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Nove1 EBV LMP2-Affibody and Affitoxin as Potential Agents for Molecular Imaging and Targeted Therapy of Nasopharyngeal Carcinoma

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Nove1 EBV LMP2-Affibody and Affitoxin as Potential Agents fo...

小鼠PD-1及其配体PD-L1胞外区基因的原核表达及亲和性

浙江省免疫学会第六次会员代表大会暨第十一次学术大会

作者： Shanli Zhu Jun Chen Yirong Xiong kamara saidu Meiping Gu Wanlin Tang Shao Chen Haiyan Dong Xiangyang Xue Lifang Zhang Institute of Molecular Virology and Immunology Department of Microbiology and Immunology School of Basic Medical Sciences Wenzhou Medical University

Epstein-Barr virus(EBV)infection is closely linked to several human malignancies including endemic Burkitt’s lymphoma,Hodgkin’s lymphoma and nasopharyngeal carcinomas(NPC).Latent membrane protein 2(LMP2)of EBV plays a pivotal role in pathogenesis of EBV-related tumors and thus,is considered to be a potential target for diagnosis and targeted therapy of EBV-related malignant *** molecules are developing as imaging probes and tumor-targeted delivery of small *** this study,four EBV LMP2-binding affibodies(Z12,Z132,Z137,and Z142)were identified by screening a phage-displayed peptide library and used for molecular imaging and targeted therapy in EBV xenograft mice model.Z affibody has high binding affinity for EBV LMP2 and accumulates in mouse tumors derived from EBV xenografts for 24 h after intravenous(Ⅳ)*** fusion of exotoxin PE38 KDEL to the Z 142 affibody led to production of Z142 X *** fused Z142 X affitoxin exhibits highly cytotoxicity specific for EBV-positive cells in vitro and significantly antitumor effect in mice bearing EBV-positive tumor xenografts by IV ***,our data suggest that EBV LMP2-specific affibody and affitoxin represent a novel agent for molecular imaging diagnosis of EBV-induced cancers and for the tumor-targeted delivery of anticancer toxins.

关键词： EBV LMP2 affibody molecules Nasopharyngeal carcinoma in vivo imaging Targeted therapy

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温州医科大学学报 2018年第6期48卷 401-407,412页

作者：吕开绩陈旭东程开王路得朱进顺 kamara saidu 张丽芳朱冠保温州医科大学附属第一医院胃肠外科浙江温州325015 温州医科大学分子病毒与免疫研究所微生物学与免疫学教研室浙江温州325035

目的:制备小鼠PD-1胞外区(mPD-1)与其配体PD-L1胞外区(mPD-L1)原核表达蛋白,并制备mPDL1兔多克隆抗体,在体外对mPD-L1和mPD-1的结合亲和性进行验证。方法:通过分子克隆方法分别构建重组质粒pET21a/mPD-L1和p GEX4T-1/mPD-1,并通过原核... 详细信息

目的:制备小鼠PD-1胞外区(mPD-1)与其配体PD-L1胞外区(mPD-L1)原核表达蛋白,并制备mPDL1兔多克隆抗体,在体外对mPD-L1和mPD-1的结合亲和性进行验证。方法:通过分子克隆方法分别构建重组质粒pET21a/mPD-L1和p GEX4T-1/mPD-1,并通过原核表达系统制备相应的重组蛋白并纯化,通过Western blot进行鉴定;将纯化的mPD-L1蛋白免疫健康日本大耳白兔制备兔多克隆血清抗体,用ELISA、Western blot分析得到的多克隆抗体的效价和特异性,并且将该多克隆抗体作为免疫荧光的阳性对照。最后应用ELISA和免疫荧光验证mPD-1与其配体mPD-L1蛋白的亲和性。