Barral-Netto M, Barral A, Brodskyn C, Carvalho E M, Reed S G
Universidade Federal da Bahia, Salvador-Bahia, Brazil.
Parasite Immunol. 1995 Jan;17(1):21-8. doi: 10.1111/j.1365-3024.1995.tb00962.x.
CD8+ T cells and lysis of parasitized macrophages seem to be important in the resistance to murine leishmaniasis. In the present study, we evaluated peripheral blood mononuclear cell (PBMC) from patients with either cutaneous (CL) or mucosal (ML) leishmaniasis in cell lysis assays using 51-Cr-labeled Daudi or K562 cells, or autologous antigen-pulsed macrophages as targets. Results are reported as lytic units (number of cells required for 30% lysis) per million PBMC. Exposure of patient PBMC (n = 12) to lysate from Leishmania amazonensis promastigotes led to an increase in cytotoxic activity compared to unstimulated patient cells against Daudi (81.8 +/- 14.9 vs 13.6 +/- 5 lytic units (LU) per million PBMC; mean +/- SEM) and K562 (65.7 +/- 8.4 vs 13.1 +/- 5 LU/10(6) PBMC). ML had higher responses than CL in both targets (80.4 +/- 11.0 vs 46.4 +/- 11.6 LU/10(6) PBMC for K562, and 104.3 +/- 23.8 vs 59.3 +/- 14.3 LU/10(6) PBMC for Daudi). Normal control PBMC, stimulated with L. amazonensis antigen had 6.32 +/- 3.72 LU/10(6) PBMC against Daudi cells and 9.06 +/- 2.78 LU/10(6) PBMC against K562. The cell responsible for lysis of the K562 cells was characterized as NK, by means of cell separation employing magnetic beads coupled to antibodies. Addition of recombinant TGF-beta or recombinant human IL-10 reduced L. amazonensis-induced cytotoxicity by 90% and 70%, respectively. Cytotoxicity of antigen-stimulated PBMC was also demonstrated against autologous L. amazonensis antigen-pulsed macrophages in the range of 6.7 to 41.7 LU/10(6) PBMC.(ABSTRACT TRUNCATED AT 250 WORDS)
CD8 + T细胞以及对被寄生巨噬细胞的裂解作用在抵抗小鼠利什曼病中似乎很重要。在本研究中，我们在细胞裂解试验中使用51 - Cr标记的Daudi或K562细胞，或以自体抗原脉冲巨噬细胞作为靶标，评估了皮肤型（CL）或黏膜型（ML）利什曼病患者的外周血单核细胞（PBMC）。结果以每百万PBMC的裂解单位（实现30%裂解所需的细胞数）报告。与未刺激的患者细胞相比，患者PBMC（n = 12）暴露于亚马逊利什曼原鞭毛体裂解物后，对Daudi细胞（每百万PBMC的裂解单位分别为81.8±14.9和13.6±5）和K562细胞（65.7±8.4和13.1±5 LU/10⁶ PBMC）的细胞毒性活性增加。在两个靶标上，ML的反应均高于CL（对K562细胞分别为80.4±11.0和46.4±11.6 LU/10⁶ PBMC，对Daudi细胞分别为104.3±23.8和59.3±14.3 LU/10⁶ PBMC）。用亚马逊利什曼原虫抗原刺激的正常对照PBMC对Daudi细胞的裂解单位为6.32±3.72 LU/10⁶ PBMC，对K562细胞为9.06±2.78 LU/10⁶ PBMC。通过使用与抗体偶联的磁珠进行细胞分离，将负责裂解K562细胞的细胞鉴定为NK细胞。添加重组TGF-β或重组人IL-10分别使亚马逊利什曼原虫诱导的细胞毒性降低了90%和70%。抗原刺激的PBMC对自体亚马逊利什曼原虫抗原脉冲巨噬细胞的细胞毒性也得到证实，范围为6.7至41.7 LU/10⁶ PBMC。（摘要截断于250字）
