OBJECTIVETo observe the protection against periodontal bone loss in the Sprague-Dawley (SD) rats periodontitis model, with the recombined plasmid pcDNA3.1+/kgpcd as DNA gene vaccine.

METHODSPcDNA3.1+/kgpcd was delivered into rats by submandibular gland-targeted injection. The anti-KGPcd sIgA in saliva was measured by indirect ELISA method. Immunohistochemistry staining was used to assess the protection in the animal model.

RESULTSThe level of specific anti-KGPcd sIgA in saliva of the experimental group was significantly higher than that of control group. HE staining showed that immunization with recombined plasmid pcDNA3.1+/kgpcd could protect or minimize tissue destruction caused by subsequent P. gingivalis challenge in the rat model.

CONCLUSIONSThe results indicate that pcDNA3.1+/kgpcd was a good candidate for anti-periodontitis gene vaccine and could provide protection against Porphyromonas gingivalis-caused periodontitis in rat lesion model.

Animals ; Bacterial Vaccines ; immunology ; therapeutic use ; Immunoglobulin A, Secretory ; analysis ; Periodontitis ; immunology ; microbiology ; prevention & control ; Porphyromonas gingivalis ; genetics ; immunology ; Rats ; Rats, Sprague-Dawley ; Vaccines, DNA ; immunology ; therapeutic use

OBJECTIVESTo evaluate if the humoral immune response of hepatitis B DNA vaccine pVAX1-S2S could be enhanced by Talpha1 and/or IFNa expression plasmid co-inoculated.

METHODSThe following mammalian expression recombinant plasmids were constructed: the plasmid pVAX1-S2S expressing hepatitis B surface antigen S2S, the plasmid pVAX1-T/I co-expressing thymosin a and IFNalpha, the plasmid pVAX1-I/S2S co-expressing IFNalpha and S2S. These plasmids were inoculated intramuscularly into several BALB/c mice groups in different combinations. In the co-immunization group 1 (pVAX1-I/S2S), each mouse was inoculated with 100 microg of pVAX1-I/S2S; in the co-immunization group 2 (pVAX1-S2S) each mouse was co-inoculated with pVAX1-S2S and 50 microg of pVAX1-TI; in the control group each mouse was inoculated with 100 microg of pVAX1-S2S. All the immunizations were boosted at 2 and 4 week intervals; then the serum samples were collected to detect the anti-HBs and anti-preS2 strengths.

RESULTS3, 5 and 8 weeks after the first inoculation, the positive rates of anti-HBs were 12.5%, 12.5%, 62.5% respectively in the co-immunization group 1 and 25%, 50%, 50% in the co-immunization group 2, while those in the control group were 0, 25%, 37.5%. The titers of anti-preS2 in co-immunization group 2 was 5 times higher than those in the other two groups.

CONCLUSIONThe data shows that Talpha1 and/or IFNalpha expression plasmid co-inoculated with pVAX1-S2S might act as an adjuvant to enhance the humoral immune response induced by pVAX1-S2S.

Adjuvants, Immunologic ; genetics ; therapeutic use ; Animals ; Female ; Hepatitis B ; immunology ; therapy ; Hepatitis B Vaccines ; immunology ; therapeutic use ; Interferon-alpha ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; immunology ; Thymosin ; genetics ; immunology ; Vaccines, DNA ; immunology

