Application of secretory IgA in preparation of medicine and diagnostic reagent for preventing and treating acute graft versus host disease

Abstract

The invention relates to the technical field of medical detection, in particular to application of secretory IgA in preparation of drugs and diagnostic reagents for preventing and treating acute graft versus host disease. The research finds that the SIgA can effectively treat the acute graft versus host disease; the content of SIgA in serum and feces of non-aGVHD and aGVHD has a significant difference (Plt; 0.05) of the substrate (1); meanwhile, SIgA in excrement is closely related to the survival rate of a patient after transplantation and the accumulation incidence rate of aGVHD, and it is indicated that the SIgA level can serve as a marker for aGVHD diagnosis and transplantation prognosis evaluation, aGVHD can be treated, and a new strategy and basis are provided for aGVHD diagnosis and treatment.

Description

Technical Field

[0001] The present invention relates to the field of medical detection technology, and in particular to the application of secretory IgA in the preparation of drugs and diagnostic reagents for preventing and treating acute graft-versus-host disease. Background Art

[0002] Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective means of treating malignant hematological tumors such as leukemia, and acute graft-versus-host disease (aGVHD) after transplantation is the most common complication and the main cause of death of allo-HSCT. The prevention and treatment of aGVHD is a key issue to improve the efficacy of allo-HSCT and is also a hot topic of research. Studies have shown that there is a close relationship between intestinal flora and the disease progression of aGVHD. Sequencing results show that the intestinal flora of patients with aGVHD showed a significant decrease in diversity (especially α diversity), a significant increase in the relative abundance of pathogenic bacteria (Enterococcus, etc.), and a significant decrease in the relative abundance of beneficial bacteria (Clostridium, etc.). Further studies revealed that Enterococcus would aggravate aGVHD, while Clostridium and Bacteroidetes could reduce the severity of aGVHD. Therefore, targeted improvement of the diversity and composition of intestinal flora will become a new option for the treatment of aGVHD.

[0003] Currently, for the prevention and treatment of graft-versus-host disease after hematopoietic stem cell transplantation, existing drugs have certain side effects, there is a lack of effective drug treatment, and there are no reports on the treatment of aGVHD with SIgA. Summary of the invention

[0004] In view of this, the present invention provides the use of secretory IgA in the preparation of drugs and diagnostic reagents for preventing and treating acute graft-versus-host disease.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The use of IgA as a marker in the preparation of products for the diagnosis of acute graft-versus-host disease and / or transplantation prognosis assessment.

[0007] In the present invention, the IgA is IgA in serum and / or feces.

[0008] In the present invention, compared with the non-aGVHD control group, the IgA content in the feces of the test group was significantly reduced (P < 0.05), and it is speculated that the IgA content may be related to the occurrence of aGVHD.

[0009] The present study showed that the serum and fecal IgA levels in the aGVHD group of clinical patients were significantly lower than those in the non-aGVHD group (P=0.005, P=0.03), indicating that the serum and fecal IgA levels are closely related to the incidence of aGVHD, can accurately distinguish aGVHD from non-aGVHD, and can be used as a marker for diagnosing aGVHD.

[0010] Furthermore, the present invention also studied the correlation between the early fecal SIgA level after transplantation and the occurrence of aGVHD and the overall prognosis of transplant patients, and found that the cumulative incidence of aGVHD after transplantation in patients with high SIgA content tended to decrease, and the overall survival of patients with low SIgA content after transplantation was slightly worse, suggesting that fecal SIgA content may be related to the disease progression of aGVHD and can be used to assess the risk of aGVHD and / or poor prognosis in transplant patients.

[0011] Based on the above content, it can be known that the reagent for detecting the IgA level can be used to prepare products for the diagnosis of acute graft-versus-host disease and / or the evaluation of transplantation prognosis.

[0012] The transplantation prognosis refers to the risk of acute graft-versus-host disease after transplantation. Specifically, the prognosis includes the prediction of the cumulative incidence of aGVHD and the survival rate of aGVHD patients.

[0013] The present invention also provides the use of IgA in preparing a medicine for preventing, treating and / or alleviating acute graft-versus-host disease.

