JP2009095340A - Method for estimating therapeutic effect to subject of autoimmune disease - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for estimating therapeutic effect, enabling to estimate in an early stage if a pharmaceutical agent is effective to a patient or not, when the pharmaceutical agent is administered to the patient suffering from an autoimmune disease. <P>SOLUTION: The therapeutic effect of the pharmaceutical agent to a subject is estimated by sampling blood of the subject of the autoimmune disease 1 week before initiation and 1 week after the initiation of administration of the pharmaceutical agent, measuring the amount of expression of a predetermined gene in each sample and using change of the amount of expression of the gene before and after the administration of the pharmaceutical agent as an indication. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a method for predicting a therapeutic effect on a subject with an autoimmune disease, a method for screening a substance having a therapeutic effect on an autoimmune disease, and an oligo / poly that can be used in the method for predicting the therapeutic effect and the screening method. It relates to a nucleotide array.

Autoimmune disease is a general term for diseases that cause symptoms when the immune system attacks and targets normal cells and tissues of the self. Various diseases such as vasculitis syndrome have been reported as autoimmune diseases. . Although autoimmune diseases are treated differently depending on the disease, abnormalities in the immune system are the cause of the disease, and in many diseases, immunosuppressants and steroidal drugs are used for treatment. For example, in the case of MPO-ANCA-related vasculitis (microscopic polyangiitis), which is one of the vasculitis syndromes, administration of cyclophosphamide, an immunosuppressant, and methylprednisolone (or prednisolone), a steroidal drug A method of performing treatment for 3 to 6 months by combination therapy with administration of is a standard treatment method (Non-patent Document 1).
"Prospective clinical study to clarify the usefulness of standard treatment protocol for MPO-ANCA-related vasculitis" Grant-in-Aid for Health and Labor Sciences Research for Overcoming Intractable Diseases: Research on Intractable Vasculitis, 2004 Summary・ Shared research report: p. 224-226, 2007.

  However, conventionally, even if an attempt is made to treat an autoimmune disease by administration of an immunosuppressive agent or the like, the therapeutic effect has been revealed after a few weeks have passed since the start of treatment.

  In other words, even if a drug that is thought to be effective for autoimmune diseases is administered to patients, the drug does not work for all patients. Sometimes it turned out to be ineffective. For example, the administration of immunosuppressants has the side effect of increasing the risk of infection, but it is not clear that some immunosuppressants are effective in that patient until more than a few weeks after the start of treatment. In addition to having no therapeutic effect, it sometimes resulted in infections.

  The present invention has been made in view of such points, and when a drug is administered to a patient with an autoimmune disease, it is predicted at an early stage whether the drug is effective for the patient. The first object of the present invention is to provide a method for predicting therapeutic effects.

  The second object of the present invention is to provide a method for screening a substance having a therapeutic effect on an autoimmune disease, which can screen a substance having a therapeutic effect on the autoimmune disease.

  Furthermore, a third object of the present invention is to provide an oligo / polynucleotide array that can be used in the prediction method and the screening method.

  As a result of earnest research, the present inventor found 74 genes that correlate with the therapeutic effect on autoimmune disease and the expression level of the gene, and further studied to complete the present invention.

That is, the first of the present invention relates to the following method for predicting therapeutic effect.
[1] A method for predicting the effect of the treatment on the subject using the expression level of the gene in blood collected from a subject with autoimmune disease treated as an index, the gene comprising the following (a- A prediction method, which is one or more genes selected from the group consisting of 1) to (a-59) and (b-1) to (b-15).
(A-1) a gene encoding CLC (a-2) a gene encoding IFIT1 (a-3) a gene encoding IFIT3 (a-4) a gene encoding CCR3 (a-5) a gene encoding HERC5 (A-6) a gene encoding NGFRAP1 (a-7) a gene encoding TNFSF10 (a-8) a gene encoding FGL2 (a-9) a gene encoding MX1 (a-10) a gene encoding CCR1 (A-11) a gene encoding EMR1 (a-12) a gene encoding XIAP-related factor-1 (a-13) a gene encoding OASL (a-14) a gene encoding OAS1 (a-15) IFIT5 (A-16) gene encoding WSB2 (a-17) gene encoding FFAR2 Child (a-18) gene encoding ISG15 (a-19) gene encoding HIST1H3H (a-20) gene encoding IFI6 (a-21) gene encoding CTSL1 (a-22) encoding GBP1 Gene (a-23) gene encoding IRF7 (a-24) gene encoding HIST2H2BE (a-25) gene encoding IFIH1 (a-26) gene encoding MMD (a-27) encoding CD36 Gene (a-28) gene encoding MT2A (a-29) gene encoding PSMB9 (a-30) gene encoding PLSCR1 (a-31) gene encoding OAS2 (a-32) encoding EIF2AK2 Gene (a-33) Gene encoding LTB (a-34) A gene encoding S4A4A (a-35) A gene encoding ADM (a-36) A gene encoding OAS3 (a-37) A gene encoding SLAMF7 (a-38) A gene encoding DDX58 (a-39) A gene encoding GNG11 (a-40) A gene encoding TSC22D1 (a-41) A gene encoding HIST1H2BD (a-42) A gene encoding IFITM3 (a-43) A gene encoding UBE2L6 (a-44) H2BFS encoding gene (a-45) CAT encoding gene (a-46) SCO2 encoding gene (a-47) PSME2 encoding gene (a-48) STAT1 encoding gene (a-49) Gene encoding MTHFD2 (a-50) MT1 (A-51) gene encoding TGFA (a-52) gene encoding GPR109B (a-53) gene encoding HIST1H3D (a-54) gene encoding IFI44L (a-55) F13A1 (A-56) gene encoding CX3CR1 (a-57) gene encoding FCGR1A (a-58) gene encoding IL1B (a-59) gene encoding ARRB1 (b-1) ADAM28 (B-2) gene encoding CEACAM8 (b-3) gene encoding COL9A2 (b-4) gene encoding DEFA4 (b-5) gene encoding LTF (b-6) SEC23B (B-7) encoding VSIG4 Gene (b-8) gene encoding DEFA1 (b-9) gene encoding CD163 (b-10) gene encoding RUNDC3A (b-11) gene encoding CAMP (b-12) encoding LCN2 Gene (b-13) Gene encoding HDAC9 (b-14) Gene encoding CD24 (b-15) Gene encoding IL1R2 [2] Selected from (a-1) to (a-59) above A method using the expression level of one or more genes as an index, wherein the expression level of a gene when a predetermined time has elapsed after receiving the treatment is a gene expression at a predetermined time before that time. The prediction method according to [1], wherein the treatment is predicted to be effective for the subject when the expression level is decreased.
[3] A method using the expression level of one or more genes selected from (b-1) to (b-15) as an index, and a predetermined time has elapsed since receiving the treatment Predicting that the treatment is effective for the subject when the expression level of the gene at that time is increased compared to the expression level of the gene at a predetermined time before that time, [1] or The prediction method according to [2].
[4] The prediction method according to any one of [1] to [3], wherein the autoimmune disease is vasculitis syndrome.
[5] The prediction method according to [4], wherein the vasculitis syndrome is MPO-ANCA-related vasculitis.
[6] The prediction method according to any one of [1] to [5], wherein the treatment is drug administration.
[7] The prediction method according to [6], wherein the drug is an immunosuppressant, a steroid drug, or a biological preparation.
[8] The administration of the drug is a combination of the administration of an immunosuppressive drug and the administration of a steroid drug, the immunosuppressive drug is cyclophosphamide, and the steroid drug is methylprednisolone or prednisolone. The prediction method according to [6].

The second aspect of the present invention relates to the following screening method.
[9] A step of administering a test substance to a non-human animal model of autoimmune disease, and a step of determining the therapeutic effect of the test substance on the autoimmune disease using the gene expression level in the blood of the model animal as an index; A method for screening a substance having a therapeutic effect on an autoimmune disease, wherein the genes are the following (a-1) to (a-59) and (b-1) to (b-15): A screening method, which is one or more genes selected from the group consisting of:
(A-1) a gene encoding CLC (a-2) a gene encoding IFIT1 (a-3) a gene encoding IFIT3 (a-4) a gene encoding CCR3 (a-5) a gene encoding HERC5 (A-6) a gene encoding NGFRAP1 (a-7) a gene encoding TNFSF10 (a-8) a gene encoding FGL2 (a-9) a gene encoding MX1 (a-10) a gene encoding CCR1 (A-11) a gene encoding EMR1 (a-12) a gene encoding XIAP-related factor-1 (a-13) a gene encoding OASL (a-14) a gene encoding OAS1 (a-15) IFIT5 (A-16) gene encoding WSB2 (a-17) gene encoding FFAR2 Child (a-18) gene encoding ISG15 (a-19) gene encoding HIST1H3H (a-20) gene encoding IFI6 (a-21) gene encoding CTSL1 (a-22) encoding GBP1 Gene (a-23) gene encoding IRF7 (a-24) gene encoding HIST2H2BE (a-25) gene encoding IFIH1 (a-26) gene encoding MMD (a-27) encoding CD36 Gene (a-28) gene encoding MT2A (a-29) gene encoding PSMB9 (a-30) gene encoding PLSCR1 (a-31) gene encoding OAS2 (a-32) encoding EIF2AK2 Gene (a-33) Gene encoding LTB (a-34) A gene encoding S4A4A (a-35) A gene encoding ADM (a-36) A gene encoding OAS3 (a-37) A gene encoding SLAMF7 (a-38) A gene encoding DDX58 (a-39) A gene encoding GNG11 (a-40) A gene encoding TSC22D1 (a-41) A gene encoding HIST1H2BD (a-42) A gene encoding IFITM3 (a-43) A gene encoding UBE2L6 (a-44) H2BFS encoding gene (a-45) CAT encoding gene (a-46) SCO2 encoding gene (a-47) PSME2 encoding gene (a-48) STAT1 encoding gene (a-49) Gene encoding MTHFD2 (a-50) MT1 (A-51) gene encoding TGFA (a-52) gene encoding GPR109B (a-53) gene encoding HIST1H3D (a-54) gene encoding IFI44L (a-55) F13A1 (A-56) gene encoding CX3CR1 (a-57) gene encoding FCGR1A (a-58) gene encoding IL1B (a-59) gene encoding ARRB1 (b-1) ADAM28 (B-2) gene encoding CEACAM8 (b-3) gene encoding COL9A2 (b-4) gene encoding DEFA4 (b-5) gene encoding LTF (b-6) SEC23B (B-7) encoding VSIG4 Gene (b-8) gene encoding DEFA1 (b-9) gene encoding CD163 (b-10) gene encoding RUNDC3A (b-11) gene encoding CAMP (b-12) encoding LCN2 Gene (b-13) Gene encoding HDAC9 (b-14) Gene encoding CD24 (b-15) Gene encoding IL1R2 [10] Selected from (a-1) to (a-59) above A method using the expression level of one or more genes as an index, wherein the expression level of a gene when a predetermined time has elapsed since the administration of the test substance is a predetermined time before that time. The screening according to [9], wherein it is determined that the test substance has a therapeutic effect on the autoimmune disease when the expression level of the gene is decreased. Method.
[11] A method using the expression level of one or more genes selected from (b-1) to (b-15) as an index, and a predetermined time after administration of the test substance When the expression level of the gene at the time of elapse of time has increased compared to the expression level of the gene at a predetermined time before that time, the test substance is determined to have a therapeutic effect on the autoimmune disease The screening method according to [9] or [10].
[12] The screening method according to any one of [9] to [11], wherein the autoimmune disease is vasculitis syndrome.
[13] The screening method according to [12], wherein the vasculitis syndrome is MPO-ANCA-related vasculitis.

