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At least 1,243 records · Page 69Linked to original sources
PubMed2001

Protection against lethal enterovirus 71 infection in newborn mice by passive immunization with subunit VP1 vaccines and inactivated virus.

Enterovirus 71 (EV71), the newest member of Enteroviridae, is notable for its etiological role in epidemics of severe neurological diseases in children. Developing effective vaccines is considered a top choice among all control measures. We compared the inactivated virus vaccine (10 microg protein/mouse) with subunit vaccines--VP1 DNA vaccine (100 microg/mouse) or recombinant VP1 protein (10 microg/mouse)--in its ability to elicit maternal antibody and to provide protection against lethal infection of EV71 in suckling mice. Prior to gestation, all three groups of vaccinated dams possessed similar levels of neutralizing antibody. With a challenge dose of 2300 LD(50) virus/mouse, suckling mice born to dams immunized with inactivated virus showed 80% survival. The subunit vaccines provided protection only at a lower challenge dosage of 230 LD(50) per mouse, with 40% survival for DNA vaccine and 80% survival for VP1 protein. The cytokine profile produced by splenocytes showed a high level of IL-4 in the inactivated virus group, high levels of IFN-gamma and IL-12 in the DNA vaccine group, and high levels of IL-10 and IFN-gamma in the VP1 protein group. Overall, the inactivated virus elicited a much greater magnitude of immune response than the subunit vaccines, including total IgG, all four IgG subtypes, and T-helper-cell responses; these antibodies were shown to be protective against lethal infection when passively transferred to susceptible newborn mice. Our data indicated that inactivated virus is the choice of vaccine preparation capable of fulfilling the demand for effective control, and that VP1 subunit vaccines remain promising vaccine strategies that require further refinement.

Animals↗
PubMed2003

In ovo delivery of DNA to the avian embryo.

We have developed a simple method of transfecting avian embryos in ovo with various plasmid vectors that results in protein expression in the embryo. Using the chloramphenicol acetyl transferease (CAT) reporter gene, we were able to show that transfecting avian embryos with a plasmid/neutral lipid/dimethylsulfoxide (DMSO) mixture delivered to the air cell, is better than transfecting naked DNA or cationic lipid encapsulated DNA, using DMSO (P < 0.05). This method resulted in CAT expression in several avian embryonic tissues of all the embryos inoculated. We found that both the cytomegalovirus (CMV) and chicken beta actin promoters worked significantly better (P < 0.05) than the Rous sarcoma virus promoter in vitro for reporter gene expression after cationic liposome-mediated transfection. However, after in ovo delivery with neutral lipid encapsulation and DMSO mediated delivery, no significant difference (P > 0.05) between the various promoters could be determined. We believe this neutral lipid encapsulation method may represent an important platform for delivery of DNA to the avian embryo.

Actins↗
PubMed2003

A comparison of DNA vaccines for the rabies-related virus, Mokola.

Mokola virus, a rabies-related virus, has been reported to date from the African continent only. Like rabies virus, it is highly pathogenic, causes acute encephalitis, and zoonotic events have been documented. Although believed to be rare, there has been an unexplained increase in the number of isolations of the virus in South Africa in recent years. We have cloned and sequenced the glycoprotein (G) and nucleoprotein (N) genes from a South African Mokola virus, and used these in the construction of different DNA vaccines for immunization against Mokola virus. Four vaccines, utilizing different promoters and DNA backbone compositions, were generated and compared for efficacy in protection against Mokola virus. In one of these, both the Mokola virus G and N genes were co-expressed. Two of the single G-expressing DNA vaccines (based on pSG5 and pCI-neo, respectively) protected laboratory mice against lethal challenge, despite major differences in their promoters. However, neither vaccine was fully protective in a single immunization only. Serological assays confirmed titers of virus-neutralizing antibodies after immunization, which increased upon booster vaccine administration. A third construct (based on pBudCE4) was less effective in inducing a protective immune response, despite employing a strong CMV enhancer/promoter also used in the pCI-neo plasmid. Dual expression of Mokola virus G and N genes in pBudCE4 did not enhance its efficacy, under the conditions described. In addition, no significant utility could be demonstrated for a combined prime-boost approach, as no cross-protective immunity was observed against rabies or Mokola viruses from the use of pSG5-mokG or vaccinia-rabies glycoprotein recombinant virus vaccines, respectively, even though both vaccines provided 60-100% protection against homologous virus challenge.

