An immunomodulatory factor, its preparation method and its application

Abstract

This invention provides an immunomodulatory factor, its preparation method, and its application. The immunomodulatory factor includes: Eotaxin-2, I-309, IL-12p40, IL-12p70, MIG, MIP-1a, TNF-α, AR, BDNF, bFGF, EGF-R, EG-VEGF, IGFBP-6, VEGF, NAP-2, PF4, Activin A, ANG-1, DAN, Follistatin, Galectin-7, ICAM-2, etc. The immunomodulatory factor can be used to prepare immunomodulators, cancer intervention agents, etc., and is of great significance for research on the mechanisms by which thymic mitochondria regulate anti-tumor immunity, including influencing immune cell metabolism, signal transduction, apoptosis, and immune escape.

Description

Technical Field

[0001] This invention relates to an immunomodulatory factor, its preparation method, and its application, belonging to the field of biotechnology. Background Technology

[0002] The thymus is a key component of the human immune system and the primary site of T cell development. T cell precursors (thymocytes) migrating from the bone marrow to the thymus undergo positive and negative selection processes in the thymic microenvironment. Approximately 90% of thymocytes undergo apoptosis, while a small percentage acquire MHC restriction and autoimmune tolerance, maturing into naïve T cells. These naïve T cells then leave the thymus and circulate through the bloodstream to peripheral immune organs. The various cytokines and thymic peptides produced by thymic stromal cells not only regulate thymocyte differentiation and development but also have regulatory effects on peripheral immune organs and immune cells.

[0003] The reticular cells of the thymus secrete thymosin, functioning as an endocrine gland. Thymosin stimulates the growth of lymphoid tissue, promoting its development into immune-functional T cells. It also inhibits the synthesis of a substance at nerve endings that excites efferent nerves, thereby suppressing motor nerve excitation. During thymus development, autoreactive T cells bind with high affinity to the self-antigen peptide-MHC complex expressed on the surface of thymic stromal cells via their antigen receptors (TCRs), triggering negative selection, initiating programmed cell death, leading to the elimination or suppression of autoreactive T cell clones, and forming central immune tolerance to self-antigens. The thymus also promotes mast cell development, regulates the body's immune balance, and maintains immune stability. The thymus plays a crucial role in the immune system. Its function is vital for maintaining health and preventing disease; it is responsible not only for the normal development of immune function but also for developing tolerance to the body's own tissues. Changes in the thymus, especially its atrophy with age, can affect immune function and may lead to decreased immunity.

[0004] Mitochondria are organelles within cells, possessing a double-membrane structure. They produce cellular energy through oxidative phosphorylation. Mitochondria play a crucial role in regulating programmed cell death (apoptosis). They absorb and release calcium ions, helping to maintain intracellular calcium homeostasis, essential for many cellular functions such as muscle contraction and cell signaling. Mitochondria participate in fatty acid synthesis and amino acid metabolism. Involved in multiple metabolic pathways, mitochondria are central to cellular metabolism. As second messengers, they participate in cell signaling processes, responding to changes in the intracellular and extracellular environment by altering their biogenesis and function, thus participating in adaptive cellular responses. Mitochondria play an indispensable role in maintaining cellular function and life activities. Mitochondrial dysfunction is closely linked to various diseases, including neurodegenerative diseases, cardiovascular diseases, and metabolic diseases.

[0005] Therefore, its application in thymus mitochondria warrants further research. Summary of the Invention

[0006] To overcome the shortcomings of the prior art, the first objective of this invention is to provide an immunomodulatory factor that is of great significance for studying the mechanism of action of thymic mitochondria in regulating anti-tumor immunity.

[0007] The second objective of this invention is to provide bovine embryonic thymus stem cells for obtaining immunomodulatory factors.

[0008] A third objective of this invention is to provide a method for preparing the aforementioned immunomodulatory factors.

[0009] The fourth objective of this invention is to provide applications of the above-mentioned immunomodulatory factors, preparing them as immunomodulatory agents or cancer intervention and therapeutic agents, which can enhance anti-tumor immunity and substantially promote the development of new immunotherapies.

