Oral Administration of Bacillus subtilis Subunit Vaccine Significantly Enhances the Immune Protection of Grass Carp against GCRV-II Infection

Grass carp reovirus (GCRV) is a severe virus that causes great losses to grass carp culture every year, and GCRV-II is the current popular and fatal strain. VP56, fibrin on the outer surface of GCRV-II, mediates cell attachment. In this study, we firstly divided the VP56 gene into four fragments to screen the optimal antigen by enzyme-linked immunosorbent assay and neutralizing antibody methods. The second fragment VP56-2 demonstrates the optimal efficiency and was employed as an antigen in the following experiments. Bacillus subtilis were used as a carrier, and VP56-2 was expressed on the surface of the spores. Then, we performed the oral immunization for grass carp and the challenge with GCRV-II. The survival rate was remarkably raised, and mRNA expressions of IgM were significantly up-regulated in spleen and head kidney tissues in the B. s-CotC-VP56-2 group. Three crucial immune indexes (complement C3, lysozyme and total superoxide dismutase) in the sera were also significantly enhanced. mRNA expressions of four important genes (TNF-α, IL-1β, IFN1 and MHC-II) were significantly strengthened. Tissue lesions were obviously attenuated by histopathological slide examination in trunk kidney and spleen tissues. Tissue viral burdens were significantly reduced post-viral challenge. These results indicated that the oral recombinant B. subtilis VP56-2 subunit vaccine is effective for controlling GCRV infection and provides a feasible strategy for the control of fish virus diseases.

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Immunomodulatory Effects of a Bioactive Compound Isolated fromDryopteris crassirhizomaon the Grass CarpCtenopharyngodon idella

Journal of Immunology Research ◽

10.1155/2016/3068913 ◽

2016 ◽

Vol 2016 ◽

pp. 1-10 ◽

Cited By ~ 7

Author(s):

Cheng Chi ◽

Sib Sankar Giri ◽

Jin Woo Jun ◽

Hyoun Joong Kim ◽

Saekil Yun ◽

...

Keyword(s):

Gene Expression ◽

Grass Carp ◽

Head Kidney ◽

Bioactive Compound ◽

Ctenopharyngodon Idella ◽

Complement C3 ◽

Immunomodulatory Activity ◽

Immune Parameters ◽

Significant Expression ◽

Immune Related Genes

In the present study, we investigated effects of compound kaempferol 3-a-L-(4-O-acetyl)rhamnopyranoside-7-a-L-rhamnopyranoside (SA) isolated fromDryopteris crassirhizomaduring immune-related gene expression inCtenopharyngodon idellahead kidney macrophages (CIHKM). The expression of immune-related genes (IL-1β,TNF-α,MyD88, andMx1) were investigated using real-time PCR at 2 h, 8 h, 12 h, and 24 h after incubation with 1, 10, and 50 μg mL−1of SA. Furthermore, fish were injected intraperitoneally with 100 μL of SA, and immune parameters such as lysozyme activity, complement C3, SOD, phagocytic activity, and IgM level were examined at 1, 2, and 3 weeks after injection. The differential expression of cytokines was observed after exposure to SA.IL-1βgenes displayed significant expression at 2 and 8 h after exposure to 1–10 μg mL−1of SA. SA also induced gene expression of cytokines such asMyD88,Mx1, andTNF-α. Furthermore, enhanced immune parameters in grass carp confirmed the immunomodulatory activity of SA. Interestingly, this compound has no toxic effect on CIHKM cells as tested by MTT assay. In addition, fish immunised with 10 μg mL−1of SA exhibited maximum resistance againstAeromonas hydrophilainfection. These results suggest that SA has the potential to stimulate immune responses in grass carp.

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Development and efficacy of a grass carp reovirus (GCRV) outer capsid protein VP7 subunit vaccine

JOURNAL OF HUNAN AGRICULTURAL UNIVERSITY ◽

10.3724/sp.j.1238.2011.00659 ◽

2012 ◽

Vol 37 (6) ◽

pp. 659-664 ◽

Cited By ~ 1

Author(s):

Shi-ying XU ◽

Jing-hui LI ◽

Yong ZOU ◽

Lin LIU ◽

Cheng-liang GONG ◽

...