Animals↗
PubMed2003

Development and large-scale use of recombinant VP2 vaccine for the prevention of infectious bursal disease of chickens.

Infectious bursal disease virus (IBDV) is the causative agent of Gumboro disease, an infectious disease of global economic importance in poultry. One of the most effective types of inactivated IBDV vaccine is produced by infecting young chickens with a virulent strain, sacrificing them and extracting the virus from the bursa of Fabricius. The goal of this study was to produce an effective subunit vaccine against IBDV thereby providing an effective means of combating the disease. In areas in which the bursa-derived vaccine is in use, this subunit vaccine would eliminate the use of live birds for the production of inactivated vaccines. The gene for viral protein 2 (VP2) of IBDV was cloned into a Pichia pastoris expression system. This efficient system allowed us to meet the need for inexpensive vaccines required by the poultry industry. Following expression and scale-up, the protein was used to vaccinate chickens, against either Gumboro disease alone or in combination with inactivated Newcastle disease virus (NDV). Full protection was conferred against IBDV following vaccination with the subunit recombinant vaccine. No untoward influence on the response to the NDV vaccine was recorded. Over 250 million birds have already been vaccinated with this vaccine. The advantages of a subunit vaccine over an inactivated one are discussed. This approach will enable rapid adjustment to new virulent strains if and when they appear.

Animals↗
PubMed1988

Use of peptides for immunization against foot-and-mouth disease.

A peptide corresponding to the major immunogenic site of the protein VP1 of foot-and-mouth disease virus (FMDV) will elicit a protective neutralizing antibody response in guinea-pigs, cattle and pigs. The response is much greater when the peptide is presented as a linear dimer or tetramer and pigs receiving as little as 40 micrograms peptide have been protected against challenge infection. An even greater response is obtained when the peptide is presented as part of the core protein of hepatitis B virus. Moreover, responsiveness to the peptide in non-responder mice can be stimulated by the simultaneous inoculation of an appropriate T-cell epitope linked to the FMDV peptide.

Animals↗
PubMedDate not supplied

Intracutaneous vaccination of rabbits with the cottontail rabbit papillomavirus (CRPV) L1 gene protects against virus challenge .

A DNA vaccine encoding the major capsid protein L1 of cottontail rabbit papillomavirus (CRPV) was constructed and administered intracutaneously (i.c.) to rabbits as supercoiled plasmids bound to gold beads using a specialized delivery device ("gene gun"). L1 DNA-vaccinated rabbits developed cellular proliferative responses to CRPV virus-like particles and developed high titered antibodies with neutralizing activity to CRPV. Following experimental challenge with CRPV, all of the L1 DNA-vaccinated rabbits, vs none of the controls, were protected from papilloma formation. These results demonstrate that i.c. vaccination of rabbits with the L1 papillomavirus capsid gene can induce antibodies that protect against subsequent papillomavirus infection.

Animals↗
PubMed1997

Study of immunogenicity and virulence of bovine herpesvirus 1 mutants deficient in the UL49 homolog, UL49.5 homolog and dUTPase genes in cattle.

We previously reported that the bovine herpesvirus 1 (BHV 1) gene homologous to herpes simplex virus gene UL49 is dispensable; nevertheless, a mutant with the UL49 homolog (UL49 h) gene deletion exhibited significantly impaired growth in cell culture. To further evaluate the role of the UL49 h in virus infectivity in the natural host of BHV 1, the pathogenesis of the UL49 h negative mutant was studied in cattle. An additional mutant with a combined defect in UL49 h, UL49.5 h and dUTPase genes was also studied in parallel. We found that both mutants were avirulent in cattle inasmuch as intranasal (i.n.) administration of either mutants induced no apparent clinical disease, nor did animals receiving the mutants shed virus. Following i.n. inoculation with the mutants animals developed low levels of serum neutralizing (SN) antibodies, and were partially protected against wild-type BHV 1 challenge. Intramuscular immunizations with either mutant induced good SN titers, and moreover, they induced nearly complete protection against respiratory challenge with wild-type virus. The results from this study establish that BHV 1 UL49 h is an important virulence factor, and also suggest that deletion of the nonessential viral genes UL49 h, UL49.5 h and dUTPase may be useful in developing recombinant BHV 1 vaccines or BHV 1-based vaccine vectors.

