KR101139237B1 - A therapeutic agent for immune disease comprising macrophage migration inhibitory factor isolated from parasite - Google Patents

A therapeutic agent for immune disease comprising macrophage migration inhibitory factor isolated from parasite Download PDF

Info

Publication number
KR101139237B1
KR101139237B1 KR1020090089005A KR20090089005A KR101139237B1 KR 101139237 B1 KR101139237 B1 KR 101139237B1 KR 1020090089005 A KR1020090089005 A KR 1020090089005A KR 20090089005 A KR20090089005 A KR 20090089005A KR 101139237 B1 KR101139237 B1 KR 101139237B1
Authority
KR
South Korea
Prior art keywords
mif
therapeutic agent
macrophage
isolated
parasite
Prior art date
Application number
KR1020090089005A
Other languages
Korean (ko)
Other versions
KR20110031663A (en
Inventor
유학선
박상균
조민경
Original Assignee
부산대학교 산학협력단
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 부산대학교 산학협력단 filed Critical 부산대학교 산학협력단
Priority to KR1020090089005A priority Critical patent/KR101139237B1/en
Publication of KR20110031663A publication Critical patent/KR20110031663A/en
Application granted granted Critical
Publication of KR101139237B1 publication Critical patent/KR101139237B1/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1767—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/68—Protozoa, e.g. flagella, amoebas, sporozoans, plasmodium or toxoplasma
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • 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
    • C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S424/00—Drug, bio-affecting and body treating compositions
    • Y10S424/81—Drug, bio-affecting and body treating compositions involving autoimmunity, allergy, immediate hypersensitivity, delayed hypersensitivity, immunosuppression, immunotolerance, or anergy

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Zoology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Public Health (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • Toxicology (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Mycology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

본 발명은 기생충으로부터 분리된 대식세포 유주저지인자를 포함하는 면역질환 치료제에 관한 것으로서, 보다 상세하게는 고래회충 제3기 유충의 분비?배설 단백질로부터 분리된 제2형 대식세포 유주억제인자를 유효 성분으로 포함하는 면역질환 치료제에 관한 것이다.The present invention relates to a therapeutic agent for immunological disease comprising macrophage-derived inhibitory factors isolated from parasites. More specifically, the present invention relates to a type 2 macrophage-suppressing inhibitor isolated from the secretion and excretory proteins of the third roundworm larvae. The present invention relates to a therapeutic agent for immune diseases, which comprises as an ingredient.

Description

기생충으로부터 분리된 대식세포 유주저지인자를 포함하는 면역질환 치료제{A THERAPEUTIC AGENT FOR IMMUNE DISEASE COMPRISING MACROPHAGE MIGRATION INHIBITORY FACTOR ISOLATED FROM PARASITE}A therapeutic agent for immunological diseases, including macrophage-derived inhibitory factors isolated from parasites {A THERAPEUTIC AGENT FOR IMMUNE DISEASE COMPRISING MACROPHAGE MIGRATION INHIBITORY FACTOR ISOLATED FROM PARASITE}

본 발명은 기생충으로부터 분리된 대식세포 유주저지인자 (Macrophage migration Inhibitory Factor, 이하, "MIF"로 명명)를 포함하는 면역질환 치료제에 관한 것으로서, 보다 상세하게는 고래회충 (Anisakis simplex)의 제3기 유충의 분비?배설 단백질로부터 분리된 제2형 MIF를 유효 성분으로 포함하는 면역질환 치료제에 관한 것이다.The present invention relates to a therapeutic agent for immune diseases comprising a macrophage migration inhibitory factor (hereinafter referred to as "MIF") isolated from parasites, and more specifically, the third phase of an anisakis simplex . The present invention relates to a therapeutic agent for immunological diseases comprising type 2 MIF isolated from the larvae secretory and excretory proteins as an active ingredient.

