• Biomolecules & Therapeutics
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• v.30 no.6
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• pp.529-539
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• 2022

Rheumatoid arthritis (RA) is a multifactorial immune-mediated disease, the pathogenesis of which involves different cell types. T-cell activation plays an important role in RA. Therefore, inhibiting T-cell activation is one of the current therapeutic strategies. Cytotoxic T-lymphocyte antigen 4-immunoglobulin (CTLA4-Ig), also known as abatacept, reduces cytokine secretion by inhibiting T-cell activation. To achieve a homeostatic therapeutic effect, CTLA4-Ig has to be administered repeatedly over several weeks, which limits its applicability in RA treatment. To overcome this limitation, we increased the number of sialic acid-capped antennas by genetically engineering the CTLA4 region to increase the therapeutic effect of CTLA4-Ig. N-acetylglucosaminyltransferase (GnT) and α2,6-sialyltransferase (α2,6-ST) were co-overexpressed in Chinese hamster ovary (CHO) cells to generate a highly sialylated CTLA4-Ig fusion protein, named ST6. The therapeutic and immunogenic effects of ST6 and CTLA4-Ig were compared. ST6 dose-dependently decreased paw edema in a mouse model of collagen-induced arthritis and reduced cytokine levels in a co-culture cell assay in a similar manner to CTLA4-Ig. ST6- and CTLA4-Ig-induced T cell-derived cytokines were examined in CD4 T cells isolated from peripheral blood mononuclear cells after cell killing through irradiation followed by flow- and magnetic-bead-assisted separation. Interestingly, compared to CTLA4-Ig, ST6 was substantially less immunogenic and more stable and durable. Our data suggest that ST6 can serve as a novel, less immunogenic therapeutic strategy for patients with RA.

• Journal of Acupuncture Research
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• v.37 no.2
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• pp.128-135
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• 2020

Background: Classical acupuncture is being used in the treatment of rheumatoid arthritis (RA). To explore the biological response to acupuncture, a network-based analysis was performed on gene expression data collected from an animal model of RA treated with acupuncture. Methods: Gene expression data were obtained from published microarray studies on blood samples from rats with collagen induced arthritis (CIA) and non-CIA rats, both treated with manual acupuncture. The weighted gene co-expression network analysis was performed to identify gene clusters expressed in association with acupuncture treatment time and RA status. Gene ontology and pathway analyses were applied for functional annotation and network visualization. Results: A cluster of 347 genes were identified that differentially downregulated expression in association with acupuncture treatment over time; specifically in rats with CIA with module-RA correlation at 1 hour after acupuncture (-0.27; p < 0.001) and at 34 days after acupuncture (-0.33; p < 0.001). Functional annotation showed highly significant enrichment of porphyrin-containing compound biosynthetic processes (p < 0.001). The network-based analysis also identified a module of 140 genes differentially expressed between CIA and non-CIA in rats (p < 0.001). This cluster of genes was enriched for antigen processing and presentation of exogenous peptide antigen (p < 0.001). Other functional gene clusters previously reported in earlier studies were also observed. Conclusion: The identified gene expression networks and their hub-genes could help with the understanding of mechanisms involved in the pathogenesis of RA, as well understanding the effects of acupuncture treatment of RA.

• Molecules and Cells
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• v.23 no.3
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• pp.398-404
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• 2007

Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are involved in various pathophysiological processes such as inflammation and carcinogenesis. In a search for inhibitors of COX-2 and iNOS production we found that extracts of Stewartia koreana strongly inhibited NO and $PGE_2$ production in LPS-treated macrophage RAW 264.7 cells. We have now shown that the mRNA and protein levels of iNOS and COX-2 are reduced by the Stewartia koreana extract (SKE). SKE inhibited expression of an NF-${\kappa}B$ reporter gene in response to LPS, and gel mobility shift assays revealed that SKE reduced NF-${\kappa}B$ DNA-binding activity. The extract also inhibited LPS-induced phosphorylation of $I{\kappa}B-{\alpha}$ and nuclear translocation of p65. Administration of the extract reduced the symptoms of arthritis in a collagen-induced arthritic mouse model. These results indicate that Stewartia extracts contain potentially useful agents for preventing and treating inflammatory diseases.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.81-89
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• 2004

Tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and lymphotoxin-$\alpha$ (LT-$\alpha$, TNF-$\beta$) can initiate and perpetuate human diseases such as multiple sclerosis (MS), rheumatoid arthritis (RA), and insulin-dependent diabetes mellitus (IDDM). TNFs can be blocked by the use of soluble TNF receptors. However, since monomeric soluble receptors generally exhibit low affinity or function as agonists, the use of monomeric soluble receptors has been limited in the case of cytokines such as TNF-$\alpha$, TNF-$\alpha$, interleukin (IL)-1, IL-4, IL-6, and IL-13, which have adapted to a multi component receptor system. For these reasons, very high-affinity inhibitors were created for the purpose of a TNFs antagonist to bind the TNFR and trigger cellular signal by using the multistep polymerase chain reaction method. First, recombinant simple TNFR-Ig fusion proteins were constructed from the cDNA sequences encoding the extracellular domain of the human p55 TNFR (CD120a) and the human p75 TNFR (CD120b), which were linked to hinge and constant regions of human $IgG_1$ heavy chain, respectively using complementary primers (CP) encoding the complementary sequences. Then, concatameric TNFR-Ig fusion proteins were constructed using recombinant PCR and a complementary primer base of recombinant simple TNFR-Ig fusion proteins. For high level expression of recombinant fusion proteins, Chinese hamster ovary (CHO) cells were used with a retroviral expression system. The transfected cells produced the simple concatameric TNFR-Ig fusion proteins capable of binding TNF and inactivating it. These soluble versions of simple concantameric TNFR-Ig fusion proteins gave rise to multiple forms such as simple dimers and concatameric homodimers. Simple TNFR-1g fusion proteins were shown to have much more reduced TNF inhibitory activity than concatameric TNFR-Ig fusion proteins. Concatameric TNFR-Ig fusion proteins showed higher affinity than simple TNFR-Ig fusion proteins in a receptor inhibitor binding assay (RIBA). Additionally, concatameric TNFR-Ig fusion proteins were shown to have a progressive effect as a TNF inhibitor compared to the simple TNFR-Ig fusion proteins and conventional TNFR-Fc in cytotoxicity assays, and showed the same results for collagen induced arthritis (CIA) in mice in vivo.

• Molecules and Cells
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• v.39 no.7
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• pp.557-565
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• 2016

The paired immunoglobulin-like type 2 receptor (PILR) family consists of two functionally opposite members, inhibitory $PILR{\alpha}$ and activating $PILR{\beta}$ receptors. PILRs are widely expressed in various immune cells and interact with their ligands, especially CD99 expressed on activated T cells, to participate in immune responses. Here we investigated whether PILR-derived agonists inhibit ${\beta}1$ integrin activity as ligands for CD99. PILR-derived peptides as well as PILR-Fc fusion proteins prevented cell adhesion to fibronectin through the regulation of ${\beta}1$ integrin activity. Especially, PILRpep3, a representative 3-mer peptide covering the conserved motifs of the PILR extracellular domain, prevented the clustering and activation of ${\beta}1$ integrin by dephosphorylating FAK and vinculin, which are major components of focal adhesion. In addition, PILRpep3 inhibited transendothelial migration of monocytes as well as endothelial cell tube formation. Furthermore, upon intraperitoneal injection of PILRpep3 into mice with collagen-induced arthritis, the inflammatory response of rheumatoid arthritis was strongly suppressed. Taken together, these results suggest that PILR-derived agonist ligands may prevent the inflammatory reactions of rheumatoid arthritis by activating CD99.

• Biomolecules & Therapeutics
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• v.23 no.1
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• pp.71-76
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• 2015

Peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) was identified as a cell-intrinsic regulator of Th17 cell differentiation. Th17 cells have been associated with several autoimmune diseases, including experimental autoimmune encephalomyelitis (EAE), inflammatory bowel disease (IBD), and collagen-induced arthritis. In this study, we confirmed $PPAR{\gamma}$-mediated inhibition of Th17 cell differentiation and cytokine production at an early stage. Treatment with ciglitazone, a $PPAR{\gamma}$ ligand, reduced both IL-$1{\beta}$-mediated enhancement of Th17 differentiation and activation of Th17 cells after polarization. For Th17 cell differentiation, we found that ciglitazone-treated cells had a relatively low proliferative activity and produced a lower amount of cytokines, regardless of the presence of IL-$1{\beta}$. The inhibitory activity of ciglitazone might be due to decrease of CCNB1 expression, which regulates the cell cycle in T cells. Hence, we postulate that a pharmaceutical $PPAR{\gamma}$ activator might be a potent candidate for treatment of Th17-mediated autoimmune disease patients.

• Herbal Formula Science
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• v.16 no.2
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• pp.115-131
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• 2008

