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Background:Secondary lymphoid tissue chemokine( SLC)is involved in lymphoid homing and anti-tumor immune response,and has a chemotactic effect on intestinal lymphocytes. Several animal studies have shown that SLC is involved in the occurrence and development of ulcerative colitis( UC). Aims:To investigate the expression and significance of SLC in UC. Methods:Forty active UC patients from Dec. 2010 to Dec. 2012 at Affiliated Hospital of Nantong University were enrolled,and 20 healthy volunteers were served as controls. Expression of SLC in the colon mucosa was detected by immunohistochemistry,and its relationship with severity of UC was analyzed. Results:SLC was positively expressed in all UC patients,while was negatively or weakly positively expressed in controls. Expression of SLC in UC patients was significantly higher than that in controls(4. 16 ± 0. 78 vs. 0. 52 ± 0. 11,P<0. 05). Expression of SLC was correlated with the severity of involvement of UC. Conclusions:Expression of SLC participates in development and progress of UC. SLC may play an important role in the induction of local damage and pathological changes of UC.
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Objective To explore the effects of SLC concentration gradient on suppression of tumor immune escape. Methods According to different SLC concentration, there were six groups. The HLA-Ⅰexpression and apoptosis of MCF-7 were detected with FCM,and intracellular BCL-2 expression was analyzed by western blot. The production of TGF-? was detected with ELISA. Results In a certain range of concentration gradient, following SLC increase, HLA-Ⅰexpression level on MCF-7 was improved, and apoptosis was induced but BCL-2 expression was enhanced. Moreover, the secretion of TGF-? was suppressed. Conclusion SLC inhibites tumor immune escape.
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Objective To construct an expression vector pET32a(+)/SLC and express the fusion protein TrxA SLC. Methods Total RNA from human inflammatory tonsil was extracted and its cDNA was generated with reverse transcription. Mature secondary lymphoid tissue chemokine (SLC) gene was amplified by RT PCR and cloned into plasmid pET32a(+) with Nco Ⅰ and Eco RⅠ sites added to the 5′ and 3′ ends respectively. E. coli DH5? was transformed with the recombinant plasmid, and the grown clones were selected. The inserted DNA was verified by enzyme digestion and DNA sequencing. The 3 amino acids between enterokinase site and target gene were deleted with mutation and the new vector was verified by sequencing. Expression of SLC was analysed by SDS PAGE. The fusion protein was purified by metal affinity chromatography and weak cation exchange chromatography, which was analysed by SDS PAGE and Western blotting. Results Trx SLC fusion protein expression vector was successfully construced, and the fusion protein was expressed with solubility. The purified fusion protein displayed the ability of binding the goat anti human SLC polyclonal antibody. Conclusion The SLC fusion protein can be expressed with stability and solubility and primary purification is performed with metal affinity chromatography and weak cation exchange chromatography.
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Objective To investigate the expression and effect of secondary lymphoid tissue chemokine(SLC)in experimental ulcerative colitis(UC)in rats.Methods Thirty Spague-Dawley rats were divided into control group and UC group.SLC expression in colon tissues was detected by RT-PCR and immunohistochemistry,respectively.Results The transcriptional level of SLC in UC tissues was significantly higher than that in the control group(0.846?0.07 vs 0.312?0.12,P＜0.01).The positive expression of SLC was concentrated mainly on submucosa,and the positive rate of SLC protein expression in UC group significantly higher than that in control group(P＜0.01).Conclusion SLC overexpression could contribute to the pathological processes in UC rats,thus SLC may be an ideal ther- apeutic target for UC.