• Journal of Plant Biotechnology
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• 제31권2호
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• pp.151-161
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• 2004

Transgenic plants have been studied as delivery system for edible vaccine against various diseases. Edible plant vaccines have several potential advantages as follows: an inexpensive source of antigen, easy administration, reduced need for medical personnel, economical to mass produce and easy transport, heat-stable vaccine without refrigerator, generation of systemic and mucosal immunity and safe antigen without fetal animal-virus contaminants. The amount of recombinant antigens in transgenic plants ranged from 0.002 to 0.8％ in total soluble protein, depending on promoters for the expression of interested genes and plants to be used for transformation. Throughout the last decade, edible plant vaccine made notable progresses that protect from challenges against virus or bacteria. However edible plant vaccines have still problems that could be solved. First, the strong promoter or inducible promoter or strategy of protein targeting could be solved to improve the low expression of antigens in transgenic plants. Second, the transformation technique of target plant should be developed to be able to eat uncooked. Third, marker-free vector could be constructed to be more safety. In this review we describe advances of edible plant vaccines, focusing on the yields depending on plants/promoters employed and the results of animal/clinical trials, and consider further research for the development of a new plant-derived vaccine.

• Journal of Plant Biotechnology
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• 제37권3호
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• pp.250-261
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• 2010

The expression of vaccine antigens in transgenic plants has the potential to provide a convenient, stable, safe approach for oral vaccination alternative to traditional parenteral vaccines. Over the past two decades, many different vaccine antigens expressed via the plant nuclear genome have elicited appropriate immunoglobulin responses and have conferred protection upon oral delivery. Up to date, efforts to produce antigen proteins in plants have focused on potato, tobacco, tomato, banana, and seed (maize, rice, soybean, etc). The choice of promoters affects transgene transcription, resulting in changes not only in concentration, but also in the stage tissue and cell specificity of its expression. Inclusion of mucosal adjuvants during immunization with the vaccine antigen has been an important step towards the success of plant-derived vaccines. In animal and Phase I clinical trials several plant-derived vaccine antigens have been found to be safe and induce sufficiently high immune response. Future areas of research should further characterize the induction of the mucosal immune response and appropriate dosage for delivery system of animal and human vaccines. This article reviews the current status of development in the area of the use of plant for the development of oral vaccines.

• Journal of Plant Biotechnology
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• 제41권1호
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• pp.50-55
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• 2014

Plant-based vaccines possess some advantages over other types of vaccine biotechnology such as safety, low cost of mass vaccination programs, and wider use of vaccines for medicine. This study was undertaken to develop the transgenic maize as edible vaccine candidates for humans. The immature embryos of HiII genotype were inoculated with A. tumefaciens strain C58C1 containing the binary vectors (V662 or V663). The vectors carrying nptII gene as selection marker and scEDIII (V662) or wCTB-scEDIII (V663) target gene, which code EIII proteins inhibite viral adsorption by cells. In total, 721 maize immature embryos were transformed and twenty-two putative transgenic plants were regenerated after 12 weeks selection regime. Of them, two- and six-plants were proved to be integrated with scEDIII and wCTB-scEDIII genes, respectively, by Southern blot analysis. However, only one plant (V662-29-3864) can express the gene of interest confirmed by Northern blot analysis. These results demonstrated that this plant could be used as a candidated source of the vaccine production.

• 대한수의학회지
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• 제53권4호
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• pp.217-224
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• 2013

Transgenic plants have been tested as an alternative host for the production and delivery of experimental oral vaccines. Here, we developed transgenic potatoes that express the major antigenic sites A and D of the glycoprotein S from transmissible gastroenteritis coronavirus (TGEV-$S_{0.7}$) under three expression vector systems. The DNA integration and mRNA expression level of the TGEV-$S_{0.7}$ gene were confirmed in transgenic plants by PCR and northern blot analysis. Antigen protein expression in transgenic potato was determined by western blot analysis. Enzyme-linked immunosorbent assay results revealed that based on a dilution series of Escherichia coli-derived antigen, the transgenic line P-2 had TGEV-$S_{0.7}$ protein at levels that were 0.015% of total soluble proteins. We then examined the immunogenicity of potato-derived TGEV-$S_{0.7}$ antigen in mice. Compared with the wild-type potato treated group and synthetic antigen treated group, mice treated with the potato-derived antigen showed significantly higher levels of immunoglobulin (Ig) G and IgA responses.