[0010] The first objective of this invention can be achieved by adopting the following technical solution: an immunomodulatory factor comprising: Eotaxin-2, I-309, IL-12p40, IL-12p70, MIG, MIP-1a, TNF-α, AR, BDNF, bFGF, EGF R, EG-VEGF, IGFBP-6, VEGF, NAP-2, PF4, Activin A, ANG-1, DAN, Follistatin, Galectin-7, ICAM-2, IL-13R1, CRP, hCGb, IGF-1R, IL-3, IL-18Rb, Leptin, MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, MMP-13, NCAM-1, NSE, Prolactin, Siglec-9, TACE, TSH, CD97, NOV, Transferrin, ACE-2, Decorin, FOLR1, Furin, and IL-17B. R, TACI, TRANCE, CTLA4, Dkk-4, IL-20, P-Cadherin, Galectin-1, LRP-6, Periostin, ADAM12, B7-H3, Cadherin-13, CD58, CD155, CD229, CystatinB, Cystatin EM, ESAM, Galectin-9, Kallikrein 5. Pref-1, TIM-3, TRAIL and ULBP-1.

[0011] Furthermore, the immunomodulatory factors include: MIG, MIP-1a, TNF RI, bFGF, EGF R, EG-VEGF, VEGF, Activin A, ANG-1, Follistatin, IL-13 R1, CRP, hCGb, IL-3, MMP-2, MMP-8, Transferrin, ACE-2, IL-17BR, TRANCE, CTLA4, ADAM12, B7-H3, Cadherin-13, CD58, ESAM, Galectin-9, Kallikrein 5, and TRAIL.

[0012] The second objective of this invention can be achieved by adopting the following technical solution: a bovine embryonic thymus stem cell, wherein the mitochondria of the bovine embryonic thymus stem cell include the immunomodulatory factors described above.

[0013] Furthermore, the bovine embryos were Holstein bovine embryos from week 3 to week 20.

[0014] The third objective of this invention can be achieved by adopting the following technical solution: a method for preparing an immunomodulatory factor, wherein mitochondria are extracted from bovine embryonic thymus stem cells, and then polypeptides are extracted from the mitochondria, the polypeptides containing the immunomodulatory factor as described above.

[0015] The fourth objective of this invention can be achieved by adopting the following technical solution: an immunomodulatory agent comprising the immunomodulatory factors as described above.

[0016] Furthermore, immunomodulators include immunomodulatory factors as described above, Cordyceps militaris, fungi, curcumin, Ganoderma lucidum, and Angelica sinensis.

[0017] Alternatively, the fourth objective of this invention can also be achieved by adopting the following technical solution: a cancer intervention therapeutic agent, which includes the immunomodulatory factors as described above.

[0018] Furthermore, the cancer is colorectal cancer.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. The immunomodulatory factor of the present invention is of great significance for the study of the mechanism by which thymic mitochondria regulate anti-tumor immunity, including its influence on the metabolism, signal transduction, apoptosis and immune escape of immune cells; 2. The bovine embryonic thymus stem cells of the present invention can be used to obtain immunomodulatory factors; 3. Application of immunomodulatory factors: Preparing immunomodulatory factors into immunomodulatory agents or cancer intervention agents can enhance anti-tumor immunity and substantially promote the development of new immunotherapies. Attached Figure Description

[0020] Figures 1-8 Images obtained from fluorescence detection; Figure 9 This is a diagram illustrating the cluster analysis of immune regulatory factors. Figure 10 Tunel staining diagram of the model group; Figure 11 This is a Tunel staining image of the experimental group. Detailed Implementation

[0021] The present invention will now be further described with reference to the accompanying drawings and specific embodiments: Example 1: 1. Source of materials: During bovine embryonic development, stem cell production occurs at a specific stage in early embryonic development. The primary stem cells are embryonic stem cells (ES cells), which appear during the blastocyst stage. The inner cell mass is the main source of ES cells; these cells are pluripotent, capable of differentiation, and ultimately develop into a complete individual. The thymus originates from the endoderm of the third and fourth pharyngeal pouches during embryonic development. These pharyngeal pouches contain components from all germ layers, thus the thymus comprises components from all three. At this stage, the thymic primordium begins to form, marking the starting point of thymic development. In the early stages of bovine embryonic development, thymic development begins with the interaction between epithelial cells from the third pharyngeal pouch region derived from the endoderm and mesenchymal cells from the neural crest. During organogenesis (E11.5-E15 days), hematopoietic stem cell lymphoid precursor cells begin to enter the thymic primordium, but at this time, the thymic epithelial cells are not yet fully capable of supporting the development and differentiation of thymic cells. The bovine thymus matures at the 20th week of embryonic development, a stage that marks the beginning of the appearance of thymic precursor cells. New Zealand Holstein Friesian embryos from weeks 3 to 20 were selected, and the embryonic thymus was isolated as the raw material for extracting mitochondria into stem cells.

