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1
Jawad, J., R. W. Astuti, A. Haryanto, and N. Wijayanti. "Antibody response to Newcastle disease virus recombinant fusion protein in post-vaccinated laying hens." Journal of the Indonesian Tropical Animal Agriculture 48, no. 1 (December 8, 2022): 20–27. http://dx.doi.org/10.14710/jitaa.48.1.20-27.
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This research was aimed to analyze antibody response in laying hens post vaccinated by Newcastle Disease Virus (NDV) recombinat Fusion (F) protein which has been succesfully expressed from the F gene of local isolates of NDV from Kulon Progo strain (0663/04/2013), Yogyakarta, Indonesia. The F gene cloned into expression vector plasmid pBT7-N-His. Two types of NDV recombinant vaccine, a concentrated and pure F recombinant protein were used for vaccination. A concentrated recombinant F protein was collected from the centrifugal ultrafiltration process and a pure recombinant F protein was collected from the electroeluted process. Recombinant F protein of NDV was successfully expressed, purified, and visualized by Sodium Dodecyl Sulphate–Polyacrylamide Gel Electrophoresis (SDS-PAGE) with Coomassie Brilliant Blue staining with a molecular weight of 28 kDa. All two types of recombinant F vaccines and a commercial live vaccine as a positive control were injected two times at 14 and 18th weeks old laying hens to analyze the antibody response in serum. In comparison with a commercial live NDV vaccine, indirect Enzyme-linked Immunosorbent Assay (ELISA) revealed that antibody responses were high in both recombinant F protein vaccinated groups. In conclusion, the recombinant F protein has the potential to be developed as a recombinant vaccine candidate to obtain a higher antibody response in laying hens compared to commercially available live NDV vaccines.
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van Diepen, Michiel, Rosamund Chapman, Nicola Douglass, Leah Whittle, Nicole Chineka, Shireen Galant, Christian Cotchobos, Akiko Suzuki, and Anna-Lise Williamson. "Advancements in the Growth and Construction of Recombinant Lumpy Skin Disease Virus (LSDV) for Use as a Vaccine Vector." Vaccines 9, no. 10 (October 4, 2021): 1131. http://dx.doi.org/10.3390/vaccines9101131.
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Attenuated vaccine strains of lumpy skin disease virus (LSDV) have become increasingly popular as recombinant vaccine vectors, to target both LSDV, as well as other pathogens, including human infectious agents. Historically, these vaccine strains and recombinants were generated in primary (lamb) testis (LT) cells, Madin–Darby bovine kidney (MDBK) cells or in eggs. Growth in eggs is a laborious process, the use of primary cells has the potential to introduce pathogens and MDBK cells are known to harbor bovine viral diarrhea virus (BVDV). In this study, data is presented to show the growth of an attenuated LSDV strain in baby hamster kidney (BHK-21) cells. Subsequently, a recombinant LSDV vaccine was generated in BHK-21 cells. Partial growth was also observed in rabbit kidney cells (RK13), but only when the vaccinia virus host range gene K1L was expressed. Despite the limited growth, the expression of K1L was enough to serve as a positive selection marker for the generation of recombinant LSDV vaccines in RK13 cells. The simplification of generating (recombinant) LSDV vaccines shown here should increase the interest for this platform for future livestock vaccine development and, with BHK-21 cells approved for current good manufacturing practice, this can be expanded to human vaccines as well.
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Prayugo, Armanda Dwi, Toto Subroto, and Wyanda Arnafia. "Efficacy of Hemagglutinin Gene of Highly Pathogenic Avian Influenza as a Vaccine Candidate in Poultry: A Review." World's Veterinary Journal 13 (March 25, 2023): 26–31. http://dx.doi.org/10.54203/scil.2023.wvj3.
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The most prevalent fatal disease in poultry that can result in high morbidity and mortality is highly pathogenic avian influenza (HPAI), subtype H5N1. A vaccination program is the most frequent way to prevent HPAI cases in poultry, especially against the H5 subtype of HPAI. There are currently a number of avian influenza vaccines available, including recombinant and inactivated whole virus vaccines. The foundation of a recombinant vaccine is possible by the expression of an avian influenza gene of interest following insertion into a carrier vector (no pathogenic virus). A recombinant HPAI vaccine is required to further challenge avian influenza cases in poultry. As a recombinant vaccine inserted into a carrier vector, the hemagglutinin (HA) gene has proven effective. The recombinant Herpes Virus Turkey (rHVT) vector vaccine for avian influenza has been discovered and is commercially available. The rHVT vaccine was developed using a hemagglutinin insert from the HPAI virus clade 2.2. Overall, studies in this review aimed to determine the efficacy of any developed recombinant avian influenza vaccine that uses the HA gene from different clades challenged with any avian influenza virus (AIV) isolate. It was found that the efficacy of hemagglutinin as a recombinant vaccine could be promising for future HPAI vaccine development. In addition, it is possible to design a recombinant vaccine using local isolates to protect poultry farms, particularly in endemic regions.
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Marra, Yasmin, and Fawziah Lalji. "Prevention of Herpes Zoster: A Focus on the Effectiveness and Safety of Herpes Zoster Vaccines." Viruses 14, no. 12 (November 29, 2022): 2667. http://dx.doi.org/10.3390/v14122667.
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Infection with varicella zoster virus typically occurs in children and it can cause primary varicella infection or “chickenpox”, or it can reactivate later in life and cause herpes zoster or “shingles”. Herpes zoster mainly occurs in older adults, causing a reduction in activities of daily living, impacting quality of life, and may lead to serious complications, including chronic pain. Two vaccines are marketed to prevent herpes zoster: the live zoster vaccine and the non-live, recombinant zoster vaccine. The pre-licensure clinical trials show the efficacy of the live zoster vaccine to be between 50 and 70% and for the recombinant vaccine to be higher at 90 to 97%. Real-world effectiveness studies, with a follow-up of approximately 10 years, were reviewed in this article. These data corroborated the efficacy studies, with vaccine effectiveness being 46% and 85% for the live and recombinant vaccines, respectively. Safety data from the effectiveness studies show similar results to the clinical trials with mostly local injection-site reactions and mild systemic reactions seen with both vaccines, although in larger proportions with the recombinant vaccine. Rare adverse events, occurring less than 1% of the time, have been seen with both vaccine types and include disseminated herpes zoster with the live zoster vaccine and Guillain–Barré syndrome with the recombinant vaccine. The wider use of preventative measures with vaccines will reduce the herpes zoster burden of illness seen in older adults.
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Ertl, H. C., and Z. Xiang. "Novel vaccine approaches." Journal of Immunology 156, no. 10 (May 15, 1996): 3579–82. http://dx.doi.org/10.4049/jimmunol.156.10.3579.
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Abstract Recent advances in immunology, molecular biology, and peptide biochemistry have allowed the construction of subunit vaccines based on viral or bacterial recombinants, peptides or plasmid vectors. Although none of these approaches is currently being used for mass vaccination (with the exception or vaccinia-rabies G protein recombinant virus for wildlife immunization); several of them are undergoing clinical trials. None of these different vaccine constructs is likely to be totally effective in either the prevention of infectious diseases or immunotherapy of cancer. Recombinant viral vaccines such as those based on vaccinia or adenovirus as a rule induce potent immune responses. Vaccinia viruses have the added advantage of being heat stable and immunogenic after oral application, making them good candidates for wildlife immunization. Recombinants based on replication-defective adenoviruses are safer compared with vaccinia virus recombinants and, as far as our data indicate, have superior efficacy. In addition, they induce excellent immunity upon application to mucosal membranes, suggesting their usefulness as vaccines for infectious agents that enter through the airways or the genital tract. Peptides are of limited benefit in infectious disease prevention but might provide custom-made vaccines for cancer therapy. Genetic vaccines that were first described less than 5 years ago have already progressed to phase I clinical trials in healthy human adults. Provided that their safety can be confirmed, they might be suited to induce immunity to numerous agents.
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Wu, Xiao-Xin, Hang-Ping Yao, Nan-Ping Wu, Hai-Nv Gao, Hai-Bo Wu, Chang-Zhong Jin, Xiang-Yun Lu, Tian-Shen Xie, and Lan-Juan Li. "Ebolavirus Vaccines: Progress in the Fight Against Ebola Virus Disease." Cellular Physiology and Biochemistry 37, no. 5 (2015): 1641–58. http://dx.doi.org/10.1159/000438531.
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Ebolaviruses are highly infectious pathogens that cause lethal Ebola virus disease (EVD) in humans and non-human primates (NHPs). Due to their high pathogenicity and transmissibility, as well as the potential to be misused as a bioterrorism agent, ebolaviruses would threaten the health of global populations if not controlled. In this review, we describe the origin and structure of ebolaviruses and the development of vaccines from the beginning of the 1980s, including conventional ebolavirus vaccines, DNA vaccines, Ebola virus-like particles (VLPs), vaccinia virus-based vaccines, Venezuelan equine encephalitis virus (VEEV)-like replicon particles, Kunjin virus-based vaccine, recombinant Zaire Ebolavirus∆VP30, recombinant cytomegalovirus (CMV)-based vaccines, recombinant rabies virus (RABV)-based vaccines, recombinant paramyxovirus-based vaccines, adenovirus-based vaccines and vesicular stomatitis virus (VSV)-based vaccines. No licensed vaccine or specific treatment is currently available to counteract ebolavirus infection, although DNA plasmids and several viral vector approaches have been evaluated as promising vaccine platforms. These vaccine candidates have been confirmed to be successful in protecting NHPs against lethal infection. Moreover, these vaccine candidates were successfully advanced to clinical trials. The present review provides an update of the current research on Ebola vaccines, with the aim of providing an overview on current prospects in the fight against EVD.
