Smith C A, Rennick D M
Proc Natl Acad Sci U S A. 1986 Mar;83(6):1857-61. doi: 10.1073/pnas.83.6.1857.
Murine mast-cell and T-cell growth factor activities, distinct from interleukins 3 and 2 (IL-3 and IL-2), have been identified and partially purified from the supernatant of the activated helper T-cell line Cl.Ly1+2-/9. This mast-cell growth factor (MCGF) activity supports only low levels of proliferation of several IL-3-dependent mast-cell lines and synergistically enhances the growth of mast cells in the presence of IL-3. The T-cell growth factor (TCGF) stimulates the proliferation of several T-cell lines, but to a lesser extent than recombinant IL-2. The MCGF and TCGF activities were not separable despite multiple biochemical fractionations, suggesting that both activities reside in the same protein. The MCGF/TCGF was separated from endogenous IL-3 by cation-exchange chromatography at neutral pH and could be distinguished from IL-2 by unique elution conditions from reverse-phase columns. Two bands of MCGF/TCGF activity were eluted from gels after sodium dodecyl sulfate/PAGE; under nonreducing conditions, the activities corresponded to molecular masses of 20 and 15 kDa, while after reduction, the molecular masses were 21 and 16 kDa. Thus, both activities may correspond to single polypeptide chains. The majority of the MCGF/TCGF activity appears to reside in the 20-kDa species, which displays a pI of 6.2 on chromatofocusing.
已从活化的辅助性T细胞系Cl.Ly1+2-/9的上清液中鉴定出并部分纯化了与白细胞介素3和2(IL-3和IL-2)不同的小鼠肥大细胞和T细胞生长因子活性。这种肥大细胞生长因子(MCGF)活性仅支持几种IL-3依赖性肥大细胞系的低水平增殖,并在IL-3存在的情况下协同增强肥大细胞的生长。T细胞生长因子(TCGF)刺激几种T细胞系的增殖,但程度低于重组IL-2。尽管进行了多次生化分级分离,MCGF和TCGF活性仍无法分离,这表明两种活性存在于同一蛋白质中。MCGF/TCGF在中性pH下通过阳离子交换色谱与内源性IL-3分离,并可通过反相柱的独特洗脱条件与IL-2区分开来。十二烷基硫酸钠/聚丙烯酰胺凝胶电泳后,从凝胶中洗脱得到两条MCGF/TCGF活性带;在非还原条件下,活性对应的分子量为20和15 kDa,而还原后分子量为21和16 kDa。因此,两种活性可能对应于单条多肽链。大部分MCGF/TCGF活性似乎存在于20-kDa的蛋白中,该蛋白在聚焦色谱上的pI为6.2。