WO2020250593A1 - Procédé de prédiction d'une réaction du greffon contre hôte (gvhd) et ordinateur permettant la prédiction d'une gvhd - Google Patents

Procédé de prédiction d'une réaction du greffon contre hôte (gvhd) et ordinateur permettant la prédiction d'une gvhd Download PDF

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WO2020250593A1
WO2020250593A1 PCT/JP2020/018225 JP2020018225W WO2020250593A1 WO 2020250593 A1 WO2020250593 A1 WO 2020250593A1 JP 2020018225 W JP2020018225 W JP 2020018225W WO 2020250593 A1 WO2020250593 A1 WO 2020250593A1
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concentration
soluble
gvhd
dnam
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渋谷　彰
和子 渋谷
祐樹 五島
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国立大学法人筑波大学
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids

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  • the present invention relates to a method for predicting graft-versus-host disease (GVHD) and a computer program for predicting GVHD.
  • GVHD graft-versus-host disease
  • Hematopoietic stem cell transplantation is a treatment for hematopoietic tumors such as leukemia and malignant lymphoma for the purpose of eradicating tumor cells or normalizing bone marrow function.
  • Hematopoietic stem cells are the mother cells of all hematopoietic cells, which have the ability to produce mature blood cells such as erythrocytes and leukocytes from one cell, that is, have pluripotency and self-renewal ability.
  • tumor cells such as leukemia and lymphoma
  • graft-versus-host disease (abbreviated as GVHD).
  • allogeneic hematopoietic cell transplantation (abbreviated as allo-HSCT) is an early complication of acute graft-versus-host disease (acute grafft-versus-host disease, acute GVHD, acute GVHD). (Omitted) may occur.
  • Acute GVHD recognizes a human leukocyte antigen (HLA: human leukocyte antigen) expressed on the leukocyte surface of a recipient (patient) by activated T cells derived from a donor (transplant donor) present in the transplanted cells.
  • HLA human leukocyte antigen
  • Non-Patent Document 1 donor-activated T cells having a cytotoxic effect damage recipient cells.
  • Patients who are said to have developed acute GVHD have gastrointestinal symptoms such as fever, skin rash on the extremities, face, and trunk, liver damage with elevated bilirubin, abdominal pain, and diarrhea.
  • Diagnosis of GVHD is usually made by pathological search of biopsy material for skin, liver, and large intestine, but pathological findings are often atypical. In addition, it is often difficult to obtain pathological materials, and in practice, there are not a few cases of diagnosis based on clinical symptoms (Non-Patent Document 2).
  • Steroid administration is generally selected as the initial treatment for acute GVHD.
  • Administration of steroids is an effective treatment for acute GVHD, but administration of steroids may cause serious side effects such as bacterial and viral infections, so the time to start treatment and its dose should be carefully determined. It must be.
  • Non-Patent Document 1 DNAM-1 is a membrane protein discovered by one of the present inventors, Shibuya, and DNAM-1 (CD226) is strongly expressed in activated T cells and NK cells having a cytotoxic effect, and is human. It has been suggested that it recognizes and adheres to ligands expressed in tumor cells and plays an important role in inducing cytotoxicity of target tumor cells (Non-Patent Document 2). Some of the DNAM-1 molecules whose expression is enhanced in activated T cells are cleaved from the cell membrane and released into the bloodstream as soluble DNAM-1. Therefore, the soluble DNAM-1 molecule that grows in the recipient's blood has been thought to be derived from the donor's leukocyte cells.
  • Patent Document 3 when the soluble DNAM-1 concentration in blood shows a value higher than the normal range, it is said that acute GVHD may be developed, close to the onset, or may reach the onset. Although it was possible to do so, the accuracy of the prediction was not satisfactory. In addition, it has been reported that the serum-soluble DNAM-1 concentration in patients who developed acute GVHD was already higher than that in patients who did not develop acute GVHD from 7 days before receiving allogeneic hematopoietic stem cell transplantation. It has been said that the -1 concentration can be an index for the onset of acute GVHD (Non-Patent Document 3).
