Therapeutic allergic rhinitis huc-msc cell suspension formulation and preparation method

Abstract

This application provides a hUC-MSC cell suspension formulation for treating allergic rhinitis and its preparation method. It comprises mammalian stem cells, a pharmaceutically acceptable suspension carrier, and a protective agent. The mammalian stem cells are mesenchymal stem cells, which can be derived from the human umbilical cord. The formulation is suitable for intravenous administration. The preparation method includes: washing the resuscitated stem cells with phosphate buffer, centrifuging, resuspending them in a suspension carrier, adding the suspension carrier and protective agent, and mixing thoroughly. Experiments show that this formulation can reduce the levels of interleukin-4 and interleukin-5 in the serum and spleen of mice with allergic rhinitis, and alleviate inflammatory cell infiltration in the nasal mucosa.

Description

Technical Field

[0001] This application belongs to the field of biopharmaceutical manufacturing technology, specifically relating to hUC-MSC cell suspension preparations for treating allergic rhinitis and their preparation methods. Background Technology

[0002] Allergic rhinitis (AR) is an IgE-mediated, non-infectious, chronic inflammatory disease of the nasal mucosa. Its global prevalence continues to rise, making it a common chronic respiratory disease affecting public health. Conventional clinical treatment primarily involves antihistamines, nasal corticosteroids, leukotriene receptor antagonists, and allergen avoidance. While these can control symptoms in mild to moderate cases to some extent, refractory allergic rhinitis patients often exhibit poor response to conventional medications, recurrent symptoms, and persistent damage to the nasal mucosal barrier. This is accompanied by severe nasal congestion, runny nose, nasal itching, sneezing, and can even trigger complications such as asthma, sinusitis, and otitis media, severely impacting patients' quality of life, sleep, and cognitive function. A safe and effective radical cure is currently lacking in clinical practice.

[0003] Current treatments for allergic rhinitis have many limitations: traditional medications only provide symptomatic relief and cannot correct immune imbalances or repair damaged nasal mucosa; long-term use carries risks of local irritation, drug resistance, and systemic adverse reactions. Specific immunotherapy has a long treatment cycle, narrow indications, and slow onset of action; some patients have poor compliance, and there is a risk of exacerbating allergies. Surgical treatment is only suitable for patients with anatomical abnormalities and cannot address the root causes of allergies and inflammation. Therefore, developing novel therapeutic agents that combine immune modulation, mucosal repair, and long-acting anti-inflammatory effects is a core unmet clinical need for refractory allergic rhinitis.

[0004] Mesenchymal stem cells possess biological characteristics such as multi-lineage differentiation, paracrine function, immunomodulation, and tissue repair. They have shown great application potential in the fields of inflammatory diseases and tissue damage repair by regulating immune cell homeostasis, secreting anti-inflammatory and repair factors, and rebuilding the mucosal barrier. However, stem cell preparations alone have limitations such as short in vivo survival time, weak local colonization ability, and insufficient anti-inflammatory and repair efficiency, which restrict their efficacy in the treatment of rhinitis through local nasal administration.

[0005] In summary, developing a stem cell compound preparation that is scientifically formulated, easy to prepare, highly safe, and has a definite curative effect, can be administered intravenously, and can simultaneously achieve immune regulation, mucosal repair, and rapid anti-inflammatory effects, is of great significance for overcoming the bottleneck in the clinical treatment of refractory allergic rhinitis. Summary of the Invention

[0006] This application addresses the shortcomings of existing medications for refractory allergic rhinitis, which only provide symptomatic relief, fail to correct immune imbalances, and are ineffective in repairing nasal mucosal damage. It provides an hUC-MSC cell suspension formulation and preparation method for treating allergic rhinitis, fundamentally correcting immune imbalances, rebuilding the nasal mucosal barrier, and inhibiting local inflammation. By using mesenchymal stem cells as the core, combined with pharmaceutically acceptable excipients, and through a triple-action pathway of immune regulation, mucosal repair, and anti-inflammatory suppression, it fundamentally blocks allergic reactions, repairs the nasal mucosal barrier, and inhibits local inflammation, achieving long-term treatment and rapid symptom relief for refractory allergic rhinitis.

