• BMB Reports
• /
• 제40권4호
• /
• pp.562-570
• /
• 2007

Soluble or cell-bound IL-1 receptor accessory protein (IL-1RAcP) does not bind IL-1 but rather forms a complex with IL-1 and IL-1 receptor type I (IL-1RI) resulting in signal transduction. Synthetic peptides to various regions in the Ig-like domains of IL-1RAcP were used to produce antibodies and these antibodies were affinity-purified using the respective antigens. An anti-peptide-4 antibody which targets domain III inhibited 70% of IL-$1\beta$-induced productions of IL-6 and PGE2 from 3T3-L1 cells. Anti-peptide-2 or 3 also inhibited IL-1-induced IL-6 production by 30%. However, antipeptide-1 which is directed against domain I had no effect. The antibody was more effective against IL-$1\beta$ compared to IL-$1\alpha$. IL-1-induced IL-6 production was augmented by coincubation with PGE2. The COX inhibitor ibuprofen blocked IL-1-induced IL-6 and PGE2 production. These results confirm that IL-1RAcP is essential for IL-1 signaling and that increased production of IL-6 by IL-1 needs the co-induction of PGE2. However, the effect of PGE2 is independent of expressions of IL-1RI and IL-1RAcP. Our data suggest that domain III of IL-1RAcP may be involved in the formation or stabilization of the IL-1RI/IL-1 complex by binding to epitopes on domain III of the IL-1RI created following IL-1 binding to the IL-1RI.

• Tuberculosis and Respiratory Diseases
• /
• 제43권3호
• /
• pp.348-358
• /
• 1996

연구배경 : Interleukin-5(IL-5)는 호산구 보이는 여러 질환들과 관련이 있으며 특히 알레르기성 천식에서 호산구의 침윤정도와 밀접한 관계가 있는 것으로 알려져 있다. 그러나 IL-2도 증상있는 천식환자의 기도에서 상승됨이 관찰되어 호산구 침윤정도와 상관관계가 있는 것이 밝혀졌다. IL-2가 호산구의 생존을 증가시키는 기전이 IL-5의 표현을 증가시킴으로써인지 또는 다른 경로를 통하여 작용하는 것인지 알기위해 다음과 같은 방법으로 호산구 생존에 미치는 IL-2의 영향을 관찰하였다. 방법 : 호산구증다증을 보인 환자의 말초혈액으로부터 호산구를 분리하여 trypan blue dye exclusion test를 이용하여 생존율을 측정하였으며 Randolp 용액을 이용하여 호산구를 계수하였다. 1) IL-2, IL-5 존재하의 호산구 생존율 및 IL-2와 anti IL-5 존재하의 호산구 생존율을 측정하였다. 2) IL-2 존재하의 말초혈액단핵구에서 IL-5 m-RNA 표현을 Reverse Transcription-Polymerase Chain Reaction(RT-PCR) 방법을 통하여 관찰하였다. 3) IL-2로 자극한 호산구의 IL-2 수용체 발현 증가를 유세포분석기(Flow cytometer)로 측정하였다. 결과 : 1) 호산구의 생존율은 IL-2 및 IL-5에 대한 농도의존성을 보이며 증가하였다. 2) IL-2에 의해 증가된 호산구의 생존율은 anti IL-5에 의해 억제되지 않았다. 3) IL-2로 자극된 말초혈액단핵구는 IL-5 m-RNA를 표현하지 않았다. 4) IL-2는 호산구에서 IL-$2{\alpha}$ 수용체의 표현을 증가시키며 IL-$2{\beta}$ 수용체의 표현은 변화가 없었다. 결론 : 사람에서는 IL-2는 IL-5 형성증가를 통하지 않고 호산구에 IL-2 수용체를 증가시킴으로써 호산구의 생존율을 증가시킨다.

