• Title/Summary/Keyword: DNA vaccines

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Virus-like Particles Containing Cytokine Plasmid DNA (사이토카인 유전자 함유 바이러스 유사입자의 제조)

  • Oh, Yu-Kyoung;Son, Tae-Jong;Sin, Kwang-Sook;Kang, Min-Jeong;Kim, Jung-Mogg;Kim, Nam-Keun;Ko, Jung-Jae;Kim, Chong-Kook
    • Journal of Pharmaceutical Investigation
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    • v.31 no.3
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    • pp.185-190
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    • 2001
  • Human papillomavirus (HPV) infection is known to cause cervical cancers. Human papillomavirus-like particles (VLP) have been studied as preventive vaccines of cervical cancers. To develop VLP as a therapeutic gene carrier, we studied the method to encapsulate cytokine genes in virus-like particles. HPV type 16 capsid L1 genes were amplified by polymerase chain reaction and cloned into T vector. L1 gene was then inserted into baculovirus transfer vector. The clone of baculovirus encoding L1 gene was isolated and used to express L1 protein in Sf 21 insect cells. VLP were purified by CsCl density gradient and ultracentrifugation. VLP were disassembled to capsomer units by treatment of a reducing agent. Given that interleukin-2 (IL-2) genes have been used in anticancer gene therapy and as a molecular adjuvant, IL-2 cytokine plasmids were chosen as a model gene. IL-2 plasmids were incubated with the disassembled capsomer suspension. To reassemble the particles, the mixture of capsomers and cytokine plasmids was dialyzed. The disassembly and reassembly of VLP were confirmed by transmission electron microscopy. The entrapment of cytokine plasmids in reassembled VLP was tested by the stability of plasmids against DNase I. After treatment of reassembled virus-like particles with DNase I, discrete IL-2 DNA band was observed. Our results indicate that IL-2 cytokine plasmid (3.5 kb size) can be encapsulated in the virus-like particles, suggesting the potential of VLP as a gene delivery system. Moreover, VLP containing the adjuvant cytokine plasmids might function as more effective subunit vaccines.

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New typhoid vaccine using sponge-like reduced protocol: development and evaluation

  • Rehab Bahy;Asmaa Gaber;Hamdallah Zedan;Mona Mabrook
    • Clinical and Experimental Vaccine Research
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    • v.12 no.1
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    • pp.70-76
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    • 2023
  • Purpose: Typhoid remains a major health problem, especially in the developing world. Furthermore, the emergence of multidrug-resistant and extensively drug-resistant strains of Salmonella typhi added a sense of urgency to develop more effective typhoid vaccines, one of which is bacterial ghosts (BGs), prepared by both genetic and chemical means. The chemical method includes incubation with numerous agents for a short time at their minimum inhibitory or minimum growth concentrations. This study included the preparation of BGs by a sponge-like reduced protocol (SLRP). Materials and Methods: Critical concentrations of sodium dodecyl sulfate, NaOH, and H2O2 were used. Moreover, high-quality BGs were visualized by scanning electron microscope (SEM). Subculturing was used to confirm the absence of vital cells. Besides, the concentrations of the released DNA and protein were estimated spectrophotometrically. In addition, the integrity of cells was proved by visualizing Gram-stained cells using a light microscope. Furthermore, a comparison between the immunogenicity and safety of the prepared vaccine and the available whole-cell killed vaccine was established. Results: Improved preparation of high-quality BGs of S. typhi, visualized by SEM, revealed punctured cells with intact outer shells. Moreover, the absence of vital cells was confirmed by subculturing. At the same time, the release of respective amounts of proteins and DNA is another evidence of BGs' production. Additionally, the challenge test provided evidence that the prepared BGs are immunogenic and have the same efficacy as the whole cell vaccine. Conclusion: The SLRP provided a simple, economical, and feasible method for BGs preparation.