Keyword(s):

Capsid Protein ◽

Grass Carp ◽

Subunit Vaccine ◽

Outer Capsid Protein ◽

Grass Carp Reovirus ◽

Outer Capsid

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Role of MDA5 in regulating CXCL10 expression induced by TLR3 signaling in human rheumatoid fibroblast-like synoviocytes

Molecular Biology Reports ◽

10.1007/s11033-020-06069-z ◽

2021 ◽

Author(s):

Tatsuro Saruga ◽

Tadaatsu Imaizumi ◽

Shogo Kawaguchi ◽

Kazuhiko Seya ◽

Tomoh Matsumiya ◽

...

Keyword(s):

Inflammatory Responses ◽

Neutralizing Antibody ◽

Poly I:C ◽

Enzyme Linked Immunosorbent Assay ◽

Dependent Manner ◽

Inflammatory Reactions ◽

Polyinosinic:Polycytidylic Acid ◽

Tlr3 Signaling ◽

Fibroblast Like Synoviocytes

AbstractC-X-C motif chemokine 10 (CXCL10) is an inflammatory chemokine and a key molecule in the pathogenesis of rheumatoid arthritis (RA). Melanoma differentiation-associated gene 5 (MDA5) is an RNA helicase that plays a role in innate immune and inflammatory reactions. The details of the regulatory mechanisms of CXCL10 production and the precise role of MDA5 in RA synovitis have not been fully elucidated. The aim of this study was to examine the role of MDA5 in regulating CXCL10 expression in cultured human rheumatoid fibroblast-like synoviocytes (RFLS). RFLS was stimulated with Toll-like receptor 3 (TLR3) ligand polyinosinic:polycytidylic acid (poly I:C), a synthetic double-stranded RNA mimetic. Expression of interferon beta (IFN-β), MDA5, and CXCL10 was measured by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blotting, and enzyme-linked immunosorbent assay. A neutralizing antibody of IFN-β and siRNA-mediated MDA5 knockdown were used to determine the role of these molecules in regulating CXCL10 expression downstream of TLR3 signaling in RFLS. Poly I:C induced IFN-β, MDA5, and CXCL10 expression in a concentration- and time-dependent manner. IFN-β neutralizing antibody suppressed the expression of MDA5 and CXCL10, and knockdown of MDA5 decreased a part of CXCL10 expression (p < 0.001). The TLR3/IFN-β/CXCL10 axis may play a crucial role in the inflammatory responses in RA synovium, and MDA5 may be partially involved in this axis.

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Characterization and protective activity of monoclonal antibodies directed against Fe (3+) ABC transporter substrate-binding protein of Glaesserella parasuis

Veterinary Research ◽

10.1186/s13567-021-00967-1 ◽

2021 ◽

Vol 52 (1) ◽

Author(s):

Kexin Zhu ◽

Dong Yu ◽

Jiahui An ◽

Yufeng Li

Keyword(s):