[0007] This application provides, on the one hand, a hUC-MSC cell suspension formulation for treating allergic rhinitis, wherein the cell suspension formulation comprises mammalian stem cells and a pharmaceutically acceptable suspension carrier and a protective agent.

[0008] Preferably, the mammalian stem cells are mesenchymal stem cells.

[0009] Preferably, the mesenchymal stem cells are derived from human umbilical cord mesenchymal stem cells.

[0010] Preferably, the suspension carrier is physiological saline.

[0011] Preferably, the protective agent is human serum albumin.

[0012] Preferably, the cell suspension preparation is suitable for intravenous injection.

[0013] This application also provides a method for preparing a hUC-MSC cell suspension formulation for treating allergic rhinitis, wherein the preparation method is as follows:

[0014] (1) Thaw and resuscitate mammalian stem cells, wash them 2-3 times with phosphate buffer, collect the cell pellet by centrifugation, add a suspension carrier to resuspend the cells, and obtain a stem cell suspension.

[0015] (2) Add a protective agent to the stem cell suspension and mix well to obtain a cell suspension preparation.

[0016] Preferably, the pH of the phosphate buffer in step (1) is 7.2 to 7.4.

[0017] This application also provides the use of any of the described cell suspension formulations and the cell suspension formulations prepared by any of the described methods in the preparation of drugs for treating refractory allergic rhinitis.

[0018] The beneficial effects of the embodiments in this application are as follows:

[0019] The essence of allergic rash (AR) is the overactivation of the Th2 immune response, disrupting the balance between Th1 / Th2 and Th17 / Treg cells, leading to a massive production of IgE. MSCs, through paracrine factors and direct contact regulation, inhibit Th2 and ILC2 activation and reduce IL-4 / IL-5 pro-inflammatory factors; simultaneously, they promote Treg cell proliferation and dendritic cell-induced immune tolerance, reducing serum IgE and eosinophil infiltration, thus blocking the amplifying cycle of allergies at its source. AR patients often experience epithelial damage, edema, congestion, and goblet cell hyperplasia in the nasal mucosa. MSCs secrete repair factors such as EGF, FGF, and HGF, promoting the proliferation and differentiation of nasal mucosal epithelial cells, accelerating ciliary structure regeneration, repairing vascular and glandular function, and rebuilding the physical barrier to reduce allergen adhesion and penetration, relieving symptoms such as nasal congestion and runny nose. MSCs also rapidly reduce local inflammation levels and quickly relieve acute attacks such as sneezing and nasal itching by inhibiting the NF-κB pathway, reducing mast cell degranulation and histamine release, and regulating macrophage polarization (promoting the M2 anti-inflammatory phenotype).

[0020] The mesenchymal stem cells in the cell suspension formulation of this application can regulate the Th1 / Th2 immune imbalance and improve the Treg cell functional defects in an allergic rhinitis model through paracrine and cell contact pathways. Experimental results showed that the levels of pro-inflammatory factors interleukin-4 (IL-4) and interleukin-5 (IL-5) in the serum and spleen of model mice were significantly reduced.

[0021] Reduces nasal mucosal inflammation and damage: In an ovalbumin-induced allergic rhinitis mouse model, after administration of the formulation of this application, histopathological examination of the nasal mucosa showed reduced infiltration of inflammatory cells and better mucosal structural integrity than the control group. The formulation of this application can alleviate local inflammation and promote mucosal repair. Attached Figure Description

[0022] Figure 1 This is a bar chart showing the interleukin-4 data in the serum of each group in the experimental cases of this application.

[0023] Figure 2 This is a bar chart showing the interleukin-5 data in the serum of each group in the experimental cases of this application. Detailed Implementation

[0024] The technical solutions of this application will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments in this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0025] Example 1

[0026] Umbilical cord mesenchymal stem cell culture

[0027] (1) Preparation of serum-free complete culture medium for umbilical cord mesenchymal stem cells: Take the culture medium additive out of the -20°C freezer, thaw it at 37°C, add it to the basal culture medium and mix thoroughly to obtain the complete culture medium; the complete culture medium should be prepared fresh and used up within 2 weeks (each time it is taken out, the time exposed to air should be minimized, and the container should be sealed in time after taking it out to avoid rapid pH increase); the complete culture medium should be preheated at room temperature for 10-30 minutes before use, and the time should not be too long, and strong light and ultraviolet radiation should be avoided.