포유동물의 MIF는 알레르기성 비염, 류마티스 관절염, 패혈쇼크 등과 같은 염증 및 면역반응을 조절하는 친염증성 사이토카인 (pro-inflammatory cytokine)으로 알려져 있다. Mammalian MIF is known as a pro-inflammatory cytokine that modulates inflammatory and immune responses such as allergic rhinitis, rheumatoid arthritis, septic shock and the like.

기생충에서도 이와 비슷한 형태의 단백질이 보고되고 있으며, 이들의 정확한 역할들에 대하여 연구가 진행 중에 있다. 이들 MIF는 두 가지 형 (type)이 보고되고 있는데, 사람 및 척추동물에서는 제1형만이 보고되고 있고, 선형동물에서는 제1형과 제2형 모두 알려져 있다. 선충 (nematode)에서 분리된 제1형 MIF의 경우, 사 람의 MIF와 구조적으로 많이 다르지만, 유사한 기능도 하는 것으로 보고되고 있다. 그러나, 기생충에서 유래된 제2형 MIF에 대한 연구는 거의 없다. Similar forms of proteins have been reported in parasites, and their exact roles are under study. Two types of these MIFs have been reported, only type 1 has been reported in humans and vertebrates, and both type 1 and type 2 are known in linear animals. Type I MIF isolated from nematodes, although structurally different from human MIF, has been reported to function similarly. However, there is little research on type 2 MIF derived from parasites.

기생충에 감염된 숙주는 과민반응 및 면역질환이 억제된다는 보고가 많이 존재하고, 이와 관련되는 여러 가지 면역조절인자들이 최근에 발표되고 있으나, 이에 대한 분자생물학적인 기전은 아직 명확하게 밝혀진 바가 없다. There are many reports that parasites are suppressed by hypersensitivity reactions and immune diseases, and various immunomodulators have been published recently, but the molecular biological mechanisms thereof have not been elucidated.

한편, 선진국에서는 뚜렷한 원인이 없어 자가면역질환 (autoimmune disease)으로 인정되고 있는 염증성 장 질환 (inflammatory bowel disease) 환자가 계속 증가 중에 있으며, 우리나라 역시 이러한 질병이 증가하고 있는 추세이다. 외국에서는 이러한 염증성 장질환의 치료제로 기생충 충란을 먹여 병을 치료하는 방법이 소개되고 있지만 정확한 인자는 찾지 못하고 있다. 이러한 염증성 장질환에서도 MIF의 기능은 염증을 일으키는 데 있어서 매우 중요한 역할을 차지하고 있기 때문에 기생충 감염에 의한 염증성 장질환의 억제현상은 감염된 기생충이 숙주의 MIF와 유사한 물질을 분비하여 숙주 MIF의 기능을 억제한 결과인 것으로 예측 가능하나, 이에 대한 정확한 세포생물학적 기전은 밝혀진 바가 없다.Meanwhile, in developed countries, inflammatory bowel disease, which is recognized as an autoimmune disease because there is no obvious cause, continues to increase, and the disease in Korea is also increasing. Foreign countries have been introduced to treat the disease by feeding parasite eggs as a treatment for inflammatory bowel disease, but the exact factors are not found. In this inflammatory bowel disease, the function of MIF plays an important role in inducing inflammation. Therefore, the inhibition of inflammatory bowel disease caused by parasite infection inhibits the function of host MIF by secreting a substance similar to host MIF. It is possible to predict this result, but the exact cell biology mechanism for this is not known.

본 발명은 상기와 같은 종래 기술의 문제점을 해결하기 위하여 안출된 것으로서, 본 발명에서 해결하고자 하는 과제는 기생충으로부터 분리된 MIF와 각종 면역질환과의 상관 관계 및 그 작용 기전을 밝힘으로써, 상기 MIF를 각종의 면역질환 치료제로서 사용할 수 있는 수단을 제공하고자 하는 것이다.The present invention has been made to solve the problems of the prior art as described above, the problem to be solved in the present invention by revealing the correlation between the MIF and various immune diseases isolated from parasites and its mechanism of action, It is to provide a means that can be used as a therapeutic agent for various immune diseases.

