• Molecules and Cells
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• 제42권10호
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• pp.721-728
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• 2019

Non-structural protein 1 (NS1) of influenza virus has been shown to inhibit the innate immune response by blocking the induction of interferon (IFN). In this study, we isolated two single-stranded RNA aptamers specific to NS1 with $K_d$ values of $1.62{\pm}0.30nM$ and $1.97{\pm}0.27nM$, respectively, using a systematic evolution of ligand by exponential enrichment (SELEX) procedure. The selected aptamers were able to inhibit the interaction of NS1 with tripartite motif-containing protein 25 (TRIM25), and suppression of NS1 enabled retinoic acid inducible gene I (RIG-I) to be ubiquitinated regularly by TRIM25. Additional luciferase reporter assay and quantitative real-time PCR (RT-PCR) experiments demonstrated that suppression of NS1 by the selected aptamers induced IFN production. It is noted that viral replication was also inhibited through IFN induction in the presence of the selected aptamers. These results suggest that the isolated aptamers are strongly expected to be new therapeutic agents against influenza infection.

• 한국양식학회지
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• 제11권4호
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• pp.495-503
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• 1998

생균(Bacillus polyfermenticus, Bacillus mesentericus, Streptococcus faecalis, Bifidobacterium breve)과 소화 요소(protease, lipase)를 함유한 비스루트의 효과를 알아보기 위해 사료 내 1% 첨가하여 나일틸라피아 치어(19.0${\pm}$0.07g)에게 60일간 공급한 다음 성장률, 체조성 및 면역반응의 효과를 조사하였다. 60일간 사육한 결과 비스루트 첨가구와 대조구 사이에 유의적인 성장 차이를 보이지 않았지만(P>0.05), 보체의 용혈 능력은 비스루트 첨가구가 대조구에 비해 용혈능이 높은 것으로 조사되고 라이소자임의 용균 효과는 대조구와 비슷하였다. 식세포의 식작용 능력과 신장 마크로파지의 respiratory burst activity 는 비스루트 첨가구가 대조구에 비해 활성이 매우 높았다. 따라서 비스루트 사료 내 첨가가, 어류의 비특이적 면역 기능중에서 세포성 면역 기능을 활성화하는데 기여하였다. 병원성 세균 Edwardsiella tarda FSW 910410 균주로 공격 실험결과 비스루트 첨가구가 누적폐사율이 59%, 대조구가 80%로 조사되었고, 그 결과 비스루트 첨가구는 병원성 세균에 대한 저항력을 증가시켰다(P<0.05). Hematocrit, hemoglobin, total protein, glucose, GOT, GPT 와 같은 혈액 성분을 분석한 결과 비스루트 첨가구와 대조구 간의 유의차는 없었지만(P>0.05), glucose, GOT, GPT는 비스루트 첨가구가 대조구에 비해 낮게 나타났다. 이상의 결과로 비스루트는 나일틸라피아의 생리 기능에 문제를 야기시키지 않고, 비특이적 면역 기능 중 세포성 면역 기능을 증강시키는 효과가 있으며, 병원성 세균에 대한 저항력의 증강 효과도 높다. 따라서, 비스루트는 나일틸라피아의 사료 첨가물로서의 역할이 충분히 기대된다.

• Journal of Periodontal and Implant Science
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• 제47권5호
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• pp.292-311
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• 2017

