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Molecular Characterization, Expression Profiles, and immunostimulation Responses of TRAF6 and TAK1 in Japanese Flounder(Paralichthys olivaceus)

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Journal of Ocean University of China 2019年第1期18卷 165-176页

作者： ZHAO Haitao CHEN Yan WANG Zhigang ZHANG Quanqi YU Haiyang College of Marine Life Sciences Ocean University of China Key Laboratory of Marine Genetics and BreedingMinistry of Education

Tumor necrosis factor receptor-associated factor 6(TRAF6) and transforming growth factor-β-activated kinase 1(TAK1) are two important adaptor molecules in Toll-like receptor(TLR) signaling pathway. In this study, TRAF6(PoTRAF6) and TAK1(PoTAK1) were cloned and characterized in Japanese flounder(Paralichthys olivaceus). The full-length cDNA sequence of Po TRAF6 is 1953 bp, with an open reading frame(ORF) of 1713 bp encoding a putative protein of 570 amino acids. PoTRAF6 contains one really interesting new gene(RING) domain, two zinc fingers, one coiled-coil region, and one meprin and TRAF homology(MATH) domain, which shows a high similarity to TRAF6 s in other species. The full-length PoTAK1 cDNA sequence is 2086 bp, with an ORF of 1728 bp that encodes a putative protein of 575 amino acids. PoTAK1 contains a conserved serine/threonine protein kinase catalytic domain and a coiled-coil region. The promoter regions of PoTRAF6 and PoTAK1 were also analyzed to predict several potential transcription factor-binding sites. In addition, the expression patterns of these two genes were examined in developmental stages, different tissues, and challenged samples. PoTRAF6 and PoTAK1 were expressed during the whole developmental stages, and the highest expressions were in intestine and heart, respectively. In challenged embryonic cells with LPS, CpG ODN, and poly I:C, the expressions of PoTRAF6 and PoTAK1 were both up-regulated significantly. These results suggest that PoTRAF6 and Po TAK1 play crucial roles in immune responses and may be involved in the developmental process of Japanese flounder.

关键词： TRAF6 TAK1 Paralichthys olivaceus cloning expression promoter analysis immunostimulation

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Application of yeast spores as β-glucan particles

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Particuology 2022年第12期71卷 34-40页

作者： Guoyu Liu Yan Yang Hiroyuki Tachikawa Xiao-Dong Gao Hideki Nakanishi Key Laboratory of Carbohydrate Chemistry and Biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi 214122China Department of Applied Biological Chemistry Graduate School of Agricultural and Life SciencesUniversity of TokyoTokyo 113-8657Japan Collaborative Research Institute for Innovative Microbiology University of TokyoTokyo 113-8657Japan

β-Glucan exhibits various beneficial biological activities including immunostimulation.This polysaccha-ride is present in the cell wall of microorganisms including the yeast Saccharomyces cerevisiae.Although vegetative yeast cells are used as dietary supplements,β-glucan in the vegetative cell wall is covered with mannan.During nutrient starvation,yeast cells differentiate into dormant cell spores.β-Glucan is also present in the spore wall,but spores have been poorly studied as application tools.The spore wall consists of four layers andβ-glucan forms the third layer from the outside.However,unlike vegeta-tive cells,theβ-glucan layer can be exposed to the cell surface in spores by genetic manipulation.For example,chs3Δmutant spores exhibit such a spore wall configuration.Dectin-1 is the primary receptor forβ-glucan in mammals and,its activation induces phagocytosis and proinflammatory cytokine secre-tions.We found that chs3Δspores are phagocytosed in mammalian cells in a manner dependent on dectin-1 and induce higher levels of TNF-αand IL-6 than vegetative cells.Our results demonstrate that chs3Δspores areβ-glucan particles with immunostimulatory activities.Thus,they would be attractive materials for application in the food industry.

关键词： Saccharomycescerevisiae Sporewall β-glucan Dectin-1 immunostimulation

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Structure-efficacy relationships of immunostimulatory activity of CpG-containing oligodeoxynucleotides on mouse spleen cells

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Acta Pharmacologica Sinica 2007年第10期28卷 1637-1644页

作者： Hai-yan DU~3,Shao-you XIA~4,Hai-feng SONG~(2,5),Na LI~2,Ming-mei SHANG~2,Jia ZOU~2,Qing-qing WANG~2,Lun OU~2,XiaoSUN~2,Ai-guo JI~3,Zhong-ming TANG~2~2 Department of Pharmacology and Toxicology,Beijing Institute of Radiation Medicine,Beijing 100850,China ~3 Institute of Biochemical andBiotech Drugs,School of Pharmacy,Shandong University,Ji-nan 250012,China ~4 Department of General Surgery,General Hospital of PLA,Beijing100853,China Department of Pharmacology and Toxicology Beijing Institute of Radiation Medicine Beijing 100850 China. songhf@***.