  • An object of the present invention is to provide a method for predicting GVHD and a computer program for predicting GVHD.
  • the present inventors have diligently studied to solve the above problems. As a result, they have found that the above problems can be solved by having the following configuration, and have completed the present invention.
  • the present invention relates to, for example, the following [Item 1] to [Item 9].
  • [Item 1] The soluble DNAM-1 concentration in the recipient's serum was measured and The soluble DNAM-1 concentration is the soluble DNAM-1 concentration transiently released from the donor-derived lymphocytes x 1 , the soluble DNAM-1 concentration constantly released from the donor-derived lymphocytes x 2 , and the recipient.
  • a method for predicting graft-versus-host disease which comprises analyzing using a mathematical model, assuming that it is the sum of soluble DNA M-1 concentrations x 3 released from lymphocytes of origin.
  • the soluble DNAM-1 concentration in the recipient's serum was measured, and the soluble DNAM-1 concentration at a certain time point (t) was the soluble DNAM-1 concentration x 1 (t) transiently released from the donor-derived lymphocytes.
  • Soluble DNAM-1 concentration constantly released from donor-derived lymphocytes x 2 (t), and soluble DNA M-1 concentration released from recipient-derived lymphocytes x 3 (t).
  • a method for predicting graft-versus-host disease which comprises calculating x 1 (t), x 2 (t), and x 3 (t) and calculating R day_n by the formula (1). ..
  • day_n is the number of days after transplantation.
  • day_n is the number of days after transplantation.
  • Item 3 Item 2. The prediction method according to Item 2, wherein the prediction method is at least one prediction method of GVHD onset and severity.
  • Item 4 Item 3. The prediction method according to any one of Items 2 and 3, wherein GVHD is a complication of allogeneic hematopoietic stem cell transplantation.
  • x 1 (t) is calculated by the formula (2).
  • ⁇ is the adjustment parameter of the gamma distribution
  • k is the shape parameter of the gamma distribution
  • ⁇ is the scale parameter of the gamma distribution
  • ⁇ is the disappearance rate of soluble DNA M-1.
  • r is the rate of production of soluble DNAM-1
  • ⁇ is the rate of elimination of soluble DNAM-1
  • N is the estimated maximum concentration of x 2 .
  • the soluble DNAM-1 concentration in the recipient's serum was measured, and the soluble DNAM-1 concentration at a certain time point (t) was the soluble DNAM-1 concentration x 1 (t) transiently released from the donor-derived lymphocytes. ), Soluble DNAM-1 concentration constantly released from donor-derived lymphocytes x 2 (t), and soluble DNA M-1 concentration released from recipient-derived lymphocytes x 3 (t).
  • GVHD graft-versus-host disease
  • day_n is the number of days after transplantation.
  • a method for predicting GVHD and a computer program for predicting GVHD can be obtained.
  • FIG. 1 is a flowchart of subject selection.
  • FIG. 2 shows the onset date (days after transplantation) of acute GVHD and its frequency.
  • FIG. 1 is a flowchart of subject selection.
  • FIG. 2 shows the onset date (days after transplantation) of acute GVHD and its frequency.
  • FIG. 6 shows the measured values of serum soluble DNAM-1 concentration in 5 GVHD patients, x 1 (t), x 2 (t), x 3 (t), and x 1 (t) + x 2 (t). ) + X 3 (t) is shown.
  • FIG. 7 shows the measured values of serum soluble DNA M-1 concentration in 5 non-GVHD patients, x 1 (t), x 2 (t), x 3 (t), and x 1 (t) + x 2 ( The value of t) + x 3 (t) is shown.
  • the present invention is roughly divided into two aspects.
  • the soluble DNAM-1 concentration in the recipient's serum is measured, and the soluble DNAM-1 concentration is the soluble DNAM-1 concentration transiently released from the donor-derived lymphocytes x 1 , the donor. It is assumed that the sum of the soluble DNAM-1 concentration x 2 constitutively released from the derived lymphocytes and the soluble DNA M-1 concentration x 3 released from the recipient-derived lymphocytes, and the analysis is performed using a mathematical model.