Purpose: Beyond the limited scope of non-specific polyclonal regulatory T cell (Treg)-based immunotherapy, which depends largely on serendipity, the present study explored a target Treg subset appropriate for the delivery of a novel epitope spreader Pep19 antigen as part of a sophisticated form of immunotherapy with defined antigen specificity that induces immune tolerance. Methods: Human polyclonal $CD4^+CD25^+CD127^{lo-}$ Tregs (127-Tregs) and $na\ddot{i}ve$ $CD4^+CD25^+CD45RA^+$ Tregs (45RA-Tregs) were isolated and were stimulated with target peptide 19 (Pep19)-pulsed dendritic cells in a tolerogenic milieu followed by ex vivo expansion. Low-dose interleukin-2 (IL-2) and rapamycin were added to selectively exclude the outgrowth of contaminating effector T cells (Teffs). The following parameters were investigated in the expanded antigen-specific Tregs: the distinct expression of the immunosuppressive Treg marker Foxp3, epigenetic stability (demethylation in the Treg-specific demethylated region), the suppression of Teffs, expression of the homing receptors CD62L/CCR7, and CD95L-mediated apoptosis. The expanded Tregs were adoptively transferred into an $NOD/scid/IL-2R{\gamma}^{-/-}$ mouse model of collagen-induced arthritis. Results: Epitope-spreader Pep19 targeting by 45RA-Tregs led to an outstanding in vitro suppressive T cell fate characterized by robust ex vivo expansion, the salient expression of Foxp3, high epigenetic stability, enhanced T cell suppression, modest expression of CD62L/CCR7, and higher resistance to CD95L-mediated apoptosis. After adoptive transfer, the distinct fate of these T cells demonstrated a potent in vivo immunotherapeutic capability, as indicated by the complete elimination of footpad swelling, prolonged survival, minimal histopathological changes, and preferential localization of $CD4^+CD25^+$ Tregs at the articular joints in a mechanistic and orchestrated way. Conclusions: We propose human $na\ddot{i}ve$ $CD4^+CD25^+CD45RA^+$ Tregs and the epitope spreader Pep19 as cellular and molecular targets for a novel antigen-specific Treg-based vaccination against collagen-induced arthritis.

• IMMUNE NETWORK
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• 제9권5호
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• pp.179-191
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• 2009

Background: The present study examines a hypothesis that short allergen-derived peptides may shift an Aspergillus fumigatus (Afu-) specific TH2 response towards a protective TH1. Five overlapping peptides (P1-P5) derived from Asp f1, a major allergen/antigen of Afu, were evaluated for prophylactic or therapeutic efficacy in BALB/c mice. Methods: To evaluate the prophylactic efficacy, peptides were intranasally administered to naive mice and challenged with Afu-allergens/antigens. For evaluation of therapeutic efficacy, the mice were sensitized with Afu-allergens/antigens followed by intranasal administration of peptides. The groups were compared for the levels of Afu-specific antibodies in sera and splenic cytokines evaluated by ELISA. Eosinophil peroxidase activity was examined in the lung cell suspensions and lung inflammation was assessed by histopathogy. Results: Peptides P1-, P2- and P3 decreased Afu-specific IgE (84.5~98.9%) and IgG antibodies (45.7~71.6%) in comparison with Afu-sensitized mice prophylactically. P1- and P2-treated ABPA mice showed decline in Afu-specific IgE (76.4~88%) and IgG antibodies (15~54%). Increased IgG2a/IgG1 and IFN-${\gamma}$/IL-4 ratios were observed. P1-P3 prophylactically and P1 therapeutically decreased IL-5 levels and eosinophil peroxidase activity. P1 decreased inflammatory cells' infiltration in lung tissue comparable to non-challenged control. Conclusion: Asp f1-derived peptide P1, prophylactically and therapeutically administered to Balb/c mice, is effective in regulating allergic response to allergens/antigens of Afu, and may be explored for immunotherapy of allergic aspergillosis in humans.

• IMMUNE NETWORK
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• 제14권3호
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• pp.164-170
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• 2014

JL1, a specific epitope on CD43, is a potential biomarker for the diagnosis of acute leukemia. Although qualitative assays for detecting leukemia-specific CD43 exist, there is a need to develop quantitative assays for the same. Here, we developed two novel monoclonal antibodies (mAbs), 2C8 and 8E10, recognizing different epitopes on CD43. These clones are capable of pairing with YG5, another mAb against JL1 epitope, because they were selectively obtained using sandwich ELISA. Antigens recognized by 2C8 and 8E10 were confirmed as CD43 by western blotting using the CD43-hFC recombinant protein. When expression on various leukemic cell lines was investigated, 2C8 and 8E10 displayed a disparity in the distribution of the epitope. Enzyme assays revealed that these mAbs recognized a sialic acid-dependent epitope on CD43. Using normal thymus and lymph node paraffin-embedded tissues, we confirmed a difference in the epitopes recognized by the two mAbs that was predicted based on the maturity of the cells in the tissue. In summary, we developed and characterized two mAbs, 2C8 and 8E10, which can be used with YG5 in a sandwich ELISA for detecting leukemia-specific CD43.

