ABSTRACT
<p><b>AIM</b>To study the effect of prothymosin alpha (Pro T alpha) as a fusion protein on secretion of IFN-gamma, IFN-alpha and TNF-alpha in vitro.</p><p><b>METHODS</b>The in vitro study was carried out on the culture of splenocytes, splenic and peritoneal macrophages isolated from Balb/c mice. Splenocytes were incubated with various concentrations of Pro T alpha (1 x 10(-7)-1 x 10(-10) mol.L-1) with or without Con A (5 micrograms.mL-1) for 72 h. Splenic and peritoneal macrophages were respectively treated with Pro T alpha (1 x 10(-7)-1 x 10(-10) mol.L-1) in the presence of LPS (10 micrograms.mL-1) for 24 h. Then IFN-gamma, IFN-alpha and TNF-alpha levels in the supernatant were detected by ELISA.</p><p><b>RESULTS</b>Pro T alpha (1 x 10(-7) mol.L-1) was found to obviously increase IFN-gamma level (P < 0.05) in the supernatant of splenocytes compared with the control group. Moreover, Pro T alpha (1 x 10(-7) mol.L-1) significantly induced the secretion of IFN-alpha (P < 0.01) and TNF-alpha (P < 0.01) in splenic and peritoneal macrophages.</p><p><b>CONCLUSION</b>In vitro, Pro T alpha could increase the secretion of IFN-gamma, IFN-alpha and TNF-alpha.</p>
Subject(s)
Animals , Female , Mice , Adjuvants, Immunologic , Pharmacology , Cell Separation , Glutathione Transferase , Pharmacology , Interferon-alpha , Bodily Secretions , Interferon-gamma , Bodily Secretions , Lymphocytes , Bodily Secretions , Macrophages , Bodily Secretions , Macrophages, Peritoneal , Bodily Secretions , Mice, Inbred BALB C , Protein Precursors , Pharmacology , Recombinant Fusion Proteins , Pharmacology , Spleen , Cell Biology , Thymosin , Pharmacology , Tumor Necrosis Factor-alpha , Bodily SecretionsABSTRACT
Objective:To analyze the polymorphism in human cDNA sequence of prothymosin-?(ProT?)by sequencing analysis.Methods:The cDNA of human ProT? was amplified from cells of peripheral blood and cord blood by RT-PCR.The product of RT-PCR was purified and linked with vector pMD18-T.After cloning and sequencing,the sequence of ProT? cDNA was compared with the standard sequence to analyze the polymorphism in the ProT? cDNA sequence.Results:The cloned ProT? cDNA sequence was different from that of the standard.We found 2 kinds of variations:(1)The nucleotide in 107 position was varied and the nucleotides in 110-121 and 191-205 positions were deleted;(2)The nucleotide in 306 position was deleted,mainly in the 60-80 years old group.Conclusion:We have identified 2 kinds of variations in human ProT? cDNA,but the first 28 amino acid in the N-terminal of cDNA of human ProT? are not involved therefore the variations do not affect the function of human ProT?.