Monoclonal Antibodies ◽

Abc Transporter ◽

Subunit Vaccine ◽

Binding Protein ◽

Substrate Binding ◽

Passive Immunization ◽

Enzyme Linked Immunosorbent Assay ◽

Control Group ◽

And Control ◽

Substrate Binding Protein

AbstractGlässer’s disease is caused by the agent Glaesserella parasuis and is difficult to prevent and control. Candidate screening for subunit vaccines contributes to the prevention of this disease. Therefore, in this study, the inactivated G. parasuis reference serovar 5 strain (G. parasuis-5) was used to generate specific monoclonal antibodies (mAbs) to screen subunit vaccine candidates. Six mAbs (1A12, 3E3, 4C6, 2D1, 3E6, and 4B2) were screened, and they all reacted with the G. parasuis serovar 5 strain according to laser confocal microscopy and flow cytometry (FCM). Indirect enzyme-linked immunosorbent assay (ELISA) showed that one mAb 2D1, can react with all 15 reference serovars of G. parasuis. Protein mass spectrometry and Western blot analysis demonstrated that mAb 2D1 specifically reacts with Fe (3+) ABC transporter substrate-binding protein. A complement killing assay found that the colony numbers of bacteria were significantly reduced in the G. parasuis-5 group incubated with mAb 2D1 (p < 0.01) in comparison with the control group. Opsonophagocytic assays demonstrated that mAb 2D1 significantly enhanced the phagocytosis of 3D4/21 cells by G. parasuis (p < 0.05). RAW264.7 cells with stronger phagocytic ability were also used for the opsonophagocytic assay, and the difference was highly significant (p < 0.01). Passive immunization of mice revealed that mAb 2D1 can eliminate the bacteria in the blood and provide protection against G. parasuis-5. Our study found one mAb that can be used to prevent and control G. parasuis infection in vivo and in vitro, which may suggest that Fe (3+) ABC transporter substrate-binding protein is an immunodominant antigen and a promising candidate for subunit vaccine development.

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Insights into the functional role of grass carp IL ‐8 in head kidney leukocytes: pro‐inflammatory effects and signaling mechanisms

Journal of Fish Biology ◽

10.1111/jfb.14934 ◽

2021 ◽

Author(s):

Minghui Zhao ◽

Yazhen Liu ◽

Yajun Gao ◽

Xinyan Wang ◽

Hong Zhou ◽

...

Keyword(s):

Grass Carp ◽

Functional Role ◽

Head Kidney ◽

Signaling Mechanisms

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Neutralization Assessments Reveal High Cardiothoracic Ratio and Old Age as Independent Predictors of Low Neutralizing Antibody Titers in Hemodialysis Patients Receiving a Single Dose of COVID-19 Vaccine

Journal of Personalized Medicine ◽

10.3390/jpm12010068 ◽

2022 ◽

Vol 12 (1) ◽

pp. 68

Author(s):

Chun-Yu Chen ◽

Kuan-Ting Liu ◽

Shin-Ru Shih ◽

Jung-Jr Ye ◽

Yih-Ting Chen ◽

...

Keyword(s):

Single Dose ◽

Old Age ◽

Neutralizing Antibodies ◽

Additive Model ◽

Humoral Response ◽

Neutralizing Antibody ◽

Enzyme Linked Immunosorbent Assay ◽

Healthy Controls ◽

Neutralization Titer ◽

Cardiothoracic Ratio

Background: Data are lacking regarding predictors of quantification of neutralizing antibodies (nAbs) based on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) 50% neutralization titer (NT50) after a single dose of COVID-19 vaccine in hemodialysis (HD) patients. Methods: This prospective single-center study enrolled 200 HD patients and 82 healthy subjects to estimate antibodies against the SARS-CoV-2 viral spike protein 1 and receptor-binding domain after a first dose of a COVID-19 vaccine (ChAdOx1 or mRNA-1273), measured by enzyme-linked immunosorbent assay and applied spline-based generalized additive model regression analysis to predict NT50 converted to international units. Results: After the first dose of ChAdOx1, multiple linear regression showed that age (p = 0.011) and cardiothoracic ratio (p = 0.002) were negatively associated with NT50. Older age (OR = 0.958, p = 0.052) and higher cardiothoracic ratio (OR < 0.001, p = 0.037) could predict negative humoral response (NT50 < 35.13 IU/mL). NT50 was lower in HD patients compared with healthy controls receiving ChAdOx1 (10.68 vs. 43.01 IU/m, p < 0.001) or mRNA-1273 (36.39 vs. 262.2 IU/mL, p < 0.001). ChAdOx1 elicited lower GMTs than mRNA-1273 in the HD cohort (10.68 vs. 36.39 IU/mL, p < 0.001) and in healthy controls (43.01 vs. 262.22 IU/mL, p < 0.001). Conclusion: High cardiothoracic ratio and old age could independently predict a decline in nAb titers in an HD cohort vaccinated with a single dose of ChAdOx1.