• Clinical and Experimental Reproductive Medicine
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• 제45권1호
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• pp.1-9
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• 2018

Objective: To determine the localization, expression, and function of Toll-like receptors (TLRs) in fallopian tube epithelial cells. Methods: The localization of TLRs in fallopian tube epithelial cells was investigated by immunostaining. Surprisingly, the intensity of staining was not equal in the secretory and ciliated cells. After primary cell culture of fallopian tube epithelial cells, ring cloning was used to isolate colonies of ciliated epithelial cells, distinct from non-ciliated epithelial cells. The expression of TLRs 1-10 was examined by quantitative real-time polymerase chain reaction, and protein localization was confirmed by immunostaining. The function of the TLRs was determined by interleukin (IL)-6 and IL-8 production in response to TLR2, TLR3, TLR5, TLR7, and TLR9 ligands. Results: Fallopian tube epithelial cells expressed TLRs 1-10 in a cell-type-specific manner. Exposing fallopian tube epithelial cells to TLR2, TLR3, TLR5, TLR7, and TLR9 agonists induced the secretion of proinflammatory cytokines such as IL-6 and IL-8. Conclusion: Our findings suggest that TLR expression in the fallopian tubes is cell-type-specific. According to our results, ciliated cells may play more effective role than non-ciliated cells in the innate immune defense of the fallopian tubes, and in interactions with gametes and embryos.

• 한국수산과학회지
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• 제34권4호
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• pp.412-418
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• 2001

[ $\beta$ ]-glucan을 시판의 배합사료에 $0.1\%$ 첨가한 다음 담수어인 잉어 (평균체중 1.0g 및 68.7g)와 해산어인 넙치 (평균 체중, 12.1 g 및 54.0g)에 2주간 첨가사료, 1주간 무첨가사료, 2주간 첨가사료 투여의 투여방법으로 5주간 경구투여한 다음 인위감염에 대한 생잔율을 조사하였다. 또 투여기간중의 말초혈액의 백혈구수와 혈중리소자임의 활성과 5주간 투여후에 채취한 백혈구의 식작용능을 조사하였다. $\beta$-glucan첨가사료 투여군에 $1\times10^6$cells의 Aeromonas hydrophila를 접종한 평균체중 68.7g의 잉어군과 $1\times10^5$cells의 Edwardsiella tarda를 접종한 평균체중 68.7g의 넙치군에서 $\beta$-glucan첨가사료 투여군이 무첨가투여군에 비해 생존율이 높았고 (P<0.05),그 외의 접종군에서는 무첨가투여군과 차이가 없었다. $\beta$-glucan첨가사료 투여기간 동안 말초혈액중의 마크로파아지와 호중구수는 투여기간에는 증가, 무투여기간에는 감소하는 경향이었으며 특히 마크로파아지의 증가가 현저하였다. 혈중 리소자임은 잉어에서는 $\beta$-glucan첨가사료 투여기간에는 상승하고 무투여기간에는 감소하였지만, $\beta$-glucan첨가사료를 다시 투여한 후 급격히 상승하여 높은 활성을 유지하였다 그러나 넙치는 잉어보다는 활성의 증가는 현저하지 않았고 무투여기간에도 변동이 없었지만 무첨가투여군보다는 항상 높은 활성을 나타내었다.

• Journal of Microbiology
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• 제40권3호
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• pp.183-192
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• 2002

Cells infected With equine herpesvirus type-1 (EHV-1) Produced both infectious and non-infectious Virus-related particles. Compared to the whole virion, non-infectious particles termed L-particles were deter-mined to lack 150 kDa protein, commonly known as nucleocapsid protein. The potential of L-particles to induce immune responses was studied in mice and foals. Intranasal immunization with L-particles or whole virions induced poor IgG antibody responses in mice. Interestingly, despite the poor antibody response, the conferred immunity protected the host from challenge infections. This was indicated by a significant reduction in virus titers in line with recovery towards normal body weight. Subsequently, the test on the usefulness of L-particles as immunizing agents was extended to foals. Immunization of specific-pathogen-free (SPF) foals resulted in similar results. As determined by a complement-fixing-antibody test (CFT), foals seroconverted when they were immunized either with inactivated L-particles or whole virions via intramuscular (i.m.) injections. The presence of the antibody correlated with the degree of protection. Beyond day 1 post challenge infection (p.i.), there was no virus shedding in the nasal mucus of foals immunized with whole EHV-1 virions. Virus shedding was observed in foals Immunized with L-particles but limited to days 6 to 8 p.i. only. In contrast, extended vim shedding was observed in non-immunized foals and it was well beyond day 14 p.i. Viremia was not detected for more than four days except in non-immunized foals. Immunization in mice via intranasal (i.n.) conferred good protection. However, compared to the i.n. route, a greater degree of protection was obtained in foals following immunization via i.m. route. Despite variation in the degree of protection due to different routes of immunization in the two animal species, our results have established significant evidence that immunization with L-particles confers protection in the natural host. It is suggested that non-infectious L-particles should be used as immunizing agents for vaccination of horses against EHV-1 infection.