Recombinant baculoviruses containing the fusion (F) and hemagglutinin-neuraminidase (HN) glycoprotein gene of the viscerotropic velogenic (vv) Newcastle disease virus (NDV) isolate, Kr-005/00, and a lentogenic La Sota strain of the NDV were constructed in an attempt to develop an effective subunit vaccine to the recent epizootic vvNDV. The level of protection was determined by evaluating the clinical signs, mortality, and virus shedding from the oropharynx and cloaca of chickens after a challenge with vvNDV Kr-005/00. The recombinant ND F (rND F) and recombinant HN (rND HN) glycoproteins derived from the velogenic strain provided good protection against the clinical signs and mortality, showing a 0.00 PI value and 100% protection after a booster immunization. On the other hand, the combined rND F + HN glycoprotein derived from the velogenic strain induced complete protection (0.00 PI value and 100% protection) and significantly reduced the amount of virus shedding even after a single immunization. The rND F and rND HN glycoproteins derived from the velogenic strain had a slightly, but not significantly, greater protective effect than the lentogenic strain. These results suggest that the combined rND F + HN glycoprotein derived from vvNDV can be an ideal subunit marker vaccine candidate in chickens in a future ND eradication program.
Animals ; Baculoviridae/genetics/*immunology ; Chickens/*virology ; DNA Primers ; Gene Amplification ; HN Protein/genetics/*therapeutic use ; Korea ; Marek Disease/immunology/prevention & control ; Newcastle Disease/immunology/*prevention & control ; Spodoptera/virology ; Vaccines, Synthetic/genetics/therapeutic use ; Viral Vaccines/genetics/therapeutic use

Cervical cancer is the fourth most lethal women's cancer worldwide. Current treatments against cervical cancer include surgery, radiotherapy, chemotherapy, and anti-angiogenic agents. However, despite the various treatments utilized for the treatment of cervical cancer, its disease burden remains a global issue. Persistent infection of human papillomavirus (HPV) has been identified as an essential step of pathogenesis of cervical cancer and many other cancers, and nation-wide HPV screening as well as preventative HPV vaccination program have been introduced globally. However, even though the commercially available prophylactic HPV vaccines, Gardasil (Merck) and Cervarix (GlaxoSmithKline), are effective in blocking the entry of HPV into the epithelium of cervix through generation of HPV-specific neutralizing antibodies, they cannot eliminate the pre-existing HPV infection. For these reason, other immunotherapeutic options against HPV-associated diseases, including therapeutic vaccines, have been continuously explored. Therapeutic HPV vaccines enhance cell-mediated immunity targeting HPV E6 and E7 antigens by modulating primarily dendritic cells and cytotoxic T lymphocyte. Our review will cover various therapeutic vaccines in development for the treatment of HPV-associated lesions and cancers. Furthermore, we will discuss the potential of immune checkpoint inhibitors that have recently been adopted and tested for their treatment efficacy against HPV-induced cervical cancer.
Dendritic Cells/immunology ; Female ; Genetic Vectors ; Humans ; *Immunotherapy ; Papillomavirus Infections/*complications/therapy ; Papillomavirus Vaccines/therapeutic use ; *Translational Medical Research ; Uterine Cervical Neoplasms/*therapy ; Vaccines, DNA/therapeutic use ; Vaccines, Subunit/therapeutic use

OBJECTIVEThe immunogenicity and protective efficacy of an experimental Campylobacter jejuni (C. jejuni) chitosan-DNA vaccines were evaluated in mice.

METHODSThe chitosan-DNA vaccines were prepared by embedding pcDNA3.1(+)-cadF and pcDNA3.1(+)-peblA with chitosan respectively. BALB/c mice were intranasally immunized in a four-dose primary series (7 d intervals) at doses of 60 microg chitosan-DNA vaccines each time. The comparative immunogenicities of nine formulations were assessed on the basis of the generation of antigen-specific antibodies in serum and intestinal secretions. Mice were attacked repeatedly through intragastric administration of C. jejuni HS:19 at the 8th week after the immunization and protective efficacy was determined by detecting the degrees of protection afforded against C. jejuni invaded.

RESULTSThe mice immunized with chitosan-DNA vaccines have generated high levels of IgA and IgG from the sera and IgA from the intestinal secretions and the P/N value went up to 20.58, 30.13 and 6.87 respectively. Meanwhile, the expression of intestinal SIgA increased correspondingly. Moreover the chitosan-DNA vaccines induced strongest level of protection in BALB/c mice against challenge with C. jejuni HS:19 strain and the protective efficacies was 93.70.

CONCLUSIONThe results of this study indicate that the chitosan-DNA vaccines could induce significant protective immunity against C. jejuni challenge in the mice model.