Aim:To study the relationship between primary structures of oligodeoxynucleo-tides （ODN） containing unmethylated deoxycytidyl-deoxyguanosine （CpG）dinucleotide motifs and their immunostimulatory activities in mouse spleen cells.Methods:A series of CpG ODN with different primary structures were synthesized.Their capabilities to stimulate mouse spleen cell proliferation were determined by[3H]thymidine incorporation assay.Cytokine （interleukin [IL]-6,IL-12,and IFN-α）secretion spectra induced by CpG ODN were assessed by ELISA.The ability ofCpG ODN to activate natural killer cells was evaluated by standard 4 h51Cr-releaseassay.Flow cytometry was utilized to examine the expressions of various lympho-cyte surface molecules on diverse immunocytes.An effective CpG ODN for murine,ODN1826,was set as the template of modification and the positive control.Results:The immunostimulatory activities of CpG ODN with different sequencesand compositions varied markedly,both in character and in extent.It was uselessfor improving the immunostimulatory activity of ODN1826 by simply increasingthe functional hexameric CpG motif number,modifying the site of CpG motifs,orchanging the distance between multi-CpG motifs.However,an addition of a self-complementary palindrome structure at the 3'-end,but not the 5'-end of CpG ODN,aroused marked improvement in its activity.Several designed ODN had superiorcomprehensive immunostimulatory properties compared to ODN 1826.Conclusion:The immunostimulatory activity of a CpG ODN was relevant to its primary structure.It was useless for promoting immunostimulatory activity to simply change CpGmotif number,space,or distance.The 3'-end palindrome structure of CpG ODN isassociated with enhanced immunostimulatory activity.

关键词： CpG oligodeoxynucleotides immunostimulation structure-activity relationship

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Phytochemical bioprospecting, antioxidant, antimicrobial and cytotoxicity activities of saline extract from Tithonia diversifolia(Hemsl) A.Gray leaves

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Asian Pacific Journal of Tropical Biomedicine 2018年第5期8卷 245-253页

作者： Bruno Rafael Barboza Bárbara Rafaela da Silva Barros Bárbara de Azevedo Ramos Maiara Celine de Moura Thiago Henrique Napoleao Maria Tereza dos Santos Correia Luana Cassandra Breitenbach Barroso Coelho Iranildo José da Cruz Filho Ana Maria Souto Maior Túlio Diego da Silva Leylianne de Cássia Rodrigues Nerys Edson Renan Barros de Santana Cláudia Sampaio de Andrade Lima Virgínia Maria Barros de Lorena Cristiane Moutinho Lagos de Melo Department of Antibiotics Biosciences Centre Federal University of Pernambuco Department of Biochemistry Biosciences Centre Federal University of Pernambuco Center for Strategic Techmologies of the Northeast Department of Biophysics Biosciences Centre Federal University of Pernambuco Aggeu Magalhaes Institute IAM/FIOCRUZ

Objective:To evaluate antimicrobial and antioxidant properties of saline extract from Tithonia diversifolia leaves by phytochemical bioprospecting,and investigate its safety against animal cells.Methods:The saline extract was prepared,with NaCl(0.15 M),by constant stirring of the dried and pulverized leaves,followed by volume reduction by lyophilization.The extract was phytochemical characterized using ultra-performance liquid chromatography,and total phenol and flavonoid analysis also was performed.The antioxidant capacity was determined through DPPH*radical,the antimicrobial property was evaluated against standard bacteria and fungi,and the viability assays were performed against mice splenocytes.Results:Fifteen compounds were identified belonging to two main classes terpenoids and phenolics.The extract showed 22.185 mg GAE/g of total phenolic compounds and 3.220 mg QE/g of flavonoid.Moreover,extract showed higher antioxidant ability similar to butylated hydroxytoluene a standard molecule[(3.042±0.019)mg AAE/g and(4.12±0.10)mg AAE/g to saline extract and butylated hydroxytoluene,respectively].The antimicrobial assays demonstrated that the extract had a significant antifungal potential against Candida species and could be used with safety against mice splenocytes,in concentrations lower than 50μg/mL,promoting higher proliferation in these cells.Conclusions:Saline extract from Tithonia diversifolia leaves presents potential antioxidant,antifungal properties and induces immunostimulation in mice splenocytes.

