ABSTRACT
In order to explore the relationship between human papilloma virus ( HPV) and upper gastrointestinal cancer(esophageal cancer), An esophageal squamous cell carcinoma(ESCC) tissue was obtained from a 76 year old Chinese female patient from Anyang city, a high-incidence area for esophageal cancer, in China. Transplanted tumor was formed through direct SCID mouse tumorigenicity experiment and cultured monolayer cells were obtained after several passages and screenings Immunofluorescence test, cell growth curve, soft agar assay, chromosome analysis and tissues HE staining were also performed to confirm the epithelial cell origin. Cell DNA STR typing results showed that no three alleles was observed,indicating no contamination of human cells. DNA analysis revealed the presence of HPV type 18 DNA in this cell line. DOLINK test found the E6 protein expression of HPV virus. We concluded that the established cell line is a new esophageal squamous cell-origincarcinoma cell line with HPV DNA positive and expression of viral oncoprotein. It provides new cytologic material for performing etiology studies on the occurrence and development of esophageal cancer.
Subject(s)
Aged , Animals , Female , Humans , Male , Mice , Carcinoma, Squamous Cell , Virology , Cell Proliferation , China , Esophageal Neoplasms , Virology , Human papillomavirus 18 , Genetics , Mice, SCID , Papillomavirus Infections , Virology , Tumor Cells, Cultured , Cell Biology , Virology , Tumor Virus Infections , VirologyABSTRACT
<p><b>OBJECTIVE</b>To study the immune effect of recombinant adeno-associated virus (rAAV) combined with recombinant adenovirus (rAdV) vaccine in BALB/c mice.</p><p><b>METHODS</b>The codon-modified HIV-1 gp120 gene was inserted into plasmid of adeno-associated virus and adenovirus vector separately. Then the rAAV and rAdV vaccines were constructed. BALB/c mice were immunized with rAAV and rAdV vaccines in different administration scheme. The IgG antibody was detected by ELISA and CTL response was detected by intracellular cytokine stain assay.</p><p><b>RESULTS</b>Both rAAV and rAdV vaccine could express gp120 gene; the mice primed with rAAV at week 0, 2 and boosted with rAdV at week 5, 14 and 20 elicited the strongest gp120 specific CTL and IgG antibody response.</p><p><b>CONCLUSION</b>The mice primed with rAAV and boosted with rAdV could elicit specific CTL response and IgG antibody.</p>
Subject(s)
Animals , Female , Mice , Adenoviridae , Genetics , CD8-Positive T-Lymphocytes , Allergy and Immunology , Dependovirus , Genetics , Genetic Vectors , HIV Envelope Protein gp120 , Genetics , Allergy and Immunology , HIV-1 , Immunoglobulin G , Blood , Interferon-gamma , Blood , Mice, Inbred BALB C , Plasmids , Recombination, Genetic , T-Lymphocytes, Cytotoxic , Allergy and Immunology , Vaccines, DNA , Allergy and Immunology , MetabolismABSTRACT
<p><b>OBJECTIVE</b>Chimeric human/bovine immunodeficiency virus (HBIV) cDNA was constructed by replacing HIV tat and LTR with bovine immunodeficiency virus (BIV) tat and LTR to study the activity of BIV tat and LTR in the chimerae.</p><p><b>METHODS</b>The target fragments of BIV tat, LTR and HIV gag, pol, env were respectively amplified by using PCR and sequentially inserted into pBluescript SK(+) vector. The chimeric clone was transfected into human MT4 cells. The transcript and gene expression of the HBIV chimeric virus were detected by using RT-PCR and a reverse transcriptase assay, respectively.</p><p><b>RESULTS</b>BIV tat mRNA and HIV gag mRNA were detected. The reverse transcriptase activity of the chimeric virus was analyzed in the fluctuation curve.</p><p><b>CONCLUSIONS</b>In chimeric HBIV cDNA transfected MT?4 cells, BIV tat and HIV gag were transcripted. The reverse transcriptase of the chimeric virus had biological activity. These data suggest that in MT4 cells, BIV LTR had promoter activity and BIV tat had the function of transactivation in the chimeric virus. The study of the chimeric virus with infectivity is in progress.</p>