상기와 같은 과제를 해결하기 위하여,In order to solve the above problems,

본 발명은 기생충으로부터 분리된 대식세포 유주저지인자를 유효 성분으로 포함하는 것을 특징으로 하는 면역질환 치료제를 제공한다.The present invention provides an agent for treating immunological diseases, which comprises macrophage hepatic inhibitory factor isolated from parasites as an active ingredient.

상기 기생충은 고래회충의 유충인 것이 바람직하며, 상기 대식세포 유주저지인자는 제2형 대식세포 유주저지인자인 것이 바람직하다.Preferably, the parasite is a larva of a whale larvae, and the macrophage-jujube inhibitor is preferably a type 2 macrophage-jujube inhibitor.

또한, 상기 면역질환은 염증성 장질환 또는 자가면역질환인 것이 바람직하다.In addition, the immune disease is preferably an inflammatory bowel disease or an autoimmune disease.

본 발명에 따른 기생충으로부터 분리된 MIF는 면역질환, 특히 염증성 장질환 또는 자가면역질환의 치료제로서 적용할 수 있다.MIF isolated from the parasite according to the invention can be applied as a therapeutic agent for immune diseases, in particular inflammatory bowel disease or autoimmune disease.

본 발명의 면역질환 치료제는 기생충으로부터 분리된 대식세포 유주저지인자를 유효 성분으로 포함하는 것을 특징으로 한다.The therapeutic agent for immunological diseases of the present invention is characterized in that it comprises a macrophage hepatic inhibitory factor isolated from parasites as an active ingredient.

본 발명의 대식세포 유주저지인자는 기생충으로 분리되는 것으로서, 바람직하게는 고래회충으로부터 분리되는 것이며, 가장 바람직하게는 고래회충의 제3기 유충의 분비?배설 단백질로부터 분리되는 것이다.The macrophage hepatic inhibitor of the present invention is isolated from parasites, preferably from whale larvae, and most preferably from the secretion and excretory proteins of the third larvae of whale worms.

상기 대식세포 유주저지인자는 제1형 대식세포 유주저지인자 또는 제2형 대식세포 유주저지인자일 수 있으나, 바람직하게는 제2형 대식세포 유주저지인자이다.The macrophage hepatic jersey factor may be a type 1 macrophage hepatic jersey factor or a type 2 macrophage hepatic jersey factor, but is preferably a type 2 macrophage hepatic jersey factor.

본 발명의 바람직한 구체예에 의하여, 고래회충의 제3기 유충의 분비?배설 단백질로부터 분리되는 제2형 대식세포 유주저지인자는 고래회충의 제3기 유충의 분비?배설 단백질로부터 분리된 DNA로부터 통상적인 클로닝 방법에 의하여 얻어진 재조합 제2형 대식세포 유주저지인자 (이하, "rAs-MIF"로 명명)로서, 전체 121 개의 아미노산을 갖는 366 bp의 뉴클레오티드로 이루어지며, 본래 단백질 크기는 13 KDa이지만, 다른 MIF와 마찬가지로 생체 내에서는 호모트리머 (homotrimer) 형태로서 39 KDa으로도 존재한다. 또한, 상기 rAs-MIF는 사람 혹은 다른 척추동물에서 분비되는 MIF의 아미노산 서열 중 Ser-64 부분이 Ala-64로 치환되어 구조적으로 다른데, 이는 다른 동물에서 분비되는 MIF와 기능적으로 다르다는 것을 시사한다. According to a preferred embodiment of the present invention, the type 2 macrophage hepatic inhibitor is isolated from the secretion and excretory protein of the tertiary larvae of whale larvae from the DNA isolated from the secretion and excretory protein of the tertiary larvae of the Recombinant type 2 macrophage hepatic inhibitor (hereinafter referred to as "rAs-MIF") obtained by a conventional cloning method, consisting of 366 bp of nucleotides having a total of 121 amino acids, originally having a protein size of 13 KDa. In vivo, as with other MIFs, it is present as 39 KDa as a homotrimer. In addition, the rAs-MIF is structurally different from Ser-64 of the amino acid sequence of MIF secreted in humans or other vertebrates by Ala-64, suggesting that it is functionally different from MIF secreted in other animals.