关键词： Tithonia diversifolia Extract Antioxidant Antifungal properties immunostimulation

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Enhancements of non-specific immune response in Mugil cephlus by seaweed extract against Vibrio alginolyticus(BRTR07)

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Journal of Coastal Life Medicine 2015年第10期 776-786页

作者： Rajasekar Thirunavukkarasu Priyadharshini Pandiyan Deivasigamani Balaraman Ilamathi Jayaraman Kumaran Subaramaniyan Edward Gnana Jothi George Center for Drug Discovery and Development Sathyabama UniversityJeppiaar NagarOld Mamallapuram RoadChennai-600119India CAS in Marine Biology Annamalai UniversityParangipettaiChidambaramTamil Nadu-608502India Department of Microbiology Sri Sankara Arts and Science College(Affiliated to Madras University)EnathurKanchipuram-631561Tamil NaduIndia

Objective:To focus on the growth rate and feed utilization of fish by using trash fish feeds supplement with marine seaweeds.Methods:Selected seaweed was extracted using hot-water and its extract was mixed with trash fish feed at different concentrations(0.5%,1%and 2%for 1-30 days)and the non-specific immune response in fish was studied and challenged with Vibrio alginolyticus at 1×106 CFU/fish.The hot-water extract of seaweeds was analysed by gas chromatography-mass spectrometry.Results:The average body weight(5.320±0.018),percent weight gain(227.66±0.28),specific growth rate(2.080±0.015),hepatosomatic index(1.197±0.00)and viscerosomatic index(4.421±0.150)were significantly increased in the fish feed with seaweed containing 5%of Sargassum wightii(S.wightii)when compared with other seaweeds and control diet.Hot-water extract of S.wightii(1%)was significantly enhanced the immune response in fish when compared with other diets(0.5%and 2%).S.wightii showed good immunostimulation properties.Gas chromatography-mass spectrometry result showed that the hot-water extract of S.wightii seaweed contained fatty acids.Conclusions:Trash fish feed will reduce the production cost and also provide evidence that aqueous leaf extract of S.wightii(1%)was added to a formulated fish diet which could activate the non-specific immune response and disease resistance against Vibrio alginolyticus in Mugil cephalus.

关键词： Mugil cephlus Vibrio alginolyticus Hot-water extract Sargassum wightii immunostimulation

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A Review of Immune System Components, Cytokines, and Immunostimulants in Cultured Finfish Species

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Open Journal of Animal Sciences 2017年第3期7卷 267-288页

作者： Timothy J. Bruce Michael L. Brown Missouri Department of Conservation Lost Valley Hatchery Warsaw Missouri USA Department of Natural Resource Management South Dakota State University Brookings South Dakota USA

Aquaculture is a rapidly growing global agriculture sector and the importance of fish health has become of upmost importance as production levels and stocking densities increase. Over the past few decades, there have been a large number of immunological investigations on commonly cultured finfish species. Further, new technologies and strategies that embody use of fish immunostimulants, probiotics, and vaccinology rely heavily upon a comprehensive understanding of teleost immune system mechanics. The teleost immune system works in concert to properly recognize, control, and clear aquatic pathogens. Recent findings have exemplified the cooperative efforts of the nonspecific and adaptive branches, and have put forth an emphasis on the importance of the mucosal immune response in all aspects of a mounted immune response. This review provides a generalized overview of the innate and adaptive arms of the fish immune system, and provides highlights of recently published work in the areas of signaling networks and mucosal immune interactions.

关键词： Innate Immunity Adaptive Immunity Cytokines immunostimulation Aquaculture

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Therapeutic properties of honey

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Health 2009年第4期1卷 281-283页

作者： Giuseppe Pipicelli Patrizio Tatti 不详

Honey has been successfully used in medicine since antiquity. However, with the advent of modern medicine it has been less used, espe-cially in the English-speaking world. Its benefi-cial effects in different disorders, rediscovered in recent decades, will be discussed below on the basis of a series of international scientific studies conducted to investigate the therapeutic properties of this natural product and published on Medline. It should be noted, however, that the therapeutic use of honey in everyday clinical practice needs to be validated by relevant guidelines and should only be adopted under medical prescription, in accordance with criteria of efficacy and safety for both patients and healthcare providers.

