Allergy and Immunology Section, Institute of Genomics and Integrative Biology (CSIR), Delhi, India.
Clin Exp Allergy. 2012 Mar;42(3):412-22. doi: 10.1111/j.1365-2222.2011.03937.x.
Serine protease activity of Per a 10 from Periplaneta americana induces airway inflammation and systemic Th2 response towards self and bystander allergen.
In the present study the effect of proteolytic activity of Per a 10 allergen on dendritic cells (DCs) polarization and consequent T cell response was investigated.
Non-atopic subjects with no family history of asthma/allergy were recruited for the study. CD14(+) peripheral blood monocytes were purified, differentiated to immature DCs and stimulated with proteolytically active/inactivated native or recombinant Per a 10. DCs phenotype was analysed with flow cytometry and antigen presenting function assessed by co-culturing with autologous CD4(+) T cells. Cytokine levels were determined using ELISA.
Immature DCs differentiated into mature CD14(-)CD83(+)HLA-DR(+) cells after incubating with proteolytically active/inactivated or recombinant Per a 10. Proteolytically active Per a 10 induced significant CD86 up-regulation on DCs compared to inactivated or recombinant Per a 10 lacking enzymatic activity. Proteolytic activity of Per a 10 showed dose-dependent effect on expression of CD80, CD86, CD83, CD1a and HLA-DR. However, no significant differences were observed phenotypically in active or inactive forms except for CD86. Active Per a 10 stimulated DCs secreted significantly low IL-12 (P < 0.01) and high IL-6, compared to inactive forms of Per a 10. Naive CD4(+) T cells primed with active Per a 10 pulsed DCs also secreted significantly less IL-12 (P < 0.01) and high IL-4, IL-5 plus IL-6 (P < 0.01); in contrast to DCs pulsed with inactivated or recombinant Per a 10.
Proteolytic activity of Per a 10 modulates DCs towards type 2 by CD86 up-regulation, high IL-6 and reduced IL-12 secretions. Proteolytically inactive Per a 10 can be further explored for immunotherapy.
美洲大蠊 Per a 10 的丝氨酸蛋白酶活性会引起气道炎症和针对自身及旁观者过敏原的全身性 Th2 反应。
本研究旨在探讨 Per a 10 过敏原的蛋白水解活性对树突状细胞 (DC) 极化及其随后的 T 细胞反应的影响。
本研究招募了无哮喘/过敏家族史的非过敏性受试者。纯化外周血单核细胞，分化为未成熟的 DC，并用蛋白水解活性/失活的天然或重组 Per a 10 刺激。通过流式细胞术分析 DC 表型，并通过与自体 CD4(+)T 细胞共培养评估抗原呈递功能。采用 ELISA 法测定细胞因子水平。
用蛋白水解活性/失活或重组 Per a 10 孵育后，未成熟的 DC 分化为成熟的 CD14(-)CD83(+)HLA-DR(+)细胞。与无酶活性的失活或重组 Per a 10 相比，蛋白水解活性的 Per a 10 诱导 DC 上的 CD86 显著上调。Per a 10 的蛋白水解活性对 CD80、CD86、CD83、CD1a 和 HLA-DR 的表达具有剂量依赖性效应。然而，在活性或非活性形式下，除了 CD86 之外，表型上没有观察到显著差异。与失活的 Per a 10 相比，活性 Per a 10 刺激的 DC 分泌的 IL-12 显著减少 (P < 0.01)，而 IL-6 显著增加。与用失活或重组 Per a 10 脉冲的 DC 孵育的幼稚 CD4(+)T 细胞相比，用活性 Per a 10 脉冲的 DC 分泌的 IL-12 也显著减少 (P < 0.01)，而 IL-4、IL-5 加 IL-6 显著增加 (P < 0.01)。
Per a 10 的蛋白水解活性通过 CD86 的上调、高 IL-6 和低 IL-12 分泌来调节 DC 向 2 型分化。无蛋白水解活性的 Per a 10 可进一步用于免疫治疗。