본 발명의 기생충으로부터 분리된 대식세포 유주저지인자는 면역질환 치료제로서 사용된다.Macrophage hepatic inhibitors isolated from parasites of the present invention are used as therapeutic agents for immune diseases.

상기 면역질환의 예는 특별히 한정되지는 않으나, 바람직하게는 염증성 장질환 (inflammatory bowel disease) 또는 자가면역질환 (autoimmune disease)이다.Examples of the immune disease are not particularly limited, but are preferably inflammatory bowel disease or autoimmune disease.

본 발명의 면역질환 치료제는 기생충으로부터 분리된 대식세포 유주저지인자 를 유효 성분으로 포함하며, 상기 기생충으로부터 분리된 대식세포 유주저지인자는 약학적으로 허용되는 염의 형태를 포함한다.The therapeutic agent for immunological disorders of the present invention includes a macrophage hepatic inhibitor which is isolated from a parasite as an active ingredient, and the macrophage hepatic inhibitor which is isolated from a parasite includes a form of a pharmaceutically acceptable salt.

본 발명의 면역질환 치료제에 포함되는 기타 성분으로서, 당업자에게 알려진 성분이라면 어떠한 것이라도 포함될 수 있음은 당연하다.As other components included in the therapeutic agent for immunological diseases of the present invention, any component known to those skilled in the art may be included.

이하, 본 발명을 구체적인 실시예를 통하여 더욱 상세하게 설명한다.Hereinafter, the present invention will be described in more detail with reference to specific examples.

[실시예][Example]

1. rAs-MIF의 클로닝 및 정제1. Cloning and Purification of rAs-MIF

상기 rAs-MIF는 먼저 고래회충의 제3기 유충의 EST로부터 전체 서열을 얻어 이를 바탕으로 프라이머를 디자인한 후, 일반적인 PCR 방법을 이용하여 증폭하였다. 이 과정에서 사용된 주형가닥으로는 제3기 유충으로부터 트리아졸 (Trizol)을 이용하여 RNA를 정제한 후, 합성한 cDNA를 사용하였고, 센스 프라이머 (sense primer) (서열번호 :1)로서 5'-GGA TCC ATG CCT CTG GTT ACG-3', 안티센스 프라이머 (antisense primer) (서열번호: 2)로서 5'-CTC GAG TCA CTG TTT GGC C-3'를 디자인하였고, 어닐링 (anealing) 온도는 50℃로 지정하였다. 먼저, MIF 유전자가 정확하게 잘 증폭되었는지 확인하기 위해, 증폭된 MIF를 상용화된 pGEM T-easy에 삽입하여 서열을 확인한 후, 발현 벡터인 pET 28b에 다시 삽입하여 단백질을 발현 및 정제하였다. 단백질 발현은 E. coli BL21 내에서 이루어졌으며, IPTG를 최종 농도가 0.1mM이 되도록 처리한 후 37에서 8시간 동안 배양하는 과정으로 이루어졌다. 배양 후 박테리아를 원심분리기를 이용하여 다운시키고, PBS로 다시 풀어 분쇄기를 이용하여 박테리아를 적절히 깬 후 친화성 크로마토그래피 (Affinity chromatography)로 rAs-MIF 단백질을 정제하였다. The rAs-MIF first obtained the entire sequence from the EST of the third larva larvae of the whale larvae and designed a primer based on this, and then amplified using a general PCR method. As the template strand used in this process, RNA was purified from triazoles using triazole, and then synthesized cDNA was used, and 5 'as a sense primer (SEQ ID NO: 1). 5'-CTC GAG TCA CTG TTT GGC C-3 'was designed as -GGA TCC ATG CCT CTG GTT ACG-3', antisense primer (SEQ ID NO: 2), and the annealing temperature was 50 ° C. It was designated as. First, in order to confirm that the MIF gene was correctly amplified, the amplified MIF was inserted into a commercialized pGEM T-easy to confirm the sequence, and then inserted into the expression vector pET 28b to express and purify the protein. Protein expression was carried out in E. coli BL21, IPTG was treated to a final concentration of 0.1mM and then incubated for 37 to 8 hours. After incubation, the bacteria were down using a centrifuge, re-dissolved in PBS, and properly crushed using a grinder to purify the rAs-MIF protein by affinity chromatography.