The third aspect of the present invention relates to the following oligo / polynucleotide array.
[14] An oligo / polynucleotide array for determining the effect of a drug or candidate drug for treating an autoimmune disease, comprising a support and two or more types of oligo / polynucleotide immobilized on the support The oligonucleotide / polynucleotide array, wherein the two or more kinds of oligo / polynucleotides are selected from the group consisting of the following (c-1) to (c-74).
(C-1) Oligo / polynucleotide that can hybridize to nucleic acid encoding CLC (c-2) Oligo / polynucleotide that can hybridize to nucleic acid encoding IFIT1 (c-3) Hybridize to nucleic acid encoding IFIT3 Oligo / polynucleotide (c-4) capable of hybridizing to nucleic acid encoding CCR3 (c-5) Oligo / polynucleotide capable of hybridizing to nucleic acid encoding HERC5 (c-6) Oligo / polynucleotide hybridizable to nucleic acid encoding NGFRAP1 (c-7) Oligo / polynucleotide hybridizable to nucleic acid encoding TNFSF10 (c-8) Oligo / polynucleotide hybridizable to nucleic acid encoding FGL2 Polynucleotide (c 9) Oligo / polynucleotide hybridizable to nucleic acid encoding MX1 (c-10) Oligo / polynucleotide hybridizable to nucleic acid encoding CCR1 (c-11) Hybridizable to nucleic acid encoding EMR1 Oligo / polynucleotide (c-12) Oligo / polynucleotide hybridizable to nucleic acid encoding XIAP-related factor-1 (c-13) Oligo / polynucleotide hybridizable to nucleic acid encoding OASL (c-14) ) Oligo / polynucleotide hybridizable to nucleic acid encoding OAS1 (c-15) Oligo / polynucleotide hybridizable to nucleic acid encoding IFIT5 (c-16) Oligo hybridizable to nucleic acid encoding WSB2 / Polynucleotide c-17) Oligo / polynucleotide hybridizable to nucleic acid encoding FFAR2 (c-18) Oligo / polynucleotide hybridizable to nucleic acid encoding ISG15 (c-19) Hybridized to nucleic acid encoding HIST1H3H Possible oligo / polynucleotide (c-20) oligo / polynucleotide hybridizable to nucleic acid encoding IFI6 (c-21) oligo / polynucleotide hybridizable to nucleic acid encoding CTSL1 (c-22) GBP1 (C-23) Oligo / polynucleotide hybridizable to nucleic acid encoding IRF7 (c-24) Oligo hybridizable to nucleic acid encoding HIST2H2BE Polynucleotide (c-25) Oligo / polynucleotide hybridizable to nucleic acid encoding IFIH1 (c-26) Oligo / polynucleotide hybridizable to nucleic acid encoding MMD (c-27) Nucleic acid encoding CD36 (C-28) Oligo / polynucleotide that can hybridize to nucleic acid encoding MT2A (c-29) Oligo / polynucleotide that can hybridize to nucleic acid encoding PSMB9 (c- 30) Oligo / polynucleotide hybridizable to a nucleic acid encoding PLSCR1 (c-31) Oligo / polynucleotide hybridizable to a nucleic acid encoding OAS2 (c-32) Hybridizable to a nucleic acid encoding EIF2AK2 Oligo / polynucleotide (c-33) Oligo / polynucleotide hybridizable to the nucleic acid encoding LTB (c-34) Oligo / polynucleotide hybridizable to the nucleic acid encoding MS4A4A (c-35) Encoding ADM (C-36) Oligo / polynucleotide hybridizable to nucleic acid encoding OAS3 (c-37) Oligo / polynucleotide hybridizable to nucleic acid encoding SLAMF7 c-38) Oligo / polynucleotide hybridizable to nucleic acid encoding DDX58 (c-39) Oligo / polynucleotide hybridizable to nucleic acid encoding GNG11 (c-40) Hybridizer to nucleic acid encoding TSC22D1 Oligo / polynucleotide (c-41) Oligo / polynucleotide hybridizable to a nucleic acid encoding HIST1H2BD (c-42) Oligo / polynucleotide hybridizable to a nucleic acid encoding IFITM3 (c-43) Oligo / polynucleotide hybridizable to nucleic acid encoding UBE2L6 (c-44) Oligo / polynucleotide hybridizable to nucleic acid encoding H2BFS (c-45) Oligo / polynucleotide hybridizable to nucleic acid encoding CAT Polynucleotide (c-46) Oligo / polynucleotide that can hybridize to nucleic acid encoding SCO2 (c-47) Oligo / polynucleotide that can hybridize to nucleic acid encoding PSME2 (c-48) Encodes STAT1 Oligo / polynucleotide that can hybridize to acid (c-49) Oligo / polynucleotide that can hybridize to nucleic acid encoding MTHFD2 (c-50) Oligo / polynucleotide that can hybridize to nucleic acid encoding MT1X (c -51) Oligo / polynucleotide hybridizable to nucleic acid encoding TGFA (c-52) Oligo / polynucleotide hybridizable to nucleic acid encoding GPR109B (c-53) Hybridized to nucleic acid encoding HIST1H3D Oligo / polynucleotide (c-54) Oligo / polynucleotide hybridizable to a nucleic acid encoding IFI44L (c-55) Oligo / polynucleotide hybridizable to a nucleic acid encoding F13A1 (c-56) C Oligo / polynucleotide hybridizable to nucleic acid encoding X3CR1 (c-57) Oligo / polynucleotide hybridizable to nucleic acid encoding FCGR1A (c-58) Oligo / polynucleotide hybridizable to nucleic acid encoding IL1B Polynucleotide (c-59) Oligo / polynucleotide hybridizable to the nucleic acid encoding ARRB1 (c-60) Oligo / polynucleotide hybridizable to the nucleic acid encoding ADAM28 (c-61) Nucleic acid encoding CEACAM8 (C-62) Oligo / polynucleotide hybridizable to a nucleic acid encoding COL9A2 (c-63) Oligo / polynucleotide hybridizable to a nucleic acid encoding DEFA4 Otide (c-64) Oligo / polynucleotide hybridizable to nucleic acid encoding LTF (c-65) Oligo / polynucleotide hybridizable to nucleic acid encoding SEC23B (c-66) Nucleic acid encoding VSIG4 Hybridizable oligo / polynucleotide (c-67) Oligo / polynucleotide hybridizable to nucleic acid encoding DEFA1 (c-68) Oligo / polynucleotide hybridizable to nucleic acid encoding CD163 (c-69) ) Oligo / polynucleotide hybridizable to nucleic acid encoding RUNDC3A (c-70) Oligo / polynucleotide hybridizable to nucleic acid encoding CAMP (c-71) Oligo hybridizable to nucleic acid encoding LCN2 / Renucleotide (c-72) Oligo / polynucleotide hybridizable to nucleic acid encoding HDAC9 (c-73) Oligo / polynucleotide hybridizable to nucleic acid encoding CD24 (c-74) Nucleic acid encoding IL1R2 Oligo / polynucleotide that can hybridize to

  According to the prediction method of the therapeutic effect of this invention, when a chemical | medical agent is administered with respect to the test subject of an autoimmune disease, it can be estimated early whether the said chemical | medical agent is effective with respect to the said test subject. Therefore, the doctor can appropriately determine at an early stage whether or not to use the drug for the subject based on the prediction result.

  According to the screening method of the present invention, a substance having a therapeutic effect on an autoimmune disease can be rapidly screened.

  According to the oligo / polynucleotide array of the present invention, it is possible to simultaneously perform expression analysis on two or more types of genes in parallel, so that the above therapeutic effect prediction method and screening method can be performed rapidly.

1. The prediction method of the present invention is a method for predicting whether or not the treatment is effective for a subject when the subject is treated for an autoimmune disease, The expression level of one or more genes selected from the group consisting of the following groups a and b in the blood of the subject is predicted as an index.

