IP PRIMING WITH RIFNG AND RIL 2 IN IP ADOPTIVE IMMUNOTHERAPY OF OVARIAN CANCER

我们在UCRC赠款下进行的翻译研究的亮点， 涉及腹膜内免疫疗法的包括以下： (A)22例患者接受IFN-γ IP注射，随后接受rIL-2 对大多数患者无明显不良反应。1/2 微小残留病变患者（定义为单个肿瘤<1cm 并且腹膜中没有多个转移，目前在一个 没有临床或放射学证据的疾病。另外两名患者 他们的肿瘤稳定了6个月。 (B)HLA-I类和HLA-II类抗原的表达与对照组相比有显著性差异（P <0.05）。 在用IFN-γ IP处理后，卵巢肿瘤细胞上增加。 然而，在某些患者中，无论是HLA I类或II类减少， 注射IP IL-2后。这些变化似乎与 腹膜中IL-10和可能的TGF- B浓度增加 流体. (C)一个在我实验室工作的研究生已经发现， 其特征在于在以下患者的腹膜液中存在分泌IL-10的单核细胞： 患者（摘要提交给AACR和美国医学会 免疫学家）。这是一个重要的发现，因为它提供了一个可能的 为将来控制体内IL-10产生的努力提供了目标。 (D)我们还表明，IL-2信息存在于92%的RNA提取物中， 的PEC，而IFN-信使目前只有22%的情况下。我们 研究结果表明，T细胞在体内可能仅部分活化。 (E)因此，我们寻找其他可能解释 体内不完全T细胞活化。 我们发现了一群 然而，具有树突细胞表型特征的细胞， 这些细胞缺乏CD 80或具有低表达的CD 80，并具有其它 暗示不成熟的特征。 (F)基于我们的发现，MHC抗原可以被IFN-γ上调， 尽管B7.1共刺激抗原的表达是缺陷的， 正在探索基因修饰的自体肿瘤疫苗在 一项新的临床试验疫苗来源于自体肿瘤细胞 感染了金丝雀痘病毒载体， CD80。新鲜的肿瘤细胞已经用IFN-γ预处理，然后感染 用ALVAC-hB 7.1载体（Pasteur Merieux）。我们观察到 MHC、CD 80和ICAM增加。这些特征在自体 疫苗应该有利于体内T细胞的更完全活化。的 临床试验设计已获得RAC、NCI和FDA的批准 并开始累积。预计一些患者将 在UCRC接受治疗。 (G)在UCRC进行的另一项试验中，我们 研究IP rIL-12的作用，一种新的细胞因子，可以直接免疫 有利于TH 1效应的反应（这有可能推动 细胞毒性前CTL向细胞毒性CTL的分化）。我们目前在 第二剂量水平和药理学以及免疫学研究是 与此次审判有关的案件 (H)如果这两项试验都达到了不同的目标， IL-12与ALVAC-hB 7.1的联合应用， 审判 (I)我们还在实验室开展了几项研究， 我们可以控制免疫抑制分子TGF-β的产生， 和IL-10 这些包括反义寡核苷酸组合， 脂质体制剂，其次是使用来自 芳香族脂肪酸组，我们已经表明，可以减少生产 TGF-B (J)最后，M正在开发一个临床项目。D.安德森 癌症中心，以解决IP治疗的一般问题。此程序将 处理更基本的问题，如药物渗透，药物 它的目标是获得改善的结果， IP治疗基于对药效学原理的更好理解。

Highlights of our translational studies conducted under the UCRC grant and involving intraperitoneal immunotherapy include the following: (A) Twenty-two patients received IP injections of IFN- followed by rIL-2 without significant adverse effects in most patients. One out of 2 patients with minimal residual disease (defined as individual tumors <1cm and absence of multiple metastases in the peritoneum, currently after one year has no clinical or radiologic evidence of disease. Two other patients had stability of their tumors for 6 months. (B) Expression of HLA class I and class II antigens were significantly increased on ovarian tumor cells following the IP treatment with IFN- . However, in certain patients, either HLA class I or class II decreased following the injections of IP IL-2. These changes appear to be correlated with increased concentrations of IL-10 and possibly TGF- B in peritoneal fluids. (C) A graduate student who works in my laboratory has identified and characterized an IL-10 secreting monocyte in the peritoneal fluid of patients (abstract submitted to AACR and American Association of Immunologists). This is an important finding since it provides a possible target for future efforts to control IL-10 production in vivo. (D) We have also shown that IL-2 message is present in 92% of RNA extracts of the PEC, whereas IFN- message was present in only 22% of cases. Our findings suggest that T cells may only be partially activated in vivo. (E) We have therefore looked for other factors that may account for incomplete T cell activation in vivo. We have identified a population of cells that are phenotypically characteristic of dendritic cells, however, these cells lack CD80 or have low expression of CD80 and have other features suggesting immaturity. (F) Based on our findings that MHC antigens can be upregulated by IFN- although expression of the B7.1 costimulatory antigens are deficient, we are exploring the effects of a gene modified autologous tumor vaccine in a new clinical trial. The vaccine is derived from autologous tumor cells that have been infected with a canarypox vector that encodes the gene for CD80. Fresh tumor cells have been pretreated with IFN- and then infected with the ALVAC-hB7.1 vector (Pasteur Merieux). We have observed an increase in MHC, CD80 and ICAM. These characteristics in an autologous vaccine should favor more complete activation of T cells in vivo. The clinical trial design has been approved by the RAC, the NCI and the FDA and accrual has started. It is anticipated that some of the patients will be treated in the UCRC. (G) In a separate trial which is being conducted at the UCRC, we are studying the effects of IP rIL-12, a novel cytokine that can direct immune responses in favor of TH1 effects (this has a potential to drive differentiation of precytotoxic CTL to cytotoxic CTL). We are presently at the second dose level and pharmacologic as well as immunologic studies are being conducted in association with this trial. (H) If both of these trials attain separate objectives, consideration will be given to the combination of IL-12 with ALVAC-hB7.1 in a future clinical trial. (I) We have also initiated several studiess in our laboratory to determine of we can control the production of the immunosuppressive molecules, TGF- and IL-10. These include antisense oligonucleotides combined with liposomal formulation, and secondly the use of small molecules from the aromatic fatty acid group that which we have shown can reduce production of TGF-B . (J) Finally, a clinical program is being developed at M. D. Anderson Cancer Center to address IP therapy issues in general. This program will deal with more fundamental issues such as drug penetration, drug concentration in tissues and its goal is to obtain improved outcomes with IP therapies based on better understanding of pharmacodynamic principles.