[0028] (2) Umbilical cord collection and transportation: One cesarean section umbilical cord (from the seed bank) was collected under aseptic conditions, placed in an umbilical cord collection bottle containing physiological saline, and transported to the laboratory at 2-8℃ (time controlled within 24-48h).

[0029] (3) Primary isolation and culture of umbilical cord mesenchymal stem cells (tissue block adherence method)

[0030] 1) Place the umbilical cord in a sterile culture dish and wash it 2-3 times with physiological saline. Soak it in 75% sterile ethanol for 10-20 seconds, then immediately transfer it to a culture dish containing physiological saline and wash it twice. Cut the umbilical cord into several pieces of about 2-3 cm and add physiological saline to wash away the bloodstains.

[0031] 2) Remove the two arteries and one vein of the umbilical cord following a spiral pattern. Use tissue forceps to separate the Wharton's jelly located between the amnion and the blood vessels, transfer it to a 50ml centrifuge tube, add 1mL of prepared complete culture medium, and cut the jelly into 3mm pieces using sterile surgical scissors. 3 Divide the tissue into two pieces. Add complete culture medium to the centrifuge tube, select the centrifugation setting, and centrifuge at 2000 rpm for 5 minutes.

[0032] 3) Inoculate approximately 0.5g of tissue block into each T75 culture flask, ensuring it is evenly distributed at the bottom of the flask; after incubating at 37℃ for 1-2 hours, gently add 6-8mL of cell culture medium, taking care not to allow the tissue block to float.

[0033] 4) Add 5 ml of culture medium on days 2-3; change the medium on days 7-10, discard the supernatant, and add 10-15 ml of fresh culture medium; observe cell expansion on days 12-14 and perform digestion and passage.

[0034] 5) The earliest time for cells to emerge is 5-7 days, and a small number of scattered cells can be observed under a microscope.

[0035] (4) Passaging of umbilical cord mesenchymal stem cells (T75 flasks)

[0036] 1) After culturing cells for approximately 72 hours, remove the cells for observation. When cell confluence reaches 80-90%, they are ready for passage.

[0037] 2) Digestion: Discard the culture supernatant. Wash each T75 culture flask with physiological saline, add 2 mL of trypsin working solution, and digest for 3-5 min. Observe under a microscope that most of the cells have detached. Add 4 mL of complete culture medium to stop the digestion. Centrifuge the cell suspension at 1000 rpm for 10 min and discard the supernatant.

[0038] 3) Resuspend the cells in complete culture medium and count them using a cell counter. Adjust the cell density by adding more complete culture medium based on the counting results, aiming for 6000–7000 cells / cm³. 2 Cells were seeded at a density of 37℃ ± 0.5℃ and then placed in a carbon dioxide constant temperature and humidity incubator for culture at a carbon dioxide volume fraction of 5% ± 0.2%.

[0039] (5) Cryopreservation of umbilical cord mesenchymal stem cells: Digest the cells to be cryopreserved with trypsin working solution and terminate digestion with complete culture medium; centrifuge at 1000 rpm for 10 min, remove the supernatant, resuspend in culture medium and count; dilute the cells to a certain density (twice the cryopreservation density) with complete culture medium according to the cell count, and then slowly add an equal volume of pre-cooled serum-free cryopreservation solution; mix gently, aliquot the cells into cryopreservation tubes or cryopreservation bags, and cryopreserve using a programmed cooling box or programmed cooling instrument.

[0040] (6) Resuscitation and culture of umbilical cord mesenchymal stem cells (T75 flask): Add pre-chilled complete culture medium (2-8℃) to centrifuge tubes. Remove the cryopreservation tubes from the liquid nitrogen tank and immediately immerse them in 37℃±0.5℃ warm water, gently shaking to thaw until a small ice crystal remains. Transfer the cells to centrifuge tubes containing pre-chilled complete culture medium, centrifuge at 4℃, 1000 rpm for 10 min, and discard the supernatant. Resuspend the cells in complete culture medium, mix well, and count. Based on the counting results, centrifuge at 7000-8000 cells / cm³. 2 Cells were seeded at a density of 37℃ ± 0.5℃ and then placed in a carbon dioxide constant temperature and humidity incubator for culture at a carbon dioxide volume fraction of 5% ± 0.2%.

