Infectious bursal disease (IBD) is an acute and highly contagious disease of young chickens caused by Birnavirus. Mortality of infected birds can be best prevented if injected with antibodies. The present study was an attempt to raise specific hyper-immune polyclonal antibodies against IBD virus in Pakistan. Commercial layers divided into four groups were injected with IBD vaccine subcutaneously according to four different treatment regimens. Eggs were collected daily and antibodies were purified from yolk with dextran sulphate. Titers of antibodies in serum and yolk were evaluated with enzyme linked immunosorbant assay and agar gel precipitation test. Antibody titers were significantly higher in yolk than serum. Eggs collected at 28 days post-vaccination had maximum antibody titers. Of treatment regimens, T3 was found to be most effective for hyperimmunization. Lyophilized antibodies stored at 4oC did not lose their activity till the end of experiment. IBD virus infected birds were injected with purified antibodies which induced 92% recovery as compared to control birds. The study implicates that the purified antibodies may be useful as a therapeutic agent to cure IBD infected birds.
Animals ; Antibodies, Viral/blood ; Birnaviridae Infections/immunology/*therapy/*veterinary/virology ; *Chickens ; Egg Yolk/immunology/virology ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Immunization/methods/*veterinary ; Immunoglobulins/*immunology ; Immunotherapy/methods/veterinary ; Infectious bursal disease virus/*immunology ; Poultry Diseases/immunology/*therapy/*virology ; Precipitin Tests/veterinary ; Viral Vaccines/*immunology/therapeutic use

Classical swine fever virus (CSFV) causes a highly contagious disease among swine that has an important economic impact worldwide. CSFV strain LOM is an attenuated virus of low virulent strain of Miyagi isolated from Japan in 1956. Eight DNA fragments representing the genome of the CSFV strain LOM were obtained by RT-PCR. These were used to determine the complete nucleotide sequence and construct a full-length cDNA clone which was called Flc-LOM. Sequence analysis of the recombinant clone (Flc-LOM) revealed the presence of eight mutations, resulting in two amino acid substitutions, when compared to the parental sequence. RNA transcripts of both LOM and Flc-LOM were directly infectious in PK-15 cells. The rescued Flc-LOM virus grew more slowly than the parental virus, LOM, in the cells. Intramuscular immunization with Flc-LOM was safe and highly immunogenic in pigs; no clinical signs or virus transmission to sentinel animals were observed after 35 days. CSFV-specific neutralizing antibodies were detected 14 days post-infection. After challenge with the virulent CSFV strain SW03, pigs immunized with Flc-LOM were shown to be fully protected. Thus, our newly established infectious clone of CSFV, Flc-LOM, could serve as a vaccine candidate.
Animals ; Antibodies, Viral/blood ; Base Sequence ; Cell Line ; Classical Swine Fever/immunology/*virology ; Classical swine fever virus/*genetics/immunology/pathogenicity ; Cloning, Molecular ; DNA, Complementary/genetics/immunology ; Immunization/methods/standards/veterinary ; Molecular Sequence Data ; Neutralization Tests/veterinary ; RNA, Viral/chemistry/genetics ; Recombinant Proteins/immunology ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Sequence Analysis, DNA ; Specific Pathogen-Free Organisms ; Swine ; Virulence

Herpes simplex virus type-1 (HSV-1) amplicon vectors are versatile and useful tools for transferring genes into cells that are capable of stimulating a specific immune response to their expressed antigens. In this work, two HSV-1-derived amplicon vectors were generated. One of these expressed the full-length glycoprotein D (gD) of bovine herpesvirus 1 while the second expressed the truncated form of gD (gDtr) which lacked the trans-membrane region. After evaluating gD expression in the infected cells, the ability of both vectors to induce a specific gD immune response was tested in BALB/c mice that were intramuscularly immunized. Specific serum antibody responses were detected in mice inoculated with both vectors, and the response against truncated gD was higher than the response against full-length gD. These results reinforce previous findings that HSV-1 amplicon vectors can potentially deliver antigens to animals and highlight the prospective use of these vectors for treating infectious bovine rhinotracheitis disease.
Animals ; Antibodies, Viral/blood ; Blotting, Western/veterinary ; Cattle ; Female ; Genetic Vectors/*immunology ; Herpesvirus 1, Bovine/genetics/*immunology ; Herpesvirus 1, Human/genetics/*immunology ; Immunity, Humoral/immunology ; Immunization/methods/veterinary ; Infectious Bovine Rhinotracheitis/*immunology/prevention & control/virology ; Mice ; Mice, Inbred BALB C ; Neutralization Tests/veterinary ; Specific Pathogen-Free Organisms ; Viral Proteins/genetics/*immunology ; Viral Vaccines/immunology

