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Fumio Takaiwa - One of the best experts on this subject based on the ideXlab platform.
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efficacy of oral immunotherapy with a rice based Edible Vaccine containing hypoallergenic japanese cedar pollen allergens for treatment of established allergic conjunctivitis in mice
Allergology International, 2018Co-Authors: Ken Fukuda, Fumio Takaiwa, Waka Ishida, Yosuke Harada, Yuhya Wakasa, Hidenori Takagi, Atsuki FukushimaAbstract:This study was supported by funding from the charitable Trust Fund for Ophthalmic Research in Commemoration of Santen Pharmaceutical's Founder to K.F. as well as by “Agri-Health Translational Research Projects” of the Ministry of Agriculture, Forestry, and Fisheries of Japan to F.T. The funding sources played no role in study design; in collection, analysis, or interpretation of data; or in writing the report.
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prevention of allergic conjunctivitis in mice by a rice based Edible Vaccine containing modified japanese cedar pollen allergens
British Journal of Ophthalmology, 2015Co-Authors: Ken Fukuda, Fumio Takaiwa, Waka Ishida, Yosuke Harada, Yuhya Wakasa, Hidenori Takagi, Atsuki FukushimaAbstract:Background/aims To determine whether oral immunotherapy with transgenic rice seeds expressing hypoallergenic modified antigens suppresses cedar pollen-induced allergic conjunctivitis by eliciting immune tolerance in mice. Methods BALB/c mice were fed once a day for 20 days with 220 mg of transgenic rice expressing modified Japanese cedar pollen allergens Cry j 1 and Cry j 2 or with non-transgenic rice seeds as a control. They were then sensitised with two intraperitoneal injections of Japanese cedar pollen in alum before challenge twice with pollen in eye drops. Twenty-four hours after the second challenge, the conjunctiva, spleen, and blood were isolated for histological analysis, cytokine production assays, and measurement of serum immunoglobulin E concentrations, respectively. Results The numbers of eosinophils and total inflammatory cells in the conjunctiva were significantly lower in mice fed the transgenic rice than in those fed non-transgenic rice. The clinical score evaluated at 15 min after antigen challenge was also significantly lower in mice fed the transgenic rice than in those fed non-transgenic rice. The serum concentrations of both total and allergen-specific immunoglobulin E were also significantly lower in mice fed the transgenic rice. Oral vaccination with transgenic rice resulted in significant down-regulation of the allergen-induced production of interleukin (IL)-2, IL-4, IL-5, IL-12p70, interferon-γ, and IL-17A by splenocytes. Conclusions Oral immunotherapy with transgenic rice expressing modified Japanese cedar pollen allergens suppressed pollen-induced experimental allergic conjunctivitis in mice by eliciting immune tolerance. This novel prophylactic approach is potentially safe and effective for allergen-specific oral immunotherapy in allergic conjunctivitis.
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a whole genome analysis of a transgenic rice seed based Edible Vaccine against cedar pollen allergy
DNA Research, 2013Co-Authors: Taiji Kawakatsu, Yoshihiro Kawahara, Takeshi Itoh, Fumio TakaiwaAbstract:Genetic modification (GM) by Agrobacterium-mediated transformation is a robust and widely employed method to confer new traits to crops. In this process, a transfer DNA is delivered into the host genome, but it is still unclear how the host genome is altered by this event at single-base resolution. To decipher genomic discrepancy between GM crops and their host, we conducted whole-genome sequencing of atransgenicricelineOSCR11.Thisricelineexpressesaseed-basedEdibleVaccinecontainingtwomajorpollenallergens, Cry j 1 and Cry j 2, against Japanese cedar pollinosis. We revealed that genetic differences between OSCR11 and its host a123 were significantly less than those between a123 and its precedent cultivar Koshihikari. The pattern of nucleotide base substitution in OSCR11, relative to a123, was consistent with somaclonal variation. Mutations in OSCR11 probably occurred during the cell culture steps. In addition, strand-specific mRNA-Seq revealed similar transcriptomes of a123 and OSCR11, supporting genomic integrity between them.