• Journal of Animal Science and Technology
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• 제57권4호
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• pp.10.1-10.7
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• 2015

Control and prevention of foot and mouth disease (FMD) by vaccination remains unsatisfactory in endemic countries. Indeed, consistent and new FMD epidemics in previously disease-free countries have precipitated the need for a worldwide control strategy. Outbreaks in vaccinated animals require that a new and safe vaccine be developed against foot and mouth virus (FMDV). FMDV can be eradicated worldwide based on previous scientific information about its spread using existing and modern control strategies.

• 대한수의학회지
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• 제51권2호
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• pp.151-158
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• 2011

The study evaluated whether a transgenic carrot vaccine could induce a K88-specific immune response in sows and whether the resultant maternal antibody could protect piglets against enterotoxigenic Escherichia coli (ETEC) K88ac infection. Sows (n = 15) selected randomly from a farm in Korea were assigned to three groups (n = 5 per group: control [untreated]), group A (orally inoculated with a nontransgenic and transgenic carrot vaccines at 2 and 4 weeks ante partum, respectively), and group B (conventionally vaccinated according to the manufacturer's instructions). After 7 days of lactation, 5 piglets selected randomly from each group were challenged with $1{\times}10^{10}$ colony forming units/mL ETEC K88ac. Group C had the lowest mean fecal consistency score on post-challenge days 1 and 7. Histiologically, On post-challenge day 7, group C showed an increased duodenum and ileum villus:crypt ratio, compared to group A in the duodenum, with group B displaying the highest ratio. Groups B and C had more increased villus width than group A in the jejunum. Group C displayed the greatest increase in villus width in the ileum. The colostrums and serum from groups B and C displayed higher concentrations of IgA and IgG against ETEC K88, compared to group A. Based on the results, it was concluded that the transgenic carrot vaccine in sow per oral may have an effect on preventing piglet diarrhea as good as commercial recombinant vaccine.

• KSBB Journal
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• 제26권6호
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• pp.491-504
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• 2011

This review article provides an overview of the evolution of diphtheria vaccine, its value and its future. Diphtheria is an infectious illness caused by diphtheria toxin produced by pathogenic strains of Corynebacterium diphtheriae. It is characterized by a sore throat with membrane formation due to local tissue necrosis, which can lead to fatal airway obstruction; neural and cardiac damage are other common complications. Diphtheria vaccine was first brought to market in the 1920s, following the discovery that diphtheria toxin can be detoxified using formalin. However, conventional formalin-inactivated toxoid vaccines have some fundamental limitations. Innovative technologies and approaches with the potential to overcome these limitations are discussed in this paper. These include genetic inactivation of diphtheria toxoid, innovative vaccine delivery systems, new adjuvants (both TLR-independent and TLR-dependent adjuvants), and heat- and freeze-stable agents, as well as novel platforms for producing improved conventional vaccine, DNA vaccine, transcutaneous (microneedle-mediated) vaccine, oral vaccine and edible vaccine expressed in transgenic plants. These innovations target improvements in vaccine quality (efficacy, safety, stability and consistency), ease of use and/or thermal stability. Their successful development and use should help to increase global diphtheria vaccine coverage.

• 생명과학회지
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• 제19권1호
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• pp.123-128
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• 2009

식물 생명공학 기술을 이용해 인간에게 유용한 치료단백질 및 백신을 생산하는 것은 최근에 각광받고 있는 연구 분야이다. 식물을 이용한 유용 단백질 생산은 다른 시스템에 비하여 경제적일 뿐만 아니라 병원성 인자에 대한 안전성이 있어서 유용하다고 할 수 있다. 암세포 표면에 특이적으로 발현하고 있는 분자 와 당 구조를 각각 인지할 수 있는 두 종류의 항체를 동시에 투여하는 면역 치료는 질병의 치료를 유도하는 데 있어서 효과적일 수 있다. 본 연구는 기존에 본 연구팀에서 확보하고 있었던 두 종류의 항체 단백질(mAb CO17-1A, mAb BR55) 생산 형질전환 식물체를 이용하여 상호교배를 통하여 한 식물에서 두 종류의 항체 단백질을 모두 생산하는 식물 발현 시스템 구축에 관한 연구이다. 각기 다른 유전자를 갖고 있는 식물체로부터 수분을 유도하여 씨앗을 얻고 이 씨앗을 배양하여 완벽한 식품 개체로 성장시켰으며, 그 식물체로부터 DNA, RNA, 단백질을 분리하여 형질전환 유전자를 포함하고 있는지 여부를 확인하였다. 그 결과, 개체에 차이는 있지만, 한 식물에서 두 항체 유전자를 갖고 있음을 확인할 수 있었고, 이 유전자는 식물체 내에서 안정적으로 transcription 되었음을 확인하였다. 또한, 두 종류의 항체를 동시 생산하는 식물체에서 분리한 단백질은 한 종류의 항체 단백질만 생산하는 식물체에 비하여 수용성 단백질 단위당 항체 발현률이 높게 나타나는 것을 확인하였다. 따라서 본 연구를 통하여 식물을 이 용한 유용 단백질 생산 효율을 높일 수 있는 시스템을 확립하였으며 앞으로 추가적으로 생산한 항체의 생물학적 활성 및 항암 효능, 당 구조 분석 등에 대한 연구용 수행한다면, 식물 생명공학적 방법을 통한 항체 생산에 대한 새로운 가능성을 제시할 수 있을 것으로 기대된다.

• Journal of Plant Biotechnology
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• 제37권3호
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• pp.275-282
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• 2010

고등식물의 엽록체 형질전환은 핵 형질전환에서 기대 할 수 없는 여러 가지 이점을 가진다. 외래 단백질의 발현율을 획기적으로 높일 수 있고, 여러 유전자를 동시에 발현시킬 수 있으며, 상동재조합에 의한 부위-특이적 유전자 삽입으로 인해 유전자 침묵 및 위치효과가 없다. 더욱이, 대부분 작물은 화분을 통한 도입된 유전자의 전이가 불가능한 모계 유전을 하기 때문에 엽록체 형질전환은 환경 친화적이다. 엽록체 형질전환 시스템은 핵 형질 전환과 달리 작물에서의 성공에 제한적이었으나 지난 10년 동안 이런 한계가 극복되어 콩, 당근, 상추 및 유채 등의 작물에서도 성공하게 되었다. 그러므로 이제 작물의 엽록체 형질전환은 농업적 형질의 개선뿐 만 아니라, 고부가가치 백신과 의료용 단백질 생산을 통한 의약품 산업의 성장에 활용될 수 있을 것이다.

• Journal of Microbiology and Biotechnology
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• 제16권12호
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• pp.1919-1927
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• 2006

Tumor cells genetically engineered to secrete cytokines are effective in tumor therapy, but various unexpected side effects are observed, which may result from the bulk activation of various bystander cells. In this study, we tested tumor vaccines expressing various membrane-bound forms of IL-2 (mbIL-2) on MethA fibrosarcoma cells to focus antitumor immune responses to CTL. Chimeric forms of IL-2 with whole CD4, deletion forms of CD4, and TNF were expressed on the tumor cell surface, respectively. Tumor clones expressing mbIL-2 or secretory form of IL-2 were able to support the cell growth of CTLL-2, an IL-2-dependent T cell line, and the proliferation of spleen cells from 2C TCR transgenic mice that are responsive to the $p2Ca/L^d$ MHC class I complex. Expression of mbIL-2 on tumor cells reduced the tumorigenicity of tumor cells, and the mice that once rejected the live IL-2/TNF tumor clone acquired systemic immunity against wild-type MethA cells. The IL-2/TNF clone was inferior to other clones in tumor formation, and superior in the stimulation of the CD8+ T cell population in vitro. These results suggest that the IL-2/TNF clone is the best tumor vaccine, and may stimulate CD8+ T cells by direct priming. Expression of IL-2/TNF on tumor cells may serve as an effective gene therapy method to ameliorate the side effects encountered in the recombinant cytokine therapy and the conventional cytokine gene therapy using the secretory form of IL-2.