1 Tumor Immunity Medical Research Center, Cancer Research Institute and Department of Pathology, College of Medicine, Seoul National University, Seoul, Republic of Korea.
2 Centre for Chronic Disease, College of Health and Biomedicine, Victoria University, Footscray, VIC, Australia.
Int J Immunopathol Pharmacol. 2018 Jan-Dec;31:394632017742504. doi: 10.1177/0394632017742504. Epub 2017 Dec 18.
Inducing cancer-specific cellular immune responses has become an attractive strategy in cancer treatment. In this study, we investigated the role of several adjuvants in eliciting T/Tn-specific cellular immunity and protection against T/Tn expressing tumor challenge. T/Tn (9:1) antigen was purified from blood type "O" erythrocytes donated from healthy Korean volunteers. Immunization was performed using: T/Tn only, T/Tn mixed with Freund's adjuvant (T/Tn + FA), keyhole limpet hemocyanin (KLH)-conjugated T/Tn mixed with FA (KLH-T/Tn + FA), and oxidized mannan-conjugated T/Tn mixed with FA (ox-M-T/Tn + FA). Mice immunized with ox-M-T/Tn + FA generated T/Tn-specific CD3, helper T (Th) cells, major histocompatibility complex (MHC) II, and MHC I; T/Tn presentation was significantly high and tolerogenic CD11b was the lowest among the tumor models. To verify Th type, we stained intracellular cytokines (interferon gamma (IFN-γ), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-4, and IL-10) using CD3 co-staining. Th1 (IFN-γ and GM-CSF) cytokines were highly expressed and showed high FasL/Fas ratios, cytotoxic T lymphocyte (CTL) activity, and cytotoxic T lymphocyte precursor (CTLp) activity in mice immunized with ox-M-T/Tn + FA. Lymphocyte infiltration was highest in mice immunized with ox-M-T/Tn + FA. Additionally, we monitored FasL, MHC I, CD301, and T/Tn expression levels using immunohistochemistry (IHC) on macrophage and tumor sites. The expression of all markers was highest in the ox-M-T/Tn + FA group. Furthermore, tumor retardation and survival rate were highest in the ox-M-T/Tn + FA group. These results demonstrate that a vaccine formulation of T/Tn conjugated with ox-M and mixed with FA-induced cellular immunity and sustained a humoral immune response without over-activating the immune system, thus effectively inhibiting tumor growth.
诱导针对癌症的特异性细胞免疫反应已成为癌症治疗的一种有吸引力的策略。在这项研究中，我们研究了几种佐剂在引发 T/Tn 特异性细胞免疫和保护 T/Tn 表达肿瘤挑战中的作用。T/Tn（9:1）抗原从来自健康韩国志愿者的“O”型血红细胞中纯化。免疫接种采用以下方法进行：仅 T/Tn、T/Tn 与弗氏佐剂混合（T/Tn+FA）、KLH 缀合的 T/Tn 与 FA 混合（KLH-T/Tn+FA）和氧化甘露聚糖缀合的 T/Tn 与 FA 混合（ox-M-T/Tn+FA）。用 ox-M-T/Tn+FA 免疫的小鼠产生了 T/Tn 特异性 CD3、辅助 T（Th）细胞、主要组织相容性复合体（MHC）II 和 MHC I；在肿瘤模型中，T/Tn 的呈递显著增加，而耐受性 CD11b 最低。为了验证 Th 型，我们使用 CD3 共染色对细胞内细胞因子（干扰素 γ（IFN-γ）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素（IL）-4 和 IL-10）进行染色。用 ox-M-T/Tn+FA 免疫的小鼠中，Th1（IFN-γ 和 GM-CSF）细胞因子表达水平较高，FasL/Fas 比值较高，细胞毒性 T 淋巴细胞（CTL）活性和 CTLp 活性较高。用 ox-M-T/Tn+FA 免疫的小鼠中淋巴细胞浸润最高。此外，我们使用免疫组织化学（IHC）在巨噬细胞和肿瘤部位监测 FasL、MHC I、CD301 和 T/Tn 的表达水平。在 ox-M-T/Tn+FA 组中，所有标志物的表达均最高。此外，在 ox-M-T/Tn+FA 组中，肿瘤抑制率和存活率最高。这些结果表明，T/Tn 与 ox-M 缀合并与 FA 混合的疫苗制剂可诱导细胞免疫并持续产生体液免疫反应，而不会过度激活免疫系统，从而有效抑制肿瘤生长。