  • This is a method for predicting graft-versus-host disease (GVHD), including the above.
  • the first aspect is shown, the soluble DNAM-1 concentration in the recipient's serum is measured, and the soluble DNAM-1 concentration at a certain time point (t) is transient from the donor-derived lymphocytes. Soluble DNAM-1 concentration released x 1 (t), soluble DNAM-1 concentration constantly released from donor-derived lymphocytes x 2 (t), and soluble DNAM-1 released from recipient-derived lymphocytes Assuming that it is the sum of the concentrations x 3 (t), x 1 (t), x 2 (t), and x 3 (t) are calculated, and R day_n is calculated by the above formula (1).
  • a method for predicting graft-versus-host disease (GVHD) including.
  • the soluble DNAM-1 concentration in the recipient's serum is measured, and the soluble DNAM-1 concentration at a certain time point (t) is the soluble DNAM-1 that is transiently released from the donor-derived lymphocytes.
  • 1 concentration x 1 (t) soluble DNAM-1 concentration constantly released from donor-derived lymphocytes x 2 (t)
  • soluble DNAM-1 concentration released from recipient-derived lymphocytes x 3 (t) Graft-versus-host, including calculating x 1 (t), x 2 (t), and x 3 (t) and calculating R day_n by equation (1) above, assuming the sum of A computer program that predicts illness (GVHD).
  • the term recipient means a patient undergoing a transplant.
  • donor means a transplant donor.
  • the serum in the present invention is prepared from blood collected from a recipient according to a conventional method.
  • the blood collection site is not particularly limited.
  • the required blood volume may be as long as the serum volume required for measuring the soluble DNAM-1 concentration can be secured, and is usually preferably 1 to 10 ml.
  • the time and frequency of blood collection is not particularly limited as long as it is multiple times from before allogeneic hematopoietic stem cell transplantation (hereinafter, also simply referred to as “transplantation”) to after transplantation, but 8 times or more between 2 weeks before transplantation and 1 year after transplantation. It is preferable to carry out, and it is more preferable to carry out 8 times or more between 7 days before transplantation and 100 days after transplantation. Further, it is more preferable to collect blood at least once within 7 days after transplantation.
  • Blood is preferably stored in the blood collection tube, and it is more preferable that the blood collection tube uses a serum separating agent and a coagulation promoting film because serum can be easily prepared.
  • the method for preparing serum from blood is not particularly limited, and a known method for obtaining a supernatant after agglutinating a blood clot can be used, but it is preferable to use a serum separating agent or a coagulation promoter.
  • the serum may be stored until the soluble DNAM-1 concentration is measured, and the temperature at the time of storage is preferably a low temperature of 4 ° C. or lower, and frozen at -20 ° C. or lower (for example, -100 to -20 ° C.). It is more preferable to store it.
  • DNAM-1 is an adhesion molecule belonging to the immunoglobulin family and is expressed in lymphocytes and monocytes such as T cells and natural killer cells. Donor-derived T cells in the transplanted tissue or cells consider the recipient's leukocytes to be non-self and they adhere to each other. DNAM-1 is involved in this adhesion and the ligand expressed in the target cell. Recognize and glue. At that time, an activation signal is transmitted to induce a cytotoxic effect on T cells.
  • Soluble DNAM-1 is cleaved from the membrane-bound DNAM-1 expressed on the surface of lymphocytes and released into the blood.
  • the method for measuring the soluble DNAM-1 concentration is not particularly limited, and methods such as ELISA and Western blotting can be used.
  • a sandwich ELISA using an anti-DNAM-1 antibody is preferable, and TX25, which is an anti-DNAM-1 monoclonal antibody, is more preferably used as an anti-DNAM-1 antibody because the measurement is simple and the sensitivity is good.
  • x 1 is the concentration of soluble DNAM-1 transiently released from donor-derived lymphocytes in the recipient's serum, and x 1 (t) is the recipient's serum at a point in time (t). Is the concentration of soluble DNAM-1 transiently released from donor-derived lymphocytes.
