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Flow Cytometric Determination of Cellular Sources and Frequencies of Key Cytokine-Producing Lymphocytes Directed against Recombinant LACK and Soluble Leishmania Antigen in Human Cutaneous Leishmaniasis

机译:流式细胞仪测定人皮肤利什曼病中针对重组LACK和可溶性利什曼原虫抗原的主要细胞因子产生淋巴细胞的细胞来源和频率

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摘要

Leishmaniasis, caused by infection with the protozoan parasite Leishmania, affects millions of individuals worldwide, causing serious morbidity and mortality. This study directly determined the frequency of cells producing key immunoregulatory cytokines in response to the recombinant antigen Leishmania homolog of receptors for activated kinase C (LACK) and soluble leishmania antigen (SLA), and it determined relative contributions of these antigens to the overall cytokine profile in individuals infected for the first time with Leishmania braziliensis. All individuals presented with the cutaneous clinical form of leishmaniasis and were analyzed for proliferative responses to LACK antigen and SLA, frequency of lymphocyte subpopulations (analyzed ex vivo), and antigen-induced (LACK and SLA) cytokine production at the single-cell level (determined by flow cytometry). The following were determined. (i) The Th1-type response previously seen in patients with cutaneous leishmaniasis is due to gamma interferon (IFN-γ) production by several different sources, listed in order of contribution: CD4+ T lymphocytes, CD4−, CD8− lymphocytes, and CD8+ T lymphocytes. (ii) SLA induced a higher frequency of lymphocytes producing IFN-γ and tumor necrosis factor alpha (TNF-α) than did LACK. (iii) LACK induced an activation of monocyte populations as reflected by an increased percentage of CD14-positive cells. (iv) Neither SLA nor LACK induced detectable frequencies of cells producing interleukin-4 (IL-4) or IL-5. These data demonstrated a multifaceted immune response to SLA in human leishmaniasis involving Th1 CD4+ T lymphocytes (IFN-γ+ and IL-10−/IL-4−), Tc1 CD8+ T cells (IFN-γ+, and IL-10−/IL-4−), and a high frequency of TNF-α-producing lymphocytes. Moreover, it was determined that the recombinant antigen LACK acts as a weak inducer of Th1-type lymphocyte responses compared to SLA.
机译:由原生动物寄生虫利什曼原虫感染引起的利什曼病,影响全世界数以百万计的个体,导致严重的发病率和死亡率。这项研究直接确定了细胞响应活化激酶C(LACK)和可溶性利什曼原虫抗原(SLA)受体的重组抗原利什曼原虫同源物而产生关键免疫调节细胞因子的频率,并且确定了这些抗原对总体细胞因子谱的相对贡献首次感染巴西利什曼原虫的个体。所有表现出利什曼病皮肤病临床表现的个体均在单细胞水平上分析了对LACK抗原和SLA的增殖反应,淋巴细胞亚群的频率(离体分析)以及抗原诱导的(LACK和SLA)细胞因子的产生(通过流式细胞仪测定)。确定了以下内容。 (i)先前在皮肤利什曼病患者中观察到的Th1型反应是由于几种不同来源产生的γ-干扰素(IFN-γ),按贡献顺序列出:CD4 + T淋巴细胞,CD4-,CD8-淋巴细胞和CD8 + T淋巴细胞。 (ii)SLA诱导淋巴细胞产生IFN-γ和肿瘤坏死因子α(TNF-α)的频率高于LACK。 (iii)LACK诱导了单核细胞群体的激活,这表现为CD14阳性细胞百分比的增加。 (iv)SLA和LACK均未诱导产生白介素4(IL-4)或IL-5的细胞的可检测频率。这些数据表明,在人类利什曼病中,针对SLA的多方面免疫反应涉及Th1 CD4 + T淋巴细胞(IFN-γ+和IL-10- / IL-4-),Tc1 CD8 + T细胞(IFN-γ+和IL-10- / IL-4-)和高频率的产生TNF-α的淋巴细胞。此外,已确定与SLA相比,重组抗原LACK充当Th1型淋巴细胞应答的弱诱导物。

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