2. rAs-MIF 처리 및 염증성 장 질환 유도2. rAs-MIF Treatment and Inflammatory Bowel Disease Induction

도 1과 같은 계획으로 rAs-MIF를 처리한 실험군과 무처리한 대조군 마우스에 덱스트란 소듐 설페이트 (Dextran sodium sulfate, 이하, "DSS"로 명명)를 투여하여 염증성 장 질환 (이하, "IBD"로 명명)을 유도하였다. 이 때 rAs-MIF는 마우스 한 마리 당 2 μg의 농도로 PBS에 녹인 상태로 복강 주사법을 이용하여 투여 하였으며, 도 1에 기재된 기간은 실험 시작 후 경과된 날짜(일)를 말한다. Inflammatory bowel disease (hereinafter referred to as "IBD") by administering dextran sodium sulfate (hereinafter referred to as "DSS") to experimental and untreated control mice treated with rAs-MIF according to the scheme shown in FIG. Naming). At this time, rAs-MIF was administered by intraperitoneal injection in a state dissolved in PBS at a concentration of 2 μg per mouse, and the period described in FIG. 1 refers to the date (days) elapsed after the start of the experiment.

3. DSS에 의한 염증성 장 질환이 유도된 마우스에서의 장 길이 및 몸무게 변화와 분변상태 및 건강상태 검사3. Intestinal Length and Weight Changes, Fecal Condition and Health Status in Mice Induced by Inflammatory Bowel Disease by DSS

상기 항목 2에 따라 처리된 실험군과 대조군 마우스에 대하여 몸무게 변화, 분변의 상태(경도, 혈의 유무), 대장의 길이 변화를 측정 및 관찰하였다.Body weight change, fecal state (hardness, presence of blood), and change in length of the colon were measured and observed for the experimental and control mice treated according to the above Item 2.

DSS에 의하여 염증이 유도되면 설사 및 혈변이 있게 되고 몸무게가 감소한다. 또한, 심한 탈수로 인해 장이 위축되고 전체적인 대장의 길이가 특징적으로 짧아진다. 도 2에 나타난 결과와 같이, rAs-MIF를 처리한 실험군은 이를 처리하지 않은 대조군에 비하여 장의 길이가 유의적으로 길게 관찰되었고, 몸무게와 분변 상태도 대조군에 비해 심하지 않았으며, 시간이 지날수록 거의 정상 수준으로 돌아왔다. 질병 지수 (diffuse axonal injury, DAI)에서도 실험군은 대조군에 비해 낮은 질병 수치를 보였다.Inflammation induced by DSS results in diarrhea and bloody stools, and weight loss. In addition, severe dehydration leads to atrophy of the intestine and a characteristic shortening of the overall colon. As shown in FIG. 2, the experimental group treated with rAs-MIF was observed to have a significantly longer intestinal length compared to the control group not treated with it, and the weight and fecal state were not as severe as those of the control group. It returned to normal level. Even in the disease index (DAI), the experimental group showed lower disease levels than the control group.