Animals ; Antibodies, Bacterial ; immunology ; Campylobacter Infections ; immunology ; prevention & control ; Campylobacter jejuni ; immunology ; Chitosan ; immunology ; therapeutic use ; Disease Models, Animal ; Immunoglobulin A, Secretory ; immunology ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; immunology ; therapeutic use

OBJECTIVETo develop human papillomavirus (HPV) 16 DNA vaccine for the treatment of HPV16 infection and its related tumors.

METHODSHPV16 oncogene E7 was modified by combined approaches including insertion and replication of specific region of E7 gene, murine codon optimization, and point-mutation at transforming regions of the E7 protein. The resulting artificial gene, named as mE7, was obtained by gene synthesis. The mE7 gene was then genetically fused to murine CD40 ligand (CD40L) by overlapping PCR to form the mE7/CD40L fusion gene. The mE7/CD40L gene was inserted into pVR1012 plasmid and then immunized C57/BL6 mice intramuscularly. The E7-specific IFN-gamma-secreting CD8+ T cells were analyzed with EIISPOT, and E7-specific antibody was measured by indirect ELISA. FACS assays were performed to analyze the activation of E7-specific Th cells. Mice were vaccinated, followed by tumor challenged or challenged before immunization. Tumor growth was observed.

RESULTSThe mE7 DNA vaccine elicited an increased E7-specific antibody level (P < 0.01), E7-specific IFN-gamma-secreting CD8+ T (P < 0.01), and CD4+ T cells number (P < 0.05), compared with those of mice immunized with wE7 gene. Furthermore, the mE7/CD40L DNA vaccine elicited an increased number of E7-specific IFN-gamma secreting CD8+ T cell compared with that of mice immunized with mE7 gene (P < 0.01); however, no significant differences were found between mice immunized with the mE7 gene and mE7/CD40L fusion gene in the E7-specific antibody production and Th cell activation. In the preventive experiment, all mice received the mE7 or mE7/CD40L remained tumor-free 7 weeks after challenges with TC-1 tumor cells, while the wE7 group exhibited tumor growth within 2 weeks. In the therapeutic experiment, all the mice in the wE7 group exhibited tumor growth within 8 days, while among mice receiving the mE7 and mE7/CD40L, 30% and 45% of mice remained tumor-free after TC-1 challenge, respectively. HE staining of tumor tissues showed copious lymphocytes infiltration around tumor cells in mE7 and mE7/CD40L mice with regression of tumor growth.

CONCLUSIONSThe mE7 DNA vaccine increases the E7-specific humoral and cellular immune responses, and the fusion of CD40L to mE7 gene enhances the specific immune responses and anti-tumor effects against HPV16 E7-expressing murine tumors. mE7/CD40L may therefore be a suitable and promising target for HPV16 therapeutic vaccine.

Animals ; CD40 Antigens ; genetics ; immunology ; Cancer Vaccines ; genetics ; immunology ; therapeutic use ; Cell Line, Tumor ; Gene Fusion ; Human papillomavirus 16 ; immunology ; Immunity, Cellular ; Immunity, Humoral ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Papillomavirus E7 Proteins ; genetics ; immunology ; Papillomavirus Vaccines ; genetics ; immunology ; therapeutic use ; Vaccines, DNA ; genetics ; immunology ; therapeutic use

OBJECTIVETo observe the efficacy of treating intrauterine infected chronic hepatitis B virus (HBV) carrier children with a combination of granulocyte-macrophage colony-stimulating factor (GM-CSF) or hepatitis B immunoglobulin (HBIG) plus recombinant hepatitis B vaccine (rHBvac).

METHODSA total of 27 chronic HBV infected children, who were born to HBV carrier mothers and received hepatitis B immunoprophylaxis at birth, were randomized into 2 groups: one receiving a combined therapy of 50 micro g of GM-CSF plus 10 micro g of rHBvac injected intramuscularly at the same location (GM-CSF group, 14 children) or 200 IU HBIG and 10 micro g rHBvac in different muscles (HBIG group, 13 children) on a monthly four-dose schedule. HBV-DNA quantification and other HBV serological markers were tested before and after the four-dose therapy.

