T cell activation and proliferation via CD3-TCR complex were investigated by lymphocyte DNA synthesis in vitro.Several interfering factors were also discussed.The result indicated that lymphocyte activation and prolif...T cell activation and proliferation via CD3-TCR complex were investigated by lymphocyte DNA synthesis in vitro.Several interfering factors were also discussed.The result indicated that lymphocyte activation and proliferation are calciumdependent.A rise of cytoplasmic free Ca2+ quickly following activation with CD3 McAb is mainly due to intracellular mobilization of Ca2+,while lymphocyte proliferation needs both intracellular mobilization of Ca2+ as well as influx of extracellular Ca2+, It was confirmed that CTX sensitive G protein plays a role in regulating T cell proliferation by pretreatment with CTX suppressing lymphocyte H-TdR incorporation obviously.PLC and PKC inhibitor neomycin and P.S.S could also decrease T cell proliferation.展开更多
Objective: Despite the existence of several therapeutic strategies, the management of cervical cancer remains challenging. Our region has very little data on the interaction between the immune system and the clinical ...Objective: Despite the existence of several therapeutic strategies, the management of cervical cancer remains challenging. Our region has very little data on the interaction between the immune system and the clinical response to chemotherapy. This work examines plasma levels of galectin-3 (Gal-3) and percentages of activated T cells in patients with cervical cancer treated with chemotherapy and investigates if there is a relationship between the rates of these two elements. Methods: We compared data from 37 patients with cervical cancer undergoing chemotherapy and 42 controls with normal cervical cytology. Plasma Gal-3 concentrations were assessed by ELISA and expression of activation markers by T cells (CD69 and HLA-DR) was assessed by flow cytometry at three different time points during chemotherapy. Results: Our results showed that patients had a significantly higher concentration of Gal-3 compared to controls (4.025 vs. 1.340, p 0.001), similarly, they had a significantly high percentage of activated lymphocytes (2.610 vs. 0.731;p 0.0001). According to the response to treatment, patients with no response to treatment had a lower concentration of circulating Gal-3 but had approximately the same percentage of activated CD4 and CD8 lymphocytes as patients with a partial or total response. In addition, we found a positive correlation between the Gal-3 level and CD4 T cells expressing the activation marker CD69 (p 0.05;rho = 0.44). Conclusion: In conclusion, our results show that there would be a relationship between circulating galectin-3 and the percentage of peripheral CD4+</sup>CD69+</sup> cells in cervical cancer.展开更多
Objective To establish a lymphocyte line capable of long survival and expressing human NT-3 to lay a foundation for future animal and human cochlear gene transfection research. Methods We collected lymphocytes from no...Objective To establish a lymphocyte line capable of long survival and expressing human NT-3 to lay a foundation for future animal and human cochlear gene transfection research. Methods We collected lymphocytes from normal human blood via Ficoll fluid and added IL-2 into the serum culture medium to promote lymphocyte growth. The NT3 cDNA was obtained by RT-PCR and ligated with the eukaryon vector which is pIRES-DsRed2 using T4DNA enzyme. The NT3 cDNA gene was transfected into the lymphocyte line using cationic liposome (LP2000). The lymphocytes transfected with NT3-cDNA were examined by RT-PCR and Western-blot methods. Results We established a new method to extend in vitro lymphocytes survival time and to transfect NT3 into lymphocytes. The genetically engineered lymphocytes were capable of surviving over relatively long time. Positive protein signals were obtained by Western blot. Conclusions Using lymphocytes as the intermediary, recombined plasmid pIRES-DsRed2NT3 is used to establish a lymphocyte line that expresses and secretes NT3. This cell line can be used in future animal gene cochlear transfection research and may help find an intermediary cell line for gene therapy for human deafness.展开更多
文摘T cell activation and proliferation via CD3-TCR complex were investigated by lymphocyte DNA synthesis in vitro.Several interfering factors were also discussed.The result indicated that lymphocyte activation and proliferation are calciumdependent.A rise of cytoplasmic free Ca2+ quickly following activation with CD3 McAb is mainly due to intracellular mobilization of Ca2+,while lymphocyte proliferation needs both intracellular mobilization of Ca2+ as well as influx of extracellular Ca2+, It was confirmed that CTX sensitive G protein plays a role in regulating T cell proliferation by pretreatment with CTX suppressing lymphocyte H-TdR incorporation obviously.PLC and PKC inhibitor neomycin and P.S.S could also decrease T cell proliferation.
文摘Objective: Despite the existence of several therapeutic strategies, the management of cervical cancer remains challenging. Our region has very little data on the interaction between the immune system and the clinical response to chemotherapy. This work examines plasma levels of galectin-3 (Gal-3) and percentages of activated T cells in patients with cervical cancer treated with chemotherapy and investigates if there is a relationship between the rates of these two elements. Methods: We compared data from 37 patients with cervical cancer undergoing chemotherapy and 42 controls with normal cervical cytology. Plasma Gal-3 concentrations were assessed by ELISA and expression of activation markers by T cells (CD69 and HLA-DR) was assessed by flow cytometry at three different time points during chemotherapy. Results: Our results showed that patients had a significantly higher concentration of Gal-3 compared to controls (4.025 vs. 1.340, p 0.001), similarly, they had a significantly high percentage of activated lymphocytes (2.610 vs. 0.731;p 0.0001). According to the response to treatment, patients with no response to treatment had a lower concentration of circulating Gal-3 but had approximately the same percentage of activated CD4 and CD8 lymphocytes as patients with a partial or total response. In addition, we found a positive correlation between the Gal-3 level and CD4 T cells expressing the activation marker CD69 (p 0.05;rho = 0.44). Conclusion: In conclusion, our results show that there would be a relationship between circulating galectin-3 and the percentage of peripheral CD4+</sup>CD69+</sup> cells in cervical cancer.
基金supported by National Natural Science Foundation of China (30973302)Medical Important People Project of Jiangsu Province (RC2007010)Medical Important Developing Project of Nanjing (ZKX06019)
文摘Objective To establish a lymphocyte line capable of long survival and expressing human NT-3 to lay a foundation for future animal and human cochlear gene transfection research. Methods We collected lymphocytes from normal human blood via Ficoll fluid and added IL-2 into the serum culture medium to promote lymphocyte growth. The NT3 cDNA was obtained by RT-PCR and ligated with the eukaryon vector which is pIRES-DsRed2 using T4DNA enzyme. The NT3 cDNA gene was transfected into the lymphocyte line using cationic liposome (LP2000). The lymphocytes transfected with NT3-cDNA were examined by RT-PCR and Western-blot methods. Results We established a new method to extend in vitro lymphocytes survival time and to transfect NT3 into lymphocytes. The genetically engineered lymphocytes were capable of surviving over relatively long time. Positive protein signals were obtained by Western blot. Conclusions Using lymphocytes as the intermediary, recombined plasmid pIRES-DsRed2NT3 is used to establish a lymphocyte line that expresses and secretes NT3. This cell line can be used in future animal gene cochlear transfection research and may help find an intermediary cell line for gene therapy for human deafness.
