Background: Identification of germline and somatic BRCA1/2 mutations in ovarian cancer is important for genetic counseling and treatment decision making with poly ADP ribose polymerase inhibitors. Unfortunately, data on the frequency of BRCA1/2 mutations in Vietnamese patients are scare. Methods: We aim to explore the occurrence of BRCA1/2 mutations in 101 Vietnamese patients with ovarian cancer including serous (n = 58), endometrioid (n = 14), mucinous (n = 24), and clear cell (n = 5) carcinomas. BRCA1/2 mutations were detected from formalin-fixed parafinembedded tumor samples using the Oncomine TM BRCA Research Assay on Personal Genome Machine Platform with Ion Reporter Software for sequencing data analysis. The presence of pathogenic mutations was confirmed by Sanger sequencing. Results: We found no BRCA2 mutation in the entire cohort. Four types of pathogenic mutations in BRCA1 (Ser454Ter, Gln541Ter, Arg1751Ter, and Gln1779AsnfsTer14) were detected in 8 unrelated patients (7.9%) belonging to serous and endometrioid carcinoma groups. Except for the c.1360_1361delAG (Ser454Ter) mutation in BRCA1 exon 11 that was somatic, the other mutations in exons 11, 20, and 22 were germline. Interestingly, the recurrent Arg1751Ter mutation in BRCA1 exon 20 appeared in 4 patients, suggesting that this is a founder mutation in Vietnamese patients. Conclusion: Mutational analysis of tumor tissue using next generation sequencing allowed the detection of both germline and somatic BRCA1/2 mutations.
Miễn dịch trị liệu hiện đang là phương pháp hấp dẫn trong điều trị bệnh ung thư. Các tế bào miễn dịch được phân lập, kích hoạt, tăng sinh ex vivo, và can thiệp về chức năng để sau đó truyền trở lại người bệnh. Ở nghiên cứu này, chúng tôi phân lập tế bào T CD4+ và T CD8+ từ tế bào đơn nhân máu ngoại vi người (PBMC), kích hoạt và nuôi cấy ex vivo hai quần thể tế bào này trong điều kiện sử dụng hạt từ phủ kháng thể CD3/CD28 ở tỉ lệ 01 hạt từ: 01 tế bào, bổ sung 30 U/mL IL-2 trong môi trường nuôi cấy. Số lượng tế bào được kiểm tra mỗi ngày bằng thuốc nhuộm Trypan Blue, tỉ lệ quần thể tế bào CD3+CD4+ và CD3+CD8+ được kiểm tra bằng phương pháp phân tích tế bào theo dòng chảy flow cytometry. Kết quả sau 10 ngày nuôi cho thấy, quần thể tế bào T CD4+ có sự tăng mạnh số lượng tế bào, tăng hơn 30 lần so với ngày đầu, đạt 6.5 x 107 tế bào. Đối với tế bào T CD8+ cũng ghi nhận có sự tăng nhưng không đáng kể số lượng tế bào sau 10 ngày nuôi, đạt 3 x 106 tế bào. Kết quả flow cytometry cho thấy quần thể tế bào T CD4+ vẫn duy trì được tỉ lệ CD3+/CD4+ hơn 90% ở ngày quan sát thứ 04, trong khi đó, quần thể T CD8+ có sự tăng số tế bào tuy nhiên tỉ lệ CD3+/CD8+ chỉ đạt hơn 40% trong quần thể. Như vậy, nhóm nghiên cứu đã có thể phân lập và nuôi cấy thành công ex vivo tế bào T CD4+ và T CD8+.
Though the hybridoma technology has been widely applied in the production of monoclonal antibodies, it has existed some disadvantages including low yield and genetic instability. Therefore, an alternative approach should be taken into account. Recently, recombinant monoclonal antibody technology has emerged as the best choice to cure hybridoma related drawbacks. However, recombinant antibodies require known genes for their generation. The purpose of this study is to collect the full-length genes encoding the anti-human CD45 antibody derived from the hybridoma cell line 16E8-F2. In this research, we designed specific primer pairs to amplify the light and heavy chain genes of the antibody through the PCR method. Afterward, the genes were separately cloned into a cloning vector called pJET1.2/blunt. The generated recombinant pJET1.2 vectors will serve as the main material source to manufacture the recombinant monoclonal antibody recognizing human CD45 protein tomorrow.
Virus-infectious diseases are permanent threats and cause serious lost for chicken industry of our country. Thus, the development of a bio-product enhancing the immune system against viral-caused diseases on chicken is highly necessary. Aims of this study were to clone, to express and to investigate anti-viral activity of recombinant chicken interferon-α and interferon-γ (ChIFN) produced from yeast Pichia pastoris. Gene fragments encoding ChIFN-α and ChIFN-γ were obtained by PCR and PCR/SOE (Splicing Overlapping Extension) based on the chicken genome DNA. Expressing vectors pPIC9K bearing gene-encoding ChINF-α or ChIFN-γ were cloned and two clones of P. pastoris stably expressing ChIFN-α and ChIFN-γ were generated. The expression of ChIFN-α and ChIFN-γ in P. pastoris culture supernatant was identified by Tricine SDS-PAGE. The biological activity of recombinant ChIFN-α was investigated on the model of chicken embryo fibroblast infected with IBDV and NDV. In case of IBDV-infected cells, the survival rate of cells treated by 1.6 µg/ml ChIFN-α was 79-88%, significantly higher than that of cells treated by PBS which was 33-34%. With NDV-infected cells, the survival rate of cells treated by 1.6 µg/ml ChIFN-α was 81-90%, also obviously higher than that of cells treated by PBS which was 27-32%. In conclusion, recombinant ChIFN-α and ChIFN-γ were expressed successfully on yeast P. pastoris and ChIFN-α was functional to stimulate anti-IBDV and NDV activities of cells.
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