[0022] 2. Extraction of mitochondrial immunomodulatory factors from bovine embryonic thymus stem cells: (1) Isolation of mitochondria from bovine embryonic thymus tissue cells: Under aseptic conditions, the extracted thymic embryonic tissue fragments were placed in a solution containing specific digestive enzymes (0.1 wt% trypsin, 0.2 wt% collagenase, pH=7.4) and gently agitated. Centrifugation was then performed to remove tissue fragments. Mitochondrial extraction buffer (e.g., 10 mM Tris-HCl, pH 7.4, 250 mM sucrose, 1 mM EDTA) was added, and the supernatant was collected after centrifugation at 1000g.

[0023] (2) Extraction of mitochondrial communication polypeptide complex: Mitochondrial extract was ultrasonically disrupted to remove debris, and the supernatant was collected. Protease was added, and the temperature was adjusted. After enzymatic hydrolysis, the temperature was adjusted to 80°C. Anhydrous citric acid (pH 2.3) was added to precipitate the peptide solution, yielding a pure peptide solution containing the target polypeptide. The concentrated peptide solution was dialyzed using a dialysis bag and freeze-dried to obtain a solid polypeptide. The purified polypeptide was aliquoted and frozen for storage at -20°C or -80°C. Peptide content was determined according to the method specified in Appendix B of GB / T 22492.

[0024] (3) Detection range of molecular weight by high performance gel electrophoresis: The extracted peptides were determined using high-performance gel filtration chromatography (GPC). This method uses porous packing material as the stationary phase and separates the sample components based on differences in their relative molecular mass. Detection is performed at a UV absorption wavelength of 220 nm for peptide bonds. Dedicated data processing software (GPC software) for determining the relative molecular mass distribution of GPC was used to process the chromatograms and data, calculating the relative molecular mass and distribution range of the peptides as follows: 0-500 dal: 26.84%; 500-2000 dal: 45.01%; 2000-5000 dal: 22.03%; >5000 dal: 6.12%.

[0025] 3. Microarray detection of mitochondrial polypeptide immunomodulatory factors in bovine embryonic thymus stem cells: Protein chip reagents used (QAH-CAA-440, RayBio® L-Series Human AntibodyArray L-1 Glass Slides, RayBiotech, Inc., USA) -Follow the standard experimental procedure: sample preparation, sample dialysis, biotin-labeled sample, complete drying of glass slides and chips, sealing and incubation; 1) Add 400µL of 1× blocking solution to each chip well and incubate on a shaker at room temperature for 1 hour to avoid generating bubbles; 2) Remove the blocking solution, add 400µL of sample to each well, one sample per array, and incubate overnight at 4°C with shaking (the labeled sample is diluted 20 times with the blocking solution before loading); 3) Remove the sample, add about 1 mL of 1× washing solution I (20× washing solution diluted with deionized water) to each well, shake at room temperature, and wash the slide 3 times, 5 min each time; 4) Remove 1× washing solution I, add 1× washing solution II and shake at room temperature. Wash the slide twice, 5 min each time. 5) Remove 1× wash solution II, add 1 mL of blocking solution to Cy3 equivalent, and then dilute 5-fold. Add 400 µL to each well and incubate at room temperature in the dark with shaking for 1 h.

[0026] 6) Wash the glass slide according to steps 3) and 4).

[0027] - Fluorescence detection: A laser scanner, such as the InnoScan 300, was used to scan the signal, employing either the Cy3 or green channel (excitation frequency = 532nm). Scanning parameters for the InnoScan 300 Microarray Scanner were: Wavelength: 532nm; Resolution: 10µm. The detected images are shown below. Figures 1-8As shown, the Cy3-labeled protein emits a fluorescent signal after binding with the antibody / antigen immobilized on the chip. Each dot represents a specific protein target, and the brighter the dot, the higher the expression level of the corresponding protein or the stronger the binding.