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Oral immunogenicity and protective efficacy in mice of transgenic rice plants producing a Vaccine candidate antigen (As16) of Ascaris suum fused with cholera toxin B subunit
Transgenic Research, 2008Co-Authors: Yasunobu Matsumoto, Fumio Takaiwa, Seiko Suzuki, Tomoko Nozoye, Takashi Yamakawa, Yasuhiro Takashima, Takeshi Arakawa, Naotoshi Tsuji, Yoshihiro HayashiAbstract:Cereal crops such as maize and rice are considered attractive for Vaccine production and oral delivery. Here, we evaluated the rice Oryza sativa for production of As16—an antigen protective against the roundworm Ascaris suum . The antigen was produced as a chimeric protein fused with cholera toxin B subunit (CTB), and its expression level in the endosperm reached 50 μg/g seed. Feeding the transgenic (Tg) rice seeds to mice elicited an As16-specific serum antibody response when administered in combination with cholera toxin (CT) as the mucosal adjuvant. Although omitting the adjuvant from the Vaccine formulation resulted in failure to develop the specific immune response, subcutaneous booster immunization with bacterially expressed As16 induced the antibody response, indicating priming capability of the Tg rice. Tg rice/CT-fed mice orally administered A. suum eggs had a lower lung worm burden than control mice. This suggests that the rice-delivered antigen functions as a prophylactic Edible Vaccine for controlling parasitic infection in animals.
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generation of a transgenic rice seed based Edible Vaccine against house dust mite allergy
Biochemical and Biophysical Research Communications, 2008Co-Authors: Lijun Yang, Hiroyuki Kajiura, Kazuya Suzuki, Sakiko Hirose, Kazuhito Fujiyama, Fumio TakaiwaAbstract:Abstract As an alternative approach to conventional allergen-specific immunotherapy, transgenic rice seed expressing a major house dust mite (HDM) allergen, Der p 1, was developed as an Edible Vaccine. The C-terminal KDEL-tagged Der p 1 allergen specifically accumulated in seed endosperm tissue under the control of the endosperm-specific GluB1 promoter. Der p 1 reached a maximum concentration of 58 μg/grain and was deposited in the endoplasmic reticulum (ER)-derived protein body I (PB-I). Plant-derived Der p 1 was posttranslationally modified with high-mannose-type glycan structures. Glycosylated Der p 1 displayed reduced IgE binding capacity in comparison with its unglycosylated counterpart in vitro . Our results indicate that transgenic Der p 1 rice seeds are a safe, potential oral delivery Vaccine for the treatment of HDM allergy.
Judith Strommer - One of the best experts on this subject based on the ideXlab platform.
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Expression of a modified Mannheimia haemolytica GS60 outer membrane lipoprotein in transgenic alfalfa for the development of an Edible Vaccine against bovine pneumonic pasteurellosis.
Journal of Biotechnology, 2008Co-Authors: Raymond W. H. Lee, Judith Strommer, Mette Cornelisse, Asma Ziauddin, Penelope J. Slack, Douglas C. Hodgins, Patricia E ShewenAbstract:Abstract The GS60 antigen is one of the protective antigens of Mannheimia haemolytica A1. GS60 contains conserved domains belonging to the LppC family of bacterial outer membrane lipoproteins. A high antibody titer to GS60 has been shown to be significantly correlated with resistance to pneumonic pasteurellosis. Calves vaccinated with a commercial Vaccine (Presponse™) and demonstrating protection against M. haemolytica A1 produced antibodies directed against GS60. Alfalfa was chosen as the platform for an Edible Vaccine. Agrobacterium tumefaciens was used to mediate the transformation of alfalfa with sequences encoding a slightly shortened derivative of the GS60 antigen (GS60 54 ). Stable transgenic alfalfa lines were recovered and production of GS60 54 was examined by Western immunoblot analysis. The antigen is stable in dried transgenic plant material stored at ambient temperature for more than a year. The plant-produced GS60 54 protein was shown to be immunogenic when injected into rabbits. Feeding of the dried transgenic alfalfa expressing the GS60 54 to rabbits is capable of inducing seroconversion, suggesting that GS60 54 could be an effective oral antigen for stimulating mucosal immune responses.