[0041] (7) Select low-passage umbilical cord mesenchymal stem cells in the logarithmic growth phase with a cell confluence of 80%–90%. Discard the culture supernatant and wash the cell monolayer 1–2 times with sterile physiological saline to remove residual culture medium and metabolic impurities. Add 2 mL of trypsin working solution to each T75 culture flask and gently digest at 37°C for 3–5 min. After observing the cell shrinkage and detachment from the flask wall under a microscope, add an equal volume of serum-free complete culture medium to terminate the digestion. Gently and repeatedly pipette the bottom of the culture flask to collect a uniform single-cell suspension, transfer it to a sterile centrifuge tube, centrifuge at 1000 rpm for 10 min, and discard the supernatant. Resuspend the cell pellet with pharmaceutical-grade isotonic buffer or cell preparation preservation solution, centrifuge and wash again, and collect the cell pellet to obtain human umbilical cord mesenchymal stem cells.

[0042] Example 2

[0043] Human umbilical cord mesenchymal stem cell suspension preparation

[0044] (1) Stem cell resuscitation: Take the cryopreservation tube of human umbilical cord mesenchymal stem cells from Example 1, thaw it rapidly in a 37°C water bath, and immediately transfer it to a clean bench after it is completely thawed.

[0045] (2) Cell washing: Transfer the cell suspension to a sterile centrifuge tube, add 4 volumes of pH 7.2 PBS, and gently pipette to mix; centrifuge at 1000 r / min for 6 min, discard the supernatant; repeat the washing 3 times and collect the pure cell pellet.

[0046] (3) Preparation of stem cell suspension: Resuspend cells in Ringer's solution, count cells using a cell counter, and adjust the cell density to 1.5 × 10⁻⁶. 10 Human umbilical cord mesenchymal stem cell suspension was obtained by using cells per L.

[0047] (4) Addition and mixing of excipients: Add in sequence: physiological saline (final concentration 0.9%), protective agent (add 5% of 20% human serum albumin per 100ml).

[0048] (5) Mixing and dispensing: Gently invert and mix for 15 minutes under sterile conditions to avoid generating air bubbles; dispense into intravenous injection bottles and store in a sealed, light-proof container.

[0049] (6) Quality test: Cell viability 96%, no turbidity, no precipitation, meeting the standards for intravenous injection.

[0050] Example 3

[0051] Human umbilical cord mesenchymal stem cell suspension preparation

[0052] (1) Stem cell thawing: Take the cryopreservation tube of human umbilical cord mesenchymal stem cells from Example 1 and place it in a 37°C water bath to thaw it completely.

[0053] (2) Cell washing: Wash three times with PBS at pH 7.4, centrifuge at 850 r / min for 5 min, discard the supernatant and collect the cell pellet.

[0054] (3) Preparation of stem cell suspension: Cells were resuspended in physiological saline and the cell density was adjusted to 1.5 × 10⁻⁶. 10 per L.

[0055] (4) Addition of excipients: Add physiological saline (final concentration 0.9%) and protectant (add 5% of 20% human serum albumin per 100ml).

[0056] (5) Mixing and dispensing: Stir for 12 minutes under sterile conditions, filter aseptically, and dispense into sterile preparation bottles.

[0057] (6) Quality testing: Cell viability is 95%, uniform and free of precipitation, suitable for intravenous injection administration.