[0014] In the present invention, the prevention and treatment include: increasing the number of Th1 cells, promoting intestinal epithelial barrier repair, inhibiting weight loss, improving survival rate, prolonging survival time and / or improving target organ damage.

[0015] The present study shows that IgA can improve survival rate and weight loss, and effectively alleviate aGVHD by affecting the number of Th1 cells; at the same time, SIgA can also promote intestinal epithelial barrier repair and alleviate the disease progression of aGVHD. This shows that SIgA can effectively treat aGVHD and improve the occurrence and development of aGVHD.

[0016] The present invention also provides the use of IgA in preparing medicine for preventing, treating and / or alleviating inflammatory bowel disease.

[0017] The prevention, treatment and / or alleviation include: up-regulating the expression level of tight junction proteins in small intestinal and / or large intestinal cells, and / or promoting the integrity of the intestinal epithelial barrier.

[0018] In the present invention, the dosage form of the drug includes but is not limited to injection, spray, oral preparation, etc. The administration method of the IgA includes oral administration.

[0019] In the present invention, the acute graft-versus-host disease includes complications caused by at least one of allogeneic hematopoietic stem cell transplantation, lung transplantation, and kidney transplantation.

[0020] In the present invention, the IgA is secretory IgA, also known as SIgA.

[0021] The present invention also provides a diagnostic product for acute graft-versus-host disease, which comprises a reagent for detecting the level of IgA.

[0022] In the present invention, the reagent comprises at least one of the following I) to III):

[0023] 1) primers and / or probes for detecting IgA gene;

[0024] II), detection of antibodies at IgA levels;

[0025] III), aptamers for detecting IgA levels.

[0026] The present invention also provides a drug for preventing, treating and / or alleviating acute graft-versus-host disease, which comprises an effective dose of secretory IgA.

[0027] The present invention provides the use of secretory IgA in the preparation of drugs and diagnostic reagents for preventing and treating acute graft-versus-host disease. The present invention has found that SIgA can effectively treat acute graft-versus-host disease; there is a significant difference in the content of SIgA in the serum and feces of non-aGVHD and aGVHD (P<0.05); at the same time, IgA in serum and feces is closely related to the patient's survival rate after transplantation and the cumulative incidence of aGVHD, indicating that SIgA levels can be used as a marker for the diagnosis of aGVHD and transplantation prognosis assessment, and can treat aGVHD, providing a new strategy and basis for the diagnosis and treatment of aGVHD. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 Figure 1 is a graph showing the relationship between the level of SIgA in feces and the progression of aGVHD in clinical patients; A is the level of IgA in serum, B is the level of SIgA in feces, and compared with the non-aGVHD group, the levels of IgA in serum and feces in the aGVHD group were significantly lower (P<0.05); C is the cumulative incidence of aGVHD patients, and D is the survival rate of aGVHD patients;

[0029] Figure 2 The data graph shows the changes in serum and feces IgA in the mouse aGVHD model. Compared with the non-aGVHD group, the serum and feces IgA levels in the aGVHD group were significantly reduced (P<0.05), where * indicates P<0.05;

[0030] Figure 3 Results of gavage of SIgA to alleviate aGVHD; AC represents the survival, score and weight change of mice;

[0031] Figure 4 The results of knocking out SIgA aggravated aGVHD; AC shows the survival, score and weight change of mice; D shows the pathological damage of the target organs, lungs, small intestine and large intestine, observed by HE staining;

[0032] Figure 5 Knocking out SIgA significantly increased the number of Th1 and Tc1 cells in the spleen;

[0033] Figure 6 Data graph showing that knocking out SIgA destroys the intestinal epithelial barrier; A is the concentration of FITC-Dextran in serum after oral administration; B and C are Real-time qPCR detections of the expression of tight junction-related proteins in the small intestine (B) and large intestine (C). DETAILED DESCRIPTION

[0034] The present invention provides the use of secretory IgA in the preparation of drugs and diagnostic reagents for preventing and treating acute graft-versus-host disease. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It is particularly important to point out that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0035] In the present invention, unless otherwise specified, the raw materials can be obtained commercially, and the methods are conventional methods in the art.

[0036] The present invention provides use of SIgA in treating acute graft-versus-host disease.