4. rAs-MIF 처리에 의한 사이토카인과 CD44. Cytokines and CD4 by rAs-MIF Treatment ++ CD25CD25 ++ Foxp3Foxp3 ++ T 세포 유도 T cell induction

실험군과 대조군 마우스로부터 비장과 장간막 림프절에서 세포를 분리 후, 이를 배양하여 각 세포로부터 분비된 사이토카인 (TGF-β, INF-γ, IL-4 및 IL-5)의 농도를 측정하여 면역반응의 형태를 관찰하였으며, 대장 조직을 병리학적으로 관찰하여 그 효과를 검증하였다. 또한, rAs-MIF의 면역 조절 기전으로서 조절 T 세포 (ragulatory T cell)의 이동을 측정하기 위해 FACS 분석을 시행하였다.Cells were isolated from spleen and mesenteric lymph nodes from experimental and control mice, and cultured to measure the levels of cytokines (TGF-β, INF-γ, IL-4 and IL-5) secreted from each cell. Morphology was observed, and colonic tissue was pathologically verified. In addition, FACS analysis was performed to measure migration of regulatory T cells as an immune regulatory mechanism of rAs-MIF.

도 3에 나타난 바와 같이, rAs-MIF를 처리한 실험군은 비장과 장 간막 림프절에서 CD4+CD25+Foxp3+ T 세포(조절 T 세포)를 활성화시키는 사이토카인인 TGF-β 농도에 있어서 대조군보다 높은 경향을 보였으며, 염증성 장 질환의 유도와 관계된 사이토카인인 IFN-γ의 농도는 대조군에 비하여 감소함을 관찰할 수 있었다. 또한, 실험군에서는 타입 2 헬퍼 T 세포(Th2) 유래 사이토카인인 IL-4와 IL-5가 대조군에 비하여 증가하는 것이 관찰되었으며, 조절 T 세포가 rAs-MIF를 처리한 실험군의 장 간막 림프절에서 증가하는 것이 관찰되었다. As shown in Figure 3, the experimental group treated with rAs-MIF tended to be higher than the control group in TGF-β concentration, a cytokine activating CD4 + CD25 + Foxp3 + T cells (regulatory T cells) in the spleen and mesenteric lymph nodes. The concentration of IFN-γ, a cytokine associated with the induction of inflammatory bowel disease, was decreased compared to the control group. In addition, it was observed that IL-4 and IL-5, which are cytokines derived from type 2 helper T cells (Th2), were increased in the experimental group compared with the control group, and that the regulatory T cells were increased in the mesenteric lymph nodes of the experimental group treated with rAs-MIF. Was observed.

rAs-MIF는 조절 T 세포를 활성화시키고, 이를 염증부위로 이동시킴으로써 과민 반응을 치료할 수 있음을 보여준다.rAs-MIF has shown that it is possible to treat hypersensitivity by activating regulatory T cells and transferring them to the site of inflammation.

도 1은 본 발명의 rAs-MIF를 처리한 실험군과 미처리 대조군의 염증성 장 질환 유도 과정을 일정으로 표시한 모식도이다. 숫자는 날짜(일) 경과를 나타낸다.1 is a schematic diagram showing the inflammatory bowel disease induction process of the experimental group and the untreated control group treated with rAs-MIF of the present invention. The numbers represent the date (days).

도 2는 본 발명에 의한 실험군과 대조군의 장 길이 및 몸무게 변화와 분변 상태 및 건강상태의 지표를 나타낸 그래프이다. MIF는 실험군을 나타내고, positive는 대조군을 나타낸다.Figure 2 is a graph showing the indicators of intestinal length and weight changes and fecal state and health status of the experimental group and the control group according to the present invention. MIF represents the experimental group and positive represents the control group.

도 3은 본 발명에 의한 실험군과 대조군의 사이토카인과 조절 T 세포의 생산 유도 증가를 나타낸 그래프이다. MIF는 실험군을 나타내고, positive는 대조군을 나타낸다. *p < 0. 05, **p < 0.01, n = 3이다.Figure 3 is a graph showing the increased production induction of cytokines and regulatory T cells of the experimental group and the control group according to the present invention. MIF represents the experimental group and positive represents the control group. * p <0.05, ** p <0.01, n = 3.

<110> Pusan National University Industry-University Cooperation Foundation <120> A therapeutic agent for immune disease comprising macrophage migration inhibitory factor isolated from parasite <160> 2 <170> KopatentIn 1.71 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> MIF-sense primer <400> 1 ggatccatgc ctctggttac g 21 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> MIF-antisense primer <400> 2 ctcgagtcac tgtttggcc 19 <110> Pusan National University Industry-University Cooperation Foundation <120> A therapeutic agent for immune disease comprising macrophage          migration inhibitory factor isolated from parasite <160> 2 <170> KopatentIn 1.71 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> MIF-sense primer <400> 1 ggatccatgc ctctggttac g 21 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> MIF-antisense primer <400> 2 ctcgagtcac tgtttggcc 19  

Claims (5)

고래 회충의 유충으로부터 분리된 제2형 대식세포 유주저지인자를 유효 성분으로 포함하는 염증성 장질환 치료용 약학적 조성물.A pharmaceutical composition for treating inflammatory bowel disease, which comprises a type 2 macrophage hepatic inhibitory factor isolated from a larva of a whale worm as an active ingredient. 제1항에 있어서, 상기 고래 회충의 유충은 제3기 유충인 것을 특징으로 하는 염증성 장질환 치료용 약학적 조성물. The pharmaceutical composition for treating inflammatory bowel disease according to claim 1, wherein the larva of the whale larva is a tertiary larva. 삭제delete 삭제delete 삭제delete
KR1020090089005A 2009-09-21 2009-09-21 A therapeutic agent for immune disease comprising macrophage migration inhibitory factor isolated from parasite KR101139237B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
KR1020090089005A KR101139237B1 (en) 2009-09-21 2009-09-21 A therapeutic agent for immune disease comprising macrophage migration inhibitory factor isolated from parasite

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR1020090089005A KR101139237B1 (en) 2009-09-21 2009-09-21 A therapeutic agent for immune disease comprising macrophage migration inhibitory factor isolated from parasite

Publications (2)

Publication Number Publication Date
KR20110031663A KR20110031663A (en) 2011-03-29
KR101139237B1 true KR101139237B1 (en) 2012-05-10

Family

ID=43936620

Family Applications (1)

Application Number Title Priority Date Filing Date
KR1020090089005A KR101139237B1 (en) 2009-09-21 2009-09-21 A therapeutic agent for immune disease comprising macrophage migration inhibitory factor isolated from parasite

Country Status (1)

Country Link
KR (1) KR101139237B1 (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20210063738A (en) 2019-11-25 2021-06-02 전남대학교산학협력단 Pharmaceutical composition for treating or preventing Arthritis
KR20220158319A (en) 2021-05-24 2022-12-01 전남대학교산학협력단 Pharmaceutical composition for preventing or treating Arthritis

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101335973B1 (en) * 2011-11-02 2013-12-04 (주)엘큐어 Cosmetic compositions or pharmaceutical compositions comprising collagen type i inducing factor isolated from trichinella spiralis

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
Immunological Reviews, Vol. 204, pages 195-207 (2005) *
Immunological Reviews, Vol. 204, pages 195-207 (2005)*
J. Immunology, Vol. 182, pages 6907-6914 (2009.June 01) *
J. Immunology, Vol. 182, pages 6907-6914 (2009.June 01)*

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20210063738A (en) 2019-11-25 2021-06-02 전남대학교산학협력단 Pharmaceutical composition for treating or preventing Arthritis
KR20220158319A (en) 2021-05-24 2022-12-01 전남대학교산학협력단 Pharmaceutical composition for preventing or treating Arthritis

