Fungicidal activity of human peripheral blood leukocytes against Paracoccidioides brasiliensis yeast cells
Fungicidal activity of human peripheral blood leukocytes against Paracoccidioides brasiliensis yeast cells
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DOI：
10.1080/13693780400011088
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发表时间：
2005-08-01
期刊：
影响因子：
2.9
通讯作者：
Brummer, E
中科院分区：
文献类型：
--
作者：
Kurita, N;Oarada, M;Brummer, E
虽然流行病学研究结果表明正常人对巴氏副球藻感染具有抵抗力，但宿主对这种真菌的防御机制尚未完全清楚。在这里，我们测试了人类白细胞对这种真菌的酵母细胞的抗真菌活性，使用了一种改进的真菌培养基对酵母细胞具有高培养效率。为了尽量减少分离过程中白细胞活性的损害，将通过离心法从外周血的黄褐色涂层中取出的白细胞用于抗真菌试验，而不进行进一步的分离。用这种方法制备的白细胞在共培养的前4h内有效地杀死了巴西巴氏杆菌酵母细胞。加入干扰素-γ(37 ng/m l)、粒-巨噬细胞集落刺激因子(5 ng/m l)、白介素1-β(12 ng/m l)或白介素4(12 ng/m l)可使白细胞的抗真菌(生长抑制)活性增强48 h，而加入IL-8(50 ng/m l)则抑制白细胞的生长。肿瘤坏死因子-α(50 ng/ml)或IL-10(25 ng/ml)对此无影响。地塞米松(1 mmol/L)可降低白细胞的抗真菌活性。这是第一次证明人类白细胞能够有效地杀死巴西巴氏杆菌的酵母细胞。
Although epidemiological findings suggest that normal humans are resistant to Paracoccidioides brasiliensis infection, the host defense mechanisms against this fungus have not been fully understood. Here we examined human leukocytes for antifungal activity against yeast cells of this fungus, using an improved mycological culture medium with high plating efficiency for the yeast cell. In an attempt to minimize the impairment of leukocyte activities during the isolation process, leukocytes removed by centrifugation from a buffy coat of peripheral blood were used in the antifungal assay without further fractionation. The leukocytes thus prepared effectively killed P. brasiliensis yeast cells within the first 4 h of co-culture. Adding interferon-gamma ( 37 ng/ml), granulocyte - macrophage colony-stimulating factor ( 5 ng/ml), interleukin (IL)-1 beta ( 12 ng/ml), or IL-4 ( 12 ng/ml) to the assay system enhanced the leukocyte antifungal ( growth inhibitory) activity by 48 h. By contrast, addition of IL-8 ( 50 ng/ml) impaired the leukocyte activity. Tumor-necrosis factor-alpha ( 50 ng/ml) or IL-10 ( 25 ng/ml) had no effect in this respect. Dexamethasone ( 1 mmol/l) reduced the antifungal activity of leukocytes. This is the first demonstration that human leukocytes are able to effectively kill yeast cells of P. brasiliensis.