• Journal of Microbiology and Biotechnology
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• 제34권7호
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• pp.1484-1490
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• 2024

The gut microbiota is a key factor significantly impacting host health by influencing metabolism and immune function. Its composition can be altered by genetic factors, as well as environmental factors such as the host's surroundings, diet, and antibiotic usage. This study aims to examine how the characteristics of the gut microbiota in pigs, used as source animals for xenotransplantation, vary depending on their rearing environment. We compared the diversity and composition of gut microbiota in fecal samples from pigs raised in specific pathogen-free (SPF) and conventional (non-SPF) facilities. The 16S RNA metagenome sequencing results revealed that pigs raised in non-SPF facilities exhibited greater gut microbiota diversity compared to those in SPF facilities. Genera such as Streptococcus and Ruminococcus were more abundant in SPF pigs compared to non-SPF pigs, while Blautia, Bacteroides, and Roseburia were only observed in SPF pigs. Conversely, Prevotella was exclusively present in non-SPF pigs. It was predicted that SPF pigs would show higher levels of processes related to carbohydrate and nucleotide metabolism, and environmental information processing. On the other hand, energy and lipid metabolism, as well as processes associated with genetic information, cell communication, and diseases, were predicted to be more active in the gut microbiota of non-SPF pigs. This study provides insights into how the presence or absence of microorganisms, including pathogens, in pig-rearing facilities affects the composition and function of the pigs' gut microbiota. Furthermore, this serves as a reference for tracing whether xenotransplantation source pigs were maintained in a pathogen-controlled environment.

• Archives of Pharmacal Research
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• 제22권5호
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• pp.437-447
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• 1999

Type I diabetes, also known as insulin-dependent diabetes mellitus (IDDM) results from the destruction of insulin-producing pancreatic $\beta$ cells by a progressive $\beta$ cell-specific autoimmune process. The pathogenesis of autoimmune IDDM has been extensively studied for the past two decades using animal models such as the non-obese diabetic (NOD) mouse and the Bio-Breeding (BB) rat. However, the initial events that trigger the immune responses leading to the selective destruction of the $\beta$ cells are poorly understood. It is thought that $\beta$ cell auto-antigens are involved in the triggering of $\beta$ cell-specific autoimmunity. Among a dozen putative $\beta$ cell autoantigens, glutamic acid decarboxylase (GAD) has bee proposed as perhaps the strongest candidate in both humans and the NOD mouse. In the NOD mouse, GAD, as compared with other $\beta$ cell autoantigens, provokes the earliest T cell proliferative response. The suppression of GAD expression in the $\beta$ cells results in the prevention of autoimmune diabetes in NOD mice. In addition, the major populations of cells infiltrating the iselts during the early stage of insulitis in BB rats and NOD mice are macrophages and dendritic cells. The inactivation of macrophages in NOD mice results in the prevention of T cell mediated autoimmune diabetes. Macrophages are primary contributors to the creation of the immune environment conducive to the development and activation of $\beta$cell-specific Th1-type CD4+ T cells and CD8+ cytotoxic T cells that cause autoimmune diabetes in NOD mice. CD4+ and CD8+ T cells are both believed to be important for the destruction of $\beta$ cells. These cells, as final effectors, can kill the insulin-producing $\beta$ cells by the induction of apoptosis. In addition, CD8+ cytotoxic T cells release granzyme and cytolysin (perforin), which are also toxic to $\beta$ cells. In this way, macrophages, CD4+ T cells and CD8+ T cells act synergistically to kill the $\beta$ cells in conjunction with $\beta$ cell autoantigens and MHC class I and II antigens, resulting in the onset of autoimmune type I diabetes.