Development of an ex vivo human skin model for intradermal vaccination: tissue viability and Langerhans cell behaviour.
Development of an ex vivo human skin model for intradermal vaccination: tissue viability and Langerhans cell behaviour.
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DOI：
10.1016/j.vaccine.2009.07.088
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发表时间：
2009-10-09
期刊：
影响因子：
5.5
通讯作者：
Birchall J
中科院分区：
文献类型：
--
作者：
Ng KW;Pearton M;Coulman S;Anstey A;Gateley C;Morrissey A;Allender C;Birchall J
表皮中存在的常驻朗格汉斯细胞(LCS)使皮肤成为DNA疫苗的诱人靶点。然而，用于皮肤接种研究的可靠动物模型是有限的。我们展示了一种用于皮肤DNA疫苗接种的体外人类皮肤模型，该模型可能会弥合临床前体内动物模型和临床研究之间的差距。皮肤转基因表达被用来证明表皮组织在培养中的活性。免疫组织化学方法检测LC对培养环境的反应。在磷酸盐缓冲盐水(PBS)和全器官培养液(OCM)中，全厚和中厚皮肤在至少72小时的培养中保持遗传活性。在OCM中培养的外植体的表皮在整个培养过程中保持形态完整。与中厚皮肤相比，全厚皮肤中的LC对培养条件的反应延迟(细胞数量减少，细胞大小增加)，后者的反应是即刻的。总之，切取的人皮肤可以培养至少72小时，用于基因表达和免疫细胞激活的分析。然而，由于激活的性质，使用中厚皮肤进行疫苗配方研究可能并不合适。全厚皮肤移植是一种更适合于评估皮肤接种的体外模型。
The presence of resident Langerhans cells (LCs) in the epidermis makes the skin an attractive target for DNA vaccination. However, reliable animal models for cutaneous vaccination studies are limited. We demonstrate an ex vivo human skin model for cutaneous DNA vaccination which can potentially bridge the gap between pre-clinical in vivo animal models and clinical studies. Cutaneous transgene expression was utilised to demonstrate epidermal tissue viability in culture. LC response to the culture environment was monitored by immunohistochemistry. Full-thickness and split-thickness skin remained genetically viable in culture for at least 72 h in both phosphate-buffered saline (PBS) and full organ culture medium (OCM). The epidermis of explants cultured in OCM remained morphologically intact throughout the culture duration. LCs in full-thickness skin exhibited a delayed response (reduction in cell number and increase in cell size) to the culture conditions compared with split-thickness skin, whose response was immediate. In conclusion, excised human skin can be cultured for a minimum of 72 h for analysis of gene expression and immune cell activation. However, the use of split-thickness skin for vaccine formulation studies may not be appropriate because of the nature of the activation. Full-thickness skin explants are a more suitable model to assess cutaneous vaccination ex vivo.
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DOI：
10.1097/00004630-199107000-00005
发表时间：
1991-01-01
期刊：
Journal of Burn Care and Rehabilitation
影响因子：
--
作者：
JOHNSON L F;DESERRES S;MEYER A A
通讯作者：
MEYER A A
影响因子：
6.5
作者：
Bauer, J;Bahmer, FA;Fartasch, M
通讯作者：
Fartasch, M
影响因子：
1.5
作者：
Hautier, A.;Sabatier, F.;Magalon, G.
通讯作者：
Magalon, G.
DOI：
10.1186/1479-0556-1-2
发表时间：
2003-09-16
期刊：
Genetic vaccines and therapy
影响因子：
--
作者：
Garmory, Helen S;Brown, Katherine A;Titball, Richard W
通讯作者：
Titball, Richard W
影响因子：
4.4
作者：
de Gruijl, TD;Sombroek, CC;Pinedo, HM
通讯作者：
Pinedo, HM