  • the calculation method of x 1 is not particularly limited, and examples thereof include the following methods.
  • the method of calculating the x 1 (t) is not particularly limited, x 1 (t) from being suspected of having a characteristic that increases transiently, it is preferable to use a function representing the distribution of the unimodal .. That is, x 1 (t) is preferably represented by the formula (2).
  • x 1 (t) is a monomodal function
  • x 1 (t) is represented by the formula (8).
  • x 1 (t) is represented by the formula (6).
  • ⁇ is the adjustment parameter of the gamma distribution
  • k is the shape parameter of the gamma distribution
  • ⁇ is the scale parameter of the gamma distribution
  • ⁇ is the disappearance rate of soluble DNA M-1.
  • x 1 (0) is preferably 0.
  • x 2 is the concentration of soluble DNAM-1 constitutively released from donor-derived lymphocytes, and x 2 (t) is constitutive from donor-derived lymphocytes in the recipient's serum at a time point (t). Is the concentration of soluble DNAM-1 released.
  • the calculation method of x 2 is not particularly limited, and examples thereof include the following methods.
  • the method of calculating the x 2 (t) is not particularly limited, x 2 (t), since it is assumed to have a feature that will level off over time, it is preferable to use a logistic function. That is, x 2 (t) is preferably represented by the formula (3).
  • x 2 (t) is represented by the formula (9).
  • r is the rate of production of soluble DNAM-1
  • ⁇ is the rate of elimination of soluble DNAM-1
  • N is the estimated maximum concentration of x 2 .
  • ⁇ Soluble DNAM-1 concentration released from recipient-derived lymphocytes x 3 > x 3 is the concentration of soluble DNAM-1 released from the recipient-derived lymphocytes, and x 3 (t) is released from the recipient-derived lymphocytes in the recipient's serum at a time point (t). Soluble DNAM-1 concentration.
  • the calculation method of x 3 is not particularly limited, and examples thereof include the following methods.
  • the method of calculating the x 3 (t) is not particularly limited, x 3 (t) is soluble DNAM-1 newly produced Sarezu recipient-derived, because the only remaining exponentially It is preferable to use the function of exponential decay because it is presumed to have the characteristic of disappearing. That is, x 3 (t) is preferably represented by the formula (5).
  • the soluble DNAM-1 concentration in the serum of the recipient is the soluble DNAM-1 concentration transiently released from the donor-derived lymphocytes x 1 , the soluble DNAM-1 constantly released from the donor-derived lymphocytes. It is assumed to be the sum of -1 concentration x 2 and soluble DNA M-1 concentration x 3 released from recipient-derived lymphocytes.
  • the soluble DNAM-1 concentration in the recipient's serum at a certain time point (t) is the soluble DNAM-1 concentration transiently released from the donor-derived lymphocyte x 1 (t), which is constant from the donor-derived lymphocyte. It is preferable to assume that it is the sum of the soluble DNA M-1 concentration x 2 (t) released from the recipient and the soluble DNA M-1 concentration x 3 (t) released from the recipient-derived lymphocytes.
  • ⁇ is the adjustment parameter of the gamma distribution
  • k is the shape parameter of the gamma distribution
  • ⁇ is the scale parameter of the gamma distribution
  • r is the production rate of soluble DNA M-1
  • N is the estimation of x 2 .
  • the maximum concentration, ⁇ is the rate of disappearance of soluble DNAM-1.
  • the mathematical model in the present invention is not particularly limited, and known mathematical formulas can be used. It is preferable to use a differential equation because it is suitable for describing a time-varying phenomenon.
  • day_n is the number of days after transplantation.
  • R day_n is, x 1 (t) the value of the integrated value from 0 days post-implantation to n date, x 1 (t) values and x 2 which is integrated from 0 days post-implantation to n date (t) was divided by the sum of the values integrated from 0 days to n days after transplantation, and multiplied by 100.
  • R day_n is the ratio of soluble DNAM-1 transiently released from donor-derived lymphocytes to the ratio of soluble DNAM-1 released from donor-derived lymphocytes between 0 and n days after transplantation. Is.
