 18-21g, were divided into two groups, including a PBS treatment group and a β-glucan treatment group. β-glucan was injected intraperitoneally (1mg / mouse), and PBS was injected intraperitoneally (100ul / mouse).

[0047] (2) The mice were sacrificed and bone marrow was extracted from the PBS group and β-glucan group and their respective mononuclear cells (BM-MOs) were sorted out for later use.

[0048] Example 2: Construction of a colorectal cancer liver metastasis model MC38 colorectal cancer cells were cultured in DMEM medium and placed in a cell culture incubator under a humid environment of 37 °C and 5% CO2. When the cells reached the stationary logarithmic phase, they were digested and diluted with PBS. Under sterile conditions, 5 x 10⁵ MC38 colorectal cancer cells were aspirated using a syringe and injected into mice via the spleen to establish a mouse model of colorectal cancer liver metastasis.

[0049] Example 3: Mononuclear cell adoptive transplantation (1) Donor mice in the β-glucan or PBS group were euthanized by cervical dislocation. The leg bones from both sides of the mice were removed, and the skin and attached muscle tissue were gently removed. The bones were then immersed in sterile PBS and transferred to a biosafety cabinet. DMEM medium was added to a new grinding mortar, and the cleaned leg bones were placed in it. The bones were repeatedly ground to release the stem cells in the bone marrow into the medium. Impurities were removed by filtering through a 70-mesh filter. Then, 5 mL of medium was added to the grinding mortar again, and the process was repeated twice. The collected cell suspension was centrifuged at 1500 rpm for 5 minutes, and the supernatant was removed. 2 mL of erythrocyte lysis buffer was added to each leg bone, and the mixture was thoroughly mixed and allowed to stand for 4 minutes. After the erythrocytes were lysed, double the volume of DMEM medium containing 10% FBS was added to terminate the reaction. The mixture was centrifuged at 1500 rpm for 5 minutes, and the supernatant was discarded.

[0050] (2) Resuspend the cells in 1 mL of FACS buffer, calculate the total cell count, and add Biotin-anti-mouse CD3, Biotin-anti-mouse CD19, Biotin-anti-mouse NK1.1, and Biotin-anti-mouse Ly6G antibodies at a ratio of 1 μL of antibody per 1 × 10⁸ cells. Mix thoroughly and incubate at 4 °C for 20 minutes. Then centrifuge at 800 g for 5 minutes at 4 °C and discard the supernatant. Add magnetic beads at a ratio of 10 μL of streptavidin per 1 × 10⁸ cells and mix thoroughly. Place the sample tube on a magnetic rack and let it stand for 2 minutes. When the magnetic beads adhere tightly to the sample tube wall, aspirate the supernatant and transfer it to a new sample tube, repeating twice. Centrifuge at 800 g for 5 minutes at 4 °C, discard the supernatant, and wash once with FACS buffer.

[0051] (3) Resuspend the cells in 1 mL of FACS, then add a flow cytometry sorting antibody cocktail (containing anti-mouse CD45, anti-mouse CD11b, anti-mouse Ly6C, and Live / Dead). Mix thoroughly and incubate at 4 °C for 25 minutes. Centrifuge at 1000 g for 5 minutes at 4 °C and discard the supernatant. Resuspend the cells in 1 mL of PBS containing 2% fetal bovine serum, place on ice, and sort the CD45+CD11b+Ly6Chi bone marrow mononuclear cells using a flow cytometer. Then centrifuge at 1000 g for 5 minutes at 4 °C and discard the supernatant. Resuspend again in sterile PBS, centrifuge and discard the supernatant, repeating twice.

[0052] (4) Add an appropriate amount of sterile PBS, count the cells, and adjust the cell density to 1 × 10⁻⁶. 7 Bone marrow mononuclear cells / mL. Place the cells on ice and transfer them to the SPF operating room in sterilized tubing.

[0053] (5) On the same day after colorectal cancer metastasizes to the liver, 1 x 10 7 One mononuclear cell was adopted into a mouse with colorectal cancer liver metastases.

[0054] Example 4: Sample Collection and Statistical Analysis (1) After 28 days, mice were injected intraperitoneally with D-Luciferin (potassium luciferin) solution at a dose of 150 mg / kg body weight. The mice were then anesthetized in an air-anesthesia chamber with isoflurane. After about 10 minutes, the anesthetized mice were placed in an IVIS Lumina II small animal in vivo imaging system for imaging analysis. After obtaining the results, the fluorescence values ​​of each mouse were standardized and then analyzed as a whole.

[0055] (2) Weigh the liver and body weight, calculate the liver to body weight ratio and perform statistical analysis.

[0056] (3) Take mouse liver specimens, fix, embed and section them: Immerse the largest lobe of the liver in a solution containing 4% paraformaldehyde overnight on a shaker; then transfer the liver tissue from the sampling tube to the embedding box and rinse with running water for 6 hours to remove fixative residue; dehydrate with different concentrations of alcohol (50% ethanol for 30 minutes; 60% ethanol for 30 minutes; 70% ethanol for 30 minutes; 80% ethanol for 30 minutes; 90% ethanol for 30 minutes; 95% ethanol I and 95% ethanol II for 30 minutes each; anhydrous ethanol I and anhydrous ethanol II for 20 minutes each; xylene I and xylene II for 5 minutes each); keep at a constant temperature of 60℃, immerse in paraffin for 40 minutes and then cool and solidify; cut 4µm tissue sections using a microtome, use a glass slide to receive the flattened sections, and dry for later use.