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Edible Vaccine development: stability of Mannheimia haemolytica A1 leukotoxin 50 during post-harvest processing and storage of field-grown transgenic white clover
Molecular Breeding, 2003Co-Authors: Amanda N. Pool, Reggie Y C Lo, Patricia E Shewen, Asma Ziauddin, Judith StrommerAbstract:Mannheimia haemolytica is the principal microorganism responsible for bovine pneumonic pasteurellosis, or shipping fever. We have previously expressed a fragment of leukotoxin, a major virulent factor of M. haemolytica A1, as a fusion protein with green fluorescent protein (GFP) in transgenic white clover and demonstrated that this antigen was immunogenic and elicited toxin neutralizing antibodies in rabbits. These previous results showed that using plants to produce M. haemolytica antigen for use as a Vaccine against this disease is a viable strategy. In this present study, we examined the stability of the truncated leukotoxin GFP-fusion protein (Lkt50-GFP) in field-grown transgenic white clover. Transgenic clover expressing Lkt50-GFP was clonally propagated and a confined field trial was established. Western immunoblotting showed that the level of Lkt50-GFP expression in field plants was the same as in transgenic plants maintained under optimal conditions in the greenhouse. We also observed that after harvesting and oven drying at 50 °C, the antigen was still present in the dried clover after 1 year of storage at ambient temperature. As special post-harvest conditions (e.g., refrigeration) are not required, the use of transgenic plants to deliver an oral Vaccine against shipping fever appears to be economically feasible.
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towards development of an Edible Vaccine against bovine pneumonic pasteurellosis using transgenic white clover expressing a mannheimia haemolytica a1 leukotoxin 50 fusion protein
Infection and Immunity, 2001Co-Authors: Judith Strommer, Doug Hodgins, Patricia E Shewen, Reggie Y C LoAbstract:Development of Vaccines against bovine pneumonia pasteurellosis, or shipping fever, has focused mainly on Mannheimia haemolytica A1 leukotoxin (Lkt). In this study, the feasibility of expressing Lkt in a forage plant for use as an Edible Vaccine was investigated. Derivatives of the M. haemolytica Lkt in which the hydrophobic transmembrane domains were removed were made. Lkt66 retained its immunogenicity and was capable of eliciting an antibody response in rabbits that recognized and neutralized authentic Lkt. Genes encoding a shorter Lkt derivative, Lkt50, fused to a modified green fluorescent protein (mGFP5), were constructed for plant transformation. Constructs were screened by Western immunoblot analysis for their ability to express the fusion protein after agroinfiltration in tobacco. The fusion construct pBlkt50-mgfp5, which employs the cauliflower mosaic virus 35S promoter for transcription, was selected and introduced into white clover by Agrobacterium tumefaciens-mediated transformation. Transgenic lines of white clover were recovered, and expression of Lkt50-GFP was monitored and confirmed by laser confocal microscopy and Western immunoblot analysis. Lkt50-GFP was found to be stable in clover tissue after drying of the plant material at room temperature for 4 days. An extract containing Lkt50-GFP from white clover was able to induce an immune response in rabbits (via injection), and rabbit antisera recognized and neutralized authentic Lkt. This is the first demonstration of the expression of an M. haemolytica antigen in plants and paves the way for the development of transgenic plants expressing M. haemolytica antigens as an Edible Vaccine against bovine pneumonic pasteurellosis.
Reggie Y C Lo - One of the best experts on this subject based on the ideXlab platform.
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Edible Vaccine development: stability of Mannheimia haemolytica A1 leukotoxin 50 during post-harvest processing and storage of field-grown transgenic white clover
Molecular Breeding, 2003Co-Authors: Amanda N. Pool, Reggie Y C Lo, Patricia E Shewen, Asma Ziauddin, Judith StrommerAbstract:Mannheimia haemolytica is the principal microorganism responsible for bovine pneumonic pasteurellosis, or shipping fever. We have previously expressed a fragment of leukotoxin, a major virulent factor of M. haemolytica A1, as a fusion protein with green fluorescent protein (GFP) in transgenic white clover and demonstrated that this antigen was immunogenic and elicited toxin neutralizing antibodies in rabbits. These previous results showed that using plants to produce M. haemolytica antigen for use as a Vaccine against this disease is a viable strategy. In this present study, we examined the stability of the truncated leukotoxin GFP-fusion protein (Lkt50-GFP) in field-grown transgenic white clover. Transgenic clover expressing Lkt50-GFP was clonally propagated and a confined field trial was established. Western immunoblotting showed that the level of Lkt50-GFP expression in field plants was the same as in transgenic plants maintained under optimal conditions in the greenhouse. We also observed that after harvesting and oven drying at 50 °C, the antigen was still present in the dried clover after 1 year of storage at ambient temperature. As special post-harvest conditions (e.g., refrigeration) are not required, the use of transgenic plants to deliver an oral Vaccine against shipping fever appears to be economically feasible.