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Koeberling, Oliver, Isabel Delany, and Dan M. Granoff. "A Critical Threshold of Meningococcal Factor H Binding Protein Expression Is Required for Increased Breadth of Protective Antibodies Elicited by Native Outer Membrane Vesicle Vaccines." Clinical and Vaccine Immunology 18, no. 5 (March 2, 2011): 736–42. http://dx.doi.org/10.1128/cvi.00542-10.
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ABSTRACTNative outer membrane vesicles (NOMV) (not detergent treated), which are prepared from recombinant strains with attenuated endotoxin activity and overexpressed factor H binding protein (fHbp), elicited broad serum bactericidal antibody responses in mice. The amount of overexpressed fHbp required for optimal immunogenicity is not known. In this study we prepared NOMV vaccines from LpxL1 knockout (ΔLpxL1) mutants with penta-acylated lipooligosaccharide and attenuated endotoxin activity. The recombinant strains had wild-type (1×) fHbp expression or were engineered for 3-fold- or 10-fold-increased fHbp expression (3× or 10× fHbp). Control vaccines included NOMV from ΔLpxL1/ΔfHbp mutants or recombinant fHbp. In mice, only the 10× fHbp NOMV vaccine elicited significantly higher serum IgG anti-fHbp antibody titers than the corresponding 1× fHbp NOMV or recombinant fHbp vaccine. The 10× fHbp NOMV vaccine also elicited higher bactericidal responses (P< 0.05) against five group B strains with heterologous PorA than the recombinant fHbp or 1× fHbp NOMV vaccine. The 3× fHbp NOMV vaccine gave higher bactericidal titers against only one strain. Serum bactericidal titers in mice immunized with the control ΔfHbp NOMV vaccines were <1:10, and bactericidal titers in mice immunized with the 10× fHbp NOMV vaccine were <1:10 after adsorption of anti-fHbp antibodies. Mixing antiserum to NOMV vaccines from fHbp knockout mutants with antiserum to recombinant fHbp did not increase anti-fHbp bactericidal titers. Thus, a critical threshold of increased fHbp expression is required for NOMV vaccines to elicit broad serum bactericidal responses, and the antibodies conferring protection are directed primarily at fHbp.
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Hudu, Shuaibu Abdullahi, Saadatu Haruna Shinkafi, and Shuaibu Umar. "AN OVERVIEW OF RECOMBINANT VACCINE TECHNOLOGY, ADJUVANTS AND VACCINE DELIVERY METHODS." International Journal of Pharmacy and Pharmaceutical Sciences 8, no. 11 (October 28, 2016): 19. http://dx.doi.org/10.22159/ijpps.2016v8i11.14311.
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Development of an effective vaccine is of paramount important in disease prevention and control. As such, recombinant technology can serve as a gateway for the development of safe and effective vaccines that can be delivered effectively with an appropriate adjuvant. Therefore, this paper aimed to review the role of recombinant vaccine technology, new adjuvants and the challenge of vaccine delivery. Related peer-reviewed journal article searches were conducted using a subscribed database at the Universiti Putra Malaysia library, involving areas of Health Sciences and Medicine via Medline, SCOPUS and Google Scholar. New generation vaccines include highly purified synthetic or recombinant antigens that stimulate effective cell-mediated immune and mucosal immunity. In order to enhance their efficacy, a number of adjuvants are used. Efforts have also been made to explore the usage of non-invasive routes of administration, devices and equipment for optimized antigen and immune-potentiator delivery of the immune system. Recombinant vaccine technology is rapid, compared to the traditional method of vaccine development and does not require the handling of live viruses. It is, therefore, a promising technology for developing a future vaccine to curb emerging and re-emerging viral infections that may be life-threatening or teratogenic.
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Dewidar, Abdelmonem A. A., Walid H. Kilany, Azza A. El-Sawah, Salama A. S. Shany, Al-Hussien M. Dahshan, Islam Hisham, Magdy F. Elkady, and Ahmed Ali. "Genotype VII.1.1-Based Newcastle Disease Virus Vaccines Afford Better Protection against Field Isolates in Commercial Broiler Chickens." Animals 12, no. 13 (June 30, 2022): 1696. http://dx.doi.org/10.3390/ani12131696.
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This study evaluated the efficacy of live and inactivated conventional GII LaSota and recombinant GVII Newcastle disease vaccines in commercial broilers. The experimental groups (G2–G7) were vaccinated on day 7 and day 21 of age with live vaccines from the same vaccine type “GII LaSota, GVII vaccine (A), GVII vaccine (B)” via eye drop; however, G3, G5, and G7 received a single dose from inactivated counterpart vaccines subcutaneously on day 7 of age. Vaccine efficacy was evaluated based on elicited humoral immunity, clinical protection, and reduction in virus shedding after challenge with virulent GVII 1.1. strain. Results demonstrated that live and inactivated recombinant GVII vaccine based on VG/GA strain backbone elicited superior protection parameters (100% protection). Although the conventional GII LaSota live and inactivated vaccination regime protected 93.3% of vaccinated birds, the virus shedding continued until 10 DPC. The post-vaccination serological monitoring was consistent with protection results. The study concludes that conventional GII ND vaccines alone are probably insufficient due to the current epidemiology of the GVII 1.1 NDV strains. Our findings further support that protection induced by recombinant GVII 1.1. ND vaccines are superior. Interestingly, the efficacy of recombinant ND vaccines seemed to be influenced by the backbone virus since the VG/GA backbone-based vaccine provided better protection and reduced virus shedding.
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Baldo, Aline, Amaya Leunda, Nicolas Willemarck, and Katia Pauwels. "Environmental Risk Assessment of Recombinant Viral Vector Vaccines against SARS-Cov-2." Vaccines 9, no. 5 (May 3, 2021): 453. http://dx.doi.org/10.3390/vaccines9050453.
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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the coronavirus disease 2019 (COVID-19) pandemic. Over the past months, considerable efforts have been put into developing effective and safe drugs and vaccines against SARS-CoV-2. Various platforms are being used for the development of COVID-19 vaccine candidates: recombinant viral vectors, protein-based vaccines, nucleic acid-based vaccines, and inactivated/attenuated virus. Recombinant viral vector vaccine candidates represent a significant part of those vaccine candidates in clinical development, with two already authorised for use in the European Union and one currently under rolling review by the European Medicines Agency (EMA). Since recombinant viral vector vaccine candidates are considered as genetically modified organisms (GMOs), their regulatory oversight includes besides an assessment of their quality, safety and efficacy, also an environmental risk assessment (ERA). The present article highlights the main characteristics of recombinant viral vector vaccine (candidates) against SARS-CoV-2 in the pipeline and discusses their features from an environmental risk point of view.
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Park, Seong Yeon. "What is Different about Recombinant Herpes Zoster Vaccine?" Korean Journal of Medicine 99, no. 4 (August 1, 2024): 180–88. http://dx.doi.org/10.3904/kjm.2024.99.4.180.
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Herpes zoster (HZ) affects about one in three persons in their life time. Compared with the general population, older adults with immune senescence and individuals who are immunocompromised therapy are at increased risk for HZ, and its debilitating complications. To prevent HZ, two HZ vaccines, zoster vaccine live (ZVL) and recombinant zoster vaccine (RZV) are available. RZV is The Korean Society of Infectious Diseases revised guidelines for HZ vaccine in 2023, and recommended to vaccinate with RZV for adults ≥ aged 50 years and for severely immunocompromised adults aged ≥ 18 years. RZV is more effective for prevention of HZ than ZVL. RZV is nonreplicating and is thus safe in immunocompromised patients. RZV has clinically acceptable safety profile. This review will help clinicians update knowledge about RZV and identify eligible subjects who may benefit from HZ vaccinations.
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Kumar, Vivek, Anuj Verma, Riddhi Singh, Priyanshi Garg, Santosh Kumar Sharma, Himanshu Narayan Singh, Santosh Kumar Mishra, and Sanjay Kumar. "Recombinant vaccines: Current updates and future prospects." Asian Pacific Journal of Tropical Medicine 17, no. 8 (August 2024): 338–50. http://dx.doi.org/10.4103/apjtm.apjtm_854_23.
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Recombinant technology-based vaccines have emerged as a highly effective way to prevent a wide range of illnesses. The technology improved vaccine manufacturing, rendering it more efficient and economical. These vaccines have multiple advantages compared to conventional vaccines. The pandemic has heightened awareness of the advantages of these vaccine technologies; trust and acceptance of these vaccines are steadily growing globally. This work offers an overview of the prospects and advantages associated with recombinant vaccines. Additionally, it discusses some of the challenges likely to arise in the future. Their ability to target diverse pathogen classes underscores their contributions to preventing previously untreatable diseases (especially vector-borne and emerging diseases) and hurdles faced throughout the vaccine development process, especially in enhancing the effectiveness of these vaccines. Moreover, their compatibility with emerging vaccination platforms of the future like virus-like particles and CRISPR/Cas9 for the production of next-generation vaccines may offer many prospects. This review also reviewed the hurdles faced throughout the vaccine development process, especially in enhancing the effectiveness of these vaccines against vector-borne diseases, emerging diseases, and untreatable diseases with high mortality rates like AIDS as well as cancer.
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Trujillo, Edgar, Abel Ramos-Vega, Elizabeth Monreal-Escalante, Consuelo Almazán, and Carlos Angulo. "Overview of Recombinant Tick Vaccines and Perspectives on the Use of Plant-Made Vaccines to Control Ticks of Veterinary Importance." Vaccines 12, no. 10 (October 17, 2024): 1178. http://dx.doi.org/10.3390/vaccines12101178.