[0057] (4) HE staining: Immerse the sections in xylene I and xylene II for 10 minutes each; hydrate (perform anhydrous ethanol I and anhydrous ethanol II for 5 minutes each; 95% ethanol I and 95% ethanol II for 3 minutes each; 90% ethanol for 3 minutes; 80% ethanol for 3 minutes; 70% ethanol for 3 minutes; 60% ethanol for 3 minutes; 50% ethanol for 3 minutes); immerse the sections in hematoxylin staining solution for 3-5 minutes, rinse thoroughly with running tap water, and observe the staining depth under a microscope; immerse the sections in eosin staining solution for 2-5 minutes; dehydrate (95% ethanol for 5 minutes, anhydrous ethanol for 5 minutes, xylene I and xylene II for 5 minutes each, and air dry the sections in a fume hood); mount the sections with neutral resin, and then acquire images using a panoramic digital slide scanner.

[0058] (5) Ki67 staining: The hydration process is the same as above; then place the slides in boiling citric acid working solution for high-temperature retrieval for 20 minutes, followed by washing with PBS three times for 5 minutes each time; use an immunohistochemistry pen to circle the tissue to be stained; add 50 μL of 3% PBS to each slide. Incubate with H2O2 at room temperature for 10 minutes, wash with PBS for 5 minutes, and repeat the washing process twice to remove endogenous alkaline phosphatase. Add 50 μL of 10% goat serum to each slide and incubate at room temperature for 30 minutes. Incubate with primary antibody overnight at 4°C. Wash with PBST working solution three times for 5 minutes each time. Add 50 μL of goat anti-rabbit secondary antibody to each slide and incubate at room temperature for 30 minutes. Wash with PBST working solution three times for 5 minutes each time. Add 50 μL of DAB working solution and incubate at room temperature for 30-60 seconds. Wash with PBST three times for 5 minutes each time. Stain the nuclei with hematoxylin and rinse thoroughly with running tap water. Dehydrate (sequentially soak in 95% ethanol for 5 minutes, anhydrous ethanol for 5 minutes, and xylene I and xylene II for 5 minutes each), and air-dry the sections in a fume hood. Add 2 drops of neutral resin and mount the slides. After air-drying, acquire images using a panoramic digital slide scanner.

[0059] The results indicated that, compared with adoptive transfer of bone marrow mononuclear cells from PBS-injected mice, adoptive transfer of bone marrow mononuclear cells from β-glucan-trained mice significantly reduced tumor fluorescence intensity in the liver region. Statistical results of liver appearance and liver weight ratio showed that the amount of liver tumor tissue decreased after receiving bone marrow mononuclear cells from β-glucan-trained mice. H&E staining and Ki67 immunohistochemical staining results showed that the invasiveness and proliferative activity of liver tumor cells in the β-glucan group were significantly reduced. These results demonstrate that β-glucan-trained mouse bone marrow mononuclear cells can effectively inhibit liver metastasis of colorectal cancer.

[0060] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The application of mononuclear cells prepared by the following method in immunocellular therapy for the prevention and / or treatment of colorectal cancer liver metastases, characterized in that, The method includes the following steps: a. Train donors using β-glucan to obtain bone marrow cells from donors; b. Flow cytometry sorts the bone marrow cells obtained in step a to obtain mononuclear cells.

2. The use of the mononuclear cells according to claim 1 in immunocellular therapy for the prevention and / or treatment of liver metastases from colorectal cancer, wherein the mononuclear cells are administered by injection.

3. The use of mononuclear cells according to claim 1 or 2 in immunocellular therapy for the prevention and / or treatment of colorectal cancer liver metastases, characterized in that, The donor is a mammal, selected from mice, rabbits, dogs, or humans.

4. The use of mononuclear cells according to any one of claims 1 to 3 in immunocellular therapy for the prevention and / or treatment of colorectal cancer liver metastases, characterized in that, The monocytes expressed Ly6C hi CD45 + CD11b + Bone marrow mononuclear cells.

5. The use of mononuclear cells according to any one of claims 1 to 4 in immunocellular therapy for the prevention and / or treatment of colorectal cancer liver metastases, characterized in that, The β-glucan is β-glucan or a composition containing β-glucan; and / or, the β-glucan is derived from yeast, fungi, algae or grains.

6. The use of mononuclear cells according to any one of claims 1 to 5 in immunocellular therapy for the prevention and / or treatment of colorectal cancer liver metastases, characterized in that, The adoptive transplantation of β-glucan-trained mononuclear cells was used to reduce the liver-to-body weight ratio in colorectal cancer metastases.

7. The use of mononuclear cells according to any one of claims 1 to 6 in immunocellular therapy for the prevention and / or treatment of colorectal cancer liver metastases, characterized in that, The adoptive transplantation of β-glucan-trained mononuclear cells was used to alleviate the infiltration and proliferation of colorectal cancer cells in the liver.

8. A therapeutic composition comprising isolated mononuclear cells induced by β-glucan.

9. The therapeutic composition according to claim 8, characterized in that, It also includes a pharmaceutically acceptable carrier, diluent, excipient and / or additive; preferably, the composition further includes a chemotherapeutic drug, immune checkpoint inhibitor or targeted drug that works synergistically with the monocytes; and / or, the therapeutic composition is administered by intravenous injection, portal vein injection, hepatic artery infusion or intraperitoneal injection; and / or, the therapeutic composition is used as adjuvant postoperative treatment, inhibition of advanced colorectal cancer liver metastases or prevention of liver metastasis recurrence.

10. The therapeutic composition according to any one of claims 8 or 9, characterized in that, The monocytes induced by this method exhibit enhanced chemotaxis, phagocytic activity, and expression of pro-inflammatory factors.

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