[Group a]
(A-1) Gene encoding CLC (Charcot-Leyden crystal protein) (a-2) Gene encoding IFIT1 (interferon-induced protein with tetratricopeptide repeats 1) (a-3) IFIT3 (interferon-induced protein with tetratricopeptide gene encoding repeats 3) (a-4) gene encoding CCR3 (chemokine (CC motif) receptor 3) (a-5) gene encoding HERC5 (hect domain and RLD 5) (a-6) NGFRAP1 ( Gene encoding nerve growth factor receptor (TNFRSF16) associated protein 1) (a-7) Gene encoding TNFSF10 (tumor necrosis factor (ligand) superfamily, member 10) (a-8) FGL2 (fibrinogen-like 2) Encoding gene (a-9) Gene encoding MX1 (myxovirus (influenza virus) resistance 1, interferon-inducible protein p78 (mouse)) (a-10) Gene encoding CCR1 (chemokine (CC motif) receptor 1) (a-11) Gene encoding EMR1 (egf-like module containing, mucin-like, hormone receptor-like 1) (a-12) XIAP-related factor Gene encoding 1 (XIAP associated factor-1) (a-13) Gene encoding OASL (2'-5'-oligoadenylate synthetase-like) (a-14) OAS1 (2 ', 5'-oligoadenylate synthetase 1 (A-15) gene encoding IFIT5 (interferon-induced protein with tetratricopeptide repeats 5) (a-16) gene encoding WSB2 (WD repeat and SOCS box-containing 2) a-17) gene encoding FFAR2 (free fatty acid receptor 2) (a-18) gene encoding ISG15 (ISG15 ubiquitin-like modifier) (a-19) HIST1H3H (histone cluster 1, H3h (A-20) gene encoding IFI6 (interferon, alpha-inducible protein 6) (a-21) gene encoding CTSL1 (cathepsin L1) (a-22) GBP1 (guanylate binding protein 1, interferon -inducible, 67 kDa) gene (a-23) IRF7 (interferon regulatory factor 7) gene (a-24) HIST2H2BE (histone cluster 2, H2be) gene (a-25) IFIH1 (interferon) gene encoding induced with helicase C domain 1) (a-26) gene encoding monocyte to macrophage differentiation-associated (a-27) gene encoding CD36 molecule (thrombospondin receptor) (a- 28) Gene encoding MT2A (metallothionein 2A) (a-29) PSMB9 (proteasome (prosome, macropain) su gene encoding bunit, beta type, 9 (large multifunctional peptidase 2)) (a-30) gene encoding PLSCR1 (phospholipid scramblase 1) (a-31) OAS2 (2'-5'-oligoadenylate synthetase 2, 69 (A-32) gene encoding EIF2AK2 (eukaryotic translation initiation factor 2-alpha kinase 2) (a-33) gene encoding LTB (lymphotoxin beta (TNF superfamily, member 3)) a-34) gene encoding MS4A4A (membrane-spanning 4-domains, subfamily A, member 4) (a-35) gene encoding ADM (adrenomedullin) (a-36) OAS3 (2'-5'-oligoadenylate gene encoding synthetase 3, 100 kDa) (a-37) gene encoding SLAMF7 (SLAM family member 7) (a-38) DDX58 (DEAD (Asp-Glu-Ala-Asp) box polypeptide 58) (A-39) Gene encoding GNG11 (guanine nucleotide binding protein (G protein), gamma 11) (a-40) Gene encoding TSC22D1 (TSC22 domain family, member 1) (a-41) HIST1H2BD A gene encoding (histone cluster 1, H2bd) (a-42) a gene encoding IFITM3 (interferon induced transmembrane protein 3 (1-8U)) (a-43) encoding UBE2L6 (ubiquitin-conjugating enzyme E2L 6) Gene (a-44) Gene encoding H2BFS (H2B histone family, member S) (a-45) Gene encoding CAT (catalase) (a-46) SCO2 (SCO cytochrome oxidase deficient homolog 2 (yeast)) Encoding gene (a-47) Gene encoding PSME2 (proteasome (prosome, macropain) activator subunit 2 (PA28 beta)) (a-48) S A gene encoding TAT1 (signal transducer and activator of transcription 1, 91 kDa) (a-49) A gene encoding MTHFD2 (methylenetetrahydrofolate dehydrogenase (NADP + dependent) 2, methenyltetrahydrofolate cyclohydrolase) (a-50) MT1X (metallothionein 1X) (A-51) Gene encoding TGFA (transforming growth factor, alpha) (a-52) Gene encoding GPR109B (G protein-coupled receptor 109B) (a-53) HIST1H3D (histone cluster 1, H3d) (A-54) gene encoding IFI44L (interferon-induced protein 44-like) (a-55) gene encoding F13A1 (coagulation factor XIII, A1 polypeptide) (a-56) CX3CR1 (chemokine ( C-X3-C motif) gene encoding receptor 1) (a-57) FCGR A gene encoding A (Fc fragment of IgG, high affinity Ia, receptor (CD64)) (a-58) gene encoding IL1B (interleukin 1, beta) (a-59) encoding ARRB1 (arrestin, beta 1) Genes

[Group b]
(B-1) gene encoding ADAM28 (ADAM metallopeptidase domain 28) (b-2) gene encoding CEACAM8 (carcinoembryonic antigen-related cell adhesion molecule 8) (b-3) COL9A2 (collagen, type IX, alpha 2 (B-4) gene encoding DEFA4 (defensin, alpha 4, corticostatin) (b-5) gene encoding LTF (lactotransferrin) (b-6) SEC23B (Sec23 homolog B (S. cerevisiae) )) Encoding gene (b-7) gene encoding VSIG4 (V-set and immunoglobulin domain containing 4) (b-8) gene encoding DEFA1 (defensin, alpha 1) (b-9) CD163 (CD163 gene encoding molecule) (b-10) gene encoding RUNDC3A (RUN domain containing 3A (also known as RPIP8)) (b-11) CAM (B-12) Gene encoding LCN2 (lipocalin 2 (oncogene 24p3)) (b-13) Gene encoding HDAC9 (histone deacetylase 9) (b-14) CD24 (CD24 gene encoding molecule) (b-15) gene encoding IL1R2 (interleukin 1 receptor, type II)

  These genes have significant correlation between the prognosis of patients and changes in gene expression after the start of treatment (immunosuppression therapy) in patients with autoimmune diseases (MPO-ANCA-related vasculitis). Gene. The procedure for extracting these genes will be described below.

[Comprehensive gene expression analysis]
Blood samples from 20 patients with vasculitis syndrome (MPO-ANCA-related vasculitis) were collected from each patient before starting standard treatment for MPO-ANCA-related vasculitis and one week after starting treatment. (Peripheral blood 10 ml) was obtained. Here, “standard treatment of MPO-ANCA-related vasculitis” means methylprednisolone pulse therapy (0.5 to 1.0 g / day, intravenous therapy, 3 days) or oral administration of prednisolone (0.6 to 1. 0 mg / kg / day or 40-60 mg / day) and cyclophosphamide pulse therapy (0.5-0.75 / m ² , intravenous therapy, once / month) or cyclophosphamide oral administration (0 0.5 to 2.0 mg / kg / day or 50 to 100 mg / day) (see Non-Patent Document 1), and the first day of methylprednisolone pulse therapy or oral administration of prednisolone was defined as the “treatment start date” .

  RNA was extracted from each of these 40 samples using the PAXgene Blood RNA system (Nippon Becton Dickinson Co., Ltd.). Exhaustive gene expression analysis was performed on each of the 40 specimens using GeneChip (registered trademark) Human Genome Focus Array (8797 gene loaded: Affymetrix Japan KK). In this chip, at least 11 pairs of perfect match and mismatch probes are designed for one gene. Therefore, the difference in signal intensity with respect to these probes (“signal intensity with respect to perfect match probe” − “signal intensity with respect to mismatch probe”) was taken as the expression value of each gene. At this time, the expression value of 0.01 or less was set to 0.01 for convenience. Next, the median value of the expression values of all the genes mounted on each chip was calculated, and the expression value of each gene in the chip was divided by the obtained median value to achieve standardization between the chips. Furthermore, the median value was calculated for each gene from the expression values after standardization of all samples, and standardization between genes was attempted by dividing the expression value of the gene of each sample by the obtained median value. In this way, the expression of each gene between chips (between samples) can be compared. All of these experimental methods were performed according to the recommended protocol.

[Extraction of expression variable genes]
When the prognosis was examined for each of the 20 cases from which samples were obtained, as shown in FIG. 1, there were 10 cases of remission, 3 cases of improvement (improvement), 3 cases of death, and 4 cases of others. Therefore, genes that showed a significant (p <0.05) expression fluctuation 1.5 times or more before and after treatment were extracted in 13 cases (26 samples) in remission or improvement (improvement). As a result, a total of 74 genes were extracted from 8797 genes, 59 genes (group a) whose expression level decreased after treatment and 15 genes (group b) whose expression level increased (see FIG. 2). When gene clustering was performed on the extracted 74 genes, as shown in FIG. 3, genes (a group) whose expression level decreased after treatment and genes (b group) whose expression level increased after treatment were clearly separated. The names, expression changes (decrease or increase), expression fluctuation amount, and significance (p value) of the 74 genes extracted are shown in FIGS.

  As described above, among the genes used as an index in the prediction method of the present invention, the gene of group a is a gene whose gene expression level has decreased after the start of treatment in a patient in remission or remission, and the gene of group b is remission. Or it is a gene whose gene expression level has increased in a patient who has improved. The base sequences of these genes are all known, and the names, GenBank accession numbers, and GenBank RefSeq of the above genes are shown in FIGS. The base sequence of each gene is not limited to the base sequence indicated by the accession number or RefSeq shown in FIGS. 7 to 11, and is not particularly limited as long as it encodes each protein.

  The gene used as an index in the prediction method of the present invention is not particularly limited as long as it is one or more genes selected from the group consisting of a group and b group, but statistical genes calculated in the extraction process are not limited. A gene serving as an index may be selected based on superiority (p value: see FIGS. 4 to 6). In this case, it is preferable to select a gene having a small p value as an index in the prediction method of the present invention. For example, a gene with p <0.03 may be selected, or a gene with p <0.01. May be selected.

For example, by using the expression level of 16 genes having a small p value (p <0.032) or less as an index, the prognosis can be predicted with very high accuracy as shown in the Examples.
(A-1) a gene encoding CLC (a-2) a gene encoding IFIT1 (a-4) a gene encoding CCR3 (a-11) a gene encoding EMR1 (a-13) a gene encoding OASL (A-15) a gene encoding IFIT5 (a-17) a gene encoding FFAR2 (a-19) a gene encoding HIST1H3H (a-21) a gene encoding CTSL1 (a-24) a gene encoding HIST2H2BE (A-31) a gene encoding OAS2 (a-33) a gene encoding LTB (a-35) a gene encoding ADM (b-1) a gene encoding ADAM28 (b-2) a gene encoding CEACAM8 (B-3) Gene encoding COL9A2

  The disease targeted by the prediction method of the present invention is not particularly limited as long as it is an autoimmune disease. Examples of autoimmune diseases include vasculitis syndrome, and examples of vasculitis syndrome include MPO-ANCA associated vasculitis.