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Ticks are obligate hematophagous ectoparasites that affect animals, and some of them transmit a wide range of pathogens including viruses, bacteria, and protozoa to both animals and humans. Several vaccines have shown immunogenicity and protective efficacy against ticks in animal models and definitive hosts. After several decades on anti-tick vaccine research, only a commercial vaccine based on a recombinant antigen is currently available. In this context, plants offer three decades of research and development on recombinant vaccine production to immunize hosts and as a delivery vehicle platform. Despite the experimental advances in plant-made vaccines to control several parasitosis and infectious diseases, no vaccine prototype has been developed against ticks. This review examines a panorama of ticks of veterinary importance, recombinant vaccine experimental developments, plant-made vaccine platforms, and perspectives on using this technology as well as the opportunities and limitations in the field of tick vaccine research.
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Muravyova, N. V., and B. S. Belov. "Vaccination against Herpes zoster in patients with immune-mediated inflammatory rheumatic diseases: new data." Modern Rheumatology Journal 18, no. 4 (August 21, 2024): 115–20. http://dx.doi.org/10.14412/1996-7012-2024-4-115-120.
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Patients with immune-mediated inflammatory rheumatic diseases (IIRD) are more likely to develop herpes zoster (HZ) than individuals in the general population. Live attenuated vaccines and inactivated recombinant vaccines with adjuvant are available to prevent the disease and its complications. Live attenuated vaccine can be used in patients with IIRD if certain conditions are met, although these cannot always be fulfilled. The advantage of the inactivated recombinant adjuvant vaccine is that it can be used against a background of anti-rheumatic therapy. The review analyzes foreign studies on the safety, immunogenicity and efficacy of recombinant adjuvant vaccine against HZ in patients with IIRD.
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Yan, Lin, Jin Qiu, Jianbo Chen, Bridgett Ryan-Payseur, Dan Huang, Yunqi Wang, Lijun Rong, Jody A. Melton-Witt, Nancy E. Freitag, and Zheng W. Chen. "Selected prfA* Mutations in Recombinant Attenuated Listeria monocytogenes Strains Augment Expression of Foreign Immunogens and Enhance Vaccine-Elicited Humoral and Cellular Immune Responses." Infection and Immunity 76, no. 8 (May 12, 2008): 3439–50. http://dx.doi.org/10.1128/iai.00245-08.
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ABSTRACT While recombinant Listeria monocytogenes strains can be explored as vaccine candidates, it is important to develop attenuated but highly immunogenic L. monocytogenes vaccine vectors. Here, prfA* mutations selected on the basis of upregulated expression of L. monocytogenes PrfA-dependent genes and proteins were assessed to determine their abilities to augment expression of foreign immunogens in recombinant L. monocytogenes vectors and therefore enhance vaccine-elicited immune responses (a prfA* mutation is a mutation that results in constitutive overexpression of PrfA and PrfA-dependent virulence genes; the asterisk distinguishes the mutation from inactivation or stop mutations). A total of 63 recombinant L. monocytogenes vaccine vectors expressing seven individual viral or bacterial immunogens each in nine different L. monocytogenes strains carrying wild-type prfA or having prfA* mutations were constructed and investigated. Mutations selected on the basis of increased PrfA activation in recombinant L. monocytogenes prfA* vaccine vectors augmented expression of seven individual protein immunogens remarkably. Consistently, prime and boost vaccination studies with mice indicated that the prfA(G155S) mutation in recombinant L. monocytogenes ΔactA prfA* strains enhanced vaccine-elicited cellular immune responses. Surprisingly, the prfA(G155S) mutation was found to enhance vaccine-elicited humoral immune responses as well. The highly immunogenic recombinant L. monocytogenes ΔactA prfA* vaccine strains were as attenuated as the recombinant parent L. monocytogenes ΔactA vaccine vector. Thus, recombinant attenuated L. monocytogenes ΔactA prfA* vaccine vectors potentially are better antimicrobial and anticancer vaccines.
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Stovba, L. F., N. K. Chernikova, A. L. Khmelev, and S. V. Borisevich. "Anti-Vector Immune Response Formed after Immunization with Recombinant Vaccines Based on the Vaccinia Virus, MVA Strain." Problems of Particularly Dangerous Infections, no. 1 (April 7, 2025): 105–11. https://doi.org/10.21055/0370-1069-2025-1-105-111.
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The search for safe approaches to primary immunization of the adult population under the absence of herd immunity to orthopoxviruses, when re-initiation of smallpox vaccination campaign is required, is currently very relevant. Thereat, the clinical trials of recombinant vaccines based on the vaccinia virus, MVA strain, against different illnesses confirm that they are safe for humans and in addition to target efficiency (capacity to induce immunity to proteins expressed by embedded foreign genes), show immunogenicity to vector – vaccinia virus. The aim of the review was to evaluate anti-vector immunity level in people immunized by recombinant viral vaccines, based on vaccinia virus, MVA strain. Explicit experimental data on the level of anti-vector immunity in response to immunization with recombinant vaccines in different countries of the world are presented. Those studies were mainly carried out with recombinants containing embedded immunodominant genes of human immunodeficiency virus (HIV), as the number of works on the creation of recombinant vaccines expressing the antigen determinants of HIV significantly exceeds the number of those on recombinant preparations based on vaccinia virus; the vaccines are successfully used in medical practice and are safe even for people with immunodeficiency conditions. The results obtained indicated an increase in anti-vector immunity with escalation of vaccine dose and peak indicators after two immunizations. Further injections of the vaccine did not lead to increase in the virus neutralizing antibodies, their production gradually decreased over a period of one year or more. In addition to the humoral immune response, cellular anti-vector immunity, represented mainly by CD8+ T-cells, was induced. The insertion of foreign genes did not affect the formation of anti-vector immunity, just as its level did not affect the development of humoral and cellular immune responses to proteins expressed by the embedded genes. Comparative characterization of the anti-vector immunity indices after immunization with recombinant vaccines and specific immunity in response to the IMVAMUNE® vaccine showed that their levels either corresponded to each other, or in the first case the values were even higher.
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Shollenberger, Lisa, Rafaella Grenfell, E. Farah Samli, and Donald Harn. "Vaccine self-assembling immune matrix (VacSIM) is a non-viral delivery platform that augments responses to recombinant protein vaccines (VAC6P.940)." Journal of Immunology 192, no. 1_Supplement (May 1, 2014): 140.1. http://dx.doi.org/10.4049/jimmunol.192.supp.140.1.
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Abstract Vaccination remains the most effective public health tool to prevent infectious disease. However, many vaccines remain marginally effective, especially for immune-compromised populations. To enhance vaccine immunogenicity, we exploited the biphasic property of certain synthetic oligopeptides to create a new vaccine delivery method, VacSIM (vaccine self-assembling immune matrix). Vaccine components are easily mixed with the VacSIM solution for injection, after which the peptides self-assemble into hydrated nanofiber gel-matrices, forming a vaccine depot. Thus, we have a non-viral, inert, biodegradable delivery system, not requiring formulation, which we can use to deliver a multitude of vaccines. Analogous to a treat-dispensing dog toy, VacSIM is a carrier for vaccine components, allowing for gradual release, rather than immediate administration. We believe this depot attracts antigen presenting cells, driving enhanced vaccine-specific responses that improve both the quantity and quality of the response. We have tested this delivery platform with live bacterial vectors, inactivated virus and multiple recombinant protein vaccines. Shown here, VacSIM augments vaccine responses to the recombinant Hepatitis B surface antigen and recombinant HIV Envelope in 2 strains of mice. Enhancement of vaccine responses by VacSIM administration could represent a fundamental paradigm shift in how vaccines are delivered.
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Hongying, Fan, Wu Xianbo, Yu Fang, Bai Yang, and Long Beiguo. "Oral Immunization with Recombinant Lactobacillus acidophilus Expressing the Adhesin Hp0410 of Helicobacter pylori Induces Mucosal and Systemic Immune Responses." Clinical and Vaccine Immunology 21, no. 2 (November 27, 2013): 126–32. http://dx.doi.org/10.1128/cvi.00434-13.
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ABSTRACTHelicobacter pyloriinfection is relatively common worldwide and is closely related to gastric mucosa-associated lymphoid tissue (MALT) lymphoma, chronic gastritis, and stomach ulcers. Therefore, a safe and effective method for preventingH. pyloriinfection is urgently needed. Given that developing an effective vaccine againstH. pyloriis one of the best alternatives,H. pyloriadhesin Hp0410 was expressed in the food-grade bacteriumLactobacillus acidophilus. The recombinant live bacterial vaccine was then used to orally vaccinate mice, and the immunoprotective effects of Hp0410-producing strains were investigated.H. pyloricolonization in the stomach of mice immunized with the recombinantL. acidophiluswas significantly reduced, in comparison with that in control groups. Furthermore, mucosal secretory IgA antibodies were elicited in the mucosal tissue of mice immunized with the recombinant bacteria, and specific anti-Hp0410 IgG responses were also detected in mouse serum. There was a significant increase in the level of protection against gastricHelicobacterinfection following a challenge withH. pyloriSydney strain 1 (SS1). Our results collectively indicate that adhesin Hp0410 is a promising candidate vaccine antigen, and recombinantL. acidophilusexpressing Hp0410 is likely to constitute an effective, low-cost, live bacterial vaccine againstH. pylori.