[0028] - Data was extracted using chip analysis software, and data analysis was performed using (AAH-BLG-8000) data analysis software.

[0029] The peptides extracted from the mitochondria of bovine embryonic thymus stem cells contain 73 immunomodulatory factors, as shown in Table 1: Table 1. Immunomodulatory Factors

[0030] Cluster analysis results as follows Figure 9 As shown, protein interaction network analysis based on 73 immunomodulatory factors in bovine embryonic thymus stem cell factors reveals the biological functional correlations among these factors, demonstrating the synergistic work, pathway formation, and ultimate driving of biological functions among them.

[0031] Data analysis revealed that immunomodulatory factors are involved in 185 immunomodulatory molecular processes, mechanisms that control and regulate the immune system's response to pathogens, tumor cells, and other foreign substances. The biological processes related to immune regulation are as follows: Negative regulation of monocyte chemotaxis Modulation of natural killer cell-mediated cytotoxicity against tumor cell targets Positive regulation of receptor binding Toll-like receptor 3 signaling pathway Positive regulation of neuroinflammatory response Active regulation of the defense response against bacteria Negative regulation of CD4-positive α-β T cell proliferation Positive regulation of humoral immune response Cells that kill another organism Monocyte chemotaxis Positive regulation of receptor signaling pathways via JAK-STAT Myeloid leukocyte migration Regulation of natural killer cell-mediated cytotoxicity Positive regulation of endothelial cell proliferation JNK Cascade White blood cell migration Negative regulation of extrinsic apoptosis signaling pathway Positive regulation of myeloid leukocyte differentiation Positive regulation of interferon-gamma production Regulation of interferon-γ production Regulation of NIK / NF-kappaB signal transduction Positive regulation of inflammatory response Positive regulation of IL-6 production Regulation of monocyte migration Positive regulation of leukocyte migration Regulation of exogenous apoptosis signaling pathways Regulation of leukocyte chemotaxis Organizational restructuring Positive regulation of tumor necrosis factor production Regulation of angiogenesis Defense response against Gram-positive bacteria Negative regulation of angiogenesis Cellular response to interferon-γ Positive regulation of ERK1 and ERK2 cascade Regulation of T cell proliferation Positive regulation of endothelial cell migration Regulation of monocyte proliferation Positive regulation of T cell proliferation Cellular response to lipopolysaccharide Regulation of endothelial cell migration Regulation of reactive oxygen species metabolism Regulation of leukocyte migration Cellular response to tumor necrosis factor Positive regulation of chemotaxis Monocyte migration Positive regulation of MAP kinase activity Positive regulation of defense response Regulation of epithelial cell differentiation Regulation of lymphocyte proliferation Regulation of IL-6 production Positive regulation of MAPK cascade Positive regulation of responses to external stimuli 14 Total 453 0.96 1.17e-07 Negative regulation of apoptosis signaling pathway Regulation of chemotaxis Regulation of epithelial cell migration Regulation of tumor necrosis factor production Regulation of epithelial cell proliferation Regulation of inflammatory response Regulation of ERK1 and ERK2 cascade Positive regulation of angiogenesis Myeloid leukocyte differentiation Chemotaxis Lymphocyte-mediated immune regulation Intercellular adhesion Angiogenesis Negative regulation of cytokine production Positive regulation of cytokine production Regulation of intercellular adhesion of leukocytes Cytokine-mediated signaling pathways MAPK Cascade Regulation of peptide hormone secretion Cell adhesion Humoral immune response Positive regulation of kappaB kinase / NF-kappaB signaling Positive regulation of peptide tyrosine phosphorylation Positive regulation of cell movement Response to lipopolysaccharide Positive regulation of