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Research of recombinant influenza A virus as a vector for Mycoplasma pneumoniae P1a and P30a.
Yu, Liang; Yongbo, Wang; Shengjun, Yang; Jia, Tan; Ya, Xu; Guoyang, Liao; Linna, Ma.
Afiliación
  • Yu L; Department of Clinical Laboratory, The First People's Hospital of Yunnan Province, Kunming, China.
  • Yongbo W; Department of Clinical Laboratory, The First People's Hospital of Yunnan Province, Kunming, China.
  • Shengjun Y; Department of Clinical Laboratory, The First People's Hospital of Yunnan Province, Kunming, China.
  • Jia T; Department of Clinical Laboratory, The First People's Hospital of Yunnan Province, Kunming, China.
  • Ya X; Department of Clinical Laboratory, The First People's Hospital of Yunnan Province, Kunming, China.
  • Guoyang L; The Fifth Department of Biological Products, Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming, China.
  • Linna M; Department of Medical Laboratory Technique, Kunming Medical University Haiyuan College, Kunming, China.
Immun Inflamm Dis ; 12(9): e70021, 2024 Sep.
Article en En | MEDLINE | ID: mdl-39291404
ABSTRACT

BACKGROUND:

Mycoplasma pneumoniae (MP) is a common respiratory pathogen affecting the longevity of the elderly and the health of children. However, the human vaccine against MP has not been successfully developed till now due to the poor immunogenicity and side effects of MP inactivated or attenuated vaccine. Therefore, it is necessary to develop a MP genetic engineering vaccine with influenza virus strain as vector.

METHODS:

In this study, the major antigen genes P1a of MP adhesion factor P1(3862-4554 bases) and P30a of P30(49-822 bases) were inserted into the nonstructural protein (NS) gene of Influenza A virus strain A/Puerto Rio/8/34(H1N1), PR8 for short, to construct the recombinant vectors NS-P1a or NS-P30a. The recombinant pHW2000 plasmids containing NS-P1a or NS-P30a were cotransfected with the rest 7 fragments of PR8 into HEK293T cells. After inoculating chicken embryos, the recombinant influenza viruses rFLU-P1a and rFLU-P30a were rescued. RT-PCR and sequencing were used to identify the recombinant viruses. The hemagglutination titers of rFLU-P1a and rFLU-P30a were determined after five successive generations in chicken embryos so as to indicate the genetic stability of the recombinant viruses. The morphology of recombinant influenza viruses was observed under electron microscopy.

RESULTS:

P1a or P30a was designed to be inserted into the modified NS gene sequence separately and synthesized successfully. RT-PCR identification of the recombinant viruses rFLU-P1a and rFLU-P30a showed that P1a (693 bp), P30a (774 bp), NS-P1a (1992bp) and NS-P30a (2073 bp) bands were found, and the sequencing results were correct. After five successive generations, each virus generation has a certain hemagglutination titer (from 132 to 164), and the band of P1a or P30a can be seen in the corresponding positions. The virus particles under the electron microscope appeared as spheres or long strips connected by several particles, revealing a complete viral membrane structure composed of virus lipid bilayer, hemagglutinin, neuraminidase, and matrix proteins.

CONCLUSION:

The recombinant viruses rFLU-P1a and rFLU-P30a which carried the advantaged immune regions of the P1 and P30 genes in MP were successfully constructed and identified. And the genetic stability of rFLU-P1a or rFLU-P30a was relatively high. The typical and complete morphology of influenza virus was observed under the electron microscope. Our research provided a foundation for the further development of MP vaccines for human.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Vectores Genéticos / Mycoplasma pneumoniae Límite: Animals / Humans Idioma: En Revista: Immun Inflamm Dis Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Vectores Genéticos / Mycoplasma pneumoniae Límite: Animals / Humans Idioma: En Revista: Immun Inflamm Dis Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido