Studues of cytokines that keratinocytes in epidermis and fibroblast cells.

为了寻找上皮细胞和骨髓源性细胞的突起，我们选择了皮肤、直肠和肺作为死亡性疾病的标本。为了了解细胞因子在各个表达部位的作用，我们尝试建立了用RT-PCR方法检测mRNA表达的系统。在Bigning，我们选择了特应性皮炎和寻常型牛皮癣进行研究。从皮肤和直肠标本中提取RNA。为制备对照RNA，用适当的限制性内切酶切下每个细胞因子的cDNA，制成新的DNA，克隆到RNA表达载体中，制备合成RNA。将每一条mRNA加入同一试管中，经逆转录酶扩增后，用聚合酶链式反应合成c DNA。经过这一步骤，每个涉及原始和合成的cDNA都被制成PCR.TNFa是更受限制的炎症细胞因子，而IL-8在寻常型银屑病中有更广泛的表达。通过本研究，我们可以在定量水平上检测到细胞因子基因的表达。在下一步中，我们选择了AD和…更多的HMA。RNatES是一种对各种重要的炎症细胞，如嗜酸性粒细胞以及记忆T细胞和单核细胞的有效趋化剂，可能会将这些细胞从循环中招募到炎症灶中。为了进一步了解嗜酸性粒细胞和T细胞的参与与RANTES在AD患者皮肤中的可能参与，我们采用RT-PCR方法同时检测RANTES的mRNAs，以明确AD皮肤中是否存在RANTES。检测RANTES在皮肤中的表达，在AD患者的皮肤和结肠组织中检测到RANTES基因的表达，在哮喘患者的BAL中检测RANTES基因的表达。结果表明，RANTES在皮肤和结肠的T细胞和嗜酸性粒细胞中起重要作用。这些研究表明，皮肤相关淋巴组织、肠道相关淋巴组织和支气管结合淋巴组织在嗜酸性粒细胞和上皮细胞中的表达方式更为简单。这些研究将有助于找出AD、哮喘等过敏性疾病来源特异性的异同。较少

To find the coliration of epitherial cell and bone marrow derived cells, we selected skin, rectum and lung for Atoppic desease. To know the role of cytokines in local of each resion, we try to make the system that measures the mRNA expression by RT-PCR method. AT the bigning, we selected atopic dermatitis and psoriasis vulgaris for study. From the skin and rectum samples, RNA were prepared. To make the control RNA,each cytokine cDNA was cut by appropriate restriction enz.and makes new DNA and cloned into the RNA expression vector and synthesis RNA was prepared. Each mRNA was added to the same sample tube and makes cDNA by reversed transcriptase with primer. After the step, each cDNA that has involved original cDNA and synthesis cDNA has been made PCR.TNFa is more restricted cytokine for inflammation but IL-8 has more broad expression in psoriasis vulgaris. From this study, we colud detect the cytokine gene expression in the level of quantitation. In the next step, we selected AD and at … More hma. RNATES is a potent chemoattractant for various important inflammatory cells such as eosinophils as well as memory T cells and monocytes, potentially recruting these cells from the circulation to an inflamed focus. To extent our understanding of the participation of eosinophils and T cells in relation to the possible involvment of RANTES in the skin of patisnts with AD.We performed RT-PCR assay for simultaneous detection of mRNAS for RANTES to clarify whether RANTES is presence in AD skin. We examined expression of RANTES in the skin, obtained at biopcy, RASNTES mRNA was detected in skin and colon of AD.And also in BAL of asthma, RANTES mRNA was detected. By contrast, normal skin and colon samples were negative for RANTES.Our finding suggest that RANTES plays a part in T cell and eosinophils in skin and colon for AD.These study showde that skin asociated lymphoid tissue, gut associated lymphoid tissue and broncho assocated lymphoid tissue were very simmirer manner in the eosinophils and epithelia. These study will be very useful to find the difference and simirality of orgin specifity of allergic disaese such as AD,Asthma. Less