OBJECTIVE: To determine the immune-protective effect of Japan Schistosoma (Chinese mainland strain) 23 kD membrane protein-heat shock protein (SjC23-Hsp70) DNA vaccine plus adjuvantinduced interleukin-12 (IL-12) plasmid DNA on Schistosoma japonicum infection in water buffalos. METHODS: Forty-five health water buffalos (8-10 months old) in non-endemic area of schistosomiasis were randomly assigned into group A (SjC23-Hsp70+IL-12, 300 μg), group B (SjC23+IL-12, 300 μg) and group C (pVAX+IL-12, 300 μg), 15 in each group. Each buffalo was immuned by shoulder intramuscular injection for 3 times, at an interval of 28 days. Twenty-eight days after the last immunization, each buffalo was infected with 1000 Japan cercariae of Schistosoma. Fecal examinations were conducted 2 days and 1 day before the perfusion, and on the day of perfusion. The number of hatching miracidia and eggs per gram feces was recorded. Fifty-six days after the infection, the buffalos were sacrificed and perfused via the descending aorta. The recovered adult worms and eggs in the liver tissue were counted. RESULTS: We compared group A and B with group C: the estrogen reduction rate was 45.7% and 26.61%; bug reduction rate was 44.51% and 25.84%; the fecal egg reduction rate was 41.1% and 31.63%; the miracidium reduction rate was 48.11% and 38.07%; and the liver egg reduction rate was 43.39% and 31.95%. The above rates in group A were higher than those in group B (P<0.05). CONCLUSION SjC23-Hsp70 DNA vaccine combined with IL-12 may have a significant immunoprotective effect on buffalos.
Animals ; Antigens, Helminth ; immunology ; Buffaloes ; Cattle ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Helminth Proteins ; immunology ; Immunization ; methods ; Interleukin-12 ; genetics ; immunology ; Membrane Proteins ; immunology ; Schistosomiasis japonica ; immunology ; prevention & control ; veterinary ; Vaccines, DNA ; administration & dosage ; immunology ; Vaccines, Synthetic ; immunology

This study examined the adjuvant effects of dimethyl dioctadecyl ammonium bromide (DDA), CpG oligodeoxynucleotides (CpG-ODN), and chicken interferon-gamma (ChIFN-gamma) on a DNA vaccine (pcDNA-VP243) against the infectious bursal disease virus (IBDV). A plasmid encoding chicken IFN-atilde was constructed. Twice at 2-week intervals, twoweek-old chickens were injected intramuscularly and intraperitoneally with either a DNA vaccine alone or a DNA vaccine together with the respective adjuvants. On week 2 after the second immunization, the chickens were orally challenged with the highly virulent IBDV. The groups that received the DNA vaccines plus either DDA or CpG-ODN showed significantly lower survival rates than the group that received the DNA vaccine alone. However, the survival rates for the DNA vaccine alone and for the DNA vaccine plus ChIFN-gamma were similar. The chickens had no detectable antibodies to the IBDV before the challenge but all the surviving chickens in all groups except for the normal control group showed the induction of antibodies to the IBDV at day 10 after the challenge. As judged by the lymphocyte proliferation assays using the a WST-8 solution performed on the peripheral blood and splenic lymphocytes, the stimulation indices (SI) of the peripheral blood lymphocytes in all groups except for the normal control group were similar immediately before the challenge. At 10 days post-challenge, the SI for DNA vaccine plus either CpG-ODN or ChIFN-gamma was similar to that of the DNA vaccine control group. For splenic lymphocytes, the SI in the DNA vaccine plus CpG-ODN and DNA vaccine plus ChIFN-gamma groups were higher than for the DNA vaccine control. These results suggest that DDA actually compromises the protection against the IBDV by DNA vaccine, and CpG-ODN and IFN-gamma had no significant effect.
Adjuvants, Immunologic ; Animals ; Antibodies, Viral/blood ; Birnaviridae Infections/*immunology/*prevention & control/virology ; Bursa of Fabricius/immunology/virology ; Cell Proliferation ; Chickens ; CpG Islands/immunology ; Enzyme-Linked Immunosorbent Assay/veterinary ; Immunization/methods/*veterinary ; Infectious bursal disease virus/*immunology ; Interferon-gamma/immunology/therapeutic use ; Lymphocytes/cytology/immunology ; Oligonucleotides/immunology ; Poultry Diseases/immunology/*prevention & control/*virology ; Specific Pathogen-Free Organisms ; Vaccines, DNA/immunology/therapeutic use ; Viral Vaccines/*immunology/therapeutic use