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Chang, Pengxiang, Faisal Ameen, Joshua E. Sealy, Jean-Remy Sadeyen, Sushant Bhat, Yongqing Li, and Munir Iqbal. "Application of HDR-CRISPR/Cas9 and Erythrocyte Binding for Rapid Generation of Recombinant Turkey Herpesvirus-Vectored Avian Influenza Virus Vaccines." Vaccines 7, no. 4 (November 22, 2019): 192. http://dx.doi.org/10.3390/vaccines7040192.
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Avian influenza viruses (AIVs) are highly contagious and have caused huge economical loss to the poultry industry. AIV vaccines remain one of the most effective methods of controlling this disease. Turkey herpesvirus (HVT) is a commonly used live attenuated vaccine against Marek’s disease; it has also been used as a viral vector for recombinant AIV vaccine development. The clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 system is a gene editing tool which, in vaccinology, has facilitated the development of recombinant DNA viral-vectored vaccines. Here, we utilize homology-directed repair (HDR) for the generation of a HVT–H7N9 HA bivalent vaccine; a H7N9 HA expression cassette was inserted into the intergenic region between UL45 and UL46 of HVT. To optimize the selection efficiency of our bivalent vaccine, we combined CRISPR/Cas9 with erythrocyte binding to rapidly generate recombinant HVT–H7HA candidate vaccines.
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McAllister, Andrés, Alejandra E. Arbetman, Stefanie Mandl, Claudia Peña-Rossi, and Raul Andino. "Recombinant Yellow Fever Viruses Are Effective Therapeutic Vaccines for Treatment of Murine Experimental Solid Tumors and Pulmonary Metastases." Journal of Virology 74, no. 19 (October 1, 2000): 9197–205. http://dx.doi.org/10.1128/jvi.74.19.9197-9205.2000.
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ABSTRACT We have genetically engineered an attenuated yellow fever (YF) virus to carry and express foreign antigenic sequences and evaluated the potential of this type of recombinant virus to serve as a safe and effective tumor vaccine. Live-attenuated YF vaccine is one of the most effective viral vaccines available today. Important advantages include its ability to induce long-lasting immunity, its safety, its affordability, and its documented efficacy. In this study, recombinant live-attenuated (strain 17D) YF viruses were constructed to express a cytotoxic T-lymphocyte epitope derived from chicken ovalbumin (SIINFEKL). These recombinant viruses replicated comparably to the 17D vaccine strain in cell culture and stably expressed the ovalbumin antigen, and infected cells presented the antigen in the context of major histocompatibility complex class I. Inoculation of mice with recombinant YF virus elicited SIINFEKL-specific CD8+lymphocytes and induced protective immunity against challenge with lethal doses of malignant melanoma cells expressing ovalbumin. Furthermore, active immunotherapy with recombinant YF viruses induced regression of established solid tumors and pulmonary metastases. Thus, recombinant YF viruses are attractive viral vaccine vector candidates for the development of therapeutic anticancer vaccines.
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Saito, Takeshi, Rachel A. Reyna, Satoshi Taniguchi, Kirsten Littlefield, Slobodan Paessler, and Junki Maruyama. "Vaccine Candidates against Arenavirus Infections." Vaccines 11, no. 3 (March 13, 2023): 635. http://dx.doi.org/10.3390/vaccines11030635.
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The viral family Arenaviridae contains several members that cause severe, and often lethal, diseases in humans. Several highly pathogenic arenaviruses are classified as Risk Group 4 agents and must be handled in the highest biological containment facility, biosafety level-4 (BSL-4). Vaccines and treatments are very limited for these pathogens. The development of vaccines is crucial for the establishment of countermeasures against highly pathogenic arenavirus infections. While several vaccine candidates have been investigated, there are currently no approved vaccines for arenavirus infection except for Candid#1, a live-attenuated Junin virus vaccine only licensed in Argentina. Current platforms under investigation for use include live-attenuated vaccines, recombinant virus-based vaccines, and recombinant proteins. We summarize here the recent updates of vaccine candidates against arenavirus infections.
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DAČIĆ, M., R. RESANOVIC, Z. RASIC, M. VALCIC, A. MILOVANOVIC, and M. VELHNER. "Efficacy of recombinant VAXXITEK HVT-IBDv vaccine against very virulent Infectious bursal disease virus (vvIBDv) challenge in layer chicks: A pilot study." Journal of the Hellenic Veterinary Medical Society 69, no. 1 (May 2, 2018): 823. http://dx.doi.org/10.12681/jhvms.16434.
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The infectious bursal disease virus (IBDv) is widespread in poultry flocks all around the world. Various biotypes have emerged and because of that, adequate management practices and vaccination of chicks are of paramount importance for the protection against field strains. One day old Lohmann Brown chicks were vaccinated with intermediate vaccines and the recombinant VAXXITEK HVT-IBDv vaccine formulation, and challenged at 48 days of life with the very virulent IBDv (vvIBDv) strain CH/99. The best protection (100%) was achieved with the recombinant vaccine administered by the subcutaneous or intramuscular route at a day old, while intermediate and intermediate plus vaccines protected 80% of birds from clinical symptoms. The highest bursa body ratio (5.33, 3.50 and 4.12) was accomplished in non- vaccinated and non-challenged birds and birds vaccinated with the VAXXITEK HVT-IBDv vaccine. The recombinant VAXXITEK HVT-IBDv vaccine has provided protection for commercial chicks against challenge with the vvIBDv strain in this experiment. Under field conditions, additional vaccination is possibly needed with supplementary application of live attenuated vaccines. However, the recombinant vector vaccines are providing significant aid against clinical signs and immunosupression caused by the vvIBDv.
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Pearl, Karlyn K., Ana A. Ortiz, and William Pearl. "Efficacy of Immunization with a Combination of Serum and Recombinant Hepatitis B Vaccines." Infection Control & Hospital Epidemiology 14, no. 8 (August 1993): 476–78. http://dx.doi.org/10.1086/646783.
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AbstractObjective:To evaluate the efficacy of giving a third dose of recombinant hepatitis B vaccine to healthcare workers who already had received two doses of serum-derived vaccine, which is no longer available in the United States.Design:Volunteers who already had received two standard doses of serum-derived vaccine were given a third dose of either serum or recombinant vaccine in a double-blind fashion. Antibodies to hepatitis B surface antigen were measured at the time of the third immunization, three months later, and one year after the third immunization.Setting:U.S. Army Medical Center, El Paso, Texas.Patients:One hundred healthy healthcare workers.Results:Three months after receiving the third immunization, the serum vaccine group had significantly higher titers than the recombinant vaccine group (P= 0.018). One year after receiving the third immunization, those who received the combined regimen had a mean hepatitis B surface antibody titer less than half that of those who received three doses of serum-derived vaccine. However, both regimens resulted in titers that are considered to confer immunity.Conclusions:A regimen that combines serum and recombinant hepatitis B vaccines may not produce as high an antibody level as three doses of the same vaccine. Those who began immunization with serum vaccine and concluded with recombinant vaccine should be monitored for an accelerated drop in serum antibodies.
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Levin, Myron J., Neda Al Rawashdh, Liliane Mofor, Pablo Anaya, Richard M. Zur, Emily B. Kahn, Daniel Yu, and Joaquin F. Mould-Quevedo. "A Clinical and Economic Comparison of Cell-Based Versus Recombinant Influenza Vaccines in Adults 18–64 Years in the United States." Vaccines 12, no. 11 (October 26, 2024): 1217. http://dx.doi.org/10.3390/vaccines12111217.
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Background: This analysis compares the cost-effectiveness of a cell-based influenza vaccine to a recombinant influenza vaccine, and each to no vaccination. The analysis is based on United States (US) commercial and societal perspectives. Methods: A Susceptible–Exposed–Infectious–Recovered (SEIR) transmission model of the total US population followed with a cost-effectiveness model for 18–64-year-olds was used to estimate the clinical and economic impact of vaccination over one influenza season (2018–2019). Deterministic and probabilistic sensitivity analyses were conducted. Results: Both enhanced vaccines prevented a substantial number of influenza cases and influenza-related deaths compared to no vaccination. The cell-based vaccine was associated with higher quality-adjusted life years (QALYs) gained compared to the recombinant vaccine or no vaccination. The cell-based vaccine had a 36% lower vaccination cost, amounting to $2.8 billion in cost savings, compared to the recombinant vaccine. The incremental cost-effectiveness ratios (ICERs) for the cell-based vaccine, compared to the recombinant vaccine or no vaccination, were dominant from all payer perspectives, regardless of risk groups. Conclusions: Overall, the cell-based vaccine was cost-saving compared to the recombinant vaccine for subjects aged 18–64 years in the US, achieving comparable health outcomes with a significant reduction in associated costs.
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Weaver, Eric A. "Dose Effects of Recombinant Adenovirus Immunization in Rodents." Vaccines 7, no. 4 (October 10, 2019): 144. http://dx.doi.org/10.3390/vaccines7040144.
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Recombinant adenovirus type 5 (rAd) has been used as a vaccine platform against many infectious diseases and has been shown to be an effective vaccine vector. The dose of the vaccine varies significantly from study to study, making it very difficult to compare immune responses and vaccine efficacy. This study determined the immune correlates induced by serial dilutions of rAd vaccines delivered intramuscularly (IM) and intranasally (IN) to mice and rats. When immunized IM, mice had substantially higher antibody responses at the higher vaccine doses, whereas, the IN immunized mice showed a lower response to the higher rAd vaccine doses. Rats did not show dose-dependent antibody responses to increasing vaccine doses. The IM immunized mice and rats also showed significant dose-dependent T cell responses to the rAd vaccine. However, the T cell immunity plateaued in both mice and rats at 109 and 1010 vp/animal, respectively. Additionally, the highest dose of vaccine in mice and rats did not improve the T cell responses. A final vaccine analysis using a lethal influenza virus challenge showed that despite the differences in the immune responses observed in the mice, the mice had very similar patterns of protection. This indicates that rAd vaccines induced dose-dependent immune responses, especially in IM immunized animals, and that immune correlates are not as predictive of protection as initially thought.