  • GVHD ⁇ Graft-versus-host disease
  • GVHD means that lymphocytes in a graft (Graft) transferred to a host (Host) having a different histocompatibility antigen engraft in the host and recognize the host tissue as non-self.
  • the "host” is the recipient who received the transfusion or transplant.
  • the present invention is intended to be mainly intended for humans, and can be suitably applied when the patient is a human. However, it can also be applied to at least non-human animals in which antibody production by an immune reaction is observed.
  • acute GVHD is classical acute GVHD that develops within 100 days after transplantation and atypical that develops 100 days after transplantation. Includes with acute GVHD.
  • Atypical acute GVHD is a persistent type in which the clinical condition of classical acute GVHD persists after 100 days, a relapse type in which acute GVHD that has once relieved relapses after 100 days, and a delayed onset of de novo after 100 days. Includes type.
  • the method for predicting GVHD of the present invention is preferably used for GVHD which is a complication of allogeneic hematopoietic stem cell transplantation, among which acute GVHD is more preferable and classical acute GVHD is further preferable. ..
  • the method for predicting GVHD of the present invention is a method for performing analysis using a sample (serum) collected from an object such as the human body, and is a method for diagnosing a human even when the recipient is a human (medical treatment).
  • a method that includes a step of determining a physical or mental condition such as a human's medical condition or health condition for the purpose) is not included.
  • the method for predicting GVHD of the present invention provides a test result that can be used as a judgment material for a doctor to diagnose GVHD of a recipient and perform appropriate treatment.
  • the first aspect of the present invention measures the soluble DNAM-1 concentration in the serum of the recipient, and the soluble DNAM-1 concentration is the soluble DNAM-1 transiently released from the donor-derived lymphocytes.
  • Mathematical model assuming the sum of concentration x 1 , soluble DNAM-1 concentration x 2 constitutively released from donor-derived lymphocytes, and soluble DNA M-1 concentration x 3 released from recipient-derived lymphocytes. It is also a test method for GVHD that provides data on graft-versus-host disease (GVHD), including analysis using.
  • the present invention measures the soluble DNAM-1 concentration in the recipient's serum, and the soluble DNAM-1 concentration is the soluble DNAM-1 concentration transiently released from the donor-derived lymphocytes x 1 , the donor-derived lymph. It is assumed that the sum of the soluble DNAM-1 concentration x 2 constantly released from the sphere and the soluble DNA M-1 concentration x 3 released from the recipient-derived lymphocytes, and the analysis is performed using a mathematical model. It may also be a test method for GVHD that provides data on at least one of GVHD onset and severity, including.
  • the method for predicting GVHD in the present invention includes prediction of the onset of GVHD in the recipient, estimation of GVHD morbidity, estimation of GVHD classification or subclassification, prediction or estimation of severity of GVHD (stage prediction or estimation, stage prediction of GVHD or Includes estimation), predicting the prognosis of GVHD, or estimating the therapeutic effect on GVHD.
  • the GVHD prediction method can also be used when continuously monitoring a GVHD that has already been predicted or diagnosed.
  • the method for predicting GVHD is preferably a method for predicting at least one of GVHD onset and severity because the prediction can be performed more accurately.
  • the second aspect of the present invention measures the soluble DNAM-1 concentration in the recipient's serum, and the soluble DNAM-1 concentration at a certain time point (t) is transiently released from the donor-derived lymphocytes.
  • Transplantation comprising calculating x 1 (t), x 2 (t), and x 3 (t), and calculating R day_n by the above equation (1), assuming the sum of (t).
  • a computer program that predicts one-to-one host disease (GVHD).
  • the computer program may take the form of a hardware embodiment, a software embodiment executed on the hardware, or a combination thereof. Further, the computer program may be a computer program on a computer-usable storage medium (having a computer-usable coding program embodied in the medium). Storage media that can be used with the computer include any suitable computer-readable medium, including discs, CD-ROMs, optical storage devices, magnetic storage devices, and the like.
  • the myeloablative pretreatment regimen included cyclophosphamide alone, or cyclophosphamide in combination with cytarabine, and total body irradiation (12 Gy) or busulfan.