intercellular adhesion of leukocytes Amoeboid cell migration Positive regulation of cell migration Leukocyte-mediated immune regulation Regulation of cytokine production Inflammatory response Negative regulation of cell adhesion Negative regulation of cell population proliferation Regulation of defense response Vascular morphogenesis Regulation of T cell activation Positive regulation of T cell activation Regulation of kappaB kinase / NF-kappaB signaling Cell migration Negative regulation of immune system processes Reaction to peptides Defense response against bacteria Intercellular adhesion via plasma membrane adhesion molecules connective tissue development Cellular response to lipids Regulation of apoptosis signaling pathway Regulating the cell's response to growth factor stimulation Gonadal development Vascular development Cellular response to abiotic stimuli Regulation of transmembrane receptor protein serine / threonine kinase signaling pathway Regulation of responses to external stimuli Regulation of immune effector processes Positive regulation of leukocyte activation Negative regulation of cell migration Negative regulation of endopeptidase activity Viral process Positive regulation of gene expression Negative regulation of multicellular biological processes The formation of anatomical structures involved in morphogenesis Regulation of leukocyte activation Leukocyte differentiation Cellular response to cytokine stimulation Regulation of lymphocyte activation Regulation of cell population proliferation Regulation of cell movement Regulation of cell migration Positive regulation of immune system processes Skeletal system development Positive regulation of protein kinase activity Positive regulation of intracellular signal transduction Positive regulation of cell population proliferation Response to cytokines Enzyme-linked receptor protein signaling pathway Transmembrane receptor protein tyrosine kinase signaling pathway hematopoietic function Positive regulation of cell adhesion Regulation of cell adhesion Negative regulation of response to external stimuli Positive regulation of signal transduction Immune response Immune system development Cell activation Development and growth Cell surface receptor signaling pathway Regulation of immune system processes Response to lipids Adaptive immune response Positive regulation of signal transmission Positive regulation of cell communication Response to exogenous stimuli Positive regulation of phosphorylation leukocyte activation Positive regulation of apoptosis Reaction to injury Positive regulation of multicellular biological processes Positive regulation of phosphate metabolism Positive regulation of cell death Reaction to oxygen-containing compounds Cellular response to oxygen-containing compounds Intercellular signal transduction Negative regulation of developmental processes Response to bacteria Cellular response to growth factor stimulation Positive regulation of response to stimuli Reaction of organic nitrogen compounds Positive regulation of protein phosphorylation Negative regulation of signal transduction Defense response to other organisms Regulation of multicellular biological processes Negative regulation of cell communication Negative modulation of signal transduction Immune system processes Negative regulation of stimulus response Regulation of multicellular organism development Negative regulation of cell death Regulation of protein kinase activity Negative regulation of cell differentiation Regulation of signal transduction Regulation of cell death Positive regulation of the development process Cell population proliferation Anatomical morphogenesis Response to endogenous stimuli Regulation of pressure response Positive regulation of cellular components and tissues Negative regulation of apoptosis Reactions to organic matter Regulation of intracellular signal transduction Regulation of programmed cell death defensive response Response to external stimuli Protein concentrations greater than LOD were selected to obtain the immunomodulators, as shown in Table 2. Table 2 Immune Regulatory Factors