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Berg, Michael G., Robert J. Adams, Ratish Gambhira, Mark C. Siracusa, Alan L. Scott, Richard B. S. Roden, and Gary Ketner. "Immune Responses in Macaques to a Prototype Recombinant Adenovirus Live Oral Human Papillomavirus 16 Vaccine." Clinical and Vaccine Immunology 21, no. 9 (July 2, 2014): 1224–31. http://dx.doi.org/10.1128/cvi.00197-14.
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ABSTRACTImmunization with human papillomavirus (HPV) L1 virus-like particles (VLPs) prevents infection with HPV. However, the expense and logistical demands of current VLP vaccines will limit their widespread use in resource-limited settings, where most HPV-induced cervical cancer occurs. Live oral adenovirus vaccines have properties that are well-suited for use in such settings. We have described a live recombinant adenovirus vaccine prototype that produces abundant HPV16 L1 protein from the adenovirus major late transcriptional unit and directs the assembly of HPV16 VLPs in tissue culture. Recombinant-derived VLPs potently elicit neutralizing antibodies in mice. Here, we characterize the immune response to the recombinant after dual oral and intranasal immunization of pigtail macaques, in which the virus replicates as it would in immunized humans. The immunization of macaques induced vigorous humoral responses to adenovirus capsid and nonstructural proteins, although, surprisingly, not against HPV L1. In contrast, immunization elicited strong T-cell responses to HPV VLPs as well as adenovirus virions. T-cell responses arose immediately after the primary immunization and were boosted by a second immunization with recombinant virus. T-cell immunity contributes to protection against a wide variety of pathogens, including many viruses. The induction of a strong cellular response by the recombinant indicates that live adenovirus recombinants have potential as vaccines for those agents. These studies encourage and will inform the continued development of viable recombinant adenovirus vaccines.
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Cuccui, Jon, Rebecca M. Thomas, Madeleine G. Moule, Riccardo V. D'Elia, Thomas R. Laws, Dominic C. Mills, Diane Williamson, Timothy P. Atkins, Joann L. Prior, and Brendan W. Wren. "Exploitation of bacterial N -linked glycosylation to develop a novel recombinant glycoconjugate vaccine against Francisella tularensis." Open Biology 3, no. 5 (May 2013): 130002. http://dx.doi.org/10.1098/rsob.130002.
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Glycoconjugate-based vaccines have proved to be effective at producing long-lasting protection against numerous pathogens. Here, we describe the application of bacterial protein glycan coupling technology (PGCT) to generate a novel recombinant glycoconjugate vaccine . We demonstrate the conjugation of the Francisella tularensis O-antigen to the Pseudomonas aeruginosa carrier protein exotoxin A using the Campylobacter jejuni PglB oligosaccharyltransferase . The resultant recombinant F. tularensis glycoconjugate vaccine is expressed in Escherichia coli where yields of 3 mg l −1 of culture were routinely produced in a single-step purification process. Vaccination of BALB/c mice with the purified glycoconjugate boosted IgG levels and significantly increased the time to death upon subsequent challenge with F. tularensis subsp. holarctica . PGCT allows different polysaccharide and protein combinations to be produced recombinantly and could be easily applicable for the production of diverse glycoconjugate vaccines.
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Shahriari, Amir Ghaffar, and Maziar Habibi-Pirkoohi. "Plant-Based Recombinant Vaccine: Fact or Fiction?" Galen Medical Journal 6, no. 4 (December 29, 2017): 268–80. http://dx.doi.org/10.31661/gmj.v6i4.792.
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In the era of recombinant DNA technology, production of recombinant vaccines in green plants has emerged as an effective approach addressing the problems of traditional vaccine production. Various antigens expressed in different plant species have been so far tested for the production of efficient oral vaccines against human and livestock diseases. However, recombinant vaccines have not yet found a prominent place in pharmaceutical market. There are still many challenges to be addressed to pave the road for commercial production of plant-based recombinant vaccines. Regarding increasing growth in laboratory studies and field trials for development of plant-based vaccines, this review paper provides a comprehensive overview on the topic of plant-derived vaccines and related issues. [GMJ.2017;6(4):268-80] DOI:10.22086/gmj.v6i3.792
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Sarmadi, Mahdieh, Azam Gheibi, Hossein Khanahmad, Mohammad Reza Khorramizadeh, Seyed Hossein Hejazi, Noushin Zahedi, Hamidreza Mianesaz, and Khosrow Kashfi. "Design and Characterization of a Recombinant Brucella abortus RB51 Vaccine That Elicits Enhanced T Cell-Mediated Immune Response." Vaccines 10, no. 3 (March 3, 2022): 388. http://dx.doi.org/10.3390/vaccines10030388.
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Brucella abortus vaccines help control bovine brucellosis. The RB51 strain is a live attenuated vaccine with low side effects compared with other live attenuated brucellosis vaccines, but it provides insufficient protective efficacy. Cell-mediated immune responses are critical in resistance against intracellular bacterial infections. Therefore, we hypothesized that the listeriolysin O (LLO) expression of Listeria monocytogenes, BAX, and SMAC apoptotic proteins in strain RB51 could enhance vaccine efficacy and safety. B. abortus RB51 was transformed separately with two broad-host-range plasmids (pbbr1ori-LLO and pBlu–mLLO-BAX-SMAC) constructed from our recent work. pbbr1ori-LLO contains LLO, and pBlu–mLLO-BAX-SMAC contains the mutant LLO and BAX-SMAC fusion gene. The murine macrophage-like cell line J774A.1 was infected with the RB51 recombinant strain containing pBlu-mLLO-BAX-SMAC, RB51 recombinant strain containing LLO, and RB51 strain. The bacterial cytotoxicity and survival and apoptosis of host cells contaminated with our two strain types—RB51 recombinants or the parental RB51—were assessed. Strain RB51 expressing mLLO and BAX-SMAC was tested in BALB/c mice and a cell line for enhanced modulation of IFN-γ production. LDH analysis showed that the RB51-mLLO-BAX-SMAC and RB51-LLO strains expressed higher cytotoxicity in J774A.1 cells than RB51. In addition, RB51 recombinants had lower macrophage survival rates and caused higher levels of apoptosis and necrosis. Mice vaccinated with the RB51 recombinant containing mLLO-BAX-SMAC showed an enhanced Th1 immune response. This enhanced immune response is primarily due to bacterial endosome escape and bacterial antigens, leading to improved apoptosis and cross-priming. This potentially enhanced TCD8+- and T cell-mediated immunity leads to the increased safety and potency of the RB51 recombinant (RB51 mLLO-BAX-SMAC) as a vaccine candidate against B. abortus.
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Schlom, Jeffrey. "Recombinant cancer vaccines and new vaccine targets." Expert Review of Vaccines 12, no. 10 (October 2013): 1121–24. http://dx.doi.org/10.1586/14760584.2013.836913.
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Stover, C. Kendall. "Recombinant vaccine delivery systems and encoded vaccines." Current Opinion in Immunology 6, no. 4 (August 1994): 568–71. http://dx.doi.org/10.1016/0952-7915(94)90143-0.
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Tang, Na, Yaoyao Zhang, Yashar Sadigh, Katy Moffat, Zhiqiang Shen, Venugopal Nair, and Yongxiu Yao. "Generation of A Triple Insert Live Avian Herpesvirus Vectored Vaccine Using CRISPR/Cas9-Based Gene Editing." Vaccines 8, no. 1 (February 21, 2020): 97. http://dx.doi.org/10.3390/vaccines8010097.
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Herpesvirus of turkeys (HVT), used originally as a vaccine against Marek’s disease (MD), has recently been shown to be a highly effective viral vector for generation of recombinant vaccines that deliver protective antigens of other avian pathogens. Until the recent launch of commercial HVT-vectored dual insert vaccines, most of the HVT-vectored vaccines in the market carry a single foreign gene and are usually developed with slow and less efficient conventional recombination methods. There is immense value in developing multivalent HVT-vectored vaccines capable of inducing simultaneous protection against multiple avian pathogens, particularly to overcome the interference between individual recombinant HVT vaccines. Here we demonstrate the use of a previously developed CRISPR/Cas9 gene editing protocol for the insertion of ILTV gD-gI and the H9N2 AIV hemagglutinin expression cassettes into the distinct locations of the recombinant HVT-IBDV VP2 viral genome, to generate the triple insert HVT-VP2-gDgI-HA recombinant vaccine. The insertion, protein expression, and stability of each insert were then evaluated by PCR, immunostaining and Western blot analyses. The successful generation of the first triple insert recombinant HVT vaccine with the potential for the simultaneous protection against three major avian viral diseases in addition to MD is a major innovation in vaccination-based control of major poultry diseases.
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Barouch, Dan H., Paul F. McKay, Shawn M. Sumida, Sampa Santra, Shawn S. Jackson, Darci A. Gorgone, Michelle A. Lifton, et al. "Plasmid Chemokines and Colony-Stimulating Factors Enhance the Immunogenicity of DNA Priming-Viral Vector Boosting Human Immunodeficiency Virus Type 1 Vaccines." Journal of Virology 77, no. 16 (August 15, 2003): 8729–35. http://dx.doi.org/10.1128/jvi.77.16.8729-8735.2003.