  • a palliative pretreatment regimen with busulfan or melphalan and furudabin, or a combination of busulfan or melphalan and furudabin and low-dose total body irradiation (2-4 Gy) was used for patients aged 55 years and older.
  • Anti-human thymocyte immunoglobulin was not used in the pretreatment regimen.
  • cyclosporine A or tacrolimus was continuously administered, and methotrexate was administered at 10 mg / m 2 , 3, 6 and 11 days for a short period of 7 mg / m 2 .
  • Fluoroquinone (fluconazole or itraconazole), solfamethoxazole, and trimethoprim were used to prevent infection with bacteria, fungi, and Pneumocystis jirovecii.
  • Cytomegalovirus was monitored weekly using a cytomegalovirus antigenemia test and patients with 1 cell / 50,000 cells or more were treated with ganciclovir as positive for antigenemia.
  • GVHD grade The severity of acute GVHD was determined by a partially revised standard of the conventional Glucksberg classification method in the 1994 Consensus Conference on Grafting of Acute GVHD. To outline, skin is rash, systemic erythema, liver is total bilirubin, gastrointestinal tract is diarrhea, nausea, abdominal pain, bleeding to determine the stage of organ damage, using the skin, liver, gastrointestinal stage It is a classification method that determines grades I, II, III, and IV. Grade IV is the most severe. The grade determined by this method is hereinafter referred to as GVHD grade.
  • TX25 antibody 8 ⁇ g / ml, established by Shibuya et al. Of the present inventor was added to a 96-well flat bottom microplate for ELISA at 100 ⁇ l / well, coated at 4 ° C. for 2 hours, and then 10% BSA / PBS. Blocking was performed at room temperature for 2 hours at (100 ⁇ l / well). Then, it was washed three times with washing buffer (0.05% Tween 20). Human DNAM-1 / Fc chimeric protein (standard) or patient serum was added at 100 ⁇ l / well and allowed to stand overnight at 4 ° C.
  • biotinylated rabbit anti-human DNAM-1 polyclonal antibody (0.6 ⁇ g / ml) was added at 100 ⁇ l / well, and the mixture was allowed to stand at room temperature for 1 hour.
  • 100 ⁇ l / well of ExtraAvidin peroxidase (manufactured by Sigma-Aldrich, diluted 1:250 with washing buffer) was added and allowed to stand at room temperature for 30 minutes before washing.
  • OPD manufactured by Sigma-Aldrich
  • the reaction was stopped by adding 12.5 ⁇ L / well of 6N HCl, and then the absorbance at 490 nm was measured using a Spectra Max Microplate reader (manufactured by Molecular Devices). All values were measured by Triplicate. In addition, all samples obtained from the same patient at different blood sampling times were measured at least twice at the same time.
  • Example 1 [Concentration of soluble DNAM-1 in serum] Traditionally, the soluble DNAM-1 that grows in the recipient's blood has been thought to be derived from donor-derived lymphocytes.
  • the recipient's serum-soluble DNAM-1 is soluble DNAM-1 (type 1) that is transiently released from donor-derived lymphocytes and soluble DNAM-1 that is constantly released from donor-derived lymphocytes (type 1). It was considered that it can be classified into three types: type 2) and soluble DNA M-1 (type 3) released from recipient-derived lymphocytes.
  • the recipient's serum-soluble DNAM-1 has a soluble DNAM-1 concentration x 1 (t) that is transiently released from the donor-derived lymphocytes and a soluble DNAM-1 that is constantly released from the donor-derived lymphocytes. It was assumed to be the sum of 1 concentration x 2 (t) and the soluble DNA M-1 concentration x 3 (t) released from recipient-derived lymphocytes. However, it is very difficult to experimentally quantify each of the three after identifying the origin of soluble DNA M-1 in serum. Therefore, the x 1 (t), the x 2 (t), and the x 3 (t) were estimated by simulating on a computer using a mathematical model.