[0032] Immunomodulatory effects of mitochondrial peptides in bovine thymus stem cells involve multiple signaling pathways, which play crucial roles in intercellular communication and the regulation of immune responses. The signaling pathways related to immune regulation are as follows: IL-17 signaling pathway Asthma (involving an immune response) Cytokine-cytokine receptor interactions TGF-β signaling pathway NF-κB signaling pathway Tumor necrosis factor signaling pathway Toll-like receptor signaling pathway The HIF-1 signaling pathway (which may be involved in the metabolism and function of immune cells) Natural killer cell-mediated cytotoxicity Chemokine signaling pathway MAPK signaling path PI3K-Akt signal path These signaling pathways play a central role in the activation, proliferation, and differentiation of immune cells, as well as in the initiation and regulation of immune responses. For example, the IL-17 signaling pathway is crucial in inflammation and autoimmune diseases; the NF-κB signaling pathway is a key regulator controlling immune and inflammatory responses; the Toll-like receptor signaling pathway is an important component of the innate immune response and is essential for pathogen recognition and immune responses. Chemokines signaling pathways are involved in the migration and localization of immune cells and are very important for the temporal and spatial regulation of immune responses.

[0033] Therefore, the biological functions of immunomodulatory factors are derived: immune and inflammation regulation, cell growth, repair and angiogenesis, extracellular matrix degradation and tissue remodeling, cell proliferation and apoptosis, metabolism and substance transport, signaling pathways and molecular regulation, etc. Specifically, their functions include: regulating the initiation, type, intensity and resolution of immune responses; driving cell proliferation, tissue regeneration and vascular network construction, providing the material and energy basis for repair; clearing damaged structures and activating growth factors; and constructing functional tissue structures, etc.

[0034] Example 2: Animal experiments: Anti-tumor effects of thymic stem cell mitochondrial polypeptide immunomodulatory factors: Treatment of SW620 tumor-bearing mouse models Experimental objective: To explore the therapeutic effect of the test substance's immunomodulatory factor on SW620 tumor-bearing mice.

[0035] Experimental animals: Balb / c mice, 3-4 weeks old, male, 15 mice, randomly divided into 3 groups according to tumor volume after quarantine: normal control group (n=3, no tumor cells inoculated), model group (n=6, tumor-bearing), experimental group (n=6, tumor-bearing). Modeling method: SW620 colorectal cancer cells in logarithmic growth phase were taken, and the cell ratio was adjusted to 5×106 / 0.1mL. The cells were then subcutaneously in the shoulder and back of 15 Balb / c nude mice. Intervention: The experimental group was given 1 mL / 100g body weight of immunomodulatory factor by gavage daily (concentration of 1g / mL), and drinking water was replaced with drinking water peptide powder (30g of immunomodulatory factor was dissolved in 200mL of drinking water); the normal control group and the model group were given pure water by gavage and drinking water, and were fed normally for 14 days.

[0036] result: 5.1 Body weight (g) is shown in Table 3: Table 3: Weight data (g) for days 0, 3, 6, 9, 11, 14, 17, 20, and 22.

[0037] 5.2 Tumor volume (mm) 3 As shown in Table 4: Table 4. Tumor volume data (mm) for days 0, 3, 6, 9, 11, 14, 17, 20, and 22. 3 )

[0038] 5.3 Tunel staining of tumor tissue: The positive cell rate in the model group (N=6) was 24.91%, and the TUNEL staining pattern (200×) is shown below. Figure 10 As shown; the positive cell rate in the experimental group (N=6) was 71.38%, and the TUNEL staining image (200×) is shown below. Figure 11 As shown.

[0039] Daily gavage administration of immunomodulatory factors for 3 weeks showed a significant therapeutic effect on SW620 tumor-bearing nude mice, with a tumor inhibition rate of 41.2%. After TUNEL staining of the tumors, the tumor cell apoptosis rate of 71.38% was significantly higher than that of the model group (24.91%).

[0040] The research concept of this invention is as follows: Based on existing research on the thymus and mitochondria, and considering that the possible mechanisms by which thymus and mitochondria regulate antitumor immunity involve multiple aspects, such as mitochondrial dynamics, including mitochondrial fusion and fission, which are crucial for the polarization and function of immune cells, this process may influence antitumor immune responses because it can regulate the activation and effector functions of immune cells. Mitochondria play a central role in T cell activation and proliferation. Mitochondrial biosynthesis and proteomic remodeling promote the one-carbon metabolism required for T cell activation. This means that the metabolic state of mitochondria directly affects the ability of T cells to respond to tumor antigens. Reactive oxygen species (ROS) produced by mitochondria are important signaling molecules that can affect T cell activation and function. ROS can act as second messengers, participating in the transmission of immune signals, thereby regulating antitumor immune responses. mtDNA can act as damage-associated molecular patterns (DAMPs), activating immune responses through the cGAS-STING pathway and promoting the production of type I interferon, thereby enhancing antitumor immunity. Mitochondria may also regulate antitumor immunity by influencing the expression and function of immune checkpoint molecules such as PD-L1. For example, mitochondrial oxidative stress can affect PD-L1-mediated immune escape through the STING-IFN signaling pathway. Mitochondria play a crucial role in regulating T cell metabolism, and metabolic reprogramming is essential for T cell fate and function. Alterations in mitochondrial metabolism may affect T cell recognition and killing capabilities. Mitochondria also participate in regulating apoptosis, affecting the balance between immune cell survival and death. By modulating apoptotic pathways, mitochondria may influence the stability and function of immune cells in the tumor microenvironment. Mitochondria may participate in tumor immune escape through multiple mechanisms, including regulating oxidative stress and the function of immunosuppressive cells in the tumor microenvironment. Thymic mitochondria regulate antitumor immunity through multiple mechanisms, including affecting immune cell metabolism, signal transduction, apoptosis, and immune escape. Therefore, thymic mitochondria have a substantial role to play in the development of novel antitumor immunotherapy strategies.