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towards development of an Edible Vaccine against bovine pneumonic pasteurellosis using transgenic white clover expressing a mannheimia haemolytica a1 leukotoxin 50 fusion protein
Infection and Immunity, 2001Co-Authors: Judith Strommer, Doug Hodgins, Patricia E Shewen, Reggie Y C LoAbstract:Development of Vaccines against bovine pneumonia pasteurellosis, or shipping fever, has focused mainly on Mannheimia haemolytica A1 leukotoxin (Lkt). In this study, the feasibility of expressing Lkt in a forage plant for use as an Edible Vaccine was investigated. Derivatives of the M. haemolytica Lkt in which the hydrophobic transmembrane domains were removed were made. Lkt66 retained its immunogenicity and was capable of eliciting an antibody response in rabbits that recognized and neutralized authentic Lkt. Genes encoding a shorter Lkt derivative, Lkt50, fused to a modified green fluorescent protein (mGFP5), were constructed for plant transformation. Constructs were screened by Western immunoblot analysis for their ability to express the fusion protein after agroinfiltration in tobacco. The fusion construct pBlkt50-mgfp5, which employs the cauliflower mosaic virus 35S promoter for transcription, was selected and introduced into white clover by Agrobacterium tumefaciens-mediated transformation. Transgenic lines of white clover were recovered, and expression of Lkt50-GFP was monitored and confirmed by laser confocal microscopy and Western immunoblot analysis. Lkt50-GFP was found to be stable in clover tissue after drying of the plant material at room temperature for 4 days. An extract containing Lkt50-GFP from white clover was able to induce an immune response in rabbits (via injection), and rabbit antisera recognized and neutralized authentic Lkt. This is the first demonstration of the expression of an M. haemolytica antigen in plants and paves the way for the development of transgenic plants expressing M. haemolytica antigens as an Edible Vaccine against bovine pneumonic pasteurellosis.
Moon-sik Yang - One of the best experts on this subject based on the ideXlab platform.
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Expression of Dengue virus EIII domain-coding gene in maize as an Edible Vaccine candidate
Journal of Plant Biotechnology, 2014Co-Authors: Hyun A Kim, Moon-sik Yang, Suk-yoon Kwon, Pil Son ChoiAbstract: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.
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Current trends in Edible Vaccine development using transgenic plants
Biotechnology and Bioprocess Engineering, 2010Co-Authors: Tae-geum Kim, Moon-sik YangAbstract:Immunology textbooks currently report orally administered antigens as inducing immune tolerance rather than immune stimulation. Nevertheless, current plant-based Edible Vaccine technology, if sufficiently developed, may offer several advantages. For example, it is easy to apply, store, and transport. It could also induce both mucosal and systemic immune responses, which cannot be achieved using an injection Vaccine. Plant-based Vaccines are also anticipated to prove quite useful in the animal industry, since the cost of injection is a significant burden in the animal industry. Although no commercial plant-based Edible Vaccines are currently available, several candidate Vaccines are undergoing clinical trials. Consequently, many scientists are anticipating that a commercial plant-based Edible Vaccine will be available in the near future.