  The treatment targeted by the prediction method of the present invention is not particularly limited, and includes, for example, drug administration (including combined administration of multiple types of drugs) and plasma exchange therapy. Examples of the drug include an immunosuppressant such as cyclophosphamide, a steroidal drug such as methylprednisolone or prednisolone, a biological preparation, and the like.

  The sample (specimen) for examining the expression level of each gene is not particularly limited, but blood (peripheral blood) is preferable. This is because blood can be easily collected without burdening the patient. When examining the expression level of genes in blood, white blood cells (neutrophils, eosinophils, basophils, lymphocytes and monocytes) may be separated from the collected blood, but the collected blood is expressed as it is. It may be used for quantity analysis. The amount of blood to be collected may be appropriately set according to a method for examining changes in the expression level of a gene described later.

  In order to predict the effect of treatment on a subject using the expression level of each gene as an index, for example, the expression level of a gene when a predetermined time (for example, one week) has elapsed since receiving the treatment, and before that time The gene expression level at a predetermined time (for example, immediately before the start of treatment) may be compared. The gene included in group a is a gene whose expression level decreases when the symptoms of autoimmune disease improve, and the gene included in group b is a gene whose expression level increases when symptoms of autoimmune disease improve. is there. Therefore, for the genes included in group a, when the expression level of a gene when a predetermined time has elapsed since receiving treatment is reduced compared to the expression level of the gene at a predetermined time before that time Predicts that the treatment will be effective for the patient. Similarly, for the genes included in group b, the gene expression level when a predetermined time has elapsed since receiving treatment is increased compared to the gene expression level at a predetermined time before that time. Sometimes predict that the treatment will be effective for the patient. Further, as shown in the Examples, a prediction formula may be obtained by paying attention to the influence of changes in the expression level of each gene on the prognosis prediction, and the effect of treatment may be predicted using the prediction formula.

  In order to examine the change in the expression level of each gene, the amount of a transcription product (mRNA) or a translation product (protein, polypeptide or a fragment thereof) in a sample collected from a subject may be measured.

  To measure the amount of transcript (mRNA) in a sample, use a known gene analysis technique such as a hybridization technique using a probe for each gene or a gene amplification technique using a primer for each gene. do it. Examples of hybridization techniques include the Northern hybridization method, dot blot method, DNA microarray (DNA chip) method and the like. Examples of gene amplification techniques include RT-PCR method and real-time PCR method (calibration curve method, comparative Ct method) and the like. When the expression level of each gene is measured using a hybridization technique or a gene amplification technique, the following (c-1) to (c-) are used for the genes (a-1) to (a-59). 59) The following oligos / polynucleotides (c-60) to (c-74) were used as probes or primers for the genes (b-1) to (b-15) above. Can be used.

[Group c]
(C-1) Oligo / polynucleotide that can hybridize to nucleic acid encoding CLC (c-2) Oligo / polynucleotide that can hybridize to nucleic acid encoding IFIT1 (c-3) Hybridize to nucleic acid encoding IFIT3 Oligo / polynucleotide (c-4) capable of hybridizing to nucleic acid encoding CCR3 (c-5) Oligo / polynucleotide capable of hybridizing to nucleic acid encoding HERC5 (c-6) Oligo / polynucleotide hybridizable to nucleic acid encoding NGFRAP1 (c-7) Oligo / polynucleotide hybridizable to nucleic acid encoding TNFSF10 (c-8) Oligo / polynucleotide hybridizable to nucleic acid encoding FGL2 Polynucleotide (c 9) Oligo / polynucleotide hybridizable to nucleic acid encoding MX1 (c-10) Oligo / polynucleotide hybridizable to nucleic acid encoding CCR1 (c-11) Hybridizable to nucleic acid encoding EMR1 Oligo / polynucleotide (c-12) Oligo / polynucleotide hybridizable to nucleic acid encoding XIAP-related factor-1 (c-13) Oligo / polynucleotide hybridizable to nucleic acid encoding OASL (c-14) ) Oligo / polynucleotide hybridizable to nucleic acid encoding OAS1 (c-15) Oligo / polynucleotide hybridizable to nucleic acid encoding IFIT5 (c-16) Oligo hybridizable to nucleic acid encoding WSB2 / Polynucleotide c-17) Oligo / polynucleotide hybridizable to nucleic acid encoding FFAR2 (c-18) Oligo / polynucleotide hybridizable to nucleic acid encoding ISG15 (c-19) Hybridized to nucleic acid encoding HIST1H3H Possible oligo / polynucleotide (c-20) oligo / polynucleotide hybridizable to nucleic acid encoding IFI6 (c-21) oligo / polynucleotide hybridizable to nucleic acid encoding CTSL1 (c-22) GBP1 (C-23) Oligo / polynucleotide hybridizable to nucleic acid encoding IRF7 (c-24) Oligo hybridizable to nucleic acid encoding HIST2H2BE Polynucleotide (c-25) Oligo / polynucleotide hybridizable to nucleic acid encoding IFIH1 (c-26) Oligo / polynucleotide hybridizable to nucleic acid encoding MMD (c-27) Nucleic acid encoding CD36 (C-28) Oligo / polynucleotide that can hybridize to nucleic acid encoding MT2A (c-29) Oligo / polynucleotide that can hybridize to nucleic acid encoding PSMB9 (c- 30) Oligo / polynucleotide hybridizable to a nucleic acid encoding PLSCR1 (c-31) Oligo / polynucleotide hybridizable to a nucleic acid encoding OAS2 (c-32) Hybridizable to a nucleic acid encoding EIF2AK2 Oligo / polynucleotide (c-33) Oligo / polynucleotide hybridizable to the nucleic acid encoding LTB (c-34) Oligo / polynucleotide hybridizable to the nucleic acid encoding MS4A4A (c-35) Encoding ADM (C-36) Oligo / polynucleotide hybridizable to nucleic acid encoding OAS3 (c-37) Oligo / polynucleotide hybridizable to nucleic acid encoding SLAMF7 c-38) Oligo / polynucleotide hybridizable to nucleic acid encoding DDX58 (c-39) Oligo / polynucleotide hybridizable to nucleic acid encoding GNG11 (c-40) Hybridizer to nucleic acid encoding TSC22D1 Oligo / polynucleotide (c-41) Oligo / polynucleotide hybridizable to a nucleic acid encoding HIST1H2BD (c-42) Oligo / polynucleotide hybridizable to a nucleic acid encoding IFITM3 (c-43) Oligo / polynucleotide hybridizable to nucleic acid encoding UBE2L6 (c-44) Oligo / polynucleotide hybridizable to nucleic acid encoding H2BFS (c-45) Oligo / polynucleotide hybridizable to nucleic acid encoding CAT Polynucleotide (c-46) Oligo / polynucleotide that can hybridize to nucleic acid encoding SCO2 (c-47) Oligo / polynucleotide that can hybridize to nucleic acid encoding PSME2 (c-48) Encodes STAT1 Oligo / polynucleotide that can hybridize to acid (c-49) Oligo / polynucleotide that can hybridize to nucleic acid encoding MTHFD2 (c-50) Oligo / polynucleotide that can hybridize to nucleic acid encoding MT1X (c -51) Oligo / polynucleotide hybridizable to nucleic acid encoding TGFA (c-52) Oligo / polynucleotide hybridizable to nucleic acid encoding GPR109B (c-53) Hybridized to nucleic acid encoding HIST1H3D Oligo / polynucleotide (c-54) Oligo / polynucleotide hybridizable to a nucleic acid encoding IFI44L (c-55) Oligo / polynucleotide hybridizable to a nucleic acid encoding F13A1 (c-56) C Oligo / polynucleotide hybridizable to nucleic acid encoding X3CR1 (c-57) Oligo / polynucleotide hybridizable to nucleic acid encoding FCGR1A (c-58) Oligo / polynucleotide hybridizable to nucleic acid encoding IL1B Polynucleotide (c-59) Oligo / polynucleotide hybridizable to the nucleic acid encoding ARRB1 (c-60) Oligo / polynucleotide hybridizable to the nucleic acid encoding ADAM28 (c-61) Nucleic acid encoding CEACAM8 (C-62) Oligo / polynucleotide hybridizable to a nucleic acid encoding COL9A2 (c-63) Oligo / polynucleotide hybridizable to a nucleic acid encoding DEFA4 Otide (c-64) Oligo / polynucleotide hybridizable to nucleic acid encoding LTF (c-65) Oligo / polynucleotide hybridizable to nucleic acid encoding SEC23B (c-66) Nucleic acid encoding VSIG4 Hybridizable oligo / polynucleotide (c-67) Oligo / polynucleotide hybridizable to nucleic acid encoding DEFA1 (c-68) Oligo / polynucleotide hybridizable to nucleic acid encoding CD163 (c-69) ) Oligo / polynucleotide hybridizable to nucleic acid encoding RUNDC3A (c-70) Oligo / polynucleotide hybridizable to nucleic acid encoding CAMP (c-71) Oligo hybridizable to nucleic acid encoding LCN2 / Renucleotide (c-72) Oligo / polynucleotide hybridizable to nucleic acid encoding HDAC9 (c-73) Oligo / polynucleotide hybridizable to nucleic acid encoding CD24 (c-74) Nucleic acid encoding IL1R2 Oligo / polynucleotide that can hybridize to

  In the above (c-1) to (c-74), examples of the nucleic acid encoding the protein include mRNA, cDNA, cRNA and the like. The nucleotide constituting the oligo / polynucleotide may be any of deoxyribonucleotide, ribonucleotide and non-natural nucleotide. The base length of the oligo / polynucleotide is not particularly limited, and may be appropriately set according to the method for measuring the amount of the transcript.

  The base sequence of each oligo / polynucleotide may be designed based on the base sequence of a nucleic acid to which each oligo / polynucleotide can hybridize. For example, a coding region (CDS) may be selected from each gene belonging to the above group a and group b, and the oligo / polynucleotide base sequence may be designed so as to hybridize to the region. Moreover, it hybridizes to the region extending from the coding region to the region adjacent to the 5 ′ end side or 3 ′ end side so that it can hybridize to the region adjacent to the 5 ′ end side or 3 ′ end side of the coding region. You may design so that it may. The designed oligo / polynucleotide is preferably used as a primer or probe to confirm whether or not it hybridizes to the target nucleic acid. Each oligo / polynucleotide may be labeled with a restriction enzyme recognition sequence, a tag, a fluorescent dye, a radioisotope, and the like.

  In order to measure the amount of translation product (protein, polypeptide or fragment thereof) in a sample, a known protein analysis technique such as Western blotting, immunoprecipitation or ELISA using an antibody or fragment thereof is used. Just measure. When the expression level of each gene is measured using a protein analysis technique, the following antibodies (d-1) to (d-59) against the genes (a-1) to (a-59) above: Alternatively, for the above genes (b-1) to (b-15), the following antibodies (d-60) to (d-74) or fragments thereof can be used.

[Group d]
(D-1) Antibody or fragment thereof capable of reacting with CLC (d-2) Antibody or fragment thereof capable of reacting with IFIT1 (d-3) Antibody or fragment thereof capable of reacting with IFIT3 (d-4) Reacting with CCR3 (D-5) Antibody or fragment thereof capable of reacting with HERC5 (d-6) Antibody or fragment thereof capable of reacting with NGFRAP (d-7) Antibody or fragment thereof capable of reacting with TNFSF10 (d -8) An antibody or fragment thereof that can react with FGL2 (d-9) An antibody or fragment thereof that can react with MX1 (d-10) An antibody or fragment thereof that can react with CCR1 (d-11) It can react with EMR1 Antibody or fragment thereof (d-12) Antibody or fragment thereof capable of reacting with XIAP-related factor-1 (d-13) Antibody or fragment thereof capable of reacting with OASL (D-14) Antibody or fragment thereof capable of reacting with OAS1 (d-15) Antibody or fragment thereof capable of reacting with IFIT5 (d-16) Antibody or fragment thereof capable of reacting with WSB2 (d-17) Reacting with FFAR2 (D-18) Antibody or fragment thereof capable of reacting with ISG15 (d-19) Antibody or fragment thereof capable of reacting with HIST1H3H (d-20) Antibody or fragment thereof capable of reacting with IFI6 (d -21) An antibody or fragment thereof that can react with CTSL1 (d-22) An antibody or fragment thereof that can react with GBP1 (d-23) An antibody or fragment thereof that can react with IRF7 (d-24) It can react with HIST2H2BE Antibody or fragment thereof (d-25) Antibody or fragment thereof that can react with IFIH1 (d-26) Can react with MMD (D-27) Antibody or fragment thereof reactive with CD36 (d-28) Antibody or fragment thereof reactive with MT2A (d-29) Antibody or fragment thereof reactive with PSMB9 (d-30) ) An antibody or fragment thereof capable of reacting with PLSCR1 (d-31) an antibody or fragment thereof capable of reacting with OAS2 (d-32) an antibody or fragment thereof capable of reacting with EIF2AK2 (d-33) an antibody capable of reacting with LTB or Fragment (d-34) Antibody or fragment thereof capable of reacting with MS4A4A (d-35) Antibody or fragment thereof capable of reacting with ADM (d-36) Antibody or fragment thereof capable of reacting with OAS3 (d-37) SLAMF7 Antibody or fragment thereof capable of reacting with (d-38) Antibody or fragment thereof capable of reacting with DDX58 (d-39) GNG11 Antibody or fragment thereof capable of reacting (d-40) Antibody or fragment thereof capable of reacting with TSC22D1 (d-41) Antibody or fragment thereof capable of reacting with HIST1H2BD (d-42) Antibody or fragment thereof capable of reacting with IFITM3 d-43) An antibody or fragment thereof capable of reacting with UBE2L6 (d-44) An antibody or fragment thereof capable of reacting with H2BFS (d-45) An antibody or fragment thereof capable of reacting with CAT (d-46) Reacting with SCO2 (D-47) Antibody or fragment thereof capable of reacting with PSME2 (d-48) Antibody or fragment thereof capable of reacting with STAT1 (d-49) Antibody or fragment thereof capable of reacting with MTHFD2 (d- 50) An antibody that can react with MT1X or a fragment thereof (d-51) An antibody that can react with TGFA or a fragment thereof Fragment (d-52) Antibody or fragment thereof capable of reacting with GPR109B (d-53) Antibody or fragment thereof capable of reacting with HIST1H3D (d-54) Antibody or fragment thereof capable of reacting with IFI44L (d-55) F13A1 Reactive antibody or fragment thereof (d-56) Antibody or fragment thereof capable of reacting with CX3CR1 (d-57) Antibody or fragment thereof capable of reacting with FCGR1A (d-58) Antibody or fragment thereof capable of reacting with IL1B ( d-59) An antibody or fragment thereof capable of reacting with ARRB1 (d-60) An antibody or fragment thereof capable of reacting with ADAM28 (d-61) An antibody or fragment thereof capable of reacting with CEACAM8 (d-62) Reacting with COL9A2 Antibody or fragment thereof (d-63) Antibody or fragment thereof capable of reacting with DEFA4 (d-6) ) An antibody or fragment thereof capable of reacting with LTF (d-65) An antibody or fragment thereof capable of reacting with SEC23B (d-66) An antibody or fragment thereof capable of reacting with VSIG4 (d-67) An antibody capable of reacting with DEFA1 or Fragment (d-68) Antibody reactive with CD163 or fragment thereof (d-69) Antibody reactive with RUNDC3A or fragment thereof (d-70) Antibody reactive with CAMP or fragment thereof (d-71) LCN2 (D-72) An antibody or fragment thereof capable of reacting with HDAC9 (d-73) An antibody or fragment thereof capable of reacting with CD24 (d-74) An antibody or fragment thereof capable of reacting with IL1R2

In the above (d-1) to (d-74), examples of the antibody include a monoclonal antibody and a polyclonal antibody. Examples of antibody fragments include Fab fragments, F (ab) ′ ₂ fragments, single chain antibodies (scFv), and the like. Each antibody (or fragment thereof) preferably reacts with the target protein but does not react with other proteins contained in blood. The method for preparing the antibodies (d-1) to (d-74) is not particularly limited, and may be prepared using methods known to those skilled in the art.

  When measuring the amount of translation product using an antibody or fragment thereof, for example, radioimmunoassay (RIA), enzyme immunoassay (EIA), chemiluminescence assay (CLIA), fluorescence immunoassay ( FIA) or the like may be used. Specifically, after capturing a target translation product in a sample using a solid phase carrier (for example, an immunoplate or latex particle) to which an antibody is bound by physical adsorption or chemical binding, What is necessary is just to quantify the translation product captured using a labeled antibody (an antibody labeled with an enzyme, a fluorescent substance, etc.) having a different antigen recognition site from the immobilized antibody.

  Alternatively, the amount of translation product may be measured by measuring the activity of the translation product. The activity of the translation product can be measured, for example, by a known method such as Western blotting or ELISA using an antibody or fragment thereof that can react with the translation product.

  Hereinafter, an embodiment of the method for predicting the therapeutic effect of the present invention (method using a DNA microarray) will be described, but the method for predicting the therapeutic effect of the present invention is not limited to this embodiment. For example, as shown in the examples, the method for predicting the therapeutic effect of the present invention can be carried out using a real-time PCR method.

  In the following description, it is assumed that a DNA microarray in which two or more kinds of oligo / polynucleotides selected from the oligo / polynucleotides included in the group c are fixed is prepared in advance.

  First, before starting the treatment, blood is collected from a subject suffering from an autoimmune disease. The timing of the first blood collection is preferably performed before the start of treatment, but is not particularly limited as long as the effect of the treatment appears, and may be after the start of treatment (for example, immediately after the start of treatment). . Hereinafter, the blood collected at this time is referred to as a “first sample”.

  Next, treatment is started for the subject. For example, when the subject suffers from MPO-ANCA-related vasculitis (microscopic polyangiitis), combined administration of cyclophosphamide and methylprednisolone (or prednisolone) is started (see Non-Patent Document 1).

  Then, blood is collected again from the subject one week after the start of treatment. The timing of performing the second blood collection is not particularly limited as long as the effect of the treatment appears, and may be, for example, three days after the start of the treatment. Hereinafter, the blood collected at this time is referred to as a “second sample”.

  Next, mRNA is extracted from the first sample (blood before the start of treatment), cDNA is prepared using the extracted mRNA as a template, and the prepared cDNA is fluorescently labeled. A solution containing fluorescently labeled cDNA is provided on the DNA microarray to hybridize the probe (oligo / polynucleotide) on the DNA microarray with the cDNA. After the completion of hybridization, the fluorescence intensity at each spot of the DNA microarray is measured using a scanner, and the expression level of each gene (genes contained in the above groups a and b) in the first sample is measured.

  Similarly, fluorescently labeled cDNA is prepared from the second sample (blood after the start of treatment), the probe (oligo / polynucleotide) on the DNA microarray is hybridized with the cDNA, and each gene in the second sample is hybridized. The expression level of the gene (the gene contained in group a and b) is measured.

  Next, the expression level of each gene is compared between the first sample and the second sample. As a result, for each of the genes included in group a, when the expression level in the second sample is reduced compared to the expression level in the first sample, the treatment performed on the subject is given to the subject. Predict that it will be effective. Similarly, when the expression level in the second sample is increased compared to the expression level in the first sample for the genes included in group b, the treatment is predicted to be effective for the subject. To do.

  As described above, the method for predicting the therapeutic effect of the present invention uses, as an index, the expression level of one or more genes selected from the group consisting of group a and group b in the blood of a subject. When a drug is administered to a subject with an autoimmune disease, it can be predicted at an early stage whether the drug is effective for the subject. Thereby, the doctor who treats the subject can appropriately determine at an early stage whether or not to use the medicine for the subject based on the prediction result.

