Primary Explosive Blast-induced Traumatic Brain Injury Model in PC12 Cell Culture
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Author:
Date:
2016-08-20
[Abstract] While it is understood that structural damage occurs at the cellular level from the traumatic brain injury event, the effect on functional activity remains largely unknown. Simplified models such as in vitro models of primary explosive blast are critically needed to deconvolute mechanisms of cellular damage. This protocol details an in vitro indoor experimental system setup (Zander et al., 2015) using real military explosive charges to more accurately represent battlefield blast exposure, and probe the effects of primary explosive blast on dissociated neurons.
[摘要] 细胞毒性CD8 + T细胞能够通过其T细胞受体(TCR)与主要组织相容性复合物(MHC)分子呈递的小免疫原性肽(抗原)之间的特异性相互作用特异性识别和杀死靶细胞。抗原特异性细胞毒性T细胞的抗原识别能力和体外裂解活性可以在所谓的铬51(<51> Cr)释放测定中功能性评估,其是几乎50年前在我们的机构中发展起来的(Brunner等人,1968年)。放射性标记的内源性抗原呈递缺陷的细胞[例如,用于抗原呈递(TAP)缺陷型T2细胞的转运蛋白],并用感兴趣的MHC稳定转染(例如 ,HLA-A2 sup + +)通常在这个4小时测定期间用作靶标。或者,内源性呈递免疫原性抗原的Cr标记的病毒感染或肿瘤细胞系可以作为靶细胞(例如,用于评估肿瘤识别)。 在肽滴定测定(部分A)中,用抗原性肽的系列稀释物对放射性标记的靶细胞进行脉冲,并在效应物(例如,CD8 + T细胞克隆)与靶细胞( Cr-T2细胞)比(E:T)为10:1的混合物在96孔V型底板中37℃温育4小时。在肿瘤杀伤试验(B部分)中,将细胞毒性CD8 51> ...
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Detection of Tumor Cell Surface-reactive Antibodies
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Author:
Date:
2014-05-05
[Abstract] Vaccine-based immunotherapy is being used to treat dogs with primary brain tumors. The vaccines are composed of a lysate of autologous tumor cells, which stimulate an immune response producing tumor specific antibodies that are capable of inducing antibody-dependent cell-mediated cytotoxicity to allogeneic, as well as autologous, tumor cells. This protocol will describe the tumor cell serum antibody-binding assay to measure the tumor-reactive IgG antibody response. Key features of this assay are that it is performed with sera collected from the canine patient prior to and following ...
[摘要] 基于疫苗的免疫疗法被用于治疗具有原发性脑肿瘤的狗。 疫苗由自体肿瘤细胞的裂解物组成,其刺激产生免疫应答的肿瘤特异性抗体,其能够诱导对同种异体以及自体的肿瘤细胞的抗体依赖性细胞介导的细胞毒性。 该方案将描述肿瘤细胞血清抗体结合测定以测量肿瘤反应性IgG抗体应答。 该测定的主要特征是,在接种疫苗之前和之后从犬患者收集的血清作为抗体的来源进行,并且使用犬脑肿瘤细胞作为靶细胞。
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