关键词： Antibacterial Oncolgy Wounds immunostimulation

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Immunostimulatory and Immunorestorative Effects of Leaf Extract and Fractions of Musanga cecropioides on Immunocompetent and Experimentally Immunocompromised Mice

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Clinical Complementary Medicine and Pharmacology 2023年第1期3卷 53-61页

作者： Cyril O.Ogbue Felix A.Onyegbule Christopher O.Ezugwu Ifebi H.M.Nchekwube Amara A.Ajaghaku Department of Pharmacognosy and Traditional Medicine Faculty of Pharmaceutical SciencesNnamdi Azikiwe UniversityAgulu 422102Anambra StateNigeria Department of Pharmaceutical and Medicinal Chemistry Faculty of Pharmaceutical SciencesNnamdi Azikiwe UniversityAgulu 422102Anambra StateNigeria Department of Pharmacognosy and Environmental Medicine University of NigeriaNsukka400221Enugu StateNigeria Department of Pharmacognosy and Traditional Medicine Faculty of Pharmaceutical SciencesChukwuemeka Odumegwu Ojukwu UniversityIgbariam432106Anambra StateNigeria

Background:Stimulation of the immune system against disease and restoration of immune functions in immun-odeficiency disease state are essential for maintaining good health.Objective:This study investigated the immunomodulatory effects of the leaf extract and fractions of Musanga cecropioides in immunocompetent and immunocompromised animal models.Methods:Humoral immune stimulatory effect of the extract and its fractions at 200 mg·kg^(−1) and 400 mg·kg^(−1) against sheep red blood cells(SRBCs)antigen was determined in both immunocompetent and cyclophosphamide(80 mg·kg^(−1))induced immunosuppressed animals.The effect of the most active fraction on leucopoiesis was also determined following high dose cyclophosphamide(300 mg·kg^(−1))suppression of the immune system.Levamisole at 2.5 mg·kg^(−1) was used as the reference drug throughout the study.Results:The extract dose dependently stimulated the humoral immune response to SRBC antigen in immuno-competent animals;however,its stimulatory effect was not significant(P>0.05).In the immunosuppressed model,the extract at 400 mg·kg^(−1) evoked significant(P<0.05)increase in antibody titer value with a corre-sponding 67%immune-restorative effect.The ethyl acetate fraction(EAF)maintained the most active fraction with significant(P<0.05)antibody stimulatory effect in immunocompetent and immunosuppressed animals.Similarly,higher recovery rate was demonstrated by the EAF as shown by steep time recovery curve in high dose immunosuppressed animals.The EAF at 200 mg·kg^(−1) and 400 mg·kg^(−1) showed above 50%leucopoiesis just like the reference drug(2.5 mg·kg^(−1) levamisole),against 33.83%of leucopoiesis shown by the immune-suppressed control group.Conclusion:M.cecropioides boosted immune stimulation in immunocompetent animals and brought about fast rate of restoration of immune status in immunocompromised animals.

关键词： Musanga cecropioides immunostimulation Immunorestoration Leucopoiesis Antibody

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The Recognition of CpG-oligodeoxynucleotides by Toll-like Receptor 9 and Cytokine Prodution are Modulated by Extracellular Acidic pH

The Recognition of CpG-oligodeoxynucleotides by Toll-like Re...

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2005全国第二届核酸疫苗研讨会

作者： HE Ying SUN Shu-Han HE Xiao-Wen LI Kai WANG Fang JIANG Lei (Department of Medical Genetics, College of Basic Medical Sciences, Second Military Medical University, 800 Xiangyin Road, Shanghai 200433, P.R.China)

Bacterial and synthetic DNAs containing CpG dinucleotides in specific sequence contexts activate the vertebrate immune system through Toll-like receptor 9 (TLR9), although the mechanism of this interaction is still unclear. In the present study, we used CpG-oligodeoxynucleotides (CpG-ODNs) as immunomers, which showed immunostimulatory activity in transfected-HEK293 cells, stably expressing human TLR9, especially in acidic pH, whereas there was no significant activity in non-transfected-HEK 293 cells. When the extracellular pH is in neutral (pH 7.4), CpG-ODN induced low levels of IFN-γ, and when it shifted to acidic (pH 6.4) ,however,the CpG-ODN-induced IFN-γproduction from transfected-293 cells markedly increased. These results suggest direct involvement of TLR9 in the recognition of CpG motif and such pH change enabled CpG-ODN bind to TLR9 to trigger CpG-related signal transduction.

关键词： CpG-oligodeoxynucleotides Toll-like receptor 9 immunostimulation

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