Cytokines in AIDS and Cancer

我们已经使用以前开发的RNA优化技术来优化IL-15细胞因子的表达，并已经证明我们可以在小鼠和猕猴体内传递DNA后过量产生生物活性细胞因子。这一时期的一个重要结论是，异源二聚体IL-15表达载体的DNA传递导致了系统活跃的细胞因子水平，并促进了NK和T细胞的增殖。因此，DNA注射现在是一种在人体内传递IL-15的实用方法。我们对IL-15的生物学进行了探索，发现只有与所谓的IL-15受体-α在同一细胞中共表达，才有可能有效地产生IL-15。我们还发现，以前在人类和啮齿动物中被鉴定为细胞内或核内的第二种形式的IL-15(SSP IL-15)，当与IL-15受体α共表达时，也可以有效地从细胞中分泌出来。这些结果为IL-15的生物学和调控提供了新的线索，并为该细胞因子的高效生产和临床应用提供了方法。已经构建了大量产生可溶性生物活性IL-15/IL-15受体α异二聚体的细胞系，并将其用于生产体内发现的真正生物活性形式的IL-15。从过量生产的人细胞中提纯的IL-15注射到小鼠体内，并显示出生物活性。在灵长类动物中的广泛研究表明，异二聚体IL-15具有良好的药代动力学和药效学特性。异二聚体IL-15延长的半衰期与其他细胞因子形成鲜明对比，并与这种细胞因子对免疫系统的动态平衡作用一致。异二聚体IL-15已被广泛研究，并且它是高度糖基化的。IL-15因其刺激包括CD8和NK细胞在内的淋巴细胞的生长、激活和存活的能力而备受关注。因此，IL-15已被考虑用于癌症免疫治疗和过继转移后支持细胞毒细胞克隆的生长。IL-15的其他拟议用途是用于淋巴细胞减少症、支持NK细胞在NK转移后的生长和激活，以及作为疫苗佐剂。我们已经证明，IL-15注射可以加速细胞毒药物治疗后淋巴细胞减少的小鼠淋巴细胞的恢复。我们利用优化的表达载体在动物体内表达IL-12细胞因子。高效的表达产生生物活性水平，从而提高DNA疫苗后的免疫应答，从而成为我们疫苗的重要分子佐剂。本工作建立了优化IL-12家族细胞因子(IL-12、IL-23、IL-27、IL-35)表达的方法。在DNA传递后高效表达IL-27与IL-2在消除小鼠神经母细胞瘤转移方面具有协同作用。

We have used the previously developed technologies of RNA optimization to optimize expression of IL-15 cytokine, and have shown that we can over-produce bioactive cytokine after DNA delivery in mice and macaques. An important conclusion this period is that DNA delivery of vectors expressing heterodimeric IL-15 leads to systemically active levels of cytokine and the increased proliferation of NK and T cells. Therefore, DNA injection is now a practical method for the delivery of IL-15 in humans. We explored the biology of IL-15 and showed that efficient production of IL-15 is possible only by co-expression in the same cell with the so-called IL-15 Receptor-alpha. We also showed that a second form of IL-15 (SSP IL-15) previously identified in humans and rodents as intracellular or nuclear IL-15 is also efficiently secreted from the cells when co-expressed with the IL-15 Receptor alpha. These results shed new light in the biology and regulation of IL-15 and provide methods for the efficient production and clinical application of this cytokine. Cell lines overproducing soluble bioactive IL-15/IL-15 Receptor alpha heterodimers have been constructed and were used for the production of the authentic bioactive form of IL-15 found in the body. IL-15 purified from over-producing human cells was injected in mice and shown to be bioactive. Extensive studies in primates have shown that heterodimeric IL-15 has favorable pharmacokinetic and pharmacodynamic properties. The extended half life of heterodimeric IL-15 is in sharp contrast to other cytokines and is in agreement with the homeostatic role of this cytokine for the immune system. Heterodimeric IL-15 has been extensively characterized and it is heavily glycosylated. IL-15 is of interest due to its ability to stimulate the growth, activation and survival of lymphocytes, including CD8 and NK cells. Thus, IL-15 has been considered for cancer immunotherapy and for support of the growth of cytotoxic cell clones after adoptive transfer. Other proposed uses of IL-15 are in lymphopenia, in supporting NK cell growth and activation after NK transfer, and as vaccine adjuvant. We have shown that IL-15 injection accelerates the recovery of lymphocytes in mice rendered lymphopenic after treatment with cytotoxic drugs. We have used optimized expression vectors to express IL-12 cytokine in animals. Efficient expression results in bioactive levels, which increase immune response after DNA vaccination, thus becoming important molecular adjuvant for our vaccines. This work established methods to optimize expression of the IL-12 family of cytokines (IL-12, IL-23, IL-27, IL-35). Efficient expression of IL-27 after DNA delivery demonstrated synergy with IL-2 in the elimination of neuroblastoma metastases in mice.