Developing New Mammalian Gene Expression Systems Using the Infectious cDNA Molecular Clone of the Japanese Encephalitis Virus

  • Yun Sang-Im;Choi Yu-Jeong;Park Jun-Sun;Kim Seok-Yong;Lee Young-Min
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2003.05a
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    • pp.83-86
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    • 2003
  • Major advances in positive-sense RNA virus research have been facilitated by the development of reverse genetics systems. These systems consist of an infectious cDNA clone that encompasses the genome of the virus in question. This clone is then used as a template for the subsequent synthesis of infectious RNA for the generation of synthetic viruses. However, the construction of infectious cDNA for the Japanese encephalitis virus (JEV) has been repeatedly thwarted by the instability of its cDNA. As JEV is an important human pathogen that causes permanent neuropsychiatric sequelae and even fatal disease, a reliable reverse genetics system for this virus is highly desirable. The availability of this tool would greatly and the development of effective vaccines as well as facilitate studies into the basic biology of the virus, including the molecular mechanisms of viral replication, neurovirulence, and pathogenesis. We have successfully constructed a genetically stable infectious JEV cDNA containing full-length viral RNA genome. Synthetic RNA transcripts generated in vitro from the cDNA were highly infectious upon transfection into susceptible cells, and the cDNA remained stable after it had been propagated in E. coli for 180 generations. Using this infectious JEV cDNA, we have successfully expressed a variety of reporter genes from the full-length genomic and various subgenomic RNAs in vitro transcribed from functional JEV cDNAS. In summary, we have developed a reverse genetics system for JEV that will greatly facilitate the research on this virus in a variety of different fields. It will also be useful as a heterologous gene expression vector and aid the development of a vaccine against JEV.

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The Aetiological Role of Human Papillomavirus in Colorectal Carcinoma: An Iranian Population- Based Case Control Study

  • Ranjbar, Reza;Saberfar, Esmaiel;Shamsaie, Alireza;Ghasemian, Ehsan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1521-1525
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    • 2014
  • Background: Human papillomavirus (HPV) is one of the most common sexually transmitted infections worldwide and the association between HPV infection and genital cancers has been well established. This study concerned the possible role of HPV infection in colorectal carcinoma (CRC) in the Iranian population. Materials and Methods: We examined 80 tissues obtained from patients with colorectal cancer consisting of 58 colon cancer samples and 22 rectal cancer samples and 80 tissues from patients with unremarkable pathologic changes as matched controls by sex, study center and anatomical sites. HPV infection and genotypes were detected using nested PCR and sequencing methods, respectively. Results: HPV DNA was detected in 5/80 (6.25%) cases including 1 of 22 (4.54%) patients with rectum cancer and 4 of 58 (6.9%) patients with colon cancer and 1/80 (1.25%) of controls. Furthermore, HPV-18 was detected as the most frequent type and we found no significant correlation between prevalence of HPV infection and anatomical sub- sites. Conclusions: Although a causal relation between human papillomavirus and colorectal cancer was not found through this study, analysis of medical records pointed to a possible role for high- risk types of HPV in increasing the potential of aggressiveness in colorectal cancer. This study shows a particular frequency of HPV genotypes in patients with colorectal cancer in Iran. Since HPV vaccines are limited to a few types of virus, using cohort studies in different geographical zones to screen for patterns of HPV infection in different organs might increase the efficacy and optimization of the current vaccines.

Genetic and Environmental Deterrents to Breeding for Disease Resistance in Dairy Cattle

  • Lin, C.Y.;Aggrey, S.E.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.9
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    • pp.1247-1253
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    • 2003
  • Selection for increased milk production in dairy cows has often resulted in a higher incidence of disease and thus incurred a greater health costs. Considerable interests have been shown in breeding dairy cattle for disease resistance in recent years. This paper discusses the limitations of breeding dairy cattle for genetic resistance in six parts: 1) complexity of disease resistance, 2) difficulty in estimating genetic parameters for planning breeding programs against disease, 3) undesirable relationship between production traits and disease, 4) disease as affected by recessive genes, 5) new mutation of the pathogens, and 6) variable environmental factors. The hidden problems of estimating genetic and phenotypic parameters involving disease incidence were examined in terms of categorical nature, non-independence, heterogeneity of error variance, non-randomness, and automatic relationship between disease and production traits. In light of these limitations, the prospect for increasing genetic resistance by conventional breeding methods would not be so bright as we like. Since the phenomenon of disease is the result of a joint interaction among host genotype, pathogen genotype and environment, it becomes essential to adopt an integrated approach of increasing genetic resistance of the host animals, manipulating the pathogen genotypes, developing effective vaccines and drugs, and improving the environmental conditions. The advances in DNA-based technology show considerable promise in directly manipulating host and pathogen genomes for genetic resistance and producing vaccines and drugs for prevention and medication to promote the wellbeing of the animals.

