The connection of innate and adaptive anti-cancer immunity

使用重组鸡痘病毒靶向递送鼠干扰素（IFN）-γ： 自然杀伤（NK）细胞募集到局部淋巴结和引发肿瘤特异性 宿主免疫IFN-γ是一种促炎细胞因子， 免疫调节剂。在本研究中，复制缺陷型重组禽（鸡痘） 将病毒工程化以表达鼠IFN-γ基因（rF-MuIFN-γ），其基本原理是 将浓缩水平的细胞因子输送到局部组织微环境。 皮下（s.c.）rF-MuIFN-γ给药导致IFN-γ产生， 局限于注射部位的组织微环境，以及（2）在生物学上 活性，如I类主要组织相容性复合物的显著增加所证明 (MHC)s.c.表达水平在rF-MuIFN-γ施用后生长的肿瘤。 用rF-MuIFN-γ感染高度致瘤性鼠细胞系MC 38， 有效的基于肿瘤细胞的疫苗，通过保护小鼠免于形成原发性肿瘤， 随后的肿瘤挑战。细胞疫苗是完全无效的， 用用rIFN-γ预处理的MC 38细胞或用 野生型鸡痘病毒（FP-WT）。局部引流淋巴结的临床分析 注射基于rF-MuIFN-γ的肿瘤细胞疫苗揭示了 肿瘤特异性细胞裂解（CTL）以及针对肿瘤细胞的大量裂解， NK敏感的YAC-1细胞。流式细胞术分析结合功能测定证实了 引流淋巴结内持续存在NK1.1（+）细胞长达5天 注射rF-MuIFN-γ后。在给药前用NK细胞耗竭抗体处理的小鼠 注射rF-MuIFN-γ感染的MC 38肿瘤细胞不能保护其免于原发性免疫缺陷。 肿瘤生长; NK耗竭小鼠中注射部位引流淋巴结的分析 揭示了伴随的肿瘤特异性CTL活性的丧失。这些发现提供 以促进宿主先天免疫而闻名的NK细胞也提供了证据， 免疫调节信号的发展所需的适应性免疫反应，其中， 反过来，保护接种疫苗的小鼠免受肿瘤生长的影响。干扰素 (IFN-α）是具有免疫调节特性的多效性细胞因子， 可以提高治疗性癌症疫苗的功效。该项目的目的是 评价重组IFN- #945联合治疗的有效性和相容性; 用痘病毒疫苗靶向人癌胚抗原（CEA）在小鼠模型中， 结肠直肠癌和胰腺腺癌，其中CEA是自身抗原。的表型和 在引流腹股沟淋巴液中评价IFN-α的功能作用 无肿瘤小鼠的淋巴结。我们研究了IFN-#945; 施用（局部与远端）对痘病毒的抗原特异性免疫应答的影响 预防针进行了机制研究以评估IFN-α和IFN-γ的功效。 CEA导向的痘病毒疫苗在荷瘤CEA转基因小鼠中的应用我们确定了剂量， 干扰素时间表-#945;导致引流局部扩大 腹股沟淋巴结和改善的细胞毒性（自然杀伤细胞和CD 8+）和抗原 演示文稿.通过施用以下药物来避免牛痘病毒的抑制： IFN-#945;接种部位远端。的组合 IFN-γ和疫苗抑制肿瘤生长，提高存活率， CEA+腺癌小鼠中CEA特异性CTL应答。在患有胰腺肿瘤的小鼠中， IFN-#945;减缓肿瘤生长，诱导CTL活性，并增加CD 8 + 肿瘤浸润淋巴细胞。这些数据表明IFN-γ可用作 一种生物反应调节剂与抗原导向的痘病毒疫苗， 治疗性抗肿瘤免疫应答。这项研究提供了理论基础和机制 支持这种免疫策略在患者中的临床试验的见解 CEA表达癌。

Targeted delivery of murine interferon (IFN)-gamma using a recombinant fowlpox virus: natural killer (NK) cell recruitment to regional lymph nodes and priming of tumor-specific host immunity. IFN-gamma is a proinflammatory cytokine that also acts as a potent immunomodulatory agent. In this study, a replication-deficient recombinant avian (fowlpox) virus was engineered to express the murine IFN-gamma gene (rF-MuIFN-gamma) with the rationale of delivering concentrated levels of the cytokine to a local tissue microenvironment. Subcutaneous (s.c.) rF-MuIFN-gamma administration resulted in IFN-gamma production that was restricted to the tissue microenvironment of the injection site and (2) was biologically active, as evidenced by a significant increase of class I major histocompatibility complex (MHC) expression levels in s.c. growing tumors following rF-MuIFN-gamma administration. Infection of a highly tumorigenic murine cell line, MC38, with rF-MuIFN-gamma functioned as an effective tumor cell-based vaccine by protecting mice from the formation of primary tumors and from subsequent tumor challenge. The cell-based vaccine was completely ineffective if mice were vaccinated with MC38 cells either pretreated with rIFN-gamma or infected with the wild-type fowlpox virus (FP-WT). Analysis of the regional lymph nodes draining the site of injection of the rF-MuIFN-gamma-based tumor cell vaccine revealed the presence of tumor-specific cell lysis (CTL) as well as a significant amount of lysis directed at NK-sensitive YAC-1 cells. Flow cytometric analyses coupled with functional assays confirmed the sustained presence of NK1.1(+) cells within those draining lymph nodes for up to 5 days after rF-MuIFN-gamma injection. Mice treated with NK cell-depleting antibodies prior to the injection of the rF-MuIFN-gamma-infected MC38 tumor cells were not protected from primary tumor growth; analysis of the lymph nodes draining the injection site in NK-depleted mice revealed an accompanying loss of the tumor-specific CTL activity. The findings provide evidence that NK cells, known for their contributions to host innate immunity, also provide immunoregulatory signals required for the development of an adaptive immune response, which, in turn, protected vaccinated mice against tumor growth. Interferon-alpha (IFN-α) is a pleiotropic cytokine possessing immunomodulatory properties that may improve the efficacy of therapeutic cancer vaccines. The aim of this project was to evaluate the effectiveness and compatibility of combining recombinant IFN-α with poxvirus vaccines targeting the human carcinoembryonic antigen (CEA) in murine models of colorectal and pancreatic adenocarcinomas, where CEA is a self-antigen. The phenotypic and functional effects of IFN-α were evaluated in the draining inguinal lymph nodes of tumor-free mice. We studied the effect of the site of IFN-α administration (local versus distal) on antigen-specific immune responses to poxvirus vaccination. Mechanistic studies were conducted to assess the efficacy of IFN-a and CEA-directed poxvirus vaccines in tumor-bearing CEA transgenic mice. We identified a dose and schedule of IFN-α that induced a locoregional expansion of the draining inguinal lymph nodes and improved cellular cytotoxicity (natural killer and CD8+) and antigen presentation. Suppression of the vaccinia virus was avoided by administering IFN-α distal to the site of vaccination. The combination of IFN-α and vaccine inhibited tumor growth, improved survival, and elicited CEA-specific CTL responses in mice with CEA+ adenocarcinomas. In mice with pancreatic tumors, IFN- α slowed tumor growth, induced CTL activity, and increased CD8+ tumor-infiltrating lymphocytes. These data suggest that IFN- α can be used as a biological response modifier with antigen-directed poxvirus vaccines to yield significant therapeutic antitumor immune responses. This study provides the rationale and mechanistic insights to support a clinical trial of this immunotherapeutic strategy in patients with CEA-expressing carcinomas.