• Tuberculosis and Respiratory Diseases
• /
• 제48권4호
• /
• pp.464-470
• /
• 2000

연구배경 : 기관지 천식은 기도의 과민성과 만성 염증반응을 보이는 질환이다. 저자들은 sIL-6R 및 IL-6 혈중치의 변화를 측정하여 이들의 변화가 증상 악화시 기관지 천식의 병인과 판련이 있는지에 대하여 연구하고자 하였다. 방 법 : 1998년 3월 부터 1999년 3월까지 기관지 천식으로 치료중인 환자중에서 증상 악화로 응급실을 경유하여 전북대학교 병원 내과에 입원한 급성 천식 환자 78예와 무증상 천식 환자 15예 그리고 정상인 10예를 대상으로 하였다. 환자의 병력과 임상 소견, 피부반응검사, 그리고 IgE 및 호산구수를 측정하고 endogen ELISA Kit ($Quantikine^{(R)}sIL$-6R, IL-6)를 이용하여 sIL-6R, IL-6을 측정하였다. 결 과 : 혈청내 IL-6와 sIL-6R농도는 급성 천식 환자에서 대조군에 비해 유의 있는 증가를 보였으며, 무증상 천식 환자와 비교할 때 IL-6의 의미있는 증가를 보였다. 또한 급성 천식 환자에서 혈청내 IL-6와 sIL-6R간에 유의한 상관 관계가 있었다. 하지만 급성 천식환자에서의 IgE와 호산구수는 IL-6 및 sIL-6R와 유의한 상관관계가 없었다. 결 론 : 혈청내 IL-6는 급성 천식의 병태 생리에 관여 할 수 있으며, IL-6와 sIL-6R의 혈청치는 기도 염증의 중증도를 예측하는 표식자로 유용할 것으로 생각된다.

• Molecules and Cells
• /
• 제42권12호
• /
• pp.869-883
• /
• 2019

Interleukin (IL)-15 is an essential immune-modulator with high potential for use in cancer treatment. Natural IL-15 has a low biological potency because of its short half-life and difficulties in mass-production. IL-15Rα, a member of the IL-15 receptor complex, is famous for its high affinity to IL-15 and its ability to lengthen the half-life of IL-15. We have double-transfected IL-15 and its truncated receptor IL-15Rα into CT26 colon cancer cells to target them for intracellular assembly. The secreted IL-15:IL-15Rα complexes were confirmed in ELISA and Co-IP experiments. IL-15:IL-15Rα secreting clones showed a higher anti-tumor effect than IL-15 secreting clones. Furthermore, we also evaluated the vaccine and therapeutic efficacy of the whole cancer-cell vaccine using mitomycin C (MMC)-treated IL-15:IL-15Rα secreting CT26 clones. Three sets of experiments were evaluated; (1) therapeutics, (2) vaccination, and (3) long-term protection. Wild-type CT26-bearing mice treated with a single dose of MMC-inactivated secreted IL-15:IL-15Rα clones prolonged survival compared to the control group. Survival of MMC-inactivated IL-15:IL-15Rα clone-vaccinated mice (without any further adjuvant) exceeded up to 100%. This protection effect even lasted for at least three months after the immunization. Secreted IL-15:IL-15Rα clones challenging trigger anti-tumor response via CD4⁺ T, CD8⁺ T, and natural killer (NK) cell-dependent cytotoxicity. Our result suggested that cell-based vaccine secreting IL-15:IL-15Rα, may offer the new tools for immunotherapy to treat cancer.

• IMMUNE NETWORK
• /
• 제24권1호
• /
• pp.1.1-1.6
• /
• 2024

IL-18 binding protein (IL-18BP) was originally discovered in 1999 while attempting to identify an IL-18 receptor ligand binding chain (also known as IL-18Rα) by subjecting concentrated human urine to an IL-18 ligand affinity column. The IL-18 ligand chromatography purified molecule was analyzed by protein microsequencing. The result revealed a novel 40 amino acid polypeptide. To isolate the complete open reading frame (ORF), various human and mouse cDNA libraries were screened using cDNA probe derived from the novel IL-18 affinity column bound molecule. The identified entire ORF gene was thought to be an IL-18Rα gene. However, IL-18BP has been proven to be a unique soluble antagonist that shares homology with a variety of viral proteins that are distinct from the IL-18Rα and IL-18Rβ chains. The IL-18BP cDNA was used to generate recombinant IL-18BP (rIL-18BP), which was indispensable for characterizing the role of IL-18BP in vitro and in vivo. Mammalian cell lines were used to produce rIL-18BP due to its glycosylation-dependent activity of IL-18BP (approximately 20 kDa). Various forms of rIL-18BP, intact, C-terminal his-tag, and Fc fusion proteins were produced for in vitro and in vivo experiments. Data showed potent neutralization of IL-18 activity, which seems promising for clinical application in immune diseases involving IL-18. However, it was a long journey from discovery to clinical use although there have been various clinical trials since IL-18BP was discovered in 1999. This review primarily covers the discovery of IL-18BP along with how basic research influences the clinical development of IL-18BP.