[0037] Specifically, compared with the non-aGVHD group, the serum and fecal IgA levels in the aGVHD group of clinical patients were significantly reduced (P<0.05); the fecal IgA level was correlated with the disease progression of aGVHD.

[0038] Furthermore, compared with the non-aGVHD group, the serum and fecal IgA levels in the aGVHD group of mice were significantly reduced;

[0039] Furthermore, oral administration of SIgA can significantly improve the weight loss and score of mice in the mouse model of acute graft-versus-host disease caused by allogeneic hematopoietic stem cell transplantation, and ultimately delay the survival time.

[0040] Furthermore, knocking out SIgA significantly aggravated the weight loss, increased the score, shortened the survival time, and aggravated the damage to the target organs in the mouse model of acute graft-versus-host disease caused by allogeneic hematopoietic stem cell transplantation.

[0041] Furthermore, knockout of SIgA significantly increased the number of Th1 and Tc1 cells in the spleen.

[0042] Furthermore, knockout of SIgA significantly disrupted the intestinal epithelial barrier.

[0043] The results of the study showed that the occurrence of aGVHD can be predicted based on the SIgA content, and SIgA can improve or alleviate the occurrence and development of aGVHD in mice. Therefore, SIgA can effectively treat acute graft-versus-host disease and can be used as a marker for the diagnosis of aGVHD and the evaluation of transplantation prognosis.

[0044] Furthermore, the SIgA is administered orally.

[0045] The present invention will be further described below in conjunction with embodiments:

[0046] Example 1: SIgA levels in serum and feces of clinical patients are correlated with aGVHD disease progression

[0047] From 2020 to 2022, allogeneic hematopoietic stem cell transplantation was performed in the First Affiliated Hospital of Soochow University. A total of 48 patients with hematological malignancies aged 18 to 65 years were included as research subjects, and serum and stool samples were collected about three weeks after transplantation. After collecting the samples, the clinical baseline characteristics of the patients were recorded and followed up for 100 days to observe the occurrence of aGVHD in the patients. According to whether aGVHD occurred within 100 days, they were divided into two groups: aGVHD group (I to IV degree aGVHD) and non-aGVHD group (no aGVHD).

[0048] ELISA was used to measure the levels of IgA in the patient's serum and SIgA in the feces. The results showed that the serum IgA level of aGVHD patients after transplantation was significantly lower than that of non-aGVHD patients (P = 0.005). At the same time, the SIgA level in the feces after transplantation was also significantly lower than that of non-aGVHD patients (P = 0.03). Figure 1 (middle AB).

[0049] According to the above results, the fecal SIgA level in aGVHD patients after transplantation was significantly lower than that in the non-aGVHD group (P=0.03), suggesting that the fecal SIgA level in the early stage after transplantation may be related to the occurrence of aGVHD.

[0050] To further analyze the correlation between early fecal SIgA levels after transplantation and the occurrence of aGVHD and overall prognosis of transplant patients, the median fecal SIgA content of patients after transplantation (Meidan: 4.59μg / g) was selected as the critical value, and the patients were divided into a high SIgA group (≥4.59μg / g, High feces SIgA group) and a low SIgA group (<4.59μg / g, Low feces SIgA group). Gray's test analysis showed that the cumulative incidence of aGVHD after transplantation in patients with high SIgA content tended to decrease (P=0.07) ( Figure 1 C). Kaplan-Meier survival analysis showed that patients with low SIgA levels had a slightly worse overall survival after transplantation (P = 0.09) ( Figure 1 D). The above results indicate that low levels of fecal SIgA may be associated with adverse aGVHD incidence and prognosis, suggesting that SIgA may play a role in the development of aGVHD.

[0051] Example 2 IgA is significantly downregulated in serum and feces of aGVHD mice

[0052] The aGVHD model of allogeneic hematopoietic stem cell transplantation in mice was constructed by using 6-8 week old C57BL / 6 (H-2b) and BALB / c (H-2d) female mice as donor and recipient animals, respectively. C57BL / 6 mice were donor mice and BALB / c mice were recipient mice. The specific method of model construction is as follows: before transplantation, the recipient mice were sent to the irradiation center to receive 6.25Gry whole body irradiation, and after irradiation, the mice were randomly divided into groups according to their weight.