2. Screening method of the present invention The screening method of the present invention comprises the steps of administering a test substance to a model animal of an autoimmune disease and determining the therapeutic effect of the test substance on the autoimmune disease. A method for screening a substance having a therapeutic effect on the sample, wherein the expression level of one or more genes selected from the group consisting of group a and group b in the sample collected from the model animal is used as an index The therapeutic effect of a test substance is determined.

The gene used as an index in the screening method of the present invention is not particularly limited as long as it is one or more genes selected from the group consisting of a group and b group, but based on the statistical superiority described above. A gene as an index may be selected. In this case, it is preferable to select a gene having a small p value as an index in the screening method of the present invention. For example, a gene with p <0.03 may be selected, or a gene with p <0.01. May be selected. For example, the expression level of 16 genes having a small p value (p <0.032) or less may be used as an index.
(A-1) a gene encoding CLC (a-2) a gene encoding IFIT1 (a-4) a gene encoding CCR3 (a-11) a gene encoding EMR1 (a-13) a gene encoding OASL (A-15) a gene encoding IFIT5 (a-17) a gene encoding FFAR2 (a-19) a gene encoding HIST1H3H (a-21) a gene encoding CTSL1 (a-24) a gene encoding HIST2H2BE (A-31) a gene encoding OAS2 (a-33) a gene encoding LTB (a-35) a gene encoding ADM (b-1) a gene encoding ADAM28 (b-2) a gene encoding CEACAM8 (B-3) Gene encoding COL9A2

  The model animal of the autoimmune disease to which the test substance is administered is not particularly limited, and a known model animal of the autoimmune disease can be used. The target disease is not particularly limited as long as it is an autoimmune disease. Examples of autoimmune diseases include vasculitis syndrome, and examples of vasculitis syndrome include MPO-ANCA associated vasculitis. The species of the model animal is not particularly limited as long as it is other than human, and rats, mice, guinea pigs, rabbits and the like can be used.

  The type of the test substance is not particularly limited, and may be, for example, a high molecular compound, a low molecular compound, an antibody, a protein, a peptide, a nucleic acid, a carbohydrate, an inorganic salt, a metal complex, or a complex thereof.

  The sample (specimen) for examining the expression level of each gene is not particularly limited, but blood (peripheral blood) is preferable. This is because blood can be easily collected without burdening the model animal. When examining the expression level of genes in blood, white blood cells (neutrophils, eosinophils, basophils, lymphocytes and monocytes) may be separated from the collected blood, but the collected blood is expressed as it is. It may be used for quantity analysis.

  In order to determine the effect of the test substance on the autoimmune disease using the expression level of each gene as an index, for example, the gene expression at the time when a predetermined time (for example, one week) has elapsed since the test substance was administered. The expression level may be compared with the expression level of the gene at a predetermined time before that time (for example, immediately before the start of administration). The gene included in group a is a gene whose expression level decreases when the symptoms of autoimmune disease improve, and the gene included in group b is a gene whose expression level increases when symptoms of autoimmune disease improve. is there. Therefore, for the genes included in group a, the expression level of the gene when a predetermined time has elapsed since the administration of the test substance is decreased compared to the expression level of the gene at a predetermined time before that time. Is determined to be effective against autoimmune diseases. Similarly, for the genes included in group b, the gene expression level when a predetermined time has elapsed since the administration of the test substance is increased compared to the gene expression level at a predetermined time before that time. The test substance is determined to be effective against autoimmune diseases. Alternatively, the determination formula may be obtained by paying attention to the influence of the change in the expression level of each gene on the determination of the therapeutic effect, and the therapeutic effect may be determined using the determination formula.

  In order to examine the change in the expression level of each gene, as described above, the amount of a transcription product (mRNA) or a translation product (protein, polypeptide or a fragment thereof) in a sample collected from a model animal may be measured. .

  Hereinafter, an embodiment of the screening method of the present invention (method using a DNA microarray) will be described, but the screening method of the present invention is not limited to this embodiment. For example, the screening method of the present invention can be carried out using a real-time PCR method.

  In the following description, it is assumed that a DNA microarray in which two or more kinds of oligo / polynucleotides selected from the oligo / polynucleotides included in the group c are fixed is prepared in advance.

  First, blood is collected from a model animal of an autoimmune disease before administering a test substance. The timing of the first blood sampling is preferably performed before administration of the test substance, but is not particularly limited as long as the effect of the test substance appears, and after administration of the test substance (for example, test substance) Immediately after administration). Hereinafter, the blood collected at this time is referred to as a “first sample”.

  Subsequently, a test substance is administered to the model animal. One week after administration of the test substance, blood is again collected from the model animal. Note that the timing of the second blood collection is not particularly limited as long as the effect of the test substance appears, and may be, for example, two weeks later. Hereinafter, the blood collected at this time is referred to as a “second sample”.

  Next, fluorescently labeled cDNA is prepared from the first sample (blood before administration of the test substance), and the probe (oligo / polynucleotide) on the DNA microarray is hybridized with the cDNA. The expression level of each gene is measured. Similarly, a fluorescently labeled cDNA is prepared from the second sample (blood after administration of the test substance), the probe (oligo / polynucleotide) on the DNA microarray is hybridized with the cDNA, and the second sample is prepared. The expression level of each gene in is measured.

  Next, the expression level of each gene is compared between the first sample and the second sample. As a result, for each of the genes contained in group a, when the expression level in the second sample is reduced compared to the expression level in the first sample, the test substance administered to the model animal is subject to the autoimmunity. Determined to be effective against the disease. Similarly, when the expression level in the second sample is increased compared to the expression level in the first sample for the gene contained in the group b, the test substance administered to the model animal is affected by the autoimmune disease. It is determined that it is effective.

  As described above, the screening method of the present invention uses the expression level of one gene or two or more genes selected from the group consisting of group a and group b in the blood of a model animal as an index, thereby providing autoimmunity. A substance having a therapeutic effect on a disease can be rapidly screened.

3. Oligo / polynucleotide array of the present invention The oligo / polynucleotide array of the present invention is an oligo / polynucleotide array comprising a support and two or more kinds of oligo / polynucleotides immobilized on the support, The oligo / polynucleotide is selected from the group c.

  The oligo / polynucleotide array of the present invention is not particularly limited as long as it is two or more kinds of oligo / polynucleotides selected from group c, but the statistical superiority of the corresponding gene (see FIGS. 4 to 6). You may choose based on: In this case, it is preferable to select an oligo / polynucleotide immobilized on the support having a small p value of the corresponding gene. For example, an oligo / polynucleotide corresponding to a gene with p <0.03 is selected. Alternatively, an oligo / polynucleotide corresponding to a gene with p <0.01 may be selected.

  The material of the support is not particularly limited as long as it is insoluble in various liquids used in the method for predicting therapeutic effects and the screening method described above, and may be, for example, glass, plastic, metal, ceramics, or a composite material thereof. That's fine. The shape of the support is not particularly limited and may be, for example, a flat plate shape.

  The oligo / polynucleotide contained in the group c is fixed on the support so that the type (whether (c-1) to (c-74)) can be identified based on the position on the support. It is preferred that The amount of the oligo / polynucleotide immobilized on the support is not particularly limited as long as the expression level of the target gene can be measured. The method for immobilizing the oligo / polynucleotide on the support is not particularly limited, and techniques known to those skilled in the art may be used. For example, the oligo / polynucleotide may be immobilized on the surface of the support using, for example, electrostatic bond, covalent bond, protein-protein interaction, protein-small molecule interaction and the like.

  As described above, in the oligo / polynucleotide array of the present invention, two or more types of oligo / polynucleotides contained in the group c are immobilized, and the expression levels of the plurality of types of genes contained in the groups a and b Therefore, the therapeutic effect prediction method and the screening method can be quickly performed.

  EXAMPLES Hereinafter, although this invention is demonstrated in detail with reference to an Example, this invention is not limited by these Examples.

  In this example, a gene (group) that seems to be optimal for predicting prognosis is extracted from 74 genes, and the prognosis of a patient with vasculitis syndrome is predicted using the gene (group) as an index.

1. Sample Preparation For 25 patients with vasculitis syndrome (MPO-ANCA-related vasculitis), each patient before starting standard treatment for MPO-ANCA-related vasculitis (described above) and one week after starting treatment A blood sample was collected from the sample to obtain a specimen (peripheral blood 10 ml). RNA was extracted from each of these 50 samples using the PAXgene Blood RNA system (Nippon Becton Dickinson Co., Ltd.).

2. Optimal gene extraction and prediction formula determination (LDA analysis)
44 genes (a-1 to a-36, b-1 to b-8: target genes) having a small p-value among 74 genes of group a and b and 4 genes (endogenous control genes) for quality control 384 well plates (TaqMan Low Density Arrays: Applied Biosystems) filled with the above primer / probe set were prepared. The endogenous control genes used are the following 4 genes.
-ACTB (Actin, beta) gene-GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) gene-Gene encoding 18S (Ribosomal RNA 18S)-HPRT1 (Hypoxanthine phosphoribosyltransferase) gene

  Using this plate, the Ct (Threshold Cycle) value of each gene was measured for each of 50 samples by the real-time PCR method. All of these experimental methods were performed according to the recommended protocol.

For each sample, the Ct value of the endogenous control gene was subtracted from the Ct value of the target gene (44 genes) to calculate the ΔCt value of each target gene. This time, the ACTB gene with the least variation between samples was selected as an endogenous control gene. Further, for each case, the ΔCt value of each target gene was calculated by subtracting the ΔCt value of the sample before treatment (reference sample) from the ΔCt value of the sample after treatment (target sample). The obtained ΔΔCt value was substituted into the following equation to calculate the signal value (relative expression level) of each target gene (comparative Ct method). This signal value means the relative expression level in the sample after treatment (target sample) when the expression level in the sample before treatment (reference sample) is “1”.

