A Genetically Modified attenuated Vaccine Expressing HPV16 E7 Kill Tumor Cells in Direct and Antigen-Specific Manner.

Attenuated (, LM) induces specific CD8 and CD4 T cell responses, and has been identified as a promising cancer vaccine vector. Cervical cancer is the third most common cancer in women worldwide, with human papillomavirus (HPV), particularly type 16, being the main etiological factor. The therapeutic HPV vaccines are urgently needed.

Evaluation of Immunogenicity and Protective Efficacy Elicited by Mycobacterium bovis BCG Overexpressing Ag85A Protein against Mycobacterium tuberculosis Aerosol Infection.

Mycobacterium bovis bacillus Calmette-Guérin (BCG) is currently the only vaccine available for preventing tuberculosis (TB), however, BCG has varying success in preventing pulmonary TB. In this study, a recombinant BCG (rBCG::Ag85A) strain overexpressing the immunodominant Ag85A antigen was constructed, and its immunogenicity and protective efficacy were evaluated. Our results indicated that the Ag85A protein was successfully overexpressed in rBCG::Ag85A, and the Ag85A peptide-MHC complexes on draining lymph node dendritic cells of C57BL/6 mice infected with rBCG::Ag85A were detectable 4 h post-infection.

[Prokaryotic expression and identification of HPV16 E7 protein].

HPV16 E7 fusion protein was expressed in E. coli BL21, and its applied value for HPV was evaluated. HPV16 E7 gene was amplified by PCR, and cloned into prokaryotic expression vector pGEX6p-1.

[Preparation and characterization of monoclonal antibodies specific for CFP-10 protein of Mycobacterium tuberculosis.].

Aim: To prepare monoclonal antibodies (mAb) against CFP-10 protein of Mycobacterium tuberculosis.

Methods: BALB/c mice were immunized with the purified His-CFP-10 expressed in BL21 (DE3)-pET-30a(+)-lhp. With the purified GST-CFP-10 as detecting antigen, mAb-produced hybridoma cells against CFP-10 were screened by indirect ELISA.

[Perparation and characterization of the monoclonal antibodies against listeriolysin O].

Aim: To prepare the monoclonal antibodies (mAbs) against listeriolysin O (LLO), which is the major virulence factor of Listeria monocytogenes.

Methods: The BALB/c mice were immunized with the SDS-PAGE product of BL21(pGEX-6p-1-hly). The purified LLO-GST protein was used as antigen for detection.

[Immunogenicity of S1 gene DNA vaccine of mouse hepatitis virus delivered by attenuated Salmonella typhimurium].

The complete S1 gene from mouse hepatitis virus (MHV) was amplified by RT-PCR and cloned into the pMD18-T vector. After confirmed by the restriction endonuclease analysis and PCR amplification, the positive clone of S1 gene was sequenced and then was transferred into eukaryotic expressing vector pVAXI. The recombinant plasmid pVAX1-S1 was transfected into COS-7 cells.

[Expression of actA gene of Listeria monocytogenes in Escherichia coli and preparation of ActA monoclonal antibodies].

The actA gene was amplified from Lm-4 strain of Listeria monocytogenes serotype 1/2a by PCR and inserted into T vector. Sequencing showed actA gene was 1833bp long and nucleotide homology was 100% compared with actA gene of Listeria monocytogenes EGD strain in GenBank. The cloned actA gene was then inserted into prokaryotic expression vector pGEX-6P-1 and pET respectively.

[Safety and efficacy of attenuated Salmonella typhimurium harbouring DNA vaccine against Newcastle disease virus].

A pair of primers were designed and synthesized according to the previously published sequence of fusion protein (F) gene of Newcastle disease virus (NDV) and used to amplify F gene by reverse-transcription polymerase chain reaction (RT-PCR) from the genomic RNA of a NDV strain JS5 isolated from goose. The PCR product was identified by sequencing. Then recombinant eukaryotic expression vector pVAX1-F was constructed through inserting F gene into MCS of pVAX1.

[Prokaryotic expression and bioactivity of the Listeria monocytogenes listeriolysin O].

To express the Listeria monocytogenes hly gene in Escherichia coli and study its primary biological characteristics, hly gene without signal peptide sequence was amplified from Listeria monocytogenes serotype 4b by PCR and inserted into T-easy vector. Sequencing showed this hly gene was 1518 bp and nucleotide homology was more than 99.8% compared with three Listeria monocytogenes hly genes in GenBank.

[Construction and characterization of Salmonella typhimurium SL7207 SifA- mutant strain].

One characteristic of Salmonella typhimurium SifA- mutant strain is to enter the cytosol of eukaryotic cells. The SifA- mutation of S. typhimurium P3H6 was transferred into S.