scholarly journals Development of Recombinant Dihydrolipoamide Dehydrogenase Subunit Vaccine against Vibrio Infection in Large Yellow Croaker

Fishes ◽  
2022 ◽  
Vol 7 (1) ◽  
pp. 17
Author(s):  
Xiaomeng Li ◽  
Yuanzhen Tan ◽  
Zheng Zhang ◽  
Yupeng Huang ◽  
Pengfei Mu ◽  
...  

Large yellow croaker (Larimichthys crocea), an economically important marine fish in China, has suffered from serious vibriosis, which has resulted in great economic losses for the large yellow croaker industry. Vaccination has been considered to be a safe and effective method to prevent and control vibriosis. However, due to the complex diversity and serotypes of the Vibrio genus, the progress of Vibrio vaccine development is still slow. In this study, we prepared recombinant Vibrio dihydrolipoamide dehydrogenase (rDLD) protein and investigated its potential as a candidate to be a subunit vaccine against Vibrio. The lysozyme activity and the rDLD-specific antibody level in sera of large yellow croakers immunized with rDLD were significantly higher than those in the control group, and the transcript levels of proinflammatory cytokines (IL-6, IL-8, IL-1β), MHC IIα/β, CD40, CD8α, IL-4/13A, and IL-4/13B were significantly up-regulated in the spleen and head kidney of large yellow croakers immunized with rDLD, suggesting that rDLD could induce both specific and nonspecific immune responses in this species. In addition, rDLD protein increased the survival rate of large yellow croakers against Vibrio alginolyticus and Vibrio parahaemolyticus, with the relative percent of survival (RPS) being 74.5% and 66.9%, respectively. These results will facilitate the development of a potential subunit vaccine against Vibrio in large yellow croaker aquaculture.

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Kexin Zhu ◽  
Dong Yu ◽  
Jiahui An ◽  
Yufeng Li

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.


2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S65-S66
Author(s):  
Joel V Chua ◽  
Charles Davis ◽  
Amy Nelson ◽  
Ka Wing J Lam ◽  
Lydiah Mutumbi ◽  
...  

Abstract Background A primary challenge for HIV vaccine development is to raise antiviral antibodies capable of recognizing highly variable viral antigens. The full-length single chain (FLSC) gp120-CD4 chimeric protein was designed to present a highly conserved CD4-induced HIV-1 envelope structure that evokes cross-reactive humoral responses (Figure 1). IHV01 is an FLSC subunit vaccine formulated in alum adjuvant. The safety and immunogenicity of IHV01 was evaluated in this first-in-human phase 1a trial. Methods This randomized, double-blind placebo-controlled study involved three dose-escalating cohorts (75 µg, 150 µg, and 300 µg doses). Eligible participants were HIV-1 uninfected healthy volunteers aged 18 to 45 years. Participants in each cohort were block randomized in groups of four in a 3:1 ratio to receive either vaccine or placebo. Intramuscular injections were given on weeks 0, 4, 8, and 24. Participants were followed for an additional 24 weeks after the last immunization. Crossreactive antibody binding titers against diverse HIV envelopes and antigens and specific CD4i epitopes on gp120 were assessed. Results Sixty-five volunteers were enrolled—49 vaccine and 16 placebo. Majority (81%) of vaccinations with IHV01 produced no localized or systemic reactions; no different from the control group. The overall incidence of adverse events (AEs) was not significantly different between groups. Majority (89%) of vaccine-related AEs were mild in severity. The most common vaccine-related AEs were injection site pain (31%), pruritus (10%), and headache (10%). There were no vaccine-related serious AE, discontinuation due to AE, or intercurrent HIV infection. By the final vaccination, all subjects in all cohorts had developed antibodies against IHV01; all placebo recipients were negative. The antibodies induced by IHV01 reacted with envelope antigens from diverse HIV-1 strains (Figure 2). Conclusion IHV01 vaccine was safe, well tolerated, and immunogenic in all doses tested. The vaccine raised broadly reactive humoral responses against multiple gp120 domains, transition state structures, and CD4i epitopes. Disclosures All Authors: No reported Disclosures.


2019 ◽  
Vol 39 (3) ◽  
pp. 168-174
Author(s):  
Luisa V.A. Otoni ◽  
Michelle P. Gabardo ◽  
Núbia R. Macêdo ◽  
Mariane M. Wagatsuma ◽  
Marina M. Pereira ◽  
...  