Administration, Intranasal↗
PubMedDate not supplied

Raccoon poxvirus live recombinant feline panleukopenia virus VP2 and rabies virus glycoprotein bivalent vaccine.

A raccoon poxvirus (RCNV) recombinant for immunizing against feline panleukopenia and rabies was developed by homologous recombination with a chimeric plasmid for insertional inactivation of the RCNV thymidine kinase gene. The recombinant, RCN-FPV/VP2-rabG, coexpressed the feline panleukopenia virus (FPV) VP2 protein and the rabies virus spike glycoprotein (rabG) under oppositely oriented vaccinia virus P11 promoters. Cats vaccinated subcutaneously with the recombinant showed relatively high neutralizing antibody responses against rabies virus and FPV, and protection against an otherwise virulent FPV challenge with no drop in white blood cell count. Because of containment constraints, no rabies virus challenges were done, but the high concentrations (> 8 IU) of rabies neutralizing antibodies were consistent with levels that usually indicate an ability to counter the infection.

Animals↗
PubMed1998

Virulence, immunogenicity and reactivation of bovine herpesvirus 1 mutants with a deletion in the gC, gG, gI, gE, or in both the gI and gE gene.

Within the framework of developing a marker vaccine against bovine herpesvirus 1 (BHV1), several mutants with deletions in non-essential glycoprotein genes were constructed. Glycoprotein gC, gG, gI and gE single deletion mutants, a gI/gE double deletion mutant and a gE frame-shift mutant were made. The virulence and immunogenicity of these mutants were evaluated in specific-pathogen-free calves. Except for the gC deletion mutant, all mutants were significantly less virulent than the parental wild-type (wt) BHV1 strain Lam. The virulence of the gI and the gI-/gE- mutants was almost completely reduced. Upon challenge infection, the calves of the control group became severely ill, whereas all other calves remained healthy. The reduction of the virus shedding after challenge infection was related to the virulence of the strain of primary inoculation. Virus shedding was almost completely reduced in calves first inoculated with Lam-wt or with gC- and the least reduced in calves inoculated with gI- or gI-/gE-. Six weeks after challenge, all calves were treated with dexamethasone to study whether mutant or challenge virus or both could be reactivated. The gC- and the gG- mutants were reactivated, whereas none of the other mutants were reisolated. Reactivation of challenge virus was reduced in all calves inoculated with mutant viruses. The gC deletion mutant was too virulent and the gI and the gI/gE deletion mutants were the least immunogenic, but based on residual virulence and immunogenicity, both the gG and the gE deletion mutants are candidates for incorporation in live BHV1 vaccines. However, it also depends on the kinetics of the anti-gG and anti-gE antibody response after wild-type virus infection, whether these deletion mutants are really suitable to be incorporated in a marker vaccine.

Animals↗
PubMed1998

Enhanced immunogenicity of a recombinant genital warts vaccine adjuvanted with monophosphoryl lipid A.

The regression of genital warts is believed to be a T-cell-mediated immune effect. We have sought to enhance the immunogenicity of a therapeutic vaccine for the treatment of genital warts with the use of the adjuvant monophosphoryl lipid A (MPL-immunostimulant), a detoxified form of the lipopolysaccharide (LPS) of Salmonella minnesota R595. The comparative immunogenicity and reactogenicity of a recombinant human papillomavirus type 6 (HPV6) L2E7 fusion protein in either aqueous, oil-in-water emulsions or Alhydrogel formulations containing MPL was evaluated. We conclude that the simple addition of MPL to the L2E7 fusion protein already adsorbed onto Alhydrogel preferentially enhances antigen specific in vitro T-cell proliferative responses, IFN gamma production and in vivo delayed type hypersensitivity responses without increasing its reactogenicity.