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ABSTRACT Heterologous “prime-boost” regimens that involve priming with plasmid DNA vaccines and boosting with recombinant viral vectors have been shown to elicit potent virus-specific cytotoxic T-lymphocyte responses. Increasing evidence, however, suggests that the utility of recombinant viral vectors in human populations will be significantly limited by preexisting antivector immunity. Here we demonstrate that the coadministration of plasmid chemokines and colony-stimulating factors with plasmid DNA vaccines markedly increases the immunogenicity of DNA prime-recombinant adenovirus serotype 5 (rAd5) boost and DNA prime-recombinant vaccinia virus (rVac) boost vaccine regimens in BALB/c mice. In mice with preexisting anti-Ad5 immunity, priming with the DNA vaccine alone followed by rAd5 boosting elicited only marginal immune responses. In contrast, cytokine-augmented DNA vaccine priming followed by rAd5 vector boosting was able to generate potent immune responses in mice with preexisting anti-Ad5 immunity. These data demonstrate that plasmid cytokines can markedly improve the immunogenicity of DNA prime-viral vector boost vaccine strategies and can partially compensate for antivector immunity.
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Kumar, Pawan, Tamer A. Sharafeldin, Rahul Kumar, Qinfeng Huang, Yuying Liang, Sagar M. Goyal, Robert E. Porter, Hinh Ly, and Sunil K. Mor. "Development of a Recombinant Pichinde Virus-Vectored Vaccine against Turkey Arthritis Reovirus and Its Immunological Response Characterization in Vaccinated Animals." Pathogens 10, no. 2 (February 13, 2021): 197. http://dx.doi.org/10.3390/pathogens10020197.
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Vaccination may be an effective way to reduce turkey arthritis reovirus (TARV)-induced lameness in turkey flocks. However, there are currently no commercial vaccines available against TARV infection. Here, we describe the use of reverse genetics technology to generate a recombinant Pichinde virus (PICV) that expresses the Sigma C and/or Sigma B proteins of TARV as antigens. Nine recombinant PICV-based TARV vaccines were developed carrying the wild-type S1 (Sigma C) and/or S3 (Sigma B) genes from three different TARV strains. In addition, three recombinant PICV-based TARV vaccines were produced carrying codon-optimized S1 and/or S3 genes of a TARV strain. The S1 and S3 genes and antigens were found to be expressed in virus-infected cells via reverse transcriptase polymerase chain reaction (RT-PCR) and the direct fluorescent antibody (DFA) technique, respectively. Turkey poults inoculated with the recombinant PICV-based TARV vaccine expressing the bivalent TARV S1 and S3 antigens developed high anti-TARV antibody titers, indicating the immunogenicity (and safety) of this vaccine. Future in vivo challenge studies using a turkey reovirus infection model will determine the optimum dose and protective efficacy of this recombinant virus-vectored candidate vaccine.
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Pastoret, P. P., D. Boulanger, and B. Brochier. "Field trials of a recombinant rabies vaccine." Parasitology 110, S1 (March 1995): S37—S42. http://dx.doi.org/10.1017/s0031182000001475.
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SummaryTo improve both safety and stability of the vaccines used in the field to vaccinate foxes against rabies by the oral route, a recombinant vaccinia virus, expressing the glycoprotein of rabies virus (VVTGgRAB) has been developed. VVTGgRAB innocuity was verified in target species and in domestic animals as well as in numerous wild animal species that could compete with the red fox in consuming vaccine baits in Europe. Oral immunization of foxes, by distributing VVTGgRAB vaccine-baits, was undertaken for the whole infected area in Belgium (10000 km2). Five campaigns of fox vaccination, were carried out from autumn 1989 until 1991. Each time, 150000 vaccine-baits were dropped by air at a mean density of 15 per km2. These campaigns induced a drastic decrease in the incidence of rabies and the elimination of the disease from 80% of the initially infected area. Regarding the geographical evolution of rabies in Belgium and in adjacent regions in neighbouring countries, new spatial strategies for bait dispersal were planned for 1992, 1993 and 1994: successive confined campaigns were carried out along political borders only. These campaigns induced a new decrease of incidence; no rabid fox could be detected in 1993 in spite of an improved epidemiological surveillance. In 1994, rabies was again confirmed in 13 foxes collected in an area close to the French border. These cases demonstrate the persistence of a border rabies focus and justify further restricted vaccination campaigns.
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Temchura, Vladimir, Jannik T. Wagner, and Dominik Damm. "Immunogenicity of Recombinant Lipid-Based Nanoparticle Vaccines: Danger Signal vs. Helping Hand." Pharmaceutics 16, no. 1 (December 23, 2023): 24. http://dx.doi.org/10.3390/pharmaceutics16010024.
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Infectious diseases are a predominant problem in human health. While the incidence of many pathogenic infections is controlled by vaccines, some pathogens still pose a challenging task for vaccine researchers. In order to face these challenges, the field of vaccine development has changed tremendously over the last few years. For non-replicating recombinant antigens, novel vaccine delivery systems that attempt to increase the immunogenicity by mimicking structural properties of pathogens are already approved for clinical applications. Lipid-based nanoparticles (LbNPs) of different natures are vesicles made of lipid layers with aqueous cavities, which may carry antigens and other biomolecules either displayed on the surface or encapsulated in the cavity. However, the efficacy profile of recombinant LbNP vaccines is not as high as that of live-attenuated ones. This review gives a compendious picture of two approaches that affect the immunogenicity of recombinant LbNP vaccines: (i) the incorporation of immunostimulatory agents and (ii) the utilization of pre-existing or promiscuous cellular immunity, which might be beneficial for the development of tailored prophylactic and therapeutic LbNP vaccine candidates.
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Rizqoh, Debie. "Genetic Engineering Technique in Virus-Like Particle Vaccine Construction." Jurnal Kesehatan Masyarakat Indonesia 16, no. 4 (December 31, 2021): 203. http://dx.doi.org/10.26714/jkmi.16.4.2021.203-211.
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Vaccine becomes a very effective strategy to deal with various infectious diseases even to the point of eradication as in the smalpox virus. At present many conventional vaccines such as inactivated and live-attenuated vaccines. However, these vaccine methods have side effects on the population. Viral-like particle (VLP) is an alternative vaccine based on recombinant DNA technology that is safe with the same immunogenicity as conventional viruses. This vaccine has been shown to induce humoral immune responses mediated by antibodies and cellular immune responses mediated by cytotoxic T cells. With these advantages, currently various types of vaccines have only been developed on a VLP basis. VLP can be produced from a variety of recombinant gene expression systems including bacterial cell expression systems, yeast cells, insect cells, mammalian cells, plant cells, and cell-free systems.
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Willadsen, P., P. Bird, G. S. Cobon, and J. Hungerford. "Commercialisation of a recombinant vaccine againstBoophilus microplus." Parasitology 110, S1 (March 1995): S43—S50. http://dx.doi.org/10.1017/s0031182000001487.
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SummaryIncreasingly, there is need for methods to control cattle tick (Boophilus microplus) infestations by the use of non-chemical technology. This need is brought about by a mixture of market forces and the failure or inadequacy of existing technology. A recombinant vaccine has now been developed against the tick. This vaccine relies on the uptake with the blood meal of antibody directed against a critical protein in the tick gut. The isolation of the vaccine antigen, Bm86, and its production as a recombinant protein is briefly described. The vaccine has been tested in the field, has been taken through the full registration process and is now in commercial use in Australia. A related development has occurred in Cuba. The potential for improvement of the current vaccine and for the development of similar vaccines against other haematophagous parasites is discussed.
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Ourmanov, Ilnour, Charles R. Brown, Bernard Moss, Miles Carroll, Linda Wyatt, Liuobov Pletneva, Simoy Goldstein, David Venzon, and Vanessa M. Hirsch. "Comparative Efficacy of Recombinant Modified Vaccinia Virus Ankara Expressing Simian Immunodeficiency Virus (SIV) Gag-Pol and/or Env in Macaques Challenged with Pathogenic SIV." Journal of Virology 74, no. 6 (March 15, 2000): 2740–51. http://dx.doi.org/10.1128/jvi.74.6.2740-2751.2000.
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ABSTRACT Prior studies demonstrated that immunization of macaques with simian immunodeficiency virus (SIV) Gag-Pol and Env recombinants of the attenuated poxvirus modified vaccinia virus Ankara (MVA) provided protection from high levels of viremia and AIDS following challenge with a pathogenic strain of SIV (V. M. Hirsch et al., J. Virol. 70:3741–3752, 1996). This MVA-SIV recombinant expressed relatively low levels of the Gag-Pol portion of the vaccine. To optimize protection, second-generation recombinant MVAs that expressed high levels of either Gag-Pol (MVA-gag-pol) or Env (MVA-env), alone or in combination (MVA-gag-pol-env), were generated. A cohort of 24 macaques was immunized with recombinant or nonrecombinant MVA (four groups of six animals) and was challenged with 50 times the dose at which 50% of macaques are infected with uncloned pathogenic SIVsmE660. Although all animals became infected postchallenge, plasma viremia was significantly reduced in animals that received the MVA-SIV recombinant vaccines as compared with animals that received nonrecombinant MVA (P = 0.0011 by repeated-measures analysis of variance). The differences in the degree of virus suppression achieved by the three MVA-SIV vaccines were not significant. Most importantly, the reduction in levels of viremia resulted in a significant increase in median (P < 0.05 by Student's t test) and cumulative (P = 0.010 by log rank test) survival. These results suggest that recombinant MVA has considerable potential as a vaccine vector for human AIDS.