ABSTRACT: Porcine proliferative enteropathy (PPE) is one of the most common enteric diseases in growing and finishing pigs. PPE is characterized by reduced growth performance, accompanied or not by diarrhea. PPE is highly prevalent in several countries of the Americas, Europe and Asia, causing high economic losses in swine herds. The most common form of PPE control in pigs is antibiotic therapy. The objective of this study was to evaluate a new product based on tylosin injectable (Eurofarma Laboratórios S.A.) to control PPE in experimentally inoculated animals. Sixty 5-week-old pigs with mean weight of 9.5kg were divided into two experimental groups of 30 animals: medication and control. All pigs were challenged with Lawsonia intracellularis, the etiologic agent of PPE, on day zero. Fecal score, body condition score, and behavior were daily evaluated. Pigs were weighted on days -2, 13 and 21 of the experiment. Pigs in the Medication Group received tylosin injectable 13 days after inoculation, in three doses with a 12-hour interval between them. Pigs in the Control Group received injectable saline solution following the same protocol. In the Control Group, 23pigs presented with diarrhea before day 13. After day 13, the number of diarrheic animals in this group was reduced to 17. In the Medication Group, 26 pigs presented with diarrhea in the initial period, and in the period after medication, only 11 animals had diarrhea. The score of gross intestinal PPE lesions in the Medication Group was lower than that in the Control Group (p=0.031). The Medication Group also showed lower score for Lawsonia intracellularis antigen-labeling by immunohistochemistry compared with that of the Control Group (p=0.032), showing lower level of infection. These results demonstrate that tylosin injectable (Eurofarma Laboratórios S.A.), administrated in three doses (1mL/20kg) every 12 hours, was effective for the control of PPE in experimentally inoculated pigs.


2012 ◽  
Vol 47 (No. 8) ◽  
pp. 213-217 ◽  
Author(s):  
B. Habrun ◽  
V. Bilič ◽  
Ž. Cvetnič ◽  
A. Humski ◽  
M. Benič

A vaccine for porcine pleuropneumonia caused by Actinobacillus pleuropneumoniae was studied in Croatia on a farm infected by agent serotypes 2 and 9. Vaccination with a commercial subunit vaccine was initiated in the second half of 1998 due to the immense economic damage caused on the farm by this disease. All prefattening and fattening pigs kept on the farm during the first three months of 1999 were allocated in two groups: vaccinated and control. In the control and vaccinated group, 226 and 35 animals (5.78% and 0.96% of the average number of prefattening and fattening pigs in control and vaccinated group), respectively, died from pleuropneumonia. The vaccine efficacy was 83.5%. Examination of the randomly selected lungs on the slaughter line revealed significant reduction in the lesions specific for the chronic form of pleuropneumonia in the vaccinated group (vaccine efficacy 78.6%). The tested vaccine significantly decreased the death rate and pulmonary lesions due to A. pleuropneumoniae.


2012 ◽  
Vol 19 (5) ◽  
pp. 740-745 ◽  
Author(s):  
André A. Grassmann ◽  
Samuel R. Félix ◽  
Carolina Ximendes dos Santos ◽  
Marta G. Amaral ◽  
Amilton C. P. Seixas Neto ◽  
...  

ABSTRACTLeptospirosis, a worldwide zoonosis, lacks an effective, safe, and cross-protective vaccine. LipL32, the most abundant, immunogenic, and conserved surface lipoprotein present in all pathogenic species ofLeptospira, is a promising antigen candidate for a recombinant vaccine. However, several studies have reported a lack of protection when this protein is used as a subunit vaccine. In an attempt to enhance the immune response, we used LipL32 coupled to or coadministered with the B subunit of theEscherichia coliheat-labile enterotoxin (LTB) in a hamster model of leptospirosis. After homologous challenge with 5× the 50% lethal dose (LD50) ofLeptospira interrogans, animals vaccinated with LipL32 coadministered with LTB and LTB::LipL32 had significantly higher survival rates (P< 0.05) than animals from the control group. This is the first report of a protective immune response afforded by a subunit vaccine using LipL32 and represents an important contribution toward the development of improved leptospirosis vaccines.


Parasitology ◽  
1986 ◽  
Vol 92 (S1) ◽  
pp. S119-S137 ◽  
Author(s):  
J. Scaife ◽  
N. Bone ◽  
M. Goman ◽  
R. Hall ◽  
I. A. Hope ◽  
...  

The human malaria parasite,Plasmodium falciparum, is currently being actively studied by molecular biologists. It is hoped that the use of recombinant DNA techniques in this area will give new insights into the biology of the organism and, at the same time, provide new approaches to diagnosis and vaccine development.Our own studies employ the blood stages of the parasite and cover three main areas: enzymes of importance in parasite metabolism; antigens of potential use in a subunit vaccine; and repetitive DNA as a probe able to distinguish genetically different isolates ofP. falciparumand as a species-specific diagnostic tool in human and mosquito infections.


Viruses ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 262 ◽  
Author(s):  
Gang Li ◽  
Guanliu Yu ◽  
Yujuan Niu ◽  
Yumei Cai ◽  
Sidang Liu

Serotype 4 fowl adenovirus (FAdV-4) is the main pathogen for hydropericardium syndrome (HPS) in chickens. It has caused major economic losses in the global poultry industry. Currently, FAdV-4′s transmission routes in chickens remain unclear. Here we investigate the airborne transmission routes of FAdV-4 in chickens. A total of 45 ten-day-old chickens were equally divided into three groups (infected group/isolator A, airborne group/isolator B, and control group/isolator C). Of note, isolators A and B were connected by a leak-free pipe. The results showed that the virus could form a viral aerosol, detected in isolators two days post infection (dpi). The viral aerosol reached a peak at 4 dpi in the infected group. Healthy chickens in the airborne group were infected by the virus at 8 dpi. The chickens of the airborne group demonstrated subclinical symptoms capable of shedding the virus for some time. This finding suggests that FAdV-4 can be efficiently transmitted among chickens by aerosol transmission. These findings have significant implications for developing strategies to control this infectious disease epidemic.