[0058] Comparative Example 1

[0059] Mouse adipose-derived mesenchymal stem cell suspension

[0060] (1) Isolation and Culture: Ten male KM mice aged 6-8 weeks, each weighing 20-25g, were purchased from the Hebei Provincial Experimental Animal Center. After euthanasia by cervical dislocation, epididymal and perigroin adipose tissue were collected. The tissue was washed with PBS containing streptomycin and penicillin, minced with ophthalmic scissors, digested with type I collagenase, and then mixed with PBS. After filtration and centrifugation, the supernatant was discarded. The precipitate was washed with PBS to remove collagenase. The cells were resuspended in DMFM / F12 medium containing 10% fetal bovine serum and counted. After adjusting the cell density, the cells were seeded into 25 mL culture flasks and cultured in a 37 ℃, 5% CO2 incubator. The medium was changed for the first time after 24 h, and then every two to three days. When the cells reached 80% confluence, they were digested with trypsin and passaged. Human umbilical cord mesenchymal stem cells of passages P3-5 were collected, centrifuged and the supernatant was discarded. The cells were resuspended in buffer solution, and 100 μL of the cell suspension was added to Eppendorf tubes. Antibody was added and the cells were incubated in the dark for 30 hours. After a period of time, add buffer solution, centrifuge and discard the supernatant, then resuspend in buffer solution. When the cells reach 80% confluence, perform subculture. After subculture, the cells exhibit a uniform elongated spindle shape and accelerated growth rate. Wash, digest, and resuspend, then incubate with CM-Dil. Wash with PBS and resuspend in PBS, adjusting the concentration to 1.5 × 10⁻⁶. 10 A suspension of mouse adipose-derived mesenchymal stem cells was obtained by merging 1 / L of mesenchymal stem cell suspension.

[0061] (2) Addition and mixing of excipients: Add potassium chloride regulator (final concentration 0.3%) and protectant (add 5% of 20% human serum albumin per 100ml) in sequence.

[0062] (3) Mixing and dispensing: Gently invert and mix for 15 minutes under sterile conditions to avoid generating air bubbles; dispense into intravenous injection bottles and store in a sealed, light-proof container.

[0063] Comparative Example 2

[0064] Human umbilical cord mesenchymal stem cell suspension (without protectant)

[0065] (1) Stem cell thawing: Take the cryopreservation tube of human umbilical cord mesenchymal stem cells prepared in step 2, place it in a 37°C water bath, and thaw it quickly and completely.

[0066] (2) Cell washing: Wash three times with PBS at pH 7.4, centrifuge at 850 r / min for 5 min, discard the supernatant and collect the cell pellet.

[0067] (3) Preparation of stem cell suspension: Cells were resuspended in physiological saline and the cell density was adjusted to 1.5 × 10⁻⁶. 10 per L.

[0068] (4) Addition of excipients: Add physiological saline (final concentration 0.9%) and the same amount of deionized water as the protective agent in Comparative Example 2.

[0069] (5) Mixing and dispensing: Stir for 12 minutes under sterile conditions, filter aseptically, and dispense into intravenous injection bottles.

[0070] Test case

[0071] Performance verification test

[0072] (1) Experimental materials: 60 KM mice aged 6-8 weeks, weighing 20-25 g, of any sex, were used for animal modeling. All mice were purchased from the Hebei Provincial Experimental Animal Center.

[0073] (2) Experimental methods:

[0074] 1) Preparation of modeling and sensitization solution:

[0075] Preparation of sensitizing and activating solutions:

[0076] Preparation of sensitization solution: Weigh 2 mg of ovalbumin V and add it to 4 mL of PBS to completely dissolve it and prepare a 500 mg / L ovalbumin solution; weigh an appropriate amount of alum and add it to an appropriate amount of PBS to completely dissolve it and prepare a 10% alum solution; add 4 mL of alum solution to the ovalbumin solution and mix thoroughly, incubate at room temperature for 60 min, centrifuge and discard the supernatant, resuspend the precipitate in PBS to 4 mL and mix thoroughly.

[0077] Preparation of stimulation solution: Weigh 40 mg of ovalbumin II, dissolve it in 1 mL of PBS, and mix thoroughly.

