Hybridoma cells display an increase in antibody productivity following exposure to hypertonic conditions. However, the underlying mechanism is not well understood. In the present study, we hypothesize that the nuclear...Hybridoma cells display an increase in antibody productivity following exposure to hypertonic conditions. However, the underlying mechanism is not well understood. In the present study, we hypothesize that the nuclear factor of activated T cells 5 （NFAT5）/tonicity enhancer binding protein （TonEBP） functions to increase the antibody productivity of hybridoma cells. NFAT5 is an osmosensitive mammalian transcription factor. However, its ubiquitous expression in various organs that are not bathed in hypertonic milieu suggests that NFAT5 may also regulate cell growth and function under isotonic conditions. In this study, we examined the expression of NFAT5 in hybridoma cells by Western blot analysis, and found that it increased significantly in hypertonic medium. To further define the function of NFAT5 in hybridoma cells, RNA interference technique was used to downregulate the expression of NFAT5 in SGB-8 cells （a hybridoma cell line）. In isotonic medium, antibody productivity ofhybridoma cells was reduced by downregulation of NFAT5 while cell proliferation was not influenced. The results presented here demonstrate that NFAT5 not only plays an important role in osmotic stress response pathway in hybridoma cells but also is essential for optimal antibody productivity.展开更多
Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s co...Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s countries. Prevention, based on early virus detection is the only effective control measure. Monoclonal antibodies appeared to be very useful tool. The authors used for the production of monoclonal antibodies hybridomas technique, by fusing spleen cells of immunized BALB/c mice to PepMV and SP2/O cancerous cells. The aim of this work is to produce hybridomas producers of Mab that could be used for ELISA in Morocco. At the same time, these efforts will serve to decrease expenses of producers concerning phytosanitory control. We obtained 16 hybridomas lines producers of Mab specific for PepMV. They were tested for efficiencies in ELISA and two lines were retained for production of Mab on large scale （1B 11-G 10 and 5A l-G5）. Isotyping of these two lines showed that they are belonging to IgG 1 class and easily purified by affinity chromatography in agarose column by protein A. The conjugation of these two antibodies to alkaline phosphatase has been verified by DAS-ELISA. These antibodies will enable to diagnose the disease from infected tomato plants, integrating several serological tests to control it and target the actions of struggles.展开更多
Objective: To understand the function of nicotinamide N-methyltransferase (NNMT) protein as tumor biomarker in renal carcinoma. Methods: Recombinant NNMT protein was used to prepare monoclonal antibodies by hybridoma ...Objective: To understand the function of nicotinamide N-methyltransferase (NNMT) protein as tumor biomarker in renal carcinoma. Methods: Recombinant NNMT protein was used to prepare monoclonal antibodies by hybridoma technique. The diagnostic and prognostic function of NNMT protein in renal carcinoma was evaluated by analyzing 74 renal cancer tissues through immunohistochemical staining for NNMT by using the prepared antibodies. Results: Two hybridomas named 2F8 and 1E7 stably secreting the monoclonal antibodies were isolated successfully, and characters such as isotypes and specificity were determined. NNMT protein was significantly up-regulated in renal cancer and significantly associated with tumor histology and ages. The univariate survival analysis demonstrated that the pT-status, high levels of NNMT, and distant metastasis were significant prognosticators. Conclusion: NNMT is over-expressed in a large proportion in renal cell cancers. High NNMT expression is significantly associated with unfavorable prognosis. However, the prognostic value of NNMT needs further verification in larger sample sizes.展开更多
文摘Hybridoma cells display an increase in antibody productivity following exposure to hypertonic conditions. However, the underlying mechanism is not well understood. In the present study, we hypothesize that the nuclear factor of activated T cells 5 （NFAT5）/tonicity enhancer binding protein （TonEBP） functions to increase the antibody productivity of hybridoma cells. NFAT5 is an osmosensitive mammalian transcription factor. However, its ubiquitous expression in various organs that are not bathed in hypertonic milieu suggests that NFAT5 may also regulate cell growth and function under isotonic conditions. In this study, we examined the expression of NFAT5 in hybridoma cells by Western blot analysis, and found that it increased significantly in hypertonic medium. To further define the function of NFAT5 in hybridoma cells, RNA interference technique was used to downregulate the expression of NFAT5 in SGB-8 cells （a hybridoma cell line）. In isotonic medium, antibody productivity ofhybridoma cells was reduced by downregulation of NFAT5 while cell proliferation was not influenced. The results presented here demonstrate that NFAT5 not only plays an important role in osmotic stress response pathway in hybridoma cells but also is essential for optimal antibody productivity.
文摘Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s countries. Prevention, based on early virus detection is the only effective control measure. Monoclonal antibodies appeared to be very useful tool. The authors used for the production of monoclonal antibodies hybridomas technique, by fusing spleen cells of immunized BALB/c mice to PepMV and SP2/O cancerous cells. The aim of this work is to produce hybridomas producers of Mab that could be used for ELISA in Morocco. At the same time, these efforts will serve to decrease expenses of producers concerning phytosanitory control. We obtained 16 hybridomas lines producers of Mab specific for PepMV. They were tested for efficiencies in ELISA and two lines were retained for production of Mab on large scale （1B 11-G 10 and 5A l-G5）. Isotyping of these two lines showed that they are belonging to IgG 1 class and easily purified by affinity chromatography in agarose column by protein A. The conjugation of these two antibodies to alkaline phosphatase has been verified by DAS-ELISA. These antibodies will enable to diagnose the disease from infected tomato plants, integrating several serological tests to control it and target the actions of struggles.
基金Project supported by the Science Foundation of Health Bureau of Zhejiang Province (Nos. 2005A055 and 2008B114)the Science Foundation of Education Bureau of Zhejiang Province (No. 20061271), China
文摘Objective: To understand the function of nicotinamide N-methyltransferase (NNMT) protein as tumor biomarker in renal carcinoma. Methods: Recombinant NNMT protein was used to prepare monoclonal antibodies by hybridoma technique. The diagnostic and prognostic function of NNMT protein in renal carcinoma was evaluated by analyzing 74 renal cancer tissues through immunohistochemical staining for NNMT by using the prepared antibodies. Results: Two hybridomas named 2F8 and 1E7 stably secreting the monoclonal antibodies were isolated successfully, and characters such as isotypes and specificity were determined. NNMT protein was significantly up-regulated in renal cancer and significantly associated with tumor histology and ages. The univariate survival analysis demonstrated that the pT-status, high levels of NNMT, and distant metastasis were significant prognosticators. Conclusion: NNMT is over-expressed in a large proportion in renal cell cancers. High NNMT expression is significantly associated with unfavorable prognosis. However, the prognostic value of NNMT needs further verification in larger sample sizes.