2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 265-266
Author(s):  
Mauro Venturini ◽  
Demian Bellido ◽  
Josefina Baztarrica ◽  
Lucia Rocha ◽  
Viviana Parreño ◽  
...  

Abstract Bovine viral diarrhea virus (BVDV) infects ruminants, with a worldwide distribution, the virus causes a broad spectrum of clinical diseases and economic losses. Vaccination against BVDV is an important component of prevention and control programs. Currently, only modified live vaccines (MLV) and inactivated vaccines are used. Both have historical disadvantages; MLV in terms of safety and inactivated vaccines in terms of immunogenicity. We have previously reported the development and efficacy trials of the first targeted subunit vaccine against BVDV. The core of the vaccine is a fusion of the BVDV structural protein, E2, to a single-chain antibody, APCH, together termed, APCH-E2. The APCH antibody targets the E2 antigen to the MHC-II present on antigen-presenting cells. The goal of this work was to evaluate passive immunity through colostrum and active immunity of calves immunized with the novel vaccine. 24 A. angus heifers were divided into two groups, 12 immunized with the vaccine and 12 received placebo, in the last trimester of gestation. Serum samples from calves were taken at day 30 of age and analyzed by competitive ELISA. In the vaccinated group, 92% of the d30 old calves maintained medium (25%) or high (67%) antibody levels against BVDV, while 50% of the animals in the control group presented low antibody level (Pearson’s chi-squared p:0,07) (Table 1). At 5 months of age, calves of both groups were immunized with the targeted vaccine. Thirty days later, 96% of the calves had medium or high antibody levels against BVDV, which was independent of their respective heifer vaccine status. These results confirm the new targeted subunit vaccine against BVDV is safe and efficacious to be used in pregnant cattle and can passively immunize newborn calves. They also show that these maternal antibodies do not interfere with the active immunization of calves.


Vaccines ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 234 ◽  
Author(s):  
Lynnette C. Goatley ◽  
Ana Luisa Reis ◽  
Raquel Portugal ◽  
Hannah Goldswain ◽  
Gareth L. Shimmon ◽  
...  

Classical approaches to African swine fever virus (ASFV) vaccine development have not been successful; inactivated virus does not provide protection and use of live attenuated viruses generated by passage in tissue culture had a poor safety profile. Current African swine fever (ASF) vaccine research focuses on the development of modified live viruses by targeted gene deletion or subunit vaccines. The latter approach would be differentiation of vaccinated from infected animals (DIVA)-compliant, but information on which viral proteins to include in a subunit vaccine is lacking. Our previous work used DNA-prime/vaccinia-virus boost to screen 40 ASFV genes for immunogenicity, however this immunization regime did not protect animals after challenge. Here we describe the induction of both antigen and ASFV-specific antibody and cellular immune responses by different viral-vectored pools of antigens selected based on their immunogenicity in pigs. Immunization with one of these pools, comprising eight viral-vectored ASFV genes, protected 100% of pigs from fatal disease after challenge with a normally lethal dose of virulent ASFV. This data provide the basis for the further development of a subunit vaccine against this devastating disease.


2007 ◽  
Vol 51 (9) ◽  
pp. 3131-3135 ◽  
Author(s):  
Junqiang Wu ◽  
Songhua Hu ◽  
Liting Cao

ABSTRACT Bovine subclinical mastitis is an inflammation of the mammary gland caused by bacterial intramammary infection, accounting for large economic losses. Treatment of subclinical mastitis is not suggested for lactating cows due to the risk of milk contamination. The objectives of this study were to evaluate an antimicrobial peptide, nisin, in the treatment of subclinical mastitis in lactating cows. A total of 90 lactating Holstein cows with subclinical mastitis were randomly divided into nisin-treated (n = 46) and control (n = 44) groups. In the nisin-treated group, cows received an intramammary infusion of nisin at a dose of 2,500,000 IU once daily for 3 days while the control cows received no treatment. Milk samples were collected from the affected mammary quarters before treatment and 1 and 2 weeks after treatment for analyses of bacteria, somatic cells, and N-acetyl-β-d-glucosaminidase (NAGase). Results indicated that nisin therapy had bacteriological cure rates of 90.1% for Streptococcus agalactiae (10 of 11), 50% for Staphylococcus aureus (7 of 14), 58.8% for coagulase-negative staphylococci (7 of 17), and 65.2% for all cases (30 of 46). Meanwhile, only 15.9% (7 of 44) of untreated cows spontaneously recovered. NAGase activity in milk samples and the number of mammary quarters with a milk somatic cell count of ≥500,000/ml were significantly decreased after nisin treatment while no significant changes took place in the control group. Because of its therapeutic effects on bovine subclinical mastitis, as well as its safety in humans, nisin deserves further study to clarify its effects on mastitis caused by different pathogens.


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