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Tseng, Ta-Yuan, Yee-Chen Liu, Yu-Chen Hsu, Poa-Chun Chang, Ming-Kun Hsieh, Jui-Hung Shien, and Shan-Chia Ou. "Preparation of Chicken Anemia Virus (CAV) Virus-Like Particles and Chicken Interleukin-12 for Vaccine Development Using a Baculovirus Expression System." Pathogens 8, no. 4 (November 23, 2019): 262. http://dx.doi.org/10.3390/pathogens8040262.
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Chicken infectious anemia (CIA) is a poultry disease that causes huge economic losses in the poultry industry worldwide. Commercially available CIA vaccines are derived from wild-type chicken anemia viruses (CAVs) by serial passage in cells or chicken embryos. However, these vaccinal viruses are not completely attenuated; therefore, they can be transmitted vertically and horizontally, and may induce clinical symptoms in young birds. In this study, we sought to eliminate these issues by developing a subunit vaccine exploiting the CAV structural proteins, engineering recombinant baculovirus-infected Spodoptera frugiperda (Sf9) cells that contained both the viral protein 1 (VP1) and VP2 of CAV. Moreover, we produced single-chain chicken interleukin-12 (chIL-12) in the same system, to serve as an adjuvant. The recombinant VP1 was recognized by chicken anti-CAV polyclonal antibodies in Western blotting and immunofluorescence assays, and the bioactivity of the recombinant chIL-12 was confirmed by stimulating interferon-γ (IFN-γ) secretion in chicken splenocytes. Furthermore, the ability of the recombinant VP1 to generate self-assembling virus-like particles (VLPs) was confirmed by transmission electron microscopy. Specific pathogen-free (SPF) chickens inoculated with VLPs and co-administered the recombinant chIL-12 induced high CAV-specific antibodies and cell-mediated immunity. Taken together, the VLPs produced by the baculovirus expression system have the potential to be a safe and effective CIA vaccine. Finally, we demonstrated the utility of recombinant chIL-12 as an adjuvant for poultry vaccine development.
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Ma, Haoyuan, Haowen Xue, Jingfeng Fu, Yanhao Song, Kunru Zhu, and Xu Gao. "Recombination of Goose Parvovirus VP3 Gene and Goose Interferon Ɣ Gene from Fowlpox Virus Immune Protection and Its Mechanism." BIO Web of Conferences 60 (2023): 01010. http://dx.doi.org/10.1051/bioconf/20236001010.
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Gosling plague (GP) is caused by goose parvovirus (GPV) gosling acute, subacute, and septic infectious diseases, commonly known as gosling plague. At present, the prevention and control of GP at home and abroad mainly adopts the method of immunizing female geese with attenuated vaccine. Goslings can be immunized, but there are still the phenomenon of dispersive virus and virulence reverse. It, therefore, is urgent to develop a new safe and effective vaccine. The recombinant avian pox virus vaccines rFPV-GoIFNγ, rFPV-GPV-VP3 and rFPV-GoIFNγ-VP3 were used to monitor their antibody levels and evaluate the protective mechanism of the vaccine in this study. This study lays a foundation for further in vivo testing of recombinant avian pox virus vaccine for goose parvovirus and provides a reference for the development of recombinant avian pox virus vaccine for other avian infectious diseases.
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Rezende, A. F. S., A. A. Brum, F. S. B. Bezerra, D. C. Braite, G. L. Sá, H. S. Thurow, F. K. Seixas, V. A. C. Azevedo, R. W. Portela, and S. Borsuk. "Assessment of the acid phosphatase CP01850 from Corynebacterium pseudotuberculosis in DNA and subunit vaccine formulations against caseous lymphadenitis." Arquivo Brasileiro de Medicina Veterinária e Zootecnia 72, no. 1 (January 2020): 199–207. http://dx.doi.org/10.1590/1678-4162-10790.
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ABSTRACT The target cp1002_RS01850 from Corynebacterium pseudotuberculosis was used to construct a DNA and recombinant subunit vaccine against caseous lymphadenitis. Recombinant protein rCP01850 was expressed in Escherichia coli using pAE vector, and DNA vaccine was engineered with pTARGET vector. BALB/c mice were divided in five groups containing eight animals each, inoculated with: pTARGET/cp01850 as DNA vaccine (G1); rCP01850 plus Al (OH)3 as recombinant subunit vaccine (G2); pTARGET/cp01850 and a boost with rCP01850 plus Al (OH)3 (G3); pTARGET (G4); or Al (OH)3 (G5). Mice were inoculated and blood samples were collected on days 0, 21, and 42 for the analysis of total IgG, IgG1 and IgG2a by ELISA. In each group, five animals were challenged with Mic-6 C. pseudotuberculosis strain, and three were used for cytokine quantification by qPCR. Although no group has been protected by vaccines against lethal challenge, G2 showed an increase in the survival rate after challenge. Significantly higher levels of IL-4, IL-12, IFN-γ, total IgG, IgG1 and IgG2a were also detected for G2, evidencing a mixed Th1/Th2 immunological profile. In conclusion, despite no protection level provided by different vaccinal strategies using cp1002_RS01850 from C. pseudotuberculosis, G2 developed a Th1/Th2 immune response with an increase in survival rate.
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Yousefi, A., Fatemeh Fotouhi, S. Hosseinzadeh, M. T. Kheiri, B. Farahmand, S. Montazeri, and F. Mousavi. "Expression of Antigenic Determinants of the Haemagglutinin Large Subunit of Novel Influenza Virus in Insect Cells." Folia Biologica 58, no. 4 (2012): 151–56. http://dx.doi.org/10.14712/fb2012058040151.
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The global outbreak of novel A/H1N1 spread in human population worldwide has revealed an emergency need for producing a vaccine against this virus. Current influenza vaccines encounter problems with safety issues and weak response in high-risk population. It has been established that haemagglutinin is the most important viral antigen to which antibody responses are directed, and recombinant subunit vaccines, haemagglutinin of influenza A and B viruses, have been considered in order to facilitate vaccine production. In the present study, we have focused on construction of a recombinant baculovirus encoding the large subunit of novel influenza virus A/H1N1 haemagglutinin. The full genome of haemagglutinin was cloned into pGEM-TEasy vector and sequenced. The large subunit of the haemagglutinin gene was amplified by PCR using specific primers and cloned into pFastBacHTc donor plasmid, which was then confirmed by restriction enzyme analysis and sequencing and transformed into E. coli DH10Bac competent cells. The bacmid DNA was transfected into insect cells to produce recombinant baculovirus. Expression of recombinant haemagglutinin in insect cells was determined by SDS-PAGE and immunoblotting. It has been shown that the recombinant haemagglutinin (rHA) obtained from the baculovirus insect cell expression system has suitable immunogenicity in human and can be considered as a candidate flu vaccine. Here we produced large amounts of the HA1 protein of novel influenza A/H1N1 (Iranian isolate) in insect cells. The immunogenicity and efficacy of the recombinant HA1 will be evaluated as a vaccine candidate and compared to the recombinant HA1 produced in a prokaryotic system.
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Zhang, Haiyan, Arun Sridhar, Natacha Delrez, Bo He, Sophie Fourny, Yuan Gao, Owen Donohoe, and Alain F. C. Vanderplasschen. "Development Using Bioluminescence Imaging of a Recombinant Anguillid Herpesvirus 1 Vaccine Candidate Associated with Normal Replication In Vitro but Abortive Infection In Vivo." Vaccines 12, no. 12 (December 17, 2024): 1423. https://doi.org/10.3390/vaccines12121423.
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Background/Objectives: Anguillid herpesvirus 1 (AngHV-1) (recently renamed Cyvirus anguillidallo 1) is the etiologic agent of a lethal disease that affects several eel species. It is thought to be one of the main infectious agents causing a population decline in wild eels and economic loss within the eel aquaculture sector. To date, no vaccines are available against AngHV-1. Recently, we developed a safe and efficacious live attenuated recombinant vaccine against Cyprinid herpesvirus 3 (CyHV-3). This CyHV-3 recombinant vaccine encodes a deletion of ORF57. Orthologues of CyHV-3 ORF57 exist in Cyprinid herpesvirus 2 (CyHV-2, ORF57) and AngHV-1 (ORF35). Methods: In the present study, using recombinant strains and bioluminescent in vivo imaging, we investigated the effect of AngHV-1 ORF35 deletion on virus replication in vitro, virulence in vivo, and the potential of an AngHV-1 ORF35-deleted recombinant as a vaccine candidate for the mass vaccination of eels by immersion. With this goal in mind, we produced ORF35-deleted recombinants using two parental strains: a UK strain and a recombinant derived from the former strain by insertion of a Luciferase–GFP reporter cassette into a non-coding intergenic region. Results: Analyses of ORF35-deleted recombinants led to the following observations: (i) AngHV-1 ORF35 is not essential for viral growth in cell culture, and its deletion does not affect the production of extracellular virions despite reducing the size of viral plaque. (ii) In contrast to what has been observed for CyHV-3 ORF57 and CyHV-2 ORF57, in vivo bioluminescent analyses revealed that AngHV-1 ORF35 is an essential virulence factor and that its deletion led to abortive infection in vivo. (iii) Inoculation of the AngHV-1 ORF35-deleted recombinant by immersion induced a protective immune response against a wild-type challenge. This protection was shown to be dose-dependent and to rely on the infectivity of AngHV-1 ORF35-deleted virions. Conclusions: This study suggests that the AngHV-1 ORF35 protein has singular properties compared to its orthologues encoded by CyHV-2 and CyHV-3. It also supports the potential of AngHV-1 ORF35-deleted recombinants for the mass vaccination of eels by immersion.