[0053] On day 0, the donor mice were killed by cervical dislocation, and the spleen of the mice was removed in a sterile environment, ground until there were no tissue blocks, and filtered with a 200-mesh filter. At the same time, the femur and iliac bone of the donor mice were taken, and the bone marrow cells were flushed out and filtered with a 200-mesh filter. Then, the red blood cells were lysed with ammonium chloride buffer to prepare bone marrow cell and spleen cell suspensions. The recipient mice were divided into two groups, each with 15 mice. In the aGVHD group, each BALB / c (H-2d) recipient mouse was injected with 0.3 mL of cell suspension containing 4×10 C57BL / 6 (H-2b) donor mice through the tail vein. 6 Spleen cells and 1×10 7 Bone marrow cells; non-aGVHD group: Each BALB / c (H-2d) recipient mouse was injected with 0.3 mL of cell suspension containing 1×107 bone marrow cells from C57BL / 6 (H-2b) donor mice via the tail vein.

[0054] In order to further observe the expression level of IgA in allogeneic transplantation models of different species, the mouse aGVHD model and non-aGVHD model were constructed according to the above method, and the serum and feces were collected 14 days after transplantation. The content of IgA in serum and SIgA in feces was detected by ELISA. The results showed that, consistent with clinical patients, in the aGVHD group, the levels of mouse serum IgA and fecal SIgA were significantly lower than those in the non-aGVHD group (P<0.0001; P=0.048).

[0055] Example 3 Oral administration of SIgA significantly improves the disease progression of aGVHD in mice

[0056] Two types of female mice, C57BL / 6 (H-2b) and BALB / c (H-2d), aged 6-8 weeks, were used as donor and recipient animals, respectively, to construct an aGVHD model of allogeneic hematopoietic stem cell transplantation in mice, in which C57BL / 6 mice were donor mice and BALB / c mice were recipient mice.

[0057] The specific method of constructing the model is as follows: before transplantation, the recipient mice were sent to the irradiation center to receive 6.25Gry whole-body irradiation. After the irradiation, the mice were randomly divided into groups according to their weight. On day 0, the donor mice were killed by dislocation of the neck, and the spleen of the mice was removed in a sterile environment, ground into tissue-free blocks and filtered with a 200-mesh filter. At the same time, the femur and iliac bone of the donor mice were taken, the bone marrow cells were flushed out and filtered with a 200-mesh filter. Then, the red blood cells were lysed with ammonium chloride buffer to prepare bone marrow cell and spleen cell suspensions. The recipient mice were divided into 2 groups, 15 in each group, and each BALB / c (H-2d) recipient mouse was injected with 0.3mL of cell suspension containing 4×10 C57BL / 6 (H-2b) donor mice through the tail vein. 6 Spleen cells and 1×10 7 Bone marrow cells.

[0058] After the aGVHD model was successfully established, the control group was gavaged with PBS on the 7th day, and the SIgA group was gavaged with SIgA on the 7th day.

[0059] The experimental results showed that after 40 days of allogeneic transplantation, the survival rate of the control group was 0, and the survival time of the mice in the SIgA administration group was significantly delayed, and the weight recovery of the mice was significantly better than that of the control group ( Figure 3 AC).

[0060] Example 4 Knockout of SIgA significantly aggravates the disease progression of aGVHD in mice

[0061] Using CRISPR / CAS9 technology, sgRNA was designed to knock out IgA, the gene encoding the IgA heavy chain structure, in the fertilized eggs of C57BL / 6 (H-2b) mice, and IgA-KO mice with the IgA gene knocked out were obtained; then the genomic DNA of the IgA-KO mice was extracted and PCR identified, and the mice identified as homozygous were bred as breeding mice and used as donor mice to construct an aGVHD model at around 6-8 weeks.

