目前乳癌位居女性癌症發生率第一位，死亡率第四位，嚴重威脅婦女健康。介白素-19(IL-19)和介白素-20(IL-20)屬於介白素-10家族的一員，目前已經知這兩分子參與在多種發炎反應相關疾病的病程，然而有研究指出，在腫瘤微環境中的發炎介質，包括細胞激素和趨化因子，可以影響乳癌的進展過程。並且實驗室先前的研究也發現到IL-19和IL-20會在病人腫瘤組織切片中偵測到，本實驗室已分別針對這兩分子在乳癌中扮演的角色，得知IL-19和IL-20與乳癌發展有關聯性。IL-19和IL-20藉由共同受體複合體IL-20 R1/IL-20R2來傳遞訊息 : 為了發展出具有潛力的抗癌藥物，我們製造出針對人類IL-20 R1蛋白的單株抗體，其中之一抗體為51D。實驗上，首先藉由表現不同長度的IL-20 R1蛋白，以用來決定抗原和抗體(51D)的結合位置。而細胞實驗中則搭配使用老鼠乳癌細胞(4T1)和人類乳癌細胞(MDAMB231)，在細胞增生和菌落生成能力分析結果發現51D能分別抑制由IL-19所促進的MDAMB231以及IL-20所促進的4T1細胞的生長和菌落生成能力。同時IL-19也會促進MDAMB231細胞的遷徙能力及誘導細胞內發炎物質(TGF-β, IL-1β) 及與轉移相關基因 (CXCR4 MMP2, MMP9)表現量上升，而這些現象皆能藉由加入51D達到抑制的效果。相對而言，IL-20能促進與骨頭破壞有關的因子蛋白酶G、K在4T1細胞裡表達量上升，但其表達量能被51D抑制下來。在動物實驗中也可以觀察到，IL-20 R1的單株抗體，51D，可以阻斷IL-19和IL-20與其受體複合體的結合，進而抑制腫瘤細胞的生長。同時在IL-20 R1的基因剔除鼠與野生鼠的動物實驗中，也能觀察到IL-20 R1基因剔除鼠有較小的腫瘤生成。所以綜合細胞和動物實驗結果，除了再次證明IL-19和IL-20參與乳癌的致病過程，也證明了IL-20 R1的單株抗體具有中和(neutralize) IL-19和IL-20活性的能力，在治療上有極大潛力。

Breast cancer is one of the leading causes of cancer-related mortality in women. Inflammatory mediators, such as cytokines and chemokines in tumor microenvironments affect the progression of breast cancer. Interleukin-19 and Interleukin-20 are cytokines that belong to the IL-10 family. Our previous studies revealed both cytokines are involved in the pathogenesis of breast cancer. Both IL-19 and IL-20 bind to IL-20 R1/IL-20R2 heterodimer to activate signal transduction. To develop a potential drug for treating breast cancer, we have generated the anti-human IL-20 R1 monoclonal antibody to block cytokine function. 51D is one of anti-IL-20 R1 monoclonal antibody. To characterize the antibody, we used the serial deletion of the antigen to determine the binding epitope between 51D and IL-20R1. Boyden chamber assay reveled that IL-19 increased the migration of MDAMB231, a human breast cancer cell line, the activity of which was inhibited by 51D. IL-19 induced the expression of the pro-inflammatory cytokines (TGF-β, IL-1β) and metastasis-associated gene (CXCR4 MMP2, MMP9) in MDAMB231 cells. 51D also inhibited the induction of these genes by IL-19. Furthermore, MTT assays showed that IL-19 and IL-20 promoted the proliferation of MDAMB231 cells and mouse breast cancer cell line 4T1 which was neutralized by 51D. In addition, soft agar colony formation assay showed IL-19 and IL-20 enhanced colony formation of breast cancer cells the activity of which was inhibited by 51D. In vivo, anti- IL-20 R1 antibody, 51D, inhibited MDAMB231 tumor growth. Furthermore, tumor size and tumor weight in the IL-20 R1 knockout mice was smaller than those in the wild type. Our study provides the evidence that 51D can be a potent antagonist to neutralize IL-19 and IL-20 activity.

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