실험목적 : 빈소산은 11가지 생약으로 구성된 복합 한약 처방으로 관절염을 포함한 다양한 염증성 질환의 치료제로 사용되어 왔으나, 관절염에 대한 직접적인 효력평가는 찾아 보기 힘들다. 따라서 본 실험에서는 빈소산 추출물이 Freund's complete adjuvant (FCA)로 유발된 rat 류마티스성 관절염에 미치는 치료 효과를 dexamethasone (15mg/kg, 복강 투여) 의 효과와 비교 평가하였다. 실험방법 : 류마티스성 관절염은 FCA (10mg in 1ml paraffin oil 0.1ml/rats)를 좌측 후지에 피내 투여하여 유발하였다. 실험동물은 Wistar 랫트를 사용하였고, FCA 투여 14일 후 유사한 무릎관절 둘레를 나타내는 류마티스성 관절염 유발 rat와 정상 rat 및 실험군을 그룹당 9마리씩 나누었다. 실험동물은 100 또는 200mg/kg의 빈소산 추출물을 FCA 투여 14일 후부터 14일간 경구 투여하였으며, dexamethasone은 15mg/kg 농도로 복강 투여한 다음, 희생하여, 체중, 연골내 collagen 함량 및 chondroitin sulphate, heparin sulphate 및 hyaluronic acid와 같은 뼈내 glycosaminoglycan 함량의 변화를 각각 관찰하였다. 실험결과는 항염 효과가 이미 입증되어 있는 dexamethasone 15mg/kg 복강 투여군과 비교하였다. 결과 : FCA 투여는 현저한 체중, 연골내 collagen 함량 및 chondroitin sulphate, heparin sulphate 및 hyaluronic acid와 같은 뼈내 glycosaminoglycan 함량의 감소와 함께 유발 관절 둘레 및 조직내 prostaglandin $E_2$의 증가와 같은 전형적인 류마티스성 염증을 초래하였으나, 이러한 류마티스성 관절염 소견은 dexamethasone 및 모든 용량의 빈소산 추출물 투여에 의해 현저히 억제되었으며, 특히 빈소산 투여군에서는 투여 용량 의존적인 감소가 인정되었다. 결론 : 이상에서 빈소산 추출물은 투여 용량 의존적인 prostaglandin $E_2$ 억제를 매개하여 FCA 유발 류마티스성 관절염에 대한 치료 효과를 나타내는 것으로 관찰되었다. 따라서 새로운 관절염에 대한 치료제로서 개발 가능성이 있을 것으로 판단된다. 한편 빈소산 추출물은 주로 prostaglandin $E_2$ 억제작용에 의해 항염 효과를 나타내는 것으로 관찰되었으나, 금후 다른 작용기전에 대한 연구와 빈소산의 구성성분 중 유효 성분 규명을 위한 실험이 수행되어야 할 것으로 판단된다.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.6
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• pp.1509-1517
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• 2008

The flowers and buds of Lonicera Flower (LF), are used in Korean herbal medicine for latent-heat-clearing, antipyretic, detoxicant and anti-inflammatory ailments. This plant is used worldwide for the treatment of many types of inflammatory disease including respiratory infections, diabetes mellitus, rheumatoid arthritis and play an important role in immune reaction. These pharmaceutical effects of LF looks like to be related to its antioxidant capacity and phytochemicals containing in LF. In this study, the antioxidant activity of extract from LF was studied in vitro methods by measuring the antioxidant activity by TEAC, measuring the scavenging effects on reactive oxygen species (ROS) [superoxide anion, hydroxyl radical] and on reactive nitrogen species (RNS) [nitric oxide and peroxynitrite] as well as measuring the inhibitory effect on $Cu^{2+}$ induced human LDL oxidation and the inhibitory effect on collagen induced platelet aggregation. The LF extracts were found to have a potent scavenging activity, as well as an inhibitory effect on LDL oxidation and on platelet aggregation. In conclusion, the LF extracts have anti-oxidative and anti-atherosclerotic effects in vitro system, which can be used for developing pharmaceutical drug against oxidative stress and atherosclerosis.

• The Journal of Dong Guk Oriental Medicine
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• v.7 no.2
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• pp.107-120
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• 1999

Synovial joint of BALB/C mice were injeced with Lipopolysaccharide(LPS) were observed to investigate the morphological changes of synovial capsule caused by rheumatoid arthritis(RA). The RA on female Balb/c mice were induced by LPS injection, as dose of $300{\mu}{\ell}/kg$, into synovial cavity of knee joint. And then these specimen were fixed in 10% neutral buffered formalin and were decalcificated in EDTA solution for 4 weeks. The hyperplasia of synovium were appeared in synovial membrane. The filopodia of phagocytic like synoviocyte(type I synoviocyte) projected into synovial cavity and the number of fibroblast like synoviocyte(type II synoviocyte) with well-developed endoplasmic reticulum were increased in synovium. In fibrous membrane, the fibrosis induced by synthesis of collagen fiber were enlarged to all fibrous membrane, and the number of fibroblast were increased. A great number of inflammation component cell as lymphocyte and neutrophil leukocyte were infiltrated around capillary and the degranulate typed mast cell were increased. As results indicated that the hyperplasia of synovium induced by LPS, subsequently to cause the fibrosis, infiltration of imflammation component cell, and increase of degranulated type mast cell as same as symptoms of RA.

• Journal of Oral Medicine and Pain
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• v.36 no.4
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• pp.261-271
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• 2011

Osteoarthritis in patients with temporomandibular disorders(TMDs) induces pain, limitation of mouth opening, occlusal problems, and most commonly affects their life quality. Control method and progressive process of osteoarthritis are being extensively researched. The researchers focus on histologic changes, synovial changes, muscular and ligamental changes and observed reaction to pain. Therefore most of them developed the animal model for osteoarthritis in TMD patients. In this study, we applied several methods which induces osteoarthritis of temporomandibular joint(TMJ) in rats or mice. For locally induce osteoarthritis in TMJ, Monosodium iodoacetate(MIA) or interleukin-$1{\alpha}$(IL-$1{\alpha}$) were injected into TMJ joint space for 5 or 3 weeks. Other groups are chosen for osteoarthritis under systemic control including hormonal changes and aging. To observe cellular change, increased collagen, degenerative bony destruction and distribution of proteoglycans (PGs), safranin-O staining and Masson's trichrome staining were used.