[0062] C57BL / 6-WT mice aged 6-8 weeks and C57BL / 6-KO mice obtained by the above construction were used as donor mice, and BALB / c mice were used as recipient mice to construct a GVHD model of allogeneic hematopoietic stem cell transplantation in mice, wherein. The specific method of constructing the model is as follows: the recipient mice were sent to the irradiation center for 6.25Gry systemic irradiation before transplantation, and the mice were randomly grouped according to their weight after irradiation. On day 0, the donor mice were killed by dislocation of the neck, and the spleen of the mice was removed in a sterile environment, ground into tissue-free blocks and filtered with a 200-mesh filter. At the same time, the femur and iliac bone of the donor mouse were taken, the bone marrow cells were flushed out and filtered with a 200-mesh filter. Then, the red blood cells were lysed with ammonium chloride buffer to prepare a suspension of bone marrow cells and spleen cells.

[0063] The recipient mice were divided into 2 groups, 15 in each group. In the WT group, each BALB / c (H-2d) recipient mouse was injected with 0.3 mL of cell suspension containing 4×10 C57BL / 6-WT donor mice through the tail vein. 6 Spleen cells and 1×10 7 Bone marrow cells; in the KO group, each BALB / c(H-2d) recipient mouse was injected with 0.3 mL of cell suspension containing 4×10 C57BL / 6-KO donor mice through the tail vein. 6 Spleen cells and 1×10 7 Bone marrow cells.

[0064] The experimental results showed that after 28 days of allogeneic transplantation, the survival rate of the KO group was 0, and the survival time of the mice in the WT group was significantly delayed. At the same time, the weight recovery of the mice was significantly better than that of the KO group ( Figure 4AC). At the same time, 14 days after transplantation, target organs including lungs, large intestine and liver were collected for HE staining. The specific steps of HE staining are as follows: 1. Dewaxing of paraffin sections to water: sequentially put the sections into xylene I for 10 minutes, xylene II for 10 minutes, anhydrous ethanol I for 5 minutes, anhydrous ethanol II for 5 minutes, 95% alcohol for 5 minutes, 90% alcohol for 5 minutes, 80% alcohol for 5 minutes, 70% alcohol for 5 minutes, and wash once with distilled water. 2. Hematoxylin staining of cell nuclei: Put the sections into hematoxylin staining solution for staining for 3-8 minutes, wash with tap water, differentiate with 1% hydrochloric acid alcohol for a few seconds, rinse with tap water, return to blue with 0.6% ammonia water, and rinse with running water. 3. Eosin staining of cytoplasm: Put the sections into eosin staining solution for staining for 1-3 minutes, and rinse with running water. 4. Dehydration and sealing: Put the slices into 95% alcohol I for 5 min-95% alcohol II for 5 min-absolute ethanol for 5 min-absolute ethanol for 5 min-xylene I for 5 min-xylene II for 5 min in turn to dehydrate and make them transparent. Take the slices out of xylene, dry them slightly, and seal them with neutral gum.

[0065] According to the pathological sections, knocking out SIgA significantly aggravated the damage of target organs ( Figure 4 (D) The above experimental results show that SIgA can significantly improve the survival rate of aGVHD model mice and has a good therapeutic effect on aGVHD.

[0066] Example 5 Knockout of SIgA significantly increased the number of Th1 and Tc1 cells in the spleen

[0067] On the fourteenth day after aGVHD modeling, the recipient mice in each experimental group were killed by cervical dislocation, and the spleens of the mice were removed. The spleens were ground until no obvious tissue blocks were seen, and the cell suspension was filtered through a 200-mesh cell sieve. The cell suspension was centrifuged at 1300 rpm and 4°C for 8 min, and the supernatant was discarded. 5 mL of red blood cell lysis buffer was added to the cell pellet. After all the cells were fully mixed, they were placed at room temperature for 5 min, and 30 mL of RPMI-1640 medium was added to terminate the reaction. The cells were centrifuged at 1300 rpm and 4°C for 6 min, and the supernatant was discarded. The cells were resuspended in 1 mL of RPMI-1640 medium and counted for later use.

[0068] Cell surface staining. Take 1×106 cell suspension and add it to the flow tube. First add CD16 / CD32 antibody to block at 4℃ for 20min, centrifuge at 1300rpm for 5min, discard the supernatant, resuspend the cells with 100μL PBS (containing 1% FBS), and add different components of flow antibodies according to the instructions. Incubate at 4℃ in the dark for 30min, centrifuge at 1300rpm for 5min after incubation, and discard the supernatant. Wash the cell pellet twice with PBS, and finally add 200μL PBS to resuspend the cells and detect them using a flow cytometer. At the same time, make single positive tube adjustment compensation, make negative tube circle gate, and analyze the flow data with Flowjo software.

