The immunotherapeutic effects of recombinant Bacillus Calmette-Guérin resistant to antimicrobial peptides on bladder cancer cells
| Autor: | In Ho Chang, Sang-Jin Lee, Young Mi Whang, Kijeong Kim, Young Wook Choi, Min-Ji Cho, Myeong Joo Kim |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
medicine.medical_treatment Antimicrobial peptides Biophysics Cancer Vaccines Biochemistry Cathelicidin law.invention Microbiology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine law Cell Line Tumor medicine Humans Molecular Biology Vaccines Synthetic Bladder cancer Innate immune system Electroporation Cell Biology Immunotherapy medicine.disease Mycobacterium bovis Immunity Innate Recombinant Proteins 030104 developmental biology Urinary Bladder Neoplasms chemistry 030220 oncology & carcinogenesis BCG Vaccine Recombinant DNA Growth inhibition Antimicrobial Cationic Peptides |
| Zdroj: | Biochemical and Biophysical Research Communications. 509:167-174 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2018.12.097 |
| Popis: | Purpose Although Mycobacterium bovis Bacillus Calmette-Guerin (BCG) is the most widely used bladder cancer immunotherapy, innate immune responses involving antimicrobial peptides (AMPs) cause BCG failure and unwanted side effects. Here, we generated genetically modified BCG strains with improved immunotherapeutic effects by adding genes that confer evasion of AMPs. Materials and methods We constructed recombinant BCG (rBCG) strains expressing Streptococcal inhibitor of complement (Sic), which confers resistance to human α-defensin-1 and cathelicidin, and d-alanyl carrier protein ligase (dltA), which confers resistance to cationic AMPs. Sic and dltA were separately cloned into the pMV306 plasmid and introduced into BCG via electroporation. Then, the efficacy of the rBCGs was tested in a growth inhibition assay using two bladder cancer cell lines (5637, T24). Results We confirmed the presence of cDNA segments corresponding to the Sic and dltA genes in total mRNA of the rBCG strains containing Sic (rBCG-Sic) and dltA (rBCG-dltA), and these rBCGs showed higher survival against AMPs. The growth inhibitory effects of rBCGs on bladder cancer cells were significantly enhanced compared to those of the parent BCG, and THP-1 migration also increased. After 8 h of infection, the levels of internalization were higher in rBCG-infected bladder cancer cells than in BCG-infected cells, and cells infected with rBCGs showed increased release of antitumor cytokines, such as IL-6/12, TNF-α, and INF-γ, resulting in inhibition of bacterial killing and immune modulation via antimicrobial peptides. Conclusions: rBCG-Sic and rBCG-dltA can effectively evade BCG-stimulated AMPs, and may be significantly improved immunotherapeutic tools to treat bladder cancer. |
| Databáze: | OpenAIRE |
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