Clinical Benefit of Vaccinating Male Against HPV-related Disease (인유두종바이러스 관련 질환의 예방을 위한 남성 대상 백신의 임상적 유용성)

  • Lee, Sei Young
    • Korean Journal of Head & Neck Oncology
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    • v.38 no.1
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    • pp.11-16
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    • 2022
  • HHPV (Human Papillomavirus) is a DNA virus that can cause benign lesions, genitourinary cancer, and oropharyngeal cancer by penetrating the mucous membrane and skin. It is widely known to be transmitted mainly through sexual contact. As with many viral infections, vaccines have been developed to prevent infection with HPV. Currently, in many countries, HPV vaccines are mainly used for national immunization for women to prevent diseases that traditionally occur frequently in women, especially cervical cancer. However, since the vaccination rate is relatively low, many countries are struggling with ways to increase the vaccination rate. Meanwhile, the incidence of oropharyngeal cancer caused by HPV in men has been increasing recently. In the United States, the annual number of oropharyngeal cancers in men already exceeds the number of cervical cancers in women, so HPV infection in men has emerged as a major problem. Accordingly, interest in HPV vaccination in men has also increased, and studies on the effectiveness and necessity of vaccination of both women and men compared to women alone are being actively conducted. In this paper, the evidence of HPV vaccination for men will be reviewed through previous studies, and its validity and cost-effectiveness will be analyzed to bolster the clinical usefulness of HPV vaccination for men.

Multifaceted Usage of HPV Related Tests and Products in the Management of Cervical Cancer - a Review

  • Nalliah, Sivalingam;Karikalan, Barani;Kademane, Kumaraswamy
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2145-2150
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    • 2015
  • HPV viruses are integral to the development of cervical cancer. The pathogenesis has been extensively studied. To date, numerous HPV tests and products have been developed and successfully utilized in diagnosis, treatment and prevention of cervical cancer. The HPV DNA test, when combined with other routine cervical cancer screening and diagnostic tests namely exfoliative cytology, visual inspection with acetic acid (VIA) and colposcopy has increased the detection rate of cervical cancer. HPV DNA products could also be measured in other body fluids like urine, lymph node tissue, and serum. HPV association could also be quantified by measuring other parameters like HPV mRNA, viral load, viral integration and methylation status. Vaccination against HPV has been found to decrease the incidence of cervical cancer. Further, therapeutic vaccines for cervical cancer against HPV continue to evolve. All these findings pertaining to HPV could possibly decrease the incidence of cervical cancer in the near future. This review aims to give an overview of the HPV tests and products in use and those under trial currently.

Poliovirus Sabin 1 as a Live Vaccine Vector: Expression of HIV-1 p24 Core Protein

  • Jung, Hye-Rhan;Bae, Yong-Soo
    • BMB Reports
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    • v.31 no.5
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    • pp.432-443
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    • 1998
  • The poliovirus Sabin 1 strain has features that make it a particularly attractive live recombinant mucosal vaccine vehicle. Sabin 1 cDNA was manipulated to have multiple cloning sites and a viral specific 3C-protease cutting site at the N-terminal end of the polyprotein. The gene for the N-terminal 169 amino acids of the HIV-1 p24 was cloned into the multiple cloning site of the manipulated Sabin cDNA. A recombinant progeny virus was produced from HeLa cells when it was transfected with the RNA synthesized from the p24-Sabin chimeric cDNA. The recombinant progeny virus expresses substantial amounts of the HIV-1 p24 protein, which was clearly detected in the infected cell lysates and culture supernatants in Western blot experiments with rabbit anti-p24 serum and AIDS patients' sera. Differing from the Mahoney strain, the recombinant Sabin 1 poliovirus maintained the foreign gene stably during the subsequent passages. Replication capacity was about 1 to 1.5 log lower than that of the wild-type Sabin 1. Other physicochemical stability characteristics of the recombinant virus were similar to that of the wild-type Sabin 1. These results suggest that the manipulated Sabin 1 poliovirus can be used as a live viral vaccine vector for the development of mucosal vaccines.

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DNA Vaccines Encoding Toxoplasma gondii Cathepsin C 1 Induce Protection against Toxoplasmosis in Mice

  • Han, Yali;Zhou, Aihua;Lu, Gang;Zhao, Guanghui;Sha, Wenchao;Wang, Lin;Guo, Jingjing;Zhou, Jian;Zhou, Huaiyu;Cong, Hua;He, Shenyi
    • Parasites, Hosts and Diseases
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    • v.55 no.5
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    • pp.505-512
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    • 2017
  • Toxoplasma gondii cathepsin C proteases (TgCPC1, 2, and 3) are important for the growth and survival of T. gondii. In the present study, B-cell and T-cell epitopes of TgCPC1 were predicted using DNAstar and the Immune Epitope Database. A TgCPC1 DNA vaccine was constructed, and its ability to induce protective immune responses against toxoplasmosis in BALB/c mice was evaluated in the presence or absence of the adjuvant ${\alpha}-GalCer$. As results, TgCPC1 DNA vaccine with or without adjuvant ${\alpha}-GalCer$ showed higher levels of IgG and IgG2a in the serum, as well as IL-2 and $IFN-{\gamma}$ in the spleen compared to controls (PBS, pEGFP-C1, and ${\alpha}-GalCer$). Upon challenge infection with tachyzoites of T. gondii (RH), $pCPC1/{\alpha}-GalCer$ immunized mice showed the longest survival among all the groups. Mice vaccinated with DNA vaccine without adjuvant (pCPC1) showed better protective immunity compared to other controls (PBS, pEGFP-C1, and ${\alpha}-GalCer$). These results indicate that a DNA vaccine encoding TgCPC1 is a potential vaccine candidate against toxoplasmosis.

CD8+ T Cell-mediated Immunity Induced by Heterologous Prime-boost Vaccination Based on DNA Vaccine and Recombinant Vaccinia Virus Expressing Epitope (Epitope발현 DNA Vaccine과 Recombinant Vaccinia Virus를 이용한 Heterologous Prime-boost Vaccination에 의하여 유도되는 CD8+ T 세포 매개성 면역)

  • Park, Seong-Ok;Yoon, Hyun-A;Aleyas, Abi George;Lee, John-Hwa;Chae, Joon-Seok;Eo, Seong-Kug
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.89-98
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    • 2005
  • Background: DNA vaccination represents an anticipated approach for the control of numerous infectious diseases. Used alone, however, DNA vaccine is weak immunogen inferior to viral vectors. In recent, heterologous prime-boost vaccination leads DNA vaccines to practical reality. Methods: We assessed prime-boost immunization strategies with a DNA vaccine (minigene, $gB_{498-505}$ DNA) and recombinant vaccinia virus $(vvgB_{498-505})$ expressing epitope $gB_{498-505}$ (SSIEF ARL) of CD8+ T cells specific for glycoprotein B (gB) of herpes simplex virus (HSV). Animals were immunized primarily with $gB_{498-505}$ epitope-expressing DNA vaccine/recombinant vaccinia virus and boosted with alternative vaccine type expressing entire Ag. Results: In prime-boost protocols using vvgBw (recombinant vaccinia virus expressing entire Ag) and $vvgB_{498-505}$, CD8+ T cell-mediated immunity was induced maximally at both acute and memory stages if primed with vvgBw and boosted with $vvgB_{498-505}$ as evaluated by CTL activity, intracellular IFN-staining, and MHC class I tetramer staining. Similarly $gB_{498-505}$ DNA prime-gBw DNA (DNA vaccine expressing entire Ag) boost immunization elicited the strongest CD8+ T cell responses in protocols based on DNA vaccine. However, the level of CD8+ T cell-mediated immunity induced with prime-boost vaccination using DNA vaccine expressing epitope or entire Ag was inferior to those based on vvgBw and $vvgB_{498-505}$. Of particular interest CD8+ T cell-mediated immunity was optimally induced when $vvgB_{498-505}$ was used to prime and gB DNA was used as alternative boost. Especially CD7+ T cell responses induced by such protocol was longer lasted than other protocols. Conclusion: These facts direct to search for the effective strategy to induce optimal CD8+ T cell-mediated immunity against cancer and viral infection.