Signal value (relative expression level) = 2− ^ΔΔCt

(Multiple regression analysis)
Of the 48 genes, 16 genes were randomly selected from 47 genes excluding the ACTB gene used as the endogenous control gene, the signal value (2- ^ΔΔCt ) of the selected gene was used as the explanatory variable, and the prognosis of the same case was the target variable As a result, multiple regression analysis (least square method) was performed. At this time, a dummy variable in which “remission” indicating prognosis was set to “0” and “non-remission” was set to “1” was set as an objective variable.

  The degree of influence on the prognosis (target variable) was calculated for each of the selected 16 genes. The gene with the least influence was removed from the selected 16 genes, other arbitrary genes not added to the analysis were added (replaced), and multiple regression analysis was performed again. This operation was repeated until no genes were replaced. After the genes to be replaced disappeared, the multiple regression analysis was repeated again after removing the gene with the smallest influence, and the multiple regression analysis was performed until there was finally one explanatory variable (variable increase / decrease method).

For each of the obtained 46 regression equations (regression equations 1 to 46), Ru (explanatory variable selection criterion) and AIC (Akaike's information criterion) are calculated, and the optimal regression equation (prediction equation with the highest prediction accuracy) is calculated. It was determined. The optimal regression equation (maximum Ru and minimum AIC) is shown below. By substituting the signal value (2- ^ΔΔCt ) of each gene of the patient whose prognosis is to be predicted into the following prediction formula (regression formula 25), the predicted value of prognosis can be calculated. When this predicted value is close to 0, it means that the possibility of remission is high, and when it is close to 1, it means that the possibility of non-remission is high.
[Regression Formula 25]
Prognosis = 0.146798412
+ (− 0.051782469) × (2− ^{ΔΔCt of} ADAM28 gene)
+ (1.25615398) × (ADM gene 2 ^−ΔΔCt )
+ (0.231899078) × (2− ^{ΔΔCt of} CCR3 gene)
+ (0.639838691) × (2- ^{ΔΔCt of} CLC gene)
+ (0.078500847) × (2- ^{ΔΔCt of} COL9A2 gene)
+ (1.294981065) × (2- ^{ΔΔCt of} CTSL1 gene)
+ (− 0.886331118) × (2- ^{ΔΔCt of} EMR1 gene)
+ (− 0.440665266) × (2- ^{ΔΔCt of} FFAR2 gene)
+ (− 0.839871298) × (2− ^{ΔΔCt of} HIST1H3H gene)
+ (− 0.6667723272) × (2− ^{ΔΔCt of} HIST2H2BE gene)
+ (− 0.9782333) × (2− ^{ΔΔCt of} IFIT1 gene)
+ (0.656838725) × (2− ^{ΔΔCt of} IFIT5 gene)
+ (− 0.404414372) × (LTB gene 2 ^−ΔΔCt )
+ (− 0.0154777967) × (2− ^{ΔΔCt of CEACAM8} gene)
+ (0.390821135) × (2− ^{ΔΔCt of} OAS2 gene)
+ (0.5555560415) × (2- ^{ΔΔCt of} OASL gene)

3. Prediction of prognosis The prognosis was predicted for the 25 cases described above using the above prediction formula using the expression level of 16 genes as an index. When calculating the ΔΔCt value of each gene, the ACTB gene was used as an endogenous control gene, and whole blood RNA before treatment of the same case was used as a reference sample.

FIG. 12 is a graph showing the relationship between the prognostic value calculated for 25 cases (horizontal axis) and the actual prognosis (remission 0 / non-remission 1: vertical axis). From this graph, it can be seen that the prognosis can be predicted with a very high correlation with R ² (multiple determination) = 0.977.

  The present invention is useful, for example, in the manufacturing industry of DNA microarrays (DNA chips) and related equipment.

Table showing the prognosis of 20 severe patients with vasculitis syndrome from which samples were obtained Graph showing change in expression level of extracted 74 genes before and after treatment The figure which shows the result of clustering of the extracted 74 genes Table showing names, expression changes, fluctuation amounts and p-values of 30 genes (a-1 to a-30) out of 74 extracted genes Table showing names, expression changes, fluctuation amounts and p-values of 29 genes (a-31 to a-59) out of 74 extracted genes Table showing names, expression changes, fluctuation amounts and p-values of 15 genes (b-1 to b-15) out of 74 extracted genes Table showing names, accession numbers, and RefSeq of 15 genes (a-1 to a-15) out of 74 genes that can be used as an index in the therapeutic effect prediction method and screening method of the present invention Table showing names, accession numbers, and RefSeq of 15 genes (a-16 to a-30) out of 74 genes that can be used as an index in the therapeutic effect prediction method and screening method of the present invention Table showing names, accession numbers, and RefSeq of 15 genes (a-31 to a-45) out of 74 genes that can be used as an index in the therapeutic effect prediction method and screening method of the present invention Table showing the names, accession numbers, and RefSeq of 14 genes (a-46 to a-59) out of 74 genes that can be used as indices in the therapeutic effect prediction method and screening method of the present invention Table showing gene (b-1 to a-15) names, accession numbers, and RefSeq as indices in the therapeutic effect prediction method and screening method of the present invention The graph which shows the result of an Example

Claims (14)

