Abstract Objective: To construct and express anti-IL-2R? genetically engineering antibody Fab and identify its biological activity.Methods: PCR were used to construct cloning vector 5G1-pA22 of anti-IL-2R? antibody Fab,by digestion of vector 5G1-pA22, anti-IL-2R? an-tibody Fab gene fragment was obtained and then this fragment was coupled with digesting product of pET28b plasmid by EcoRI and HindIII toget a recombinant expression vector 5G1-pET28b, it was highly expressed in the E. coli BL21(DE3) in the form of inclusion bodies, compoing20% of the total protein. After the renaturation of antibody Fab, its binding activity was identified by ELISA and antibody competitive inhibitionassay in vitro. Results: Anti-IL-2R? antibody Fab was successfully constructed and highly expressed in the form of inclusion bodies, its yield wasup to 1 mg/ml.When applied to ELISA, antibody Fab exhibited good spectfic reactivity with rIL-2R, antibody competitive inhibition assay invitro indicated that antibody Fab can compete with IL-2 for binding IL-2R in activated T cell surface.As a result,the activation and proliferationof T cells were inhibited and inhibition percentage exceeded 90%. Conclusion: Had successfully constructed and expressed anti-IL-2R? geneti-cally engineering antibody. Fab,it had good biological ativity.

By using 5 G 1-sepharose 4 B immune affinity column, we recovered approximately 160 ?g purified and soluble IL-2 receptor (sIL-2 R ) from 500ml PHA-stimulated human PBL supernatant. Only one band was showed in SDS-PAGE, its molecular weight was about 44 Kd. The purified sIL-2 R, which blocked the binding of anti-IL-2 R monoclonal antibody with IL- 2 R~+ cell, inhibited (at the higher doses) but enhanced (at the lower doses) the growth of IL-2 dependent cells.

Using ~(125)I-labeled anti-IL-2R_? McAb, we investigated the effect of rIL-2 R_? expression . In addition to increasing the IL-2 R_? expression the actived-lymphoblasts, we found that the higher concentration of IL-2 also directly induced the IL-2 R_? expression on the resting PBL, but the peak was at the 5th day. After treated by a higher concentration of IL-2, there was a positive interrelation of time courses between the incorporation of ~3H-TdR and/or ~3H-UR and IL-2 R_? level on resting PBL. This result indicated that it was by means of inducing IL-2 R_? expression that the higher concentration IL-2 caused the proliferation of the resting PBL.

Arsenicals ; pharmacology ; DNA Methylation ; drug effects ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; K562 Cells ; Membrane Proteins ; genetics ; metabolism ; Oxides ; pharmacology

Aim To study the anti-inflammatory effects and mechanisms of SBM isolated from Scutellariae radix by high hydrostatic pressure.Methods The effect of SBM on ConA-induced splenocyte proliferation and LPS-induced IL-1? production from peritoneal macrophage was studied in vitro,and the therapeutic effects of SBM on adjuvant-induced arthritis,formalin-induced paw oedema and acetic acid-induced vascular permeability were also investigated.Results SBM(31.25～500 mg?L-1)significantly inhibited ConA-induced splenocyte proliferation,IL-1? production from peritoneal macrophages in vitro.SBM 20 mg?kg-1 inhibited primary and secondary inflammatory reaction,decreased the elevated level of IL-1? released from peritoneal macrophages,inhibited splenocyte proliferation in adjuvant-induced arthritis in rats.Moreover,SBM inhibited formalin-induced paw oedema and acetic acid-induced vascular permeability.Conclusions SBM could inhibit inflammatory reaction.Its mechanisms might be related to the suppression of immune function and inhibition on proinflammatory cytokines production.

Aim To study the anti-inflammatory effects and mechanisms of SBM isolated from Scutellariae radix by high hydrostatic pressure.Methods The effect of SBM on ConA-induced splenocyte proliferation,LPS-induced PGE2 production and protein expression of COX-2 were studied in vitro,and the therapeutic effects of SBM on carragineen-induced paw oedema and acetic acid-induced twisting reaction were also investigated.Results SBM(15.63～250) mg?L-1 significantly inhibited LPS-induced PGE2 production and COX-2 enzyme activity in peritoneal macrophages in mice.SBM(125 mg?L-1)also inhibited ConA-induced protein phosphorylation of p38 MAP kinase and LPS-induced COX-2 protein expression.SBM(10,20 mg?kg-1)inhibited carragineen-induced paw oedema and acetic acid-induced twisting reaction.Conclusion SBM can inhibit inflammatory reaction.Its mechanisms may be related to the suppression of inflammatory medium production.

Objective: To study the anti-tumor effect of recombinant human MUC1-MBP. Methods: The C57BL/6 mice were in oculated with MUC1-MBP by subcutaneous. MUC1 specific CTL activity of spleen were determined by MTT; The effects on prevention and treatment of tumor were observed by establishing lewis lung cancer-carrying mice. Results: The cytotoxicity of CTL from immunized mice to the MCF7 and Lewis lung cancer cells respectively was (47.7?4.3) % and (67.5 ?6.5) %; 5?10 5 lewis lung cancer cells following immunization were injected iv into C57BL/6 mice, after three weeks, the number of lung and tail tumor colonies was 51 and 5 for PBS and MUC1-MBP groups respectively and the suvival time was significantly delayed in immunized mice. The average volume of tumors in mice with MUC1-MBP was 386 mm 3 wherea control group was 4 000 mm 3 at tumor treating experiment. Conclusions: Recombinant human MUC1-MBP have significantly effects on prevention, treatment and inhibiting metastases of tumor. Our results suggested that the recombinant MUC1-MBP might be used to develop protein vaccine against human carcinoma.

Objective:To study the effects of hyperthermia on apoptosis in human colonic cacinoma cell line Lovo. Methods:Lovo cells were exposed to hyperthemia and apoptosis was measured with fluorescent Hocehst 33258 staining, electropherogram gel, flow cytometry (FCM). Results:Significant changes in cell morphology , apoptosis peak apeared before the G1 phase and DNA ladder was observed in various group cells which were exposed to different temperature and in different time. Most significent changes were observed in cells exposed to 43℃ for 3,4 h and 44℃ for 1,2 h. Also cell cycle was found changed. Conclusion:Hyperthermia can cause apoptosis in human colonic cacinoma cell line Lovo, and has phase-specificity.

Objective:To study MUC1 based cancer vaccine.Methods:MUC1 gene was inserted into pMAL-p2 vector and constructed recombinant pMAL-MUC1. MUC1-MBP fusion protein expression was induced by IPTG in E coli DH5? transformed by the recombinant pMAL-MUC1 .The fusion protein was analyzed by Western blot and purified by amylose affinity chromatography. The antiserum,T cell proliferation and CTL activity of spleen from C57 mice immunized by MUC1-MBP were determined respectively by ELISA,adding 3H-TdR and MTT.Results:Had successfully constructed pMAL-MUC1 expression vector,and purified MUC1-MBP and MBP. C57 mice immunized by MUC-MBP generated MUC1 specific antibody and CTL.The titer of polyclonal antibody to MUC1 was about 1∶5 760?3 221. CTL cytotoxicity to the MCF7 and lewis lung cancer cells respectively were at 47.7%?4.3% and 67.5%?6.5%.Conclusion:Human recombinant MUC1-MBP fusion protein activated T and B cell response in mice.The results suggested that the recombinant Muc1 may be used to develop protein vaccine against carcinoma.

0.05). Significant difference existed between the patients and endoscopy-negative (pathological reflux) and endoscopy-negative (physiological reflux) (P