  • x 1 (t) has a characteristic of transiently increasing, it is considered appropriate that it is a monomodal function, so the gamma distribution was selected. Since it is presumed that x 2 (t) has a characteristic that it reaches a plateau over time, a logistic function was selected.
  • the formula used for estimating the x 1 (t), the x 2 (t), and the x 3 (t) is the following formula (10).
  • the mathematical model according to the equation (10) is hereinafter referred to as the mathematical model of the first embodiment.
  • lambda is the adjustment parameter of the gamma distribution
  • k is shape parameter of the gamma distribution
  • theta is scale parameter of the gamma distribution
  • r is the production rate of soluble DNAM-1
  • N is the estimation of x 2
  • the maximum concentration, ⁇ is the rate of disappearance of soluble DNAM-1.
  • x 1 is the initial value of x 1 (t) (0) was set to 0.
  • x 2 (t) is the sum of the initial value of x 3 (t), x 2 (0) + x 3 (0) was set to the initial measured value.
  • the initial measured values were defined as follows (i) to (iv) and applied to each patient. When multiple definitions were applied, the definition with the smaller number among (i) to (iv) was preferentially adopted.
  • day_n is the number of days after transplantation.
  • the integral value of x 1 from the transplant day 0 to day post-transplant day n (t) is, that is, the AUC from the transplantation day 0 x 1 (t) to post-transplant n Date (Area Under the Curve).
  • ⁇ result ⁇ Patients with age, total body irradiation, pretreatment, donor conditions, HLA (human leukocyte antigen) allele match, cyclosporine A or tacrolimus administration, GVHD severity, etc., depending on the presence or absence of GVHD.
  • HLA human leukocyte antigen
  • the time when GVHD developed (the number of days after transplantation) of 48 patients who developed GVHD is shown in FIG.
  • the horizontal axis is the time when GVHD developed (Onset day of aGVHD, the number of days after transplantation), and the vertical axis is the frequency (Frequency, number of patients).
  • the mean time of onset of GVHD was 23 days after transplantation, and the median was 21 days after transplantation.
  • R day_n and the presence or absence of GVHD onset [R day_n and the presence or absence of GVHD onset]
  • R day_n (n 20, 30, 40, 50) up to 20, 30, 40, and 50 days after transplantation of each patient is calculated, classified according to the presence or absence of GVHD, and the value of R day_n and the patient.
  • GVHD (+) patients who developed GVHD
  • GVHD ( -) patients who did not develop GVHD
  • FIG. 4 shows a box plot created for R day_n classified into GVHD (+) and GVHD (-).
  • the R day_n of GVHD (+) is higher than that of GVHD (-) on all 20, 30, 40, and 50 days after transplantation.
  • Table 3 shows the evaluation results of the data suitability of the model of Example 1 and the model of Comparative Example 1.
  • the numbers indicate the optimal number of patients for each model, the percentage of the total number of patients (%), and the 95% confidence interval. The results of the binomial test are also shown.
  • Example 1 fits the data better than the model of Comparative Example 1 in about 70% of the patients.
  • a binomial test was performed to examine this difference, and it was found that the model of Example 1 was statistically significantly better suited to the data.
  • TP number of true positives, true positive number
  • TN number of true negatives, true negative number
  • FP number of false positives, false positive number
  • FN number of negative negatives, false positive number
  • FN number of negative negatives.
  • Example 1 tends to have higher AUC, sensitivity, specificity, and accuracy than Comparative Example 2.
  • Comparative Example 2 uses the maximum value of the serum-soluble DNAM-1 concentration actually measured value. The difference is that R day_n is calculated on the assumption that the measured value of the soluble DNA M-1 concentration in serum is the sum of the three components x 1 (t), x 2 (t), and x 3 (t).
  • the soluble DNAM-1 concentration in the serum of the recipient is measured, and the soluble DNAM-1 concentration is the soluble DNAM-1 concentration transiently released from the donor-derived lymphocytes x 1 (t), derived from the donor.
  • the soluble DNAM-1 concentration x 2 (t) constantly released from lymphocytes and the soluble DNA M-1 concentration x 3 (t) released from recipient-derived lymphocytes, x It was found that the onset of GVHD can be accurately evaluated by calculating 1 (t), x 2 (t), and x 3 (t), and further calculating R day_n .

