The state of hyporeactivity in interferon production: localization and partial purification of the factor repressing the interferon production

The hyporeactivity factor in interferon production by L-929 cells designated IRP (interferon repressing protein) has been studied. In particular, its localization and methods of its purification have been studied. The kinetics of IRP accumulation by producing cells correlate with the development of hyporeactivity condition. Most of IRP is localized in cell sap and in ribosomal fraction in evidence to regulatory role of repressor at the level of interferon mRNA translation. A 100-fold increase in repressor activity was achieved by IRP concentration by ammonium sulfate precipitation. IRP as well as interferon have been shown to possess high affinity to polyU sepharose. The preparations of IRP and interferon concentrated by ammonium sulfate precipitation were subsequently purified by fractioning in a polyI sepharose column. A 10,000-fold (6 x 10(4) U/mg) purification was achieved for IRP and 250-fold (10(4) U/mg) for interferon.     

Stimulation of interferon production]

Data on the effect of some factors on interferon production in vitro are presented The kinetics of interferon synthesis in response to superinduction was similar to the respective curve of the effect of UV-radiation on the cell. Possible similarity in the effect of these factors on the mechanisms controlling interferon production is noted. An increase in interferon synthesis under the effect of ascorbic acid in cells of chick embryo fibroblast and L-929 was found. Combined use of the inductors provided an increase in the tests of interferon.     

The effect of stress on interferon production in mice

In order to study the effect of stress on the interferon (IFN) production, we determined the IFN-alpha/beta and -gamma producing capacities in restraint-stressed and SART-stressed mice. Restraint-stress caused not only a significant reduction in body weight, but also a reduction in the weight of spleen. The IFN-alpha/beta producing capacity by Newcastle disease virus (NDV) was significantly lower in the stressed mice. The IFN-gamma producing capacity by both Con-A and PHA-P was also depressed by this stress. On the other hand, SART-stress, whose severity was thought to be mild because no loss of body and spleen weights was seen, did not affect IFN-alpha/beta and -gamma producing capacities. These results suggest that the suppression of IFN producing capacity is dependent on the severity of physical stress.     

大型除菌过滤器在重组干扰素生产中的应用

为适应大规模生产的需要,用Millipore293型滤器取代常用的赛氏滤器对重组干扰素α1b及γ进行除菌过滤,此种大型滤器不仅滤膜面积大,而且单位面积过滤量也有明显增加,因此,过滤速度显著加快,可适应大规模生产的应用,用此二种滤器作除菌过滤对干扰素活性无影响,过滤制品前用2%白蛋白预滤可减少滤膜吸附干扰素作用,Millipore和赛氏滤器除菌过滤后热原质均能达到肌肉注射标准,且大部分能达到静脉注射标准,有个别制品用赛氏滤器过滤后热原质达至不到静脉注射标准者,用Millipore滤器复滤后,可以合格,上述结果表明用Milli-pore293型滤器制备重组干扰素α1b及γ是切实可行的。     

Effect of mycoplasma on interferon production and interferon assay in cell cultures

Armstrong, D. (The Children's Hospital of Philadelphia, Philadelphia, Pa.), and K. Paucker. Effect of mycoplasma on interferon production and interferon assay in cell cultures. J. Bacteriol. 92:97-101. 1966.-The influence of mycoplasma on the production and action of interferon was studied in cultures of both L and human embryonic kidney (HEK) cells. Mycoplasma hominis 1, the Negroni agent, and the F12 mycoplasma were used for infection of L cells, and M. hominis 1 and M. pneumoniae for inoculation of HEK cells. All strains were capable of multiplication in the culture systems employed. None produced detectable levels of interferon, and responsiveness of the cells to induction of interferon by virus remained unaltered. Infection with mycoplasma did not impair the sensitivity of the cells to the action of interferon, nor was the replication of vesicular stomatitis virus noticeably diminished.