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Expression and Immunogenicity of Enterotoxigenic Escherichia coli Heat-Labile Toxin B Subunit in Transgenic Rice Callus
Molecular Biotechnology, 2009Co-Authors: Jae-kwon Choi, Eun-sun Jung, Moon-sik YangAbstract:Enterotoxigenic Escherichia coli is one of the leading causes of diarrhea in developing countries, and the disease may be fatal in the absence of treatment. Enterotoxigenic E. coli heat-labile toxin B subunit (LTB) can be used as an adjuvant, as a carrier of fused antigens, or as an antigen itself. The synthetic LTB (sLTB) gene, optimized for plant codon usage, has been introduced into rice cells by particle bombardment-mediated transformation. The integration and expression of the sLTB gene were observed via genomic DNA PCR and western blot analysis, respectively. The binding activity of LTB protein expressed in transgenic rice callus to G_M1-ganglioside, a receptor for biologically active LTB, was confirmed by G_M1-ELISA. Oral inoculation of mice with lyophilized transgenic rice calli containing LTB generated significant IgG antibody titers against bacterial LTB, and the sera of immunized mice inhibited the binding of bacterial LTB to G_M1-ganglioside. Mice orally immunized with non-transgenic rice calli failed to generate detectable anti-LTB IgG antibody titers. Mice immunized with plant-produced LTB generated higher IgG1 antibody titers than IgG2a, indicating a Th2-type immune response. Mice orally immunized with lyophilized transgenic rice calli containing LTB elicited higher fecal IgA antibody titers than mice immunized with non-transgenic rice calli. These experimental results demonstrate that LTB proteins produced in transgenic rice callus and given to mice by oral administration induce humoral and secreted antibody immune responses. We suggest that transgenic rice callus may be suitable as a plant-based Edible Vaccine to provide effective protection against enterotoxigenic E. coli heat-labile toxin.
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Expression of a synthetic neutralizing epitope of porcine epidemic diarrhea virus fused with synthetic B subunit of Escherichia coli heat labile enterotoxin in rice endosperm
Molecular Biotechnology, 2007Co-Authors: Maria Oszvald, Tae-geum Kim, Tae-jin Kang, Sandor Tomoskozi, Cecilia Tamas, Laszlo Tamas, Moon-sik YangAbstract:Epitopes often require co-delivery with adjuvant and targeting proteins to enable recognition by the immune system, and this approach may also increase the efficacy of the antigen. In this study, we assess and describe the ability of transgenic rice plants to express a fusion protein consisting of the B-subunit of the Escherichia coli heat-labile enterotoxin (LTB) and a synthetic core-neutralizing epitope (COE) of porcine epidemic diarrhea virus (PEDV), inducing an enteric disease that is seen most predominantly in piglets. Both components of the fusion proteins were detected with Western blot analysis. The fusion protein was determined to assemble into pentamers, as was evidenced by its ability to bind to GM_1 gangliosides, and evidenced an average level of expression in a trangenic rice endosperm. This indicates that the expression system of the plant is capable of generating a sizable amount of antigen, possibly allowing for the successful development of an Edible Vaccine.
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Expression of neutralizing epitope of porcine epidemic diarrhea virus in potato plants
Plant Cell Tissue and Organ Culture, 2005Co-Authors: Tae-jin Kang, Yong-suk Jang, Moon-sik YangAbstract:Transgenic plants expressing recombinant proteins from pathogenic microorganisms provide an inexpensive Edible Vaccine for induction of local immunity. A neutralizing epitope of porcine epidemic diarrhea virus (PEDV) gene containing SEKDEL was expressed in potato using Agrobacterium -mediated transformation system. Putative transgenic plants were regenerated, and genomic PCR confirmed the presence of PEDV epitope gene in the potato plants. Based on the ELISA results, epitope of PEDV protein made up approximately 0.1% of the total soluble tuber protein.
Patricia E Shewen - One of the best experts on this subject based on the ideXlab platform.
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Expression of a modified Mannheimia haemolytica GS60 outer membrane lipoprotein in transgenic alfalfa for the development of an Edible Vaccine against bovine pneumonic pasteurellosis.
