尿調理素（uromodulin；URO）是一種由孕婦尿中純化之醣蛋白，分子量約 90-100 kDa。在過去的研究中，發現URO 可以誘發週邊血液單核性細胞（peripheral blood mononuclear cells；PBMC）產生發炎性細胞激素（inflammatory cytokine），並且可以抑制抗原專一性的 T 細胞增殖，但其免疫調節的機轉至今仍不清楚。本篇論文分成兩個方向探討這個問題，其一是探討 URO 是如何作用在 PBMC 上。其二是 URO 對 T 細胞分泌的細胞激素與表現輔助刺激因子（costimulatory molecules）的影響。
我們利用矽藻土純化出 URO，再用蛋白酵素（protease）切除蛋白質部分製造出 UROp（URO 的醣類部份）。用 URO 與 UROp 作用在 PBMC 上，結果發現 UROp 與 URO 相似，皆能刺激 PBMC 的增殖，並且引起 CD14+ 的細胞族群相對的增加。這些相對增加的 CD14+ 細胞（單核球；monocyte）還表現輔助刺激分子CD80。除此之外，URO 與 UROp還可以活化人類 PBMC 釋出發炎性細胞激素，如腫瘤壞死因子 a（TNF-a）、介白素 1 號 b（IL-1b）及介白素 6 號（IL-6），以及趨化素（chemokine）介白素 8 號（IL-8）。同時我們發現，URO及 UROp 也可以使單核球上的TLR4（toll-like receptor 4）表現量增加，並且TLR4 抗體可以部分抑止 URO 及 UROp 所引起的 TNF-a 的產生。綜合上述實驗結果，我們認為 URO 及 UROp 可能經由 TLR4 刺激單核球引起類似發炎的免疫反應。
有趣的是，雖然 URO 及 UROp 可以引起 PBMC 產生類似發炎的免疫反應，卻無法測得 IFN-g 的產生。即使用 RT-PCR 看 IFN-g mRNA 的表現，URO 與 UROp也只能刺激 PBMC 產生微量的 IFN-g mRNA。為了進一步了解為何URO無法使PBMC產生IFN-g，我們先觀察 URO 刺激 PBMC 後T 細胞活化情形。結果發現，經由 URO 或 UROp作用後的 PBMC，其 T 細胞表面分子 CD152 （CTLA-4）可被刺激而增高。若同時用純化的 T 細胞與 URO 或 UROp 作用，發現除了可以抑制 CD3 / CD28 所引起的T細胞增殖外，URO 與 UROp還可降低已活化T 細胞（CD45RO+）表面分子 CD28 及 CD152 的表現。另外，因為 IFN-g 的產生和單核球是否產生 IL-12 相關，因此我們使用一種類似單核球的細胞株 THP-1 與 URO 及 UROp 作用。結果發現，URO 並不能抑制由 LPS 刺激 THP-1 所引起的 IL-12 之產生，但 UROp 則可些微抑制 IL-12 的產生。
總而言之，URO 和 UROp 可能可以經由 TLR4 刺激單核球增殖和產生發炎性細胞激素，但對於 T 細胞的活化則傾向抑止，進而影響 IFN-g 的產生，並且 URO 之免疫調理作用可能與其醣類的結構 UROp 有極大的關係。

Uromodulin (URO) with molecular weight about 90-100kDa is the most abundant glycoprotein in the urine of pregnant women. URO can induce human monocytes to secrete proinflammatory cytokines. On the other hand, URO can also inhibit antigen-specific T cell proliferation. The mechanism of URO to modulate these two contrasting immune responses is unclear. In this study, URO was purified from urine using diatomaceous earth filter. Both URO and URO after protease digestion (UROp) were used to understand the mechanism of URO to regulate monocytes and T cells response. URO and UROp induced human peripheral blood mononuclear cells (PBMC) to proliferate and increased the percentage of CD14+ cells (monocytes). In addition, both CD80 and toll like receptor 4 (TLR4) expression of these monocytes was increased after URO or UROp stimulation. The production of proinflammatory cytokines such as TNF-a, IL-1b and IL-6 and chemokine (IL-8) by PBMC after URO or UROp stimulation was also increased. Antibodies against TLR4 could inhibit URO and UROp induced TNF-a production of PBMC. These results indicated that TLR4 might be involved in URO and UROp induced proinflammtory cytokines production of monocytes.
Interestingly, unlike LPS, URO and UROp induced only trace amount of IFN-g production both protein and mRNA levels. In addition, using anti-CD3 and anti-CD28 coated plates, we found in the presence of URO and UROp, T cells proliferation was inhibited. In addition, active T cells (CD45RO+) were decreased and the costimulatory molecules CD28 and CD152 (CTLA-4) on these cells were down-regulated in the presence of URO or UROp. It is possible inhibition of costimulatory molecules expression of T cells by URO and UROp may contribute to the inhibition of T cell proliferation. In addition, to the costimulatory molecules, IL-12 produced by monocyte was also required for T cell to produce IFN-g. Therefore, a promonocytic cell line, THP-1 was used to understand the effect of URO and UROp on the production of IL-12 by monocytes. It was found UROp, like mannan, could inhibit LPS induced IL-12 production of THP-1 cells. However, URO was unable to do this. In summary, these results indicate the carbohydrate moiety of URO may play important roles in the immunomodulation induced by URO.

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