[0078] 2) Grouping:

[0079] Example 2 group: 0.1 mL of Example 2 suspension was injected via tail vein;

[0080] Example 3 group; 0.1 mL of Example 3 suspension was injected via tail vein;

[0081] Comparative Example 1: 0.1 mL of Comparative Example 1 suspension was injected via the tail vein;

[0082] Comparative Example 2: 0.1 mL of Comparative Example 2 suspension was injected via the tail vein;

[0083] Normal control group: 0.1 mL PBS solution was injected via tail vein;

[0084] Adaptive rhinitis model group: 0.1 mL PBS solution was injected via tail vein; the injection volume was 1×10⁻⁶ for a single administration. 6 ~2×10 6 Cells / kg, cycle is every 10 days / time, 3 consecutive times constitute one course of treatment, 2 courses / person, dilution concentration conversion.

[0085] Example 2, Example 3, Comparative Example 1, and Comparative Example 2 were sensitized by intraperitoneal injection of 200 μL of ovalbumin sensitization solution at the same time on days 0, 7, and 14. From days 15 to 19, each animal was sensitized by 20 μL of ovalbumin stimulation solution (10 μL per nostril) daily. The normal control group received no treatment.

[0086] 2) Modeling and Validation: Based on the observation of general symptoms in mice, from the first day of modeling, the behavior, morphology, and feeding of mice were observed. After each nasal stimulation, the mice were observed to scratch their noses, sneeze, and have clear nasal discharge. The total score was calculated, and a total score of more than 5 points indicated successful modeling. The control indicators are shown in Table 1. All modeled mice showed symptoms of allergic rhinitis, and all scores were >5 points, indicating successful modeling. The normal control group did not show any allergic rhinitis symptoms, proving that the modeling was successful.

[0087]

[0088] (3) Experimental results:

[0089] 1) Detection indicators:

[0090] Detection of interleukin-4 and interleukin-5 in mouse serum: 48 h after the last treatment, peripheral blood was collected by enucleation after anesthesia of mice in each group. A small amount of serum was centrifuged and protein quantification was performed. The remaining serum was used to determine the levels of interleukin-4 and interleukin-5 by ELISA.

[0091] Detection of interleukin-4 and interleukin-5 levels in mouse spleen: After euthanizing mice in the above groups by enucleating their eyeballs and collecting peripheral blood, the spleen was removed, total RNA was extracted, and 2 μg of RNA was used for reverse transcription. The expression of interleukin-4 and interleukin-5 was detected by fluorescent PCR.

[0092] Pathological examination: Mice were sacrificed 48 h after the last treatment, and nasal respiratory mucosa was taken, fixed in formaldehyde, dehydrated, embedded in paraffin, sectioned, and then routinely stained with hematoxylin and eosin. The infiltration of inflammatory cells was observed under a light microscope.

[0093] The main observation indicators were: serum interleukin-4 and interleukin-5 levels; spleen interleukin-4 and interleukin-5 levels; migration of mesenchymal stem cells in the nasal mucosa and pathological changes in the nasal mucosa.

[0094] Statistical analysis: All data were processed, and measurement data were analyzed using... ±s indicates that the comparison between groups was performed using one-way ANOVA, and P < 0.05 was considered statistically significant.

[0095] 2) Detection data: As shown in Table 2, the serum interleukin-4 data for each group are as follows: Figure 1 As shown, the interleukin-5 data are as follows: Figure 2 As shown;

[0096] Table Note: Compared with the adaptive rhinitis model group, a P < 0.05.

[0097] Through Table 2, Figure 1 and Figure 2 get:

[0098] Serum inflammatory factor levels: IL-4 and IL-5. The data of the Example 2 and Example 3 groups were significantly lower than those of the model group and close to those of the normal control group. Although the data of the comparative proportion group decreased, the effect was significantly worse than that of the Example group.

[0099] Splenic inflammatory factor mRNA expression: IL-4 and IL-5 mRNA: The data of the Example 2 and Example 3 groups were significantly lower than those of the model group and close to those of the normal control group. Although the data of the comparative group decreased, the effect was significantly worse than that of the Example group.

[0100] Pathological damage: In Examples 2 and 3, the infiltration of inflammatory cells in the nasal mucosa was significantly reduced, and the mucosal structure was more intact; in Comparative Examples 1 and 2, the inflammatory infiltration was less severe than in the model group, but more severe than in the Example groups.

