| Vaccine-induced Cytokine Production Detected by Luminex Multiplex Analysis
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Author:
Date:
2014-08-20
[Abstract] Different vaccine and adjuvant combinations are known to rapidly induce antigen presenting cell (APC) maturation and pro-inflammatory cytokine and production, which in turn play an important role in the priming of antigen-specific T cells. Measuring cytokine production systemically in the serum fails to detect localized responses in the lymph nodes draining a subcutaneous immunization site. On the other hand, stimulating APC with vaccine formulations in vitro lacks the complexity of the lymph node microenvironment and the presence of other in vivo factors. Here we analyse cytokine production directly in vaccine draining lymph nodes (dLN) extracted early after in vivo vaccination. To do this we perform cytokine multiplex analysis of supernatants from whole dLN ...
[摘要] 已知不同的疫苗和佐剂组合快速诱导抗原呈递细胞（APC）成熟和促炎细胞因子和产生，其反过来在抗原特异性T细胞的引发中起重要作用。 在血清中全身测量细胞因子产生不能检测排出皮下免疫位点的淋巴结中的局部反应。 另一方面，用疫苗制剂在体外刺激APC缺少淋巴结微环境的复杂性和其他体内因子的存在。 在这里我们分析细胞因子生产直接在疫苗引流淋巴结（dLN）提前早期在体内接种。 为此，我们在短暂的离体孵育后对来自整个dLN细胞悬浮液的上清液进行细胞因子多重分析。
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| Analysis of Tumor-infiltrating Lymphocytes Following CD45 Enrichment
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Author:
Date:
2014-08-20
[Abstract] Measuring antigen-specific T cell responses in the blood and lymphoid organs of vaccinated mice can give us a useful indication of the potency of a vaccine formulation. Unfortunately, systemic or even localized lymphoid T cell responses are not always predictive of the ability of a vaccine to induce tumor protection. Measuring the antigen-specific T cell response within the tumor infiltrating lymphocytes is a more accurate indicator or vaccine efficacy. However, multi-parameter flow cytometric analysis of T cells isolated from tumor tissue can be quite challenging due to the over-whelming number of tumor cells present in relation to the tumor infiltrating lymphocytes (TIL) and to problems associated to the large and adhesive nature of many tumor cell. Here we take advantage of a pre-flow ...
[摘要] 测量接种疫苗的小鼠的血液和淋巴器官中的抗原特异性T细胞应答可以给予我们疫苗制剂的效力的有用指示。不幸的是，全身性或甚至局部的淋巴T细胞应答并不总是预示疫苗诱导肿瘤保护的能力。测量肿瘤浸润淋巴细胞内的抗原特异性T细胞应答是更准确的指示或疫苗功效。然而，从肿瘤组织分离的T细胞的多参数流式细胞术分析可能是相当具有挑战性的，因为相对于肿瘤浸润性淋巴细胞（TIL）存在的肿瘤细胞的数量过多，以及与大的和粘附性质相关的问题的许多肿瘤细胞。在这里我们利用使用MACS磁性细胞分选系统从肿瘤组织中预流动分离CD45 +白细胞，产生更清洁的细胞制备物，进行流式细胞染色和分析。
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