[0069] The experimental results showed that compared with the aGVHD model control group, knocking out SIgA significantly increased the number of Th1 cells and Tc1 cells in the spleen ( Figure 5 The experimental results suggest that SIgA can alleviate aGVHD by reducing Th1 cells and Tc1 cells in the spleen.

[0070] Example 6 Knockout of SIgA significantly disrupts the intestinal mucosal barrier

[0071] In order to further explore the effect of SIgA on the intestinal epithelial barrier, the recipient mice of each experimental group were taken on the fourteenth day after modeling, and FITC-dextran was orally administered to the recipient mice at a dose of 0.6 mg / g. The content of FITC in the peripheral blood serum of the mice was detected by an enzyme marker to evaluate the degree of damage to the intestinal mucosal barrier function of each group of recipient mice. The specific steps are as follows: 10 mice in each group, weighing about 20 g, fasting for more than 12 hours, FITC-dextran was dissolved in PBS at a concentration of 125 mg / mL, and 60 mg FITC-dextran was orally administered per 100 g body weight. The mice were killed 4 hours later, eyeball blood was collected, centrifuged at 3500 r / min for 10 minutes, serum was collected, and the absorbance value was measured with a fluorescence spectrophotometer (excitation wavelength 480 nm, emission wavelength 530 nm), and the FITC-dextran content was calculated according to the standard curve. The experimental results showed that compared with the WT group, the KO group significantly increased the content of FITC in the peripheral blood serum of the recipient mice ( Figure 6 In addition, the expression of tight junction proteins in the intestinal tissue of mice was detected by real-time quantitative PCR. The real-time quantitative PCR instrument model was QuantStudio 3, and the reaction conditions were: 95℃ for 3 minutes, 1 cycle; 95℃ for 10 seconds, 60℃ for 1 minute, 40 cycles. The Ct value was analyzed using QuantStudioTM Design & Analysis software (Applied Biosystems). The results showed that SIgA significantly upregulated the small intestine ( Figure 6 Middle B) and large intestine ( Figure 6C) Expression of tight junction proteins in cells.

[0072] The above experimental results showed that SIgA significantly promoted the integrity of the intestinal epithelial barrier in GVHD mice.

[0073] Obviously, the above embodiments are merely examples for clear explanation and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived from these are still within the protection scope of the invention.

Claims

1. Use of IgA in the preparation of drugs for preventing, treating and / or alleviating acute graft-versus-host disease.

2. The use according to claim 1, characterized in that: The prevention and treatment include: reducing the number of Th1 cells, promoting intestinal epithelial barrier repair, inhibiting weight loss, improving survival rate, prolonging survival time and / or improving target organ damage.

3. The use according to any one of claims 1 to 2, characterized in that: The administration method of IgA includes oral administration.

4. Use of IgA as a marker in the preparation of products for the diagnosis of acute graft-versus-host disease and / or transplantation prognosis assessment.

5. The use according to claim 4, characterized in that: The IgA is IgA in serum and / or feces.

6. The use according to any one of claims 1 to 5, characterized in that: The acute graft-versus-host disease includes complications caused by at least one of allogeneic hematopoietic stem cell transplantation, lung transplantation, and kidney transplantation.

7. The use according to any one of claims 1 to 6, characterized in that: The IgA is secretory IgA.

8. A diagnostic product for acute graft-versus-host disease, characterized in that: It is characterized in that Includes reagents for measuring IgA levels.

9. The diagnostic product according to claim 8, characterized in that: The reagent comprises at least one of the following I) to III): 1) primers and / or probes for detecting IgA gene; II), detection of antibodies at IgA levels; III), aptamers for detecting IgA levels.

10. A drug, characterized in that including an effective dose of secretory IgA; The drug is a drug for preventing, treating and / or alleviating acute graft-versus-host disease, or a drug for alleviating inflammatory bowel disease.

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