[0041] Example 3: The immunomodulatory factors listed in Table 2 are combined with traditional Chinese medicine ingredients to prepare preparations that enhance immunoglobulin activity. Examples of traditional Chinese medicine ingredients include Cordyceps militaris, fungi, curcumin, Ganoderma lucidum, and Angelica sinensis. Excipients such as spirulina and resistant dextrin can also be added. These preparations are beneficial for vulnerable populations with weakened immune systems, post-operative recoveries, cancer patients, those with weak constitutions, and those exposed to special occupational environments. They promote the proliferation and activation of immune cells, provide crucial nutrients to immune organs, build an immune defense barrier, and help maintain the normal development of the immune system.

[0042] For those skilled in the art, various other corresponding changes and modifications can be made based on the technical solutions and concepts described above, and all such changes and modifications should fall within the protection scope of the claims of this invention.

Claims

1. An immunomodulatory factor, characterized in that, The immune regulatory factors include: Eotaxin-2, I-309, IL-12p40, IL-12p70, MIG, MIP-1a, TNF RI, AR, BDNF, bFGF, EGF R, EG-VEGF, IGFBP-6, VEGF, NAP-2, PF4, Activin A, ANG-1, DAN, Follistatin, Galectin-7, ICAM-2, IL-13 R1, CRP, hCGb, IGF-1R, IL-3, IL-18 Rb, Leptin, MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, MMP-13, NCAM-1, NSE, Prolactin, Siglec-9, TACE, TSH, CD97, NOV, Transferrin, ACE-2, Decorin, FOLR1, Furin, IL-17B R, TACI, TRANCE, CTLA4, Dkk-4, IL-20, P-Cadherin, Galectin-1, LRP-6, Periostin, ADAM12, B7-H3, Cadherin-13, CD58, CD155, CD229, Cystatin B, Cystatin EM, ESAM, Galectin-9, Kallikrein 5. Pref-1, TIM-3, TRAIL and ULBP-1.

2. The immunomodulatory factor as described in claim 1, characterized in that, The immunomodulatory factors include: MIG, MIP-1a, TNF-α, bFGF, EGF R, EG-VEGF, VEGF, Activin A, ANG-1, Follistatin, IL-13 R1, CRP, hCGb, IL-3, MMP-2, MMP-8, Transferrin, ACE-2, IL-17BR, TRANCE, CTLA4, ADAM12, B7-H3, Cadherin-13, CD58, ESAM, Galectin-9, Kallikrein 5, and TRAIL.

3. A bovine embryonic thymus stem cell, characterized in that, The mitochondria of the bovine embryonic thymus stem cells include the immunomodulatory factors as described in claim 1.

4. The bovine embryonic thymus stem cells as described in claim 3, characterized in that, The bovine embryos in question are Holstein bovine embryos from week 3 to week 20.

5. A method for preparing an immunomodulatory factor, characterized in that, Mitochondria were extracted from bovine embryonic thymus stem cells as described in claim 3, and then polypeptides were extracted from the mitochondria, the polypeptides containing the immunomodulatory factor as described in claim 1.

6. An immunomodulatory agent, characterized in that, The immunomodulatory agent includes the immunomodulatory factor as described in claim 1.

7. The immunomodulatory agent as described in claim 6, characterized in that, The immunomodulatory agents include the immunomodulatory factors as described in claim 1, Cordyceps militaris, fungi, curcumin, Ganoderma lucidum, and Angelica sinensis.

8. A cancer interventional therapeutic agent, characterized in that, The cancer interventional therapeutic agent includes the immunomodulatory factor as described in claim 1.

9. The cancer interventional therapeutic agent as described in claim 8, characterized in that, The cancer in question is colorectal cancer.

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