Generation of cytotoxic T cell responses directed to human leucocyte antigen Class I restricted epitopes from the Aspergillus f16 allergen
Generation of cytotoxic T cell responses directed to human leucocyte antigen Class I restricted epitopes from the Aspergillus f16 allergen
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DOI：
10.1111/j.1365-2249.2005.02738.x
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发表时间：
2005-04-01
影响因子：
4.6
通讯作者：
Keever-Taylor, CA
中科院分区：
文献类型：
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作者：
Ramadan, G;Davies, B;Keever-Taylor, CA
侵袭性曲霉病（IA）是血液系统恶性肿瘤患者感染相关死亡的主要原因，尤其是在造血干细胞移植受者中。我们已经制备了重叠的十五肽（与之前的肽重叠11个氨基酸），其跨越曲霉变应原Aspf 16的整个427个氨基酸编码区，Aspf 16之前在小鼠体内诱导Th 1型细胞应答，在人类中诱导增殖性和细胞毒性CD 4（+）T细胞应答。用完整的肽库脉冲的成熟树突状细胞（DC）用于产生T细胞系。发现来自HLA-B*3501（+）供体的两个细胞系在4-5周的初免后对自体Asp f16-肽池-和曲霉培养物提取物脉冲靶具有强烈的细胞毒性。细胞毒性T淋巴细胞（CTL）培养上清杀死曲霉分生孢子，细胞直接杀死曲霉菌丝。细胞毒性活性和干扰素（IFN）-γ的产生仅由CD 8（+）T细胞介导，以响应池脉冲靶。未检测到白细胞介素（IL）-4产生。CTL活性受HLA-B*3501限制，并且基于肽预测程序，在一个供体中最可能针对YFKYTAAAL（YFK）、LPLCSAQTW（LPL）和GTRFPQTPM（GTR），而来自第二个供体的CTL仅识别LPL。合并脉冲的B*3503（+）BLCL将肽呈递给1号供体，但B*3502（+）或B*3508（+）BLCL不呈递。B*3503（+）BLCL呈递YFK和较小程度的GTR，但不呈递肽LPL。我们的数据表明，除了我们以前确定的II类限制性肽的反应，DC脉冲与十五肽池从天冬氨酸f16能够诱导多克隆，HLA-I类限制性，巨噬细胞特异性T细胞，可能能够赋予免疫IA。
Invasive aspergillosis (IA) is a major cause of infection-related mortality in patients with haematological malignancies, especially in recipients of haematopoietic stem cell transplants. We have prepared overlapping pentadecapeptides (11-aa overlap with previous peptide) spanning the entire 427-aa coding region of the Aspergillus allergen, Asp f16 shown previously in mice to induce Th1-type cell responses in vivo and in humans to induce proliferative and cytotoxic CD4(+) T cell responses. Mature dendritic cells (DC) pulsed with a complete pool of peptides were used to generate T cell lines. Two lines from HLA-B*3501(+) donors were found to be strongly cytotoxic to autologous Asp f16-peptide pool- and Aspergillus culture extract-pulsed targets after 4-5 weekly primings. Cytotoxic T lymphocyte (CTL) culture supernatant killed Aspergillus conidia, and cells directly killed Aspergillus hyphae. Cytotoxic activity and interferon (IFN)-gamma production were mediated exclusively by CD8(+) T cells in response to pool-pulsed targets. Interleukin (IL)-4 production was not detected. CTL activity was restricted by HLA-B*3501 and based on peptide prediction programmes was most probably directed to YFKYTAAAL (YFK), LPLCSAQTW (LPL) and GTRFPQTPM (GTR) in one donor, while only LPL was recognized by CTL from the second donor. Pool-pulsed B*3503(+) BLCL but not B*3502(+) or B*3508(+) BLCL presented peptide to donor no. 1. B*3503(+) BLCL presented YFK and to a lesser extent GTR, but not peptide LPL. Our data show that in addition to our previously identified Class II restricted peptide response, DC pulsed with a pentadecapeptide pool from Asp f16 are capable of inducing polyclonal, HLA-Class I-restricted, Aspergillus-specific T cells that may be capable of conferring immunity to IA.