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Plant-derived vaccines against respiratory syncytial virus

a technology of respiratory syncytial virus and plant-derived vaccines, which is applied in the field of vaccines against respiratory syncytial virus (rsv), can solve the problems of limited immunity, limitations of such approaches, and natural infection confer, and achieve the effect of enhancing antibody production

Inactive Publication Date: 2005-06-16
KRASNYANSKI SERGEI F +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A major difficulty in developing an effective RSV vaccine has been the fact that natural infection confers, at most, only temporary protection against the disease (1).
However, there are limitations imposed by such approaches.
For example, live infection does not provide full immunity and single point mutations may revert to wild type with undesirable consequences.

Method used

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  • Plant-derived vaccines against respiratory syncytial virus
  • Plant-derived vaccines against respiratory syncytial virus
  • Plant-derived vaccines against respiratory syncytial virus

Examples

Experimental program
Comparison scheme
Effect test

example 1

Results and Discussion for Example 1

[0114] A series of chimeric nucleic acid constructs were generated by fusing an AMV RNA4 leader and an enhancer (P268) to the CaMV 35S promoter driving the expression of the RSV-F gene (FIG. 1). The recombinant RSV-F gene expression was analyzed by ELISA following PEG-mediated transfection of apple leaf mesophyll protoplasts.

[0115] A trace amount of non-specific antibody binding was observed in protoplasts transfected with the TE buffer in the absence of DNA. To analyze the results, the absorbence in protoplasts transfected with chimeric gene constructs was compared to that of the promoterless control (pJSS5) after subtracting the absorbence of the protoplasts transfected with the TE buffer. Protoplasts transfected with the promoterless RSV-F construct (pJSS5) produced a low level of RSV-F antigen (A405nm=0.002±0.001) (FIG. 2).

[0116] The mean A405nm value from protoplasts transfected with the plasmid containing the RSV-F gene inserted downstream...

example 2

Results and Discussion for Example 2

[0126] Generation of transgenic tomato plants producing RSV-F antigen. The human RSV gene encoding the F protein was placed under the transcriptional control of either the constitutively expressed CaMV 35S promoter (37) or the fruit specific E8 promoter (38) to make chimeric gene constructs pJSS3 and pJSS4 (FIG. 4). Through Agrobacterium-mediated transformation, 30 independent transgenic tomato plants were generated with each construct and confirmed by Southern blotting (data not shown). ELISA analysis demonstrated that recombinant RSV-F antigen was expressed in the tomato plant cells. Sixteen transgenic plants containing pJSS3 and 19 containing pJSS4 expressed the RSV-F antigen at levels ranging from 1.0 to 32.5 μg / g of fruit fresh weight. The amount of RSV-F in the plant tissue varied among different transgenic lines; however, the average level in transgenic plants containing the E8 and CaMV 35S promoters was similar, 12.68±2.55 and 9.01±1.87 μg...

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Abstract

A plant-derived vaccine against respiratory syncytial virus (RSV) is disclosed. The vaccine includes an immunogenic complex that includes plant cells transformed with a chimeric gene containing a nucleotide sequence adapted for protein expression in plants and an RSV coding sequence that encodes an antigenic protein of RSV. Also disclosed are methods of making the plant-derived vaccine of the invention, as well as transgenic plants, transgenic plant cells, and nucleic acid constructs useful in immunizing a mammal against RSV.

Description

[0001] The present application claims benefit of U.S. provisional application No. 60 / 133,536, filed May 11, 1999.TECHNICAL FIELD [0002] The present invention generally relates to vaccines against respiratory syncytial virus (RSV) and to plant-derived vaccines against RSV. The vaccines may be used prophylatically and / or therapeutically to treat RSV infection. BACKGROUND ART [0003] Respiratory syncytial virus (RSV) is an RNA virus of the genus Pneumovirus containing two main antigenic glycoproteins, F and G (1). Additional structural proteins include a major nucleocapsid protein (N), a nucleocapsid phosphoprotein (P), a large nucleocapsid protein (L), an envelope matrix protein (M), and a glycoprotein (22,000 daltons) (138). A virally encoded protein of about 9,500 daltons and other small proteins are also known to be in infected cells (138). Infections with RSV are referable to all segments of the respiratory tract, and are usually associated with fever, cough, runny nose and fatigue...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/00C07K14/135C12N15/45C12N15/82
CPCA61K39/00C12N2760/18522C12N15/8258C07K14/005A61P31/14
Inventor BUETOW, DENNISKORBAN, SCHUYLERSANDHU, JAGDEEPKRASNYANSKI, SERGEI
Owner KRASNYANSKI SERGEI F
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