Expression of human cytomegalovirus pp150 gene in transgenic Vicia faba L. and immunogenicity of pp150 protein in mice.
The pp150 gene of human cytomegalovirus (HCMV) was transferred into Vicia faba plants by Agrobacterium tumefaciens-mediated transformation. Three of five hygromycin resistant V. faba plants were identified as positive by PCR and dot-blot hybridization. The ELISA results indicated that pp150 protein from three plants of transformed V. faba leaves and seeds made up 0.005-0.015% of the total soluble protein. The results of detection by immunoblot and inhibition of immunofluorescent assay (IFA) showed that pp150 soluble protein had immunity activity. HCMV pp150-specific antibody (IgG, IgA) and IFN-gamma producing T cells were detected in 100% of the mice immunized with pp150 transgenic V. faba seeds by ELISA and intracellular staining and flow cytometry analysis, respectively. The transgenic V. faba plants will provide new material for the development of edible vaccination against HCMV infection.
Yan
Hua
H
Department of Microbiology and Immunology, School of Medicine, Yangzhou University, Yangzhou 225001, PR China. yzyanhua@yahoo.com.cn
Yan
Huishen
H
Li
Guocai
G
Gong
Weijuan
W
Jiao
Hongmei
H
Chen
Hongju
H
Ji
Mingchun
M
D000821:Animal Feed; D000824:Animal Nutritional Physiological Phenomena / Q000276:immunology; D000818:Animals; D000917:Antibody Formation / Q000502:physiology; D002478:Cells, Cultured; D005260:Female; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010750:Phosphoproteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D018515:Plant Leaves / Q000276:immunology / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000276:immunology; D012639:Seeds / Q000276:immunology / Q000378:metabolism; D014170:Transformation, Genetic; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D031307:Vicia faba / Q000235:genetics* / Q000276:immunology / Q000378:metabolism / Q000502:physiology; D014763:Viral Matrix Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism
D010750:Phosphoproteins; D014614:Vaccines, Synthetic; D014763:Viral Matrix Proteins; C056367:pp150 protein, Cytomegalovirus