[0101] The human umbilical cord mesenchymal stem cell suspension was far more effective than the comparative group of mouse adipose mesenchymal stem cell suspension in reducing pro-inflammatory factors, increasing anti-inflammatory factors, and alleviating nasal mucosal inflammation.

[0102] The immune imbalance between Th1 and Th2 cells is regulated by multiple factors. The Th1 / Th2 cell imbalance theory posits that allergic rhinitis is a nasal allergic reaction caused by the shift in the immune response of Th2 cells, leading to the release of cytokines and inflammatory mediators. Therefore, preventing the shift in the Th2 response and restoring the Th1 / Th2 immune balance has become a new approach to treating allergic rhinitis. Treg cells promote dendritic cell maturation, inhibit T lymphocyte proliferation, and regulate the Th1 / Th2 cell balance. In patients with allergic rhinitis, Treg cell deficiency is the main factor causing their immune imbalance. Allergic rhinitis is an inflammatory disease caused by Th2 immune imbalance and Treg cell deficiency. Mesenchymal stem cells are distributed in tissues such as bone marrow and adipose tissue, and possess the potential for self-renewal, differentiation, and proliferation. Adipose-derived mesenchymal stem cells and bone marrow mesenchymal stem cells have the ability to differentiate into multiple lineages, proliferate stably, and promote repair. They can differentiate into hematopoietic cells, skeletal muscle cells, nerve cells, and cardiomyocytes, and have the functions of reducing fibroblast apoptosis, promoting proliferation, and participating in angiogenesis. It has immunosuppressive effects. The study investigated its effects on T-cell immunity in mice with allergic rhinitis. The results showed that human umbilical cord mesenchymal stem cells can reduce the levels of interleukin-4 and interleukin-5 in serum, thereby regulating the Th1 / Th2 imbalance. The pathological sections of mice showed mild damage to the nasal mucosal epithelium and little eosinophil infiltration.

[0103] In summary, the cell suspension formulation described in this application can fundamentally correct immune imbalance, repair the nasal mucosal barrier, and rapidly inhibit local inflammation through stem cell immunomodulation, providing a safe and effective innovative treatment strategy for refractory allergic rhinitis, and has clear clinical application prospects and industrialization value.

[0104] Although embodiments of this application have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of this application. All equivalent changes and improvements made within the scope of this application shall still fall within the patent coverage of this application.

Claims

1. A hUC-MSC cell suspension preparation for treating allergic rhinitis, characterized in that, The cell suspension formulation contains mammalian stem cells and a pharmaceutically acceptable suspension carrier and protectant.

2. The hUC-MSC cell suspension preparation for treating allergic rhinitis as described in claim 1, characterized in that, The mammalian stem cells mentioned are mesenchymal stem cells.

3. The hUC-MSC cell suspension preparation for treating allergic rhinitis as described in claim 2, characterized in that, The mesenchymal stem cells are derived from human umbilical cord mesenchymal stem cells.

4. The hUC-MSC cell suspension preparation for treating allergic rhinitis as described in claim 1, characterized in that, The suspension carrier is physiological saline.

5. The hUC-MSC cell suspension preparation for treating allergic rhinitis as described in claim 1, characterized in that, The protective agent is human serum albumin.

6. The hUC-MSC cell suspension preparation for treating allergic rhinitis as described in claim 1, characterized in that, The cell suspension preparation is suitable for intravenous injection.

7. The method for preparing the hUC-MSC cell suspension formulation for treating allergic rhinitis as described in any one of claims 1-6, characterized in that, The preparation method is as follows: (1) Thaw and resuscitate mammalian stem cells, wash them 2-3 times with phosphate buffer, collect the cell pellet by centrifugation, add a suspension carrier to resuspend the cells, and obtain a stem cell suspension. (2) Add a protective agent to the stem cell suspension and mix well to obtain a cell suspension preparation.

8. The method for preparing the hUC-MSC cell suspension formulation for treating allergic rhinitis as described in claim 7, characterized in that, The pH of the phosphate buffer in step (1) is 7.2 to 7.

4.

9. The use of the cell suspension formulation according to any one of claims 1-6, and the cell suspension formulation prepared by any one of claims 7-8, in the preparation of a drug for treating refractory allergic rhinitis.

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