Methods of treating cancer using il-21
a cancer and il-21 technology, applied in the field of cancer treatment methods using il21, can solve the problems of not being cured, not being able to be treated as easily,
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example 1
Mouse IL-21 is Active in Mouse Bone Marrow Assay
A. Isolation of Non-Adherent Low Density Marrow Cells:
[0130] Fresh mouse femur aspirate (marrow) was obtained from 6-10 week old male Balb / C or C57BL / 6 mice. The marrow was then washed with RPMI+10% FBS (JRH, Lenexa Kans.; Hyclone, Logan Utah) and suspended in RPMI+10% FBS as a whole marrow cell suspension. The whole marrow cell suspension was then subjected to a density gradient (Nycoprep, 1.077, Animal; Gibco BRL) to enrich for low density, mostly mononuclear, cells as follows: The whole marrow cell suspension (About 8 ml) was carefully pipetted on top of about 5 ml Nycoprep gradient solution in a 15 ml conical tube, and then centrifuged at 600×g for 20 minutes. The interface layer, containing the low density mononuclear cells, was then removed, washed with excess RPMI+10% FBS, and pelleted by centrifugation at 400×g for 5-10 minutes. This pellet was resuspended in RPMI+10% FBS and plated in a T-75 flask at approximately 106 cell...
example 2
IL-21 Transgenic Mice
A. Generation of Transgenic Mice Expressing Human and Mouse IL-21
[0134] DNA fragments from transgenic vectors (U.S. Pat. No. 6,307,024) containing 5′ and 3′ flanking sequences of the respective promoter (MT-1 liver-specific promoter (mouse IL-21 (U.S. Pat. No. 6,307,024) or lymphoid specific LCK promoter (mouse and human IL-21 (U.S. Pat. No. 6,307,024), the rat insulin II intron, IL-21 cDNA and the human growth hormone poly A sequence were prepared and used for microinjection into fertilized B6C3fl (Taconic, Germantown, N.Y.) murine oocytes, using a standard microinjection protocol. See, Hogan, B. et al., Manipulating the Mouse Embryo. A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1994.
[0135] Eight transgenic mice expressing human IL-21 from the lymphoid-specific EμLCK promoter were identified among 44 pups. Four of these were pups that died and 4 grew to adulthood. Expression levels were fairly low in these animals. Twenty transgenic mice expre...
example 3
IL-21 Purified Recombinant Human Protein
Dose-Response Study in Normal Mice
A. Summary
[0142] Normal six week old female C57B1 / 6 (Harlan Sprague Dawley, Indianapolis, Ind.). mice were treated by intraperitoneal injection once daily for either four or eight days with one of four dose levels of purified recombinant human IL-21 (U.S. Pat. No. 6,307,024) at 0.1, 0.5, 5 or 50 μg / mouse / day or with vehicle as a control. Body weights and body temperatures were monitored daily. On either day four or day nine, four of the eight mice from each protein treatment group and five of the ten mice in the vehicle control group were sacrificed. Blood, bone marrow and tissues were harvested and analyzed. Potential perturbations in lymphoid tissues were examined, as well as general physiologic and toxicological parameters.
[0143] There was no evidence of toxicity of human IL-21 protein at any of the doses tested. Body weights and temperatures were unchanged. There were no apparent changes in clinical...
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