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Abstract

La présente invention aborde le problème consistant à fournir un procédé permettant la prédiction d'une réaction du greffon contre hôte (GVHD) et un ordinateur permettant la prédiction d'une GVHD. Ce procédé de prédiction d'une GVHD consiste : à mesurer la concentration d'un DNAM-1 soluble dans le sérum provenant d'un receveur ; et à mettre en œuvre une analyse par utilisation d'un modèle mathématique, en supposant que la concentration du DNAM-1 soluble est la somme de la concentration x1 d'un DNAM-1 soluble qui est libéré transitoirement d'un lymphocyte provenant d'un donneur, de la concentration x2 d'un DNAM-1 soluble qui est libéré en permanence d'un lymphocyte provenant d'un donneur, et de la concentration x3 d'un DNAM-1 soluble qui est libéré d'un lymphocyte provenant d'un receveur.
PCT/JP2020/018225 2019-06-11 2020-04-30 Procédé de prédiction d'une réaction du greffon contre hôte (gvhd) et ordinateur permettant la prédiction d'une gvhd WO2020250593A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007116945A1 (fr) * 2006-04-07 2007-10-18 University Of Tsukuba Marqueur de prévision de la maladie du greffon contre l'hôte et utilisation
JP2014501098A (ja) * 2011-06-20 2014-01-20 ピービーディー バイオダイアグノスティックス，エルエルシー 造血細胞移植のドナーをスクリーニング、予想、特定する方法、キット及びアレイ、並びに、造血細胞移植（ｈｃｔ）が移植片対宿主病（ｇｖｈｄ）を引き起こすリスクを予測する方法、キット及びアレイ
US9563744B1 (en) * 2014-12-03 2017-02-07 Biodesix, Inc. Method of predicting development and severity of graft-versus-host disease
WO2017183665A1 (fr) * 2016-04-20 2017-10-26 国立大学法人筑波大学 Activateur de lymphocyte t régulateur et son utilisation

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007116945A1 (fr) * 2006-04-07 2007-10-18 University Of Tsukuba Marqueur de prévision de la maladie du greffon contre l'hôte et utilisation
JP2014501098A (ja) * 2011-06-20 2014-01-20 ピービーディー バイオダイアグノスティックス，エルエルシー 造血細胞移植のドナーをスクリーニング、予想、特定する方法、キット及びアレイ、並びに、造血細胞移植（ｈｃｔ）が移植片対宿主病（ｇｖｈｄ）を引き起こすリスクを予測する方法、キット及びアレイ
US9563744B1 (en) * 2014-12-03 2017-02-07 Biodesix, Inc. Method of predicting development and severity of graft-versus-host disease
WO2017183665A1 (fr) * 2016-04-20 2017-10-26 国立大学法人筑波大学 Activateur de lymphocyte t régulateur et son utilisation

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
GOSHIMA, YUKI ET AL.: "A mathematical model for dynamics of soluble form of DNA.M-1 as a biomarker for graft-versus-host disease", PLOS ONE, vol. 15, no. 2, 10 February 2020 (2020-02-10), pages e0228508, XP055771309 *
KANAMARU, YUMI ET AL.: "Roleof DNAM-1 in immune response". Clinical immunology& allergology", DNAM, vol. 66, no. 2, 2016, pages 177 - 183 *
KANAYA, MINORU ET AL.: "Soluble DNAM-1, as a Predictive Biomarker for Acute Graft-Versus-Host Disease", PLOS ONE, vol. 11, no. 6, 2016, pages e0154173, XP055730758 *
ZHANG, DONGLIANG ET AL.: "TIGIT-Fc alleviates acute graft-versus-host disease by suppressing CTL activation via promoting the generation of immunoregulatory dendritic cells", BBA - MOLECULAR BASIS OF DISEASE, vol. 1864, 2018, pages 3085 - 3098, XP085428789, DOI: 10.1016/j.bbadis.2018.06.022 *

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