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Immunogenicity of heterologous prime/booster-inactivated and adenoviral-vectored COVID-19 vaccine: real-world data

10.21203/rs.3.rs-785693/v1 ◽

2021 ◽

Author(s):

Nasamon Wanlapakorn ◽

Nungruthai Suntronwong ◽

Harit Phowatthanasathian ◽

Ritthideach Yorsang ◽

Thanunrat Thongmee ◽

...

Keyword(s):

Neutralizing Antibody ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Activity ◽

Serum Samples ◽

Real World Data ◽

Comparison Groups ◽

Chemiluminescent Microparticle Immunoassay ◽

Specific Igg ◽

Vectored Vaccine ◽

Sera Samples

Abstract Limited data are available on the responses to heterologous vaccine regimens for SARS-CoV-2, especially among countries using inactivated and adenoviral-vectored vaccines. A total of 77 participants who received heterologous prime/booster-inactivated COVID-19 vaccine and adenoviral-vectored vaccine were enrolled in our study. There were two comparison groups vaccinated with the homologous CoronaVac (N = 79) and AZD1222 (N = 80) regimen. All sera samples were tested for SARS-CoV-2 spike receptor-binding-domain (RBD) IgG using a chemiluminescent microparticle immunoassay (CMIA). The neutralizing activity in a subset of serum samples was tested against the original Wuhan strain and variants of concern, B.1.1.7 and B.1.351, using an enzyme-linked immunosorbent assay (ELISA)-based surrogate virus neutralization test (sVNT). The CoronaVac followed by the AZD1222 vaccine induced higher levels of spike RBD-specific IgG than that of two-dose CoronaVac or AZD1222 vaccines (p < 0.001). Sera samples of the CoronaVac/AZD1222 vaccine recipients elicited higher neutralizing antibody activity against the original Wuhan and the B.1.351 strain than in the recipients of the two-dose CoronaVac or AZD1222. Following the inactivated CoronaVac/adenoviral-vectored (AZD1222) vaccination administered 14–72 days apart, participants receiving the heterologous vaccine regimen had higher spike RBD-specific IgG and neutralizing activities than the homologous CoronaVac vaccine recipients.

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Protection of Hamsters Challenged with SARS-CoV-2 Variants of Concern by Two Doses of MVC-COV1901 Vaccine Followed by a Single Dose of Beta Variant Version of MVC-COV1901

10.1101/2021.09.29.462344 ◽

2021 ◽

Author(s):

Tsun-Yung Kuo ◽

Chia-En Lien ◽

Yi-Jiun Lin ◽

Meei-Yun Lin ◽

Chung-Chin Wu ◽

...

Keyword(s):

Subunit Vaccine ◽

Virus Titer ◽

Neutralizing Antibody ◽

Protein S ◽

Protein Subunit ◽

Live Virus ◽

The Third ◽

Severity Of Disease ◽

Potent Immune Response ◽

Ancestral Strain

AbstractThe current fight against COVID-19 is compounded by the Variants of Concern (VoCs), which can diminish the effectiveness of vaccines, increase viral transmission and severity of disease. MVC-COV1901 is a protein subunit vaccine based on the prefusion SARS-CoV-2 spike protein (S-2P) adjuvanted with CpG 1018 and aluminum hydroxide. Here we used the Delta variant to challenge hamsters innoculated with S-2P based on the ancestral strain or the Beta variant in two-dose or three-dose regimens. Two doses of ancestral S-2P followed by the third dose of Beta variant S-2P was shown to induce the highest neutralizing antibody titer against live SARS-CoV-2 of the ancestral strain as well as all VoCs. All regimens of vaccination were able to protect hamsters from SARS-CoV-2 Delta variant challenge and reduce lung live virus titer. Three doses of vaccination significantly reduced lung viral RNA titer, regardless of using the ancestral or Beta variant S-2P as the third dose. Based on the immunogenicity and viral challenge data, two doses of ancestral S-2P followed by the third dose of Beta variant S-2P could induce broad and potent immune response against the variants.

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