• IMMUNE NETWORK
• /
• 제5권1호
• /
• pp.16-22
• /
• 2005

Background: IL-18 was originally cloned as a IFN-${\gamma}$ inducing factor in primed T cells. In synergy with IL-12, IL-18 has been shown to induce strikingly high levels of IFN-${\gamma}$ production by T cells and to enhance Th1 development. Also this cytokine exerts induction of Th2 development through IL-4 induction. Methods: Resting $CD4^+$ T cells were sorted by negative selection and activated by anti-CD3 plus anti-CD28 Ab. Expression of IL-12 binding sites, IL-18 binding sites, IL-18R ${\alpha}$, and GATA-3 mRNA were analysed by FACS and RT-PCR, respectively. Results: Resting $CD4^+$ T cells expressed IL-18R ${\alpha}$ chain but not IL-18 binding sites, suggesting a lack of IL-18R ${\beta}$ expression. IL-18R ${\alpha}$ was maintained on the Th1 and Th2 committed cells. IL-18 binding sites were induced on the Th1 but not Th2 cells. Exposure of these cells to IL-18 led to up-regulation of GATA-3 mRNA expression only in Th2 committed cells. To elucidate the relationship between IL-18R ${\alpha}$ expression and GATA-3 induction by IL-18, Th1 and Th2 committed cells were further cultured in medium with or without IL-12 for 2 days. IL-12 binding sites were maintained on the Th1 and Th2 cells regardless of IL-12 treatment, but IL-18R a expression was rapidly down-regulated on the IL12-untreated Th2 cells which did not induce GATA-3 mRNA expression followed by IL-18 stimulation. Conclusion: IL-12 supports expression of IL-18R ${\alpha}$ and GATA-3 mRNA expression was induced by IL-18 through IL-18R ${\alpha}$ without expression of IL-18 binding site in Th2 cells.

• IMMUNE NETWORK
• /
• 제8권1호
• /
• pp.29-37
• /
• 2008

Interleukin-23 (IL-23) is a novel pro-inflammatory cytokine which has been implicated to play a pathogenic role in rheumatoid arthritis (RA). This study was undertaken to investigate the IL-23 inductive activity of the proinflammatory cytokine IL-17, $IL-1{\beta}$ and tumor necrosis factor (TNF-${\alpha}$) in RA synovial fluid mononuclear cells (SFMC). Expression of IL-23p19, IL-17, $IL-1{\beta}$ and TNF-${\alpha}$ in joint was examined by immunohistochemistry (IHC) of patients with RA and osteoarthritis (OA). The effects of IL-17 and $IL-1{\beta}$ on expression of IL-23p19 in human SFMC from RA patients were determined by reverse transcriptase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). IL-23p19 was expressed in the RA fibroblast like synoviocyte (FLS), but not from OA FLS. Similar to the protein expression, IL-23p19 mRNA could be detected by RT-PCR in RA SFMC. IL-17 and $IL-1{\beta}$ could induce RA SFMC to produce the IL-23p19. The effects of IL-17 were much stronger than $IL-1{\beta}$ or TNF-${\alpha}$. These responses were observed in a doseresponsive manner. In addition, IL-17 or $IL-1{\beta}$ neutralizing antibody down-regulated the expression of IL-23p19 induced by LPS in RA-SFMC. Our results demonstrate that IL-23p19 is overexpressed in RA synovium and IL-17 and $IL-1{\beta}$ appears to upregulate the expression of IL-23p19 in RA-SFMC.