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Shehata, Mahmoud M., Ahmed Kandeil, Ahmed Mostafa, Sara H. Mahmoud, Mokhtar R. Gomaa, Rabeh El-Shesheny, Richard Webby, Ghazi Kayali, and Mohamed A. Ali. "A Recombinant Influenza A/H1N1 Carrying A Short Immunogenic Peptide of MERS-CoV as Bivalent Vaccine in BALB/c Mice." Pathogens 8, no. 4 (December 2, 2019): 281. http://dx.doi.org/10.3390/pathogens8040281.
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Middle East Respiratory Syndrome Coronavirus (MERS-CoV) became a global human health threat since its first documentation in humans in 2012. An efficient vaccine for the prophylaxis of humans in hotspots of the infection (e.g., Saudi Arabia) is necessary but no commercial vaccines are yet approved. In this study, a chimeric DNA construct was designed to encode an influenza A/H1N1 NA protein which is flanking immunogenic amino acids (aa) 736–761 of MERS-CoV spike protein. Using the generated chimeric construct, a novel recombinant vaccine strain against pandemic influenza A virus (H1N1pdm09) and MERS-CoV was generated (chimeric bivalent 5 + 3). The chimeric bivalent 5 + 3 vaccine strain comprises a recombinant PR8-based vaccine, expressing the PB1, HA, and chimeric NA of pandemic 2009 H1N1. Interestingly, an increase in replication efficiency of the generated vaccine strain was observed when compared to the PR8-based 5 + 3 H1N1pdm09 vaccine strain that lacks the MERS-CoV spike peptide insert. In BALB/c mice, the inactivated chimeric bivalent vaccine induced potent and specific neutralizing antibodies against MERS-CoV and H1N1pdm09. This novel approach succeeded in developing a recombinant influenza virus with potential use as a bivalent vaccine against H1N1pdm09 and MERS-CoV. This approach provides a basis for the future development of chimeric influenza-based vaccines against MERS-CoV and other viruses.
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Payton, C. D., D. A. Scarisbrick, S. Sikotra, and A. J. E. Flower. "Vaccination against hepatitis B: comparison of intradermal and intramuscular administration of plasma derived and recombinant vaccines." Epidemiology and Infection 110, no. 1 (February 1993): 177–80. http://dx.doi.org/10.1017/s0950268800050792.
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SUMMARYA retrospective analysis of levels of antibody to hepatitis B surface antigen in 1419 health care workers was carried out to compare the efficacy of intramuscular and intradermal administration of plasma derived and recombinant hepatitis B vaccines. No significant difference was detected between the response to intradermal and intramuscular plasma derived vaccine. However of those who received intramuscular recombinant vaccine 81.6%, 13.8% and 4.7% were good (> 100 miu/ml). low (10–99 miu/ml) and non-responders (< 10 miu/ml) respectively. compared with 51.1%, 29.8% and 19.2% of the intradermal group (P > 0.0001). Low dose intradermal administration of recombinant vaccine did not produce satisfactory levels of antibody to hepatitis B surface antigen.
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KOŠOROK, Stane, and Miran KASTELIC. "Antibody response following sow vaccination using cell and recombinant vaccines, single and multiple application." Acta agriculturae Slovenica, no. 2 (September 15, 2008): 167–71. http://dx.doi.org/10.14720/aas-s.2008.2.19243.
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The vaccination of pregnant sows with E. coli antigens to increase specific antibodies in colostrum and milk and subsequently the protection of suckling piglets from neonatal E. coli diarrhea is a common and efficient method, already used for many years. The vaccination to protect individual animals against erysipelas has been used even for a longer time. There are many vaccines and vaccination techniques available on the market, among them herd specific “stable” vaccines for E. coli, inactivated cell vaccines and recombinant DNA vaccines. Unlike vaccines for pet animals, which are mostly polyvalent vaccines, there are very few polyvalent pig vaccines available on the EU market. Recently, Intervet company has developed Diluvac forte (DF) adjuvant which enables mixing of different monovalent vaccines. Porcilis DF vaccine line makes it possible to combine different antigens in the syringe and only one injection of desirable combination of antigens. The difference between two different vaccines (Colisorb, Porcilis DF) was tested to find out the antibody response following vaccination with monovalent recombinant vaccine against bivalent cell vaccine, and the antibody response followed separate and mixed injection of two antigens in one syringe. Statistical evaluation of serum antibody response obtained by ELISA test confirmed the expectations. DNA recombinant vaccines gave significantly higher antibody titers compared to cell vaccine. Porcilis vaccines mixed together prior to application gave surprisingly better response compared to separated application. The result leads to reduction of injections to pregnant sows and consequently to better immune response.
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Cuervo, Nancy Stella, Sophie Guillot, Natalia Romanenkova, Mariana Combiescu, André Aubert-Combiescu, Mohamed Seghier, Valérie Caro, Radu Crainic, and Francis Delpeyroux. "Genomic Features of Intertypic Recombinant Sabin Poliovirus Strains Excreted by Primary Vaccinees." Journal of Virology 75, no. 13 (July 1, 2001): 5740–51. http://dx.doi.org/10.1128/jvi.75.13.5740-5751.2001.
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ABSTRACT The trivalent oral poliomyelitis vaccine (OPV) contains three different poliovirus serotypes. It use therefore creates particularly favorable conditions for mixed infection of gut cells, and indeed intertypic vaccine-derived recombinants (VdRec) have been frequently found in patients with vaccine-associated paralytic poliomyelitis. Nevertheless, there have not been extensive searches for VdRec in healthy vaccinees following immunization with OPV. To determine the incidence of VdRec and their excretion kinetics in primary vaccinees, and to establish the general genomic features of the corresponding recombinant genomes, we characterized poliovirus isolates excreted by vaccinees following primary immunization with OPV. Isolates were collected from 67 children 2 to 60 days following vaccination. Recombinant strains were identified by multiple restriction fragment length polymorphism assays. The localization of junction sites in recombinant genomes was also determined. VdRec excreted by vaccinees were first detected 2 to 4 days after vaccination. The highest rate of recombinants was on day 14. The frequency of VdRec depends strongly on the serotype of the analyzed isolates (2, 53, and 79% of recombinant strains in the last-excreted type 1, 2, and 3 isolates, respectively). Particular associations of genomic segments were preferred in the recombinant genomes, and recombination junctions were found in the genomic region encoding the nonstructural proteins. Recombination junctions generally clustered in particular subgenomic regions that were dependent on the serotype of the isolate and/or on the associations of genomic segments in recombinants. Thus, VdRec are frequently excreted by vaccinees, and the poliovirus replication machinery requirements or selection factors appear to act in vivo to shape the features of the recombinant genomes.
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Zhang, Guihua, Yang Fu, Yu’an Li, Quan Li, Shifeng Wang, and Huoying Shi. "Oral Immunization with Attenuated Salmonella Choleraesuis Expressing the FedF Antigens Protects Mice against the Shiga-Toxin-Producing Escherichia coli Challenge." Biomolecules 13, no. 12 (November 30, 2023): 1726. http://dx.doi.org/10.3390/biom13121726.
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Edema disease (ED) is a severe and lethal infectious ailment in swine, stemming from Shiga-toxin-producing Escherichia coli (STEC). An efficient, user-friendly, and safe vaccine against ED is urgently required to improve animal welfare and decrease antibiotic consumption. Recombinant attenuated Salmonella vaccines (RASV) administered orally induce both humoral and mucosal immune responses to the immunizing antigen. Their potential for inducing protective immunity against ED is significant through the delivery of STEC antigens. rSC0016 represents an enhanced recombinant attenuated vaccine vector designed for Salmonella enterica serotype Choleraesuis. It combines sopB mutations with a regulated delay system to strike a well-balanced equilibrium between host safety and immunogenicity. We generated recombinant vaccine strains, namely rSC0016 (pS-FedF) and rSC0016 (pS-rStx2eA), and assessed their safety and immunogenicity in vivo. The findings demonstrated that the mouse models immunized with rSC0016 (pS-FedF) and rSC0016 (pS-rStx2eA) generated substantial IgG antibody responses to FedF and rStx2eA, while also provoking robust mucosal and cellular immune responses against both FedF and rStx2eA. The protective impact of rSC0016 (pS-FedF) against Shiga-toxin-producing Escherichia coli surpassed that of rSC0016 (pS-rStx2eA), with percentages of 83.3%. These findings underscore that FedF has greater suitability for vaccine delivery via recombinant attenuated salmonella vaccines (RASVs). Overall, this study provides a promising candidate vaccine for infection with STEC.
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Husseiny, Mohamed I., and Michael Hensel. "Rapid Method for the Construction of Salmonella enterica Serovar Typhimurium Vaccine Carrier Strains." Infection and Immunity 73, no. 3 (March 2005): 1598–605. http://dx.doi.org/10.1128/iai.73.3.1598-1605.2005.
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ABSTRACT Salmonella enterica serovar Typhimurium is a versatile organism for the generation of live recombinant vaccines for mucosal immunization. Various strategies have been devised for the stable and efficient expression of heterologous antigens by attenuated S. enterica strains, but these methods often require complex manipulations. Use of phage λ Red recombinase has recently been devised for gene replacements in Escherichia coli and S. enterica after introduction of PCR products. Based on this method, we have developed an approach that allows the integration of recombinant expression cassettes for heterologous antigens in a single step. The recombinant construct is integrated into the chromosome and is devoid of any selective marker such as antibiotic resistance. We observed the stable expression of model antigens without selective pressure. In addition, the method allows the simultaneous generation of attenuating mutations by gene deletions. The novel “knock-in” approach allows the rapid and efficient construction of recombinant Salmonella strains as vaccine carriers.