Adjuvants, Immunologic↗
PubMed2003

Growth of recombinant equine herpesvirus 1 (EHV-1) replaced with passage-induced mutant gene 1 and gene 71 derived from an attenuated EHV-1 in cell cultures and in the lungs of mice.

The relationship of passage-induced mutant genes 1 and 71 of an attenuated equine herpesvirus 1 (EHV-1) with virulence was analysed by constructing nine recombinant EHV-1 viruses by homologous recombination. Gene 1 or/and gene 71 of a virulent EHV-1 strain, HH1, was replaced by a mutant gene 1 or/and 71 of an attenuated HH1 strain, BK343, respectively. The beta-galactosidase gene of Escherichia coli was inserted within the gene 1 or 71 coding sequence of HH1 to inactivate the genes. Virus replications of these recombinant viruses in cell cultures were similar, but release of the gene 71-inactivated virus from infected cells was delayed compared to that of the other viruses. Plaque sizes of the recombinant viruses were similar to those of HH1, but those of BK343 were significantly smaller, indicating an effect of some unknown factor(s) on viral cell-to-cell spread. The growth abilities of the recombinant viruses with a mutant gene 1 or/and 71 in lungs of mice were similar to those of HH1, but those of gene 71-inactivated viruses were reduced to the level of BK343 and the titers were about 100-times lower than those of the other recombinant viruses. These results indicate that the mutant genes 1 and 71 of BK343 might not confer an attenuated nature to EHV-1.

Amino Acid Sequence↗
PubMed1999

The molecular biology of hepatitis C virus. Genotypes and quasispecies.

Hepatitis C virus (HCV) is an important cause of chronic liver disease worldwide. HCV is a positive-strand genotype RNA virus with extensive genetic heterogeneity; HCV isolates define 6 major genotypes, and HCV circulates within an infected individual as a number of closely related but distinct species, termed a quasispecies. This article reviews characteristic aspects of HCV molecular biology and their implications for treatment and vaccine development.

3' Untranslated Regions↗
PubMed2006

[Construction and immunological characterization of recombinant Marek's disease virus expressing IBDV VP2 fusion protein].

A transfer plasmid vector pUC18-US10-VP2 was first constructed by inserting the gene of the enhancer green fluorescent protein(eGFP) fused to the VP2 gene of very virulent Infectious bursal disease virus (IBDV) JS strain into the US10 fragment of the Marek's disease virus (MDV) CV1988/Rispens. The recombinant virus, designated as rMDV, was developed by co-transfecting CEF with the transfer plasmid vector and simultaneously infecting with the CVI988/Rispens virus. The PCR and IFA results indicated that the rMDV is stable after 31 passages. Chickens vaccinated with rMDV were protected from challenge with 100LD50 of IBDV JS. The protection ratio of the chickens vaccinated with the 1000PFU, 2000PFU, 5000PFU of the rMDV were 50%, 60%, and 80% respectively. It is interesting that the average histopathology BF lesion scores of chicken group immunized with 5000PFU of rMDV by one-time vaccination was close to that of chicken group vaccinated with IBDV live vaccine NF8 strain for twice (2.0/1.5). There is no difference in protection between the groups (P > 0.05) but significent difference between groups immunized with 5000 PFU of rMDV and with normal MDV. This demonstrated that rMDV expressing VP2 fusion protein was effective vaccine against IBDV in SPF chickens.

Animals↗
PubMed2000

Folding defects caused by single amino acid substitutions in a subunit are not alleviated by assembly.

Significant stabilization of a protein often occurs when it is assembled into an oligomer. Bacteriophage P22 contains 420 monomers of coat protein that are stabilized by the assembly and maturation processes. The effects of eight single amino acid substitutions in coat protein that each cause a temperature-sensitive-folding defect were investigated to determine if the conformational differences previously observed in the monomers could be alleviated by assembly or maturation. Several techniques including differential scanning calorimetry, heat-induced expansion, urea denaturation, and sensitivity to protease digestion were used to explore the effects of the amino acid substitutions on the conformation of coat protein, once assembled. Each of the amino acid substitutions caused a change in the conformation as compared to wild-type coat protein, observed by at least one of the probes used. Thus, neither assembly nor expansion entirely corrected the conformational defects in the monomeric subunits of the folding mutants.