• 한국가금학회지
• /
• 제48권3호
• /
• pp.111-122
• /
• 2021

최근에 IL-34가 MCSFR에 대한 두 번째 기능적 리간드로 확인되었으며, M-CSFR에 대한 결합을 통해 IL-34는 M-CSF와 유사한 기능을 공유한다. 지금까지 닭의 IL-34에 대한 실험적 연구가 아닌 예측된 서열로만 보고되었으며, 닭의 IL-34 기능에 대한 정보는 아직 부족하다. 닭 IL-34의 잠재적 생물학적 기능을 구명하기 위하여, 다양한 품종의 닭, 세포주에 대한 괴사성 장염, 살모넬라 및 E. coli 유래의 LPS 등을 처리하여 관찰하였다. 또한 닭의 IL-34와 M-CSF 재조합 단백질 생산 및 이를 이용한 전염증성 사이토카인 유도를 위하여 클로닝 및 재조합 단백질을 발현 및 정제하였다. 특히 관절이상 육계(unknown cause)에 대한 IL-34, M-CSF, 그리고 M-CSFR 유전자의 발현이 대조구보다 유의하게 증가하였으나, 병원균 자극 조직의 경우 많은 조직에서 감소함을 보였다. M-CSFR의 발현은 in vitro에서 IL-34와 M-CSF 재조합 단백질에 의해 증가함을 보였으며, IFN-γ은 재조합 단백질 M-CSF을 제외한 IL-34에 의해 증가하였다. 하지만, IL-12 발현은 유의적인 차이가 없었으며, IL-1β의 경우는 감소하였다. 이 결과는 IL-34와 M-CSF가 고전적인 대식세포와 대체 대식세포 모두에서 역할을 한다고 할 수 있다. 결론적으로, in vitro와 in vivo 실험을 통하여 병원균 처리 시 닭의 IL-34 발현을 관찰하였으며, IL-34 재조합 단백질이 대식세포에서와 전염증성과 항염증성 반응에 중요한 역할을 함을 증명하였다. 추후에 IL-34의 사이토카인, 케모카인 그리고 염증반응 경로 등에 대한 추가 연구가 필요하다고 판단된다.

• Molecules and Cells
• /
• 제19권2호
• /
• pp.180-184
• /
• 2005

IL-17 is a major proinflammatory cytokine secreted by activated T-lymphocytes that accumulates in the inflamed joints of rheumatoid arthritis (RA) patients. Additional IL-17-related molecules and their receptors have been discovered and may also contribute to RA pathogenesis. We examined the expression of the prototypic IL-17 (IL-17A) and its homologs, IL-17B-F, by RT-PCR analyses of synovial fluid mononuclear cells (SFMCs) and peripheral blood mononuclear cells (PBMCs) from RA patients. We also tested for induction of the IL-17 receptor homologs upon stimulation of the fibroblast-like synoviocytes (FLSs) of RA patients with IL-17. The patients' SFMCs expressed IL-17C, E and F in addition to IL-17A. As in the case of IL-17, IL-15 appears to be the major inducer of these homologs in RA SFMCs. We detected transcripts of IL-17R, as well as those of IL-17RB, C and D, in the FLSs of RA patients. Whereas IL-17R expression increased upon in vitro stimulation with IL-17, expression of IL-17RB, C and D was unchanged. However the possibility of cross-interaction between other IL-17 homologs and receptor isoforms remains to be investigated. Our data suggest that these additional homologs should also be considered as targets for immune modulation in the treatment of RA joint inflammation.

• 한국축산식품학회지
• /
• 제23권1호
• /
• pp.75-79
• /
• 2003

본 연구는 몽고 마유에서 pro-inflammatory cytokine(IL-l$\beta$, IL-6, TNF-a, IL-18)과 IL-1 receptor accessory를 Western blotting방법으로 확인하여 기능성 식품과 의약품 소재로 이 용하기 위한 기초 자료를 얻고자 실시하였다. 몽고 마유의 4개군 모두에서 IL-1, TNF-a가 17 kD에서 확인되었다. IL-18은 6~7 kD에서 확인할 수 있었다. 그리고, IL-1 receptor accessory를 55 kD에서 확인하였다. 그러나 탈지분유(Difco) 에서는 IL-l$\beta$, TNF-a를 확인할 수 없었으나, IL-6는 탈지분유(Difco) 및 몽고 마유에서 모두 60 kD에서 확인하였다. 이 와 같이 몽고 마유는 다양한 cytokine을 함유하고 있어서 기능성 식품 및 의약품 소재로 이용가치가 있다고 사료된다.