以树突状细胞(DCs)作为抗原呈递细胞(APCs),表征巴拿马利什曼原虫(Leishmania (Viannia) panamensis)感染期间对动质体膜蛋白-11(KMP-11)的细胞免疫反应。

Characterizing cellular immune response to kinetoplastid membrane protein-11 (KMP-11) during Leishmania (Viannia) panamensis infection using dendritic cells (DCs) as antigen presenting cells (APCs).

作者信息

Delgado Gabriela, Parra-López Carlos A, Vargas Luis Eduardo, Hoya Rubén, Estupiñán Mónica, Guzmán Fanny, Torres Angela, Alonso Carlos, Velez Iván Dario, Spinel Clara, Patarroyo Manuel E

机构信息

Fundación Instituto de Inmunología de Colombia (FIDIC), Departamento de Farmacia, Facultad de Ciencios, Universidad Nacional de Colombia, Universidad Autonoma de Madrid, Spain.

出版信息

Parasite Immunol. 2003 Apr;25(4):199-209. doi: 10.1046/j.1365-3024.2003.00626.x.

Abstract

In vitro peptide binding assays and DCs pulsed with recombinant KMP-11 (rKMP-11) plus six 20-mer overlapping peptides covering the entire protein of Leishmania (Viannia) panamensis (L(V)p) promastigotes were used to identify T-cell epitopes in this protein. Such in vitro binding assays, using HLA DRB1* 0101, -0401, -0701 and -1101 alleles, demonstrated that two peptide sequences (DEEFNKKMQEQNAKFFADKP and FKHKFAELLEQQKAAQYPSK) exhibited high HLA DRB1* 0401 allele binding capacity. rKMP-11 specific T-cell proliferation and cytokine production, derived from 13 volunteers exposed to the parasite, suggested that using autologous DCs as APCs becomes advantageous in uncovering T-cell epitopes promoting proliferation and differences in IFN-gamma and IL-4 production in T-cells from volunteers with ACTIVE and CURED undetectable disease when other APCs were used. The two peptides which bound in vitro to the HLA DRB1* 0401 allele were immunogenic in HLA DRB1* 04 volunteers, thus validating the use of in vitro binding assays for predicting epitopes in this protein. The experimental approach used here may prove useful for characterizing T-cell epitopes in a protein useful in designing peptide-based vaccine candidates for Leishmania and other intracellular pathogens.

摘要

采用体外肽结合试验以及用重组KMP-11(rKMP-11)加覆盖巴拿马利什曼原虫(Viannia亚种)前鞭毛体整个蛋白的六条20聚体重叠肽脉冲处理的树突状细胞(DCs),来鉴定该蛋白中的T细胞表位。使用HLA DRB1* 0101、-0401、-0701和-1101等位基因的此类体外结合试验表明,两条肽序列(DEEFNKKMQEQNAKFFADKP和FKHKFAELLEQQKAAQYPSK)表现出较高的HLA DRB1* 0401等位基因结合能力。来自13名接触过该寄生虫的志愿者的rKMP-11特异性T细胞增殖和细胞因子产生情况表明,当使用其他抗原呈递细胞(APC)时,使用自体DCs作为APC在揭示促进增殖的T细胞表位以及来自患有活动性和已治愈但检测不到疾病的志愿者的T细胞中IFN-γ和IL-4产生差异方面具有优势。在体外与HLA DRB1* 0401等位基因结合的这两条肽在HLA DRB1* 04志愿者中具有免疫原性,从而验证了体外结合试验在预测该蛋白表位方面的用途。此处使用的实验方法可能证明对鉴定一种蛋白中的T细胞表位有用,该蛋白有助于设计针对利什曼原虫和其他细胞内病原体的基于肽的候选疫苗。

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