Amino Acid Substitution↗
PubMed2003

Vaccination with E. coli recombinant empty viral particles of infectious bursal disease virus (IBDV) confer protection.

The A genome segment of the highly virulent Infectious bursal disease virus (IBDV) was amplified using long and accurate-RT-PCR (LA-RT-PCR). The entire sequence region encoding VP2, VP4, and VP3 in that order was cloned and sequenced. Following subcloning into the Escherichia coli expression vector pET21a under the T7 promoter, viral proteins were expressed and processed as demonstrated by Western blot analysis. Virus-like particles could be visualized by immuno-electron microscopy in IPTG-induced cells suggesting that viral assembly can take place in E. coli. Induction of anti-IBDV antibodies was detected in chickens immunized with purified recombinant IBDV by intra muscular (i.m.) injection. Furthermore, the vaccinated chickens were protected when challenged with the Gep 5 isolate of IBDV.

Animals↗
PubMed2004

Expression of immunogenic VP2 protein of infectious bursal disease virus in Arabidopsis thaliana.

VP2 protein is the major host-protective immunogen of infectious bursal disease virus (IBDV) of chickens. Transgenic lines of Arabidopsis thaliana expressing recombinant VP2 were developed. The VP2 gene of an IBDV antigenic variant E strain was isolated, amplified by RT-PCR and introduced into a plant expression vector, pE1857, having a strong promoter for plant expression. A resulting construct with a Bar gene cassette for bialaphos selection in plant (rpE-VP2) was introduced into Agrobacterium tumefaciens by electroporation. Agrobacterium containing the rpE-VP2 construct was used to transform Ar. thaliana and transgenic plants were selected using bialaphos. The presence of VP2 transgene in plants was confirmed by PCR and Southern blot analysis and its expression was confirmed by RT-PCR. Western blot analysis and antigen-capture ELISA assay using monoclonal anti-VP2 were used to determine the expression of VP2 protein in transgenic plants. The level of VP2 protein in the leaf extracts of selected transgenic plants varied from 0.5% to 4.8% of the total soluble protein. Recombinant VP2 protein produced in plants induced antibody response against IBDV in orally-fed chickens.

Animals↗
PubMed2000

Immunotherapy of a human papillomavirus (HPV) type 16 E7-expressing tumour by administration of fusion protein comprising Mycobacterium bovis bacille Calmette-Guérin (BCG) hsp65 and HPV16 E7.

Human papillomavirus type 16 (HPV16) infection has been linked to the development of cervical and anal dysplasia and cancer. One hallmark of persistent infection is the synthesis of the viral E7 protein in cervical epithelial cells. The expression of E7 in dysplastic and transformed cells and its recognition by the immune system as a foreign antigen make it an ideal target for immunotherapy. Utilizing the E7-expressing murine tumour cell line, TC-1, as a model of cervical carcinoma, an immunotherapy based on the administration of an adjuvant-free fusion protein comprising Mycobacterium bovis BCG heat shock protein (hsp)65 linked to HPV16 E7 (hspE7) has been developed. The data show that prophylactic immunization with hspE7 protects mice against challenge with TC-1 cells and that these tumour-free animals are also protected against re-challenge with TC-1 cells. In addition, therapeutic immunization with hspE7 induces regression of palpable tumours, confers protection against tumour re-challenge and is associated with long-term survival (> 253 days). In vitro analyses indicated that immunization with hspE7 leads to the induction of a Th1-like cell-mediated immune response based on the pattern of secreted cytokines and the presence of cytolytic activity following antigenic recall. In vivo studies using mice with targeted mutations in CD8 or MHC class II or depleted of CD8 or CD4 lymphocyte subsets demonstrate that tumour regression following therapeutic hspE7 immunization is CD8-dependent and CD4-independent. These studies extend previous observations on the induction of cytotoxic T lymphocytes by hsp fusion proteins and are consistent with the clinical application of hspE7 as an immunotherapy for human cervical and anal dysplasia and cancer.

Animals↗