Journal of Biotechnology, 2008Co-Authors: Raymond W. H. Lee, Judith Strommer, Mette Cornelisse, Asma Ziauddin, Penelope J. Slack, Douglas C. Hodgins, Patricia E ShewenAbstract:Abstract The GS60 antigen is one of the protective antigens of Mannheimia haemolytica A1. GS60 contains conserved domains belonging to the LppC family of bacterial outer membrane lipoproteins. A high antibody titer to GS60 has been shown to be significantly correlated with resistance to pneumonic pasteurellosis. Calves vaccinated with a commercial Vaccine (Presponse™) and demonstrating protection against M. haemolytica A1 produced antibodies directed against GS60. Alfalfa was chosen as the platform for an Edible Vaccine. Agrobacterium tumefaciens was used to mediate the transformation of alfalfa with sequences encoding a slightly shortened derivative of the GS60 antigen (GS60 54 ). Stable transgenic alfalfa lines were recovered and production of GS60 54 was examined by Western immunoblot analysis. The antigen is stable in dried transgenic plant material stored at ambient temperature for more than a year. The plant-produced GS60 54 protein was shown to be immunogenic when injected into rabbits. Feeding of the dried transgenic alfalfa expressing the GS60 54 to rabbits is capable of inducing seroconversion, suggesting that GS60 54 could be an effective oral antigen for stimulating mucosal immune responses.
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Edible Vaccine development: stability of Mannheimia haemolytica A1 leukotoxin 50 during post-harvest processing and storage of field-grown transgenic white clover
Molecular Breeding, 2003Co-Authors: Amanda N. Pool, Reggie Y C Lo, Patricia E Shewen, Asma Ziauddin, Judith StrommerAbstract:Mannheimia haemolytica is the principal microorganism responsible for bovine pneumonic pasteurellosis, or shipping fever. We have previously expressed a fragment of leukotoxin, a major virulent factor of M. haemolytica A1, as a fusion protein with green fluorescent protein (GFP) in transgenic white clover and demonstrated that this antigen was immunogenic and elicited toxin neutralizing antibodies in rabbits. These previous results showed that using plants to produce M. haemolytica antigen for use as a Vaccine against this disease is a viable strategy. In this present study, we examined the stability of the truncated leukotoxin GFP-fusion protein (Lkt50-GFP) in field-grown transgenic white clover. Transgenic clover expressing Lkt50-GFP was clonally propagated and a confined field trial was established. Western immunoblotting showed that the level of Lkt50-GFP expression in field plants was the same as in transgenic plants maintained under optimal conditions in the greenhouse. We also observed that after harvesting and oven drying at 50 °C, the antigen was still present in the dried clover after 1 year of storage at ambient temperature. As special post-harvest conditions (e.g., refrigeration) are not required, the use of transgenic plants to deliver an oral Vaccine against shipping fever appears to be economically feasible.
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towards development of an Edible Vaccine against bovine pneumonic pasteurellosis using transgenic white clover expressing a mannheimia haemolytica a1 leukotoxin 50 fusion protein
Infection and Immunity, 2001Co-Authors: Judith Strommer, Doug Hodgins, Patricia E Shewen, Reggie Y C LoAbstract:Development of Vaccines against bovine pneumonia pasteurellosis, or shipping fever, has focused mainly on Mannheimia haemolytica A1 leukotoxin (Lkt). In this study, the feasibility of expressing Lkt in a forage plant for use as an Edible Vaccine was investigated. Derivatives of the M. haemolytica Lkt in which the hydrophobic transmembrane domains were removed were made. Lkt66 retained its immunogenicity and was capable of eliciting an antibody response in rabbits that recognized and neutralized authentic Lkt. Genes encoding a shorter Lkt derivative, Lkt50, fused to a modified green fluorescent protein (mGFP5), were constructed for plant transformation. Constructs were screened by Western immunoblot analysis for their ability to express the fusion protein after agroinfiltration in tobacco. The fusion construct pBlkt50-mgfp5, which employs the cauliflower mosaic virus 35S promoter for transcription, was selected and introduced into white clover by Agrobacterium tumefaciens-mediated transformation. Transgenic lines of white clover were recovered, and expression of Lkt50-GFP was monitored and confirmed by laser confocal microscopy and Western immunoblot analysis. Lkt50-GFP was found to be stable in clover tissue after drying of the plant material at room temperature for 4 days. An extract containing Lkt50-GFP from white clover was able to induce an immune response in rabbits (via injection), and rabbit antisera recognized and neutralized authentic Lkt. This is the first demonstration of the expression of an M. haemolytica antigen in plants and paves the way for the development of transgenic plants expressing M. haemolytica antigens as an Edible Vaccine against bovine pneumonic pasteurellosis.