RESULTSTwelve children in each group completed the study. Of them, 3 children in the GM-CSF group and 4 in the HBIG group had elevated serum alanine transaminase (ALT) before the trial, and then 2 in each group became ALT normal after the treatment. Before the therapy, hepatitis B e antigen (HBeAg) positivity was found in nine children in the GM-CSF group and 10 in the HBIG group. One from each group had an HBeAg/anti-HBe seroconversion after the treatment. The quantity of HBV-DNA was significantly lower after the treatment (P = 0.023) in GM-CSF group, but was not significantly reduced in HBIG group. No subjects were found to be negative for hepatitis B surface antigen (HBsAg) after the treatment, and no serious adverse events occurred in either group.

CONCLUSIONCombined GM-CSF and rHBvac therapy inhibit HBV replication in carrier children who were not protected after treatment with immunoprophylaxis.

Carrier State ; therapy ; Child ; Child, Preschool ; Combined Modality Therapy ; DNA, Viral ; blood ; Granulocyte-Macrophage Colony-Stimulating Factor ; therapeutic use ; Hepatitis B Vaccines ; immunology ; Hepatitis B, Chronic ; therapy ; Humans ; Immunoglobulins ; therapeutic use ; Pilot Projects ; 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

OBJECTIVETo develop an oral DNA vaccine based on MG(7)-Ag mimotope of gastric cancer using attenuated Salmonella typhimurium and evaluate its efficacy and protective effect.

METHODSThe eukaryotic expression vector including the MG(7)-Ag mimotope and a Th epitope was constructed, and then transduced into an attenuated Salmonella typhimurium to get the oral DNA vaccine. C57BL/6 J mice were orally immunized with 1 x 10(8) cfu Salmonella transfectants, with Salmonella harboring empty plasmid, with phophate buffered saline (PBS) as control. At the 6th week, serum titer of MG(7) antibody was detected by ELISA. In the 8th week, a [(3)H]-thymidine incorporation assay was performed to test the proliferation of murine spleen cells to the stimulant of MG(7)-Ag mimicry peptide. At the same time, Ehrlich ascites carcinoma cells expressing MG(7)-Ag were used in tumor challenge assay to evaluate the protective effect of the immunization.

RESULTSThe oral DNA vaccine induced MG(7) antibody in mice, while in vivo unprimed proliferation assay of the spleenocytes showed no difference among the three groups. Two weeks after tumor challenge, 2 in 7 immunized mice were tumor free, while none in the control group was protected.

CONCLUSIONOral DNA vaccine based on the MG(7)-Ag momitope is immunogenic. It is able to induce specific immunity response against tumor in mice, and the vaccine is partially protective.

Administration, Oral ; Amino Acid Sequence ; Animals ; Antigens, Neoplasm ; blood ; genetics ; immunology ; Base Sequence ; Cancer Vaccines ; genetics ; immunology ; therapeutic use ; Epitopes ; genetics ; immunology ; Female ; Mice ; Mice, Inbred C57BL ; Molecular Mimicry ; genetics ; immunology ; Molecular Sequence Data ; Plasmids ; genetics ; Polymerase Chain Reaction ; Stomach Neoplasms ; drug therapy ; immunology ; Treatment Outcome ; Vaccines, DNA ; genetics ; immunology ; therapeutic use

Animals ; Antigen-Antibody Complex ; therapeutic use ; DNA, Viral ; blood ; Dendritic Cells ; immunology ; Ducks ; Female ; Hepatitis B Antibodies ; blood ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B Vaccines ; immunology ; therapeutic use ; Hepatitis B e Antigens ; blood ; immunology ; Hepatitis B virus ; immunology ; Hepatitis B, Chronic ; blood ; immunology ; therapy ; Humans ; Male ; Mice ; T-Lymphocytes