A method for predicting the effect of the treatment on the subject, using as an index the expression level of a gene in blood collected from a subject with autoimmune disease treated,
The prediction method, wherein the gene is one or more genes selected from the group consisting of the following (a-1) to (a-59) and (b-1) to (b-15).
(A-1) a gene encoding CLC (a-2) a gene encoding IFIT1 (a-3) a gene encoding IFIT3 (a-4) a gene encoding CCR3 (a-5) a gene encoding HERC5 (A-6) a gene encoding NGFRAP1 (a-7) a gene encoding TNFSF10 (a-8) a gene encoding FGL2 (a-9) a gene encoding MX1 (a-10) a gene encoding CCR1 (A-11) a gene encoding EMR1 (a-12) a gene encoding XIAP-related factor-1 (a-13) a gene encoding OASL (a-14) a gene encoding OAS1 (a-15) IFIT5 (A-16) gene encoding WSB2 (a-17) gene encoding FFAR2 Child (a-18) gene encoding ISG15 (a-19) gene encoding HIST1H3H (a-20) gene encoding IFI6 (a-21) gene encoding CTSL1 (a-22) encoding GBP1 Gene (a-23) gene encoding IRF7 (a-24) gene encoding HIST2H2BE (a-25) gene encoding IFIH1 (a-26) gene encoding MMD (a-27) encoding CD36 Gene (a-28) gene encoding MT2A (a-29) gene encoding PSMB9 (a-30) gene encoding PLSCR1 (a-31) gene encoding OAS2 (a-32) encoding EIF2AK2 Gene (a-33) Gene encoding LTB (a-34) A gene encoding S4A4A (a-35) A gene encoding ADM (a-36) A gene encoding OAS3 (a-37) A gene encoding SLAMF7 (a-38) A gene encoding DDX58 (a-39) A gene encoding GNG11 (a-40) A gene encoding TSC22D1 (a-41) A gene encoding HIST1H2BD (a-42) A gene encoding IFITM3 (a-43) A gene encoding UBE2L6 (a-44) H2BFS encoding gene (a-45) CAT encoding gene (a-46) SCO2 encoding gene (a-47) PSME2 encoding gene (a-48) STAT1 encoding gene (a-49) Gene encoding MTHFD2 (a-50) MT1 (A-51) gene encoding TGFA (a-52) gene encoding GPR109B (a-53) gene encoding HIST1H3D (a-54) gene encoding IFI44L (a-55) F13A1 (A-56) gene encoding CX3CR1 (a-57) gene encoding FCGR1A (a-58) gene encoding IL1B (a-59) gene encoding ARRB1 (b-1) ADAM28 (B-2) gene encoding CEACAM8 (b-3) gene encoding COL9A2 (b-4) gene encoding DEFA4 (b-5) gene encoding LTF (b-6) SEC23B (B-7) encoding VSIG4 Gene (b-8) gene encoding DEFA1 (b-9) gene encoding CD163 (b-10) gene encoding RUNDC3A (b-11) gene encoding CAMP (b-12) encoding LCN2 Gene (b-13) Gene encoding HDAC9 (b-14) Gene encoding CD24 (b-15) Gene encoding IL1R2   A method using the expression level of one or more genes selected from (a-1) to (a-59) as an index, wherein a predetermined time has elapsed since receiving the treatment The prediction according to claim 1, wherein the treatment is predicted to be effective for the subject when the expression level of the gene is decreased as compared to the expression level of the gene at a predetermined time before that time. Method.   A method using the expression level of one or more genes selected from (b-1) to (b-15) as an index, when a predetermined time has elapsed since receiving the treatment The prediction according to claim 1, wherein the treatment is predicted to be effective for the subject when the expression level of the gene is increased compared to the expression level of the gene at a predetermined time before that time. Method.   The prediction method according to claim 1, wherein the autoimmune disease is vasculitis syndrome.   The prediction method according to claim 4, wherein the vasculitis syndrome is MPO-ANCA-related vasculitis.   The prediction method according to claim 1, wherein the treatment is administration of a drug.   The prediction method according to claim 6, wherein the drug is an immunosuppressant, a steroid drug, or a biological preparation. The administration of the drug is a combination of the administration of an immunosuppressive agent and the administration of a steroidal drug,
The immunosuppressive agent is cyclophosphamide;
The steroidal drug is methylprednisolone or prednisolone,
The prediction method according to claim 6. Administering a test substance to a non-human animal model of autoimmune disease;
Determining the therapeutic effect of the test substance on the autoimmune disease, using as an index the expression level of the gene in the blood of the model animal;
A method for screening a substance having a therapeutic effect against an autoimmune disease, comprising:
The screening method, wherein the gene is one or more genes selected from the group consisting of the following (a-1) to (a-59) and (b-1) to (b-15).
(A-1) a gene encoding CLC (a-2) a gene encoding IFIT1 (a-3) a gene encoding IFIT3 (a-4) a gene encoding CCR3 (a-5) a gene encoding HERC5 (A-6) a gene encoding NGFRAP1 (a-7) a gene encoding TNFSF10 (a-8) a gene encoding FGL2 (a-9) a gene encoding MX1 (a-10) a gene encoding CCR1 (A-11) a gene encoding EMR1 (a-12) a gene encoding XIAP-related factor-1 (a-13) a gene encoding OASL (a-14) a gene encoding OAS1 (a-15) IFIT5 (A-16) gene encoding WSB2 (a-17) gene encoding FFAR2 Child (a-18) gene encoding ISG15 (a-19) gene encoding HIST1H3H (a-20) gene encoding IFI6 (a-21) gene encoding CTSL1 (a-22) encoding GBP1 Gene (a-23) gene encoding IRF7 (a-24) gene encoding HIST2H2BE (a-25) gene encoding IFIH1 (a-26) gene encoding MMD (a-27) encoding CD36 Gene (a-28) gene encoding MT2A (a-29) gene encoding PSMB9 (a-30) gene encoding PLSCR1 (a-31) gene encoding OAS2 (a-32) encoding EIF2AK2 Gene (a-33) Gene encoding LTB (a-34) A gene encoding S4A4A (a-35) A gene encoding ADM (a-36) A gene encoding OAS3 (a-37) A gene encoding SLAMF7 (a-38) A gene encoding DDX58 (a-39) A gene encoding GNG11 (a-40) A gene encoding TSC22D1 (a-41) A gene encoding HIST1H2BD (a-42) A gene encoding IFITM3 (a-43) A gene encoding UBE2L6 (a-44) H2BFS encoding gene (a-45) CAT encoding gene (a-46) SCO2 encoding gene (a-47) PSME2 encoding gene (a-48) STAT1 encoding gene (a-49) Gene encoding MTHFD2 (a-50) MT1 (A-51) gene encoding TGFA (a-52) gene encoding GPR109B (a-53) gene encoding HIST1H3D (a-54) gene encoding IFI44L (a-55) F13A1 (A-56) gene encoding CX3CR1 (a-57) gene encoding FCGR1A (a-58) gene encoding IL1B (a-59) gene encoding ARRB1 (b-1) ADAM28 (B-2) gene encoding CEACAM8 (b-3) gene encoding COL9A2 (b-4) gene encoding DEFA4 (b-5) gene encoding LTF (b-6) SEC23B (B-7) encoding VSIG4 Gene (b-8) gene encoding DEFA1 (b-9) gene encoding CD163 (b-10) gene encoding RUNDC3A (b-11) gene encoding CAMP (b-12) encoding LCN2 Gene (b-13) Gene encoding HDAC9 (b-14) Gene encoding CD24 (b-15) Gene encoding IL1R2   A method using the expression level of one or more genes selected from (a-1) to (a-59) as an index, and a predetermined time has elapsed since the administration of the test substance The test substance is determined to have a therapeutic effect on the autoimmune disease when the expression level of the gene at the time is decreased compared to the expression level of the gene at a predetermined time before that time. Item 10. The screening method according to Item 9.   A method using the expression level of one or more genes selected from (b-1) to (b-15) as an index, and a predetermined time has elapsed since the administration of the test substance When the expression level of the gene at the time is increased compared to the expression level of the gene at a predetermined time before that time, the test substance is determined to have a therapeutic effect on the autoimmune disease. Item 10. The screening method according to Item 9.   The screening method according to claim 9, wherein the autoimmune disease is vasculitis syndrome.   The screening method according to claim 12, wherein the vasculitis syndrome is MPO-ANCA-related vasculitis. An oligo / polynucleotide array for determining the effect of a medicament or candidate medicament for treating an autoimmune disease comprising:
A support and two or more kinds of oligo / polynucleotides immobilized on the support;
The two or more kinds of oligo / polynucleotides are selected from the group consisting of the following (c-1) to (c-74).
(C-1) Oligo / polynucleotide that can hybridize to nucleic acid encoding CLC (c-2) Oligo / polynucleotide that can hybridize to nucleic acid encoding IFIT1 (c-3) Hybridize to nucleic acid encoding IFIT3 Oligo / polynucleotide (c-4) capable of hybridizing to nucleic acid encoding CCR3 (c-5) Oligo / polynucleotide capable of hybridizing to nucleic acid encoding HERC5 (c-6) Oligo / polynucleotide hybridizable to nucleic acid encoding NGFRAP1 (c-7) Oligo / polynucleotide hybridizable to nucleic acid encoding TNFSF10 (c-8) Oligo / polynucleotide hybridizable to nucleic acid encoding FGL2 Polynucleotide (c 9) Oligo / polynucleotide hybridizable to nucleic acid encoding MX1 (c-10) Oligo / polynucleotide hybridizable to nucleic acid encoding CCR1 (c-11) Hybridizable to nucleic acid encoding EMR1 Oligo / polynucleotide (c-12) Oligo / polynucleotide hybridizable to nucleic acid encoding XIAP-related factor-1 (c-13) Oligo / polynucleotide hybridizable to nucleic acid encoding OASL (c-14) ) Oligo / polynucleotide hybridizable to nucleic acid encoding OAS1 (c-15) Oligo / polynucleotide hybridizable to nucleic acid encoding IFIT5 (c-16) Oligo hybridizable to nucleic acid encoding WSB2 / Polynucleotide c-17) Oligo / polynucleotide hybridizable to nucleic acid encoding FFAR2 (c-18) Oligo / polynucleotide hybridizable to nucleic acid encoding ISG15 (c-19) Hybridized to nucleic acid encoding HIST1H3H Possible oligo / polynucleotide (c-20) oligo / polynucleotide hybridizable to nucleic acid encoding IFI6 (c-21) oligo / polynucleotide hybridizable to nucleic acid encoding CTSL1 (c-22) GBP1 (C-23) Oligo / polynucleotide hybridizable to nucleic acid encoding IRF7 (c-24) Oligo hybridizable to nucleic acid encoding HIST2H2BE Polynucleotide (c-25) Oligo / polynucleotide hybridizable to nucleic acid encoding IFIH1 (c-26) Oligo / polynucleotide hybridizable to nucleic acid encoding MMD (c-27) Nucleic acid encoding CD36 (C-28) Oligo / polynucleotide that can hybridize to nucleic acid encoding MT2A (c-29) Oligo / polynucleotide that can hybridize to nucleic acid encoding PSMB9 (c- 30) Oligo / polynucleotide hybridizable to a nucleic acid encoding PLSCR1 (c-31) Oligo / polynucleotide hybridizable to a nucleic acid encoding OAS2 (c-32) Hybridizable to a nucleic acid encoding EIF2AK2 Oligo / polynucleotide (c-33) Oligo / polynucleotide hybridizable to the nucleic acid encoding LTB (c-34) Oligo / polynucleotide hybridizable to the nucleic acid encoding MS4A4A (c-35) Encoding ADM (C-36) Oligo / polynucleotide hybridizable to nucleic acid encoding OAS3 (c-37) Oligo / polynucleotide hybridizable to nucleic acid encoding SLAMF7 c-38) Oligo / polynucleotide hybridizable to nucleic acid encoding DDX58 (c-39) Oligo / polynucleotide hybridizable to nucleic acid encoding GNG11 (c-40) Hybridizer to nucleic acid encoding TSC22D1 Oligo / polynucleotide (c-41) Oligo / polynucleotide hybridizable to a nucleic acid encoding HIST1H2BD (c-42) Oligo / polynucleotide hybridizable to a nucleic acid encoding IFITM3 (c-43) Oligo / polynucleotide hybridizable to nucleic acid encoding UBE2L6 (c-44) Oligo / polynucleotide hybridizable to nucleic acid encoding H2BFS (c-45) Oligo / polynucleotide hybridizable to nucleic acid encoding CAT Polynucleotide (c-46) Oligo / polynucleotide that can hybridize to nucleic acid encoding SCO2 (c-47) Oligo / polynucleotide that can hybridize to nucleic acid encoding PSME2 (c-48) Encodes STAT1 Oligo / polynucleotide that can hybridize to acid (c-49) Oligo / polynucleotide that can hybridize to nucleic acid encoding MTHFD2 (c-50) Oligo / polynucleotide that can hybridize to nucleic acid encoding MT1X (c -51) Oligo / polynucleotide hybridizable to nucleic acid encoding TGFA (c-52) Oligo / polynucleotide hybridizable to nucleic acid encoding GPR109B (c-53) Hybridized to nucleic acid encoding HIST1H3D Oligo / polynucleotide (c-54) Oligo / polynucleotide hybridizable to a nucleic acid encoding IFI44L (c-55) Oligo / polynucleotide hybridizable to a nucleic acid encoding F13A1 (c-56) C Oligo / polynucleotide hybridizable to nucleic acid encoding X3CR1 (c-57) Oligo / polynucleotide hybridizable to nucleic acid encoding FCGR1A (c-58) Oligo / polynucleotide hybridizable to nucleic acid encoding IL1B Polynucleotide (c-59) Oligo / polynucleotide hybridizable to the nucleic acid encoding ARRB1 (c-60) Oligo / polynucleotide hybridizable to the nucleic acid encoding ADAM28 (c-61) Nucleic acid encoding CEACAM8 (C-62) Oligo / polynucleotide hybridizable to a nucleic acid encoding COL9A2 (c-63) Oligo / polynucleotide hybridizable to a nucleic acid encoding DEFA4 Otide (c-64) Oligo / polynucleotide hybridizable to nucleic acid encoding LTF (c-65) Oligo / polynucleotide hybridizable to nucleic acid encoding SEC23B (c-66) Nucleic acid encoding VSIG4 Hybridizable oligo / polynucleotide (c-67) Oligo / polynucleotide hybridizable to nucleic acid encoding DEFA1 (c-68) Oligo / polynucleotide hybridizable to nucleic acid encoding CD163 (c-69) ) Oligo / polynucleotide hybridizable to nucleic acid encoding RUNDC3A (c-70) Oligo / polynucleotide hybridizable to nucleic acid encoding CAMP (c-71) Oligo hybridizable to nucleic acid encoding LCN2 / Renucleotide (c-72) Oligo / polynucleotide hybridizable to nucleic acid encoding HDAC9 (c-73) Oligo / polynucleotide hybridizable to nucleic acid encoding CD24 (c-74) Nucleic acid encoding IL1R2 Oligo / polynucleotide that can hybridize to

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