Also Published As

Publication number Publication date
KR20110031663A (en) 2011-03-29

Similar Documents

Publication Publication Date Title
Bo et al. Characterization of interleukin-1β as a proinflammatory cytokine in grass carp (Ctenopharyngodon idella)
Hong et al. Molecular cloning and characterization of chicken lipopolysaccharide-induced TNF-α factor (LITAF)
Wirtz et al. Interleukin-35 mediates mucosal immune responses that protect against T-cell–dependent colitis
Park et al. Molecular cloning and functional analysis of nucleotide-binding oligomerization domain 1 (NOD1) in olive flounder, Paralichthys olivaceus
Xu et al. Characterization and expression analysis of bactericidal permeability-increasing protein (BPI) antimicrobial peptide gene from channel catfish Ictalurus punctatus
Sun et al. Tumor necrosis factor receptor-associated factor 6 (TRAF6) participates in anti-lipopolysaccharide factors (ALFs) gene expression in mud crab
Quynh et al. The cytosolic sensor, DDX41, activates antiviral and inflammatory immunity in response to stimulation with double-stranded DNA adherent cells of the olive flounder, Paralichthys olivaceus
Nguyen et al. Identification and expression analysis of two pro-inflammatory cytokines, TNF-α and IL-8, in cobia (Rachycentron canadum L.) in response to Streptococcus dysgalactiae infection
Norouzitallab et al. Non-lethal heat shock induces HSP70 and HMGB1 protein production sequentially to protect Artemia franciscana against Vibrio campbellii
Li et al. Molecular characterization and expression pattern of X box-binding protein-1 (XBP1) in common carp (Cyprinus carpio L.): indications for a role of XBP1 in antibacterial and antiviral immunity
Ciechomska et al. Role of toll-like receptors in systemic sclerosis
Lv et al. A novel C-type lectin, Nattectin-like protein, with a wide range of bacterial agglutination activity in large yellow croaker Larimichthys crocea
Qi et al. Molecular cloning, expression analysis and functional characterization of interleukin-22 in So-iny mullet, Liza haematocheila
Huang et al. Identification and characterization of the TLR18 gene in grass carp (Ctenopharyngodon idella)
Kim et al. Molecular cloning and functional characterization of the avian macrophage migration inhibitory factor (MIF)
Wang et al. Molecular characterization and expression analysis of large yellow croaker (Larimichthys crocea) interleukin-12A, 16 and 34 after poly I: C and Vibrio anguillarum challenge
Nakharuthai et al. Molecular characterization, functional analysis, and defense mechanisms of two CC chemokines in Nile tilapia (Oreochromis niloticus) in response to severely pathogenic bacteria
Chen et al. Cloning and characterisation of natural resistance associated macrophage protein (Nramp) cDNA from red sea bream (Pagrus major)
KR101139237B1 (en) A therapeutic agent for immune disease comprising macrophage migration inhibitory factor isolated from parasite
Shen et al. Molecular cloning, characterization and expression analysis of the complement component C6 gene in grass carp
Duan et al. Antimicrobial peptide LL-37 forms complex with bacterial DNA to facilitate blood translocation of bacterial DNA and aggravate ulcerative colitis
Das et al. Molecular characterization of interleukin 15 mRNA from rohu, Labeo rohita (Hamilton): its prominent role during parasitic infection as indicated from infection studies
Jiao et al. Activated T cells are the cellular source of IL-22 that enhances proliferation and survival of lymphocytes in Nile tilapia
Shanthi et al. Alpha 2 macroglobulin gene and their expression in response to GFP tagged Vibrio parahaemolyticus and WSSV pathogens in Indian white shrimp Fenneropenaeus indicus
Wei et al. Cloning, expression prolife, and immune characterization of a novel stat family member (stat5bl) in Chinese tongue sole (Cynoglossus semilaevis)

Legal Events

Date Code Title Description
A201 Request for examination
E902 Notification of reason for refusal
E701 Decision to grant or registration of patent right
GRNT Written decision to grant
FPAY Annual fee payment

Payment date: 20160330

Year of fee payment: 5

FPAY Annual fee payment

Payment date: 20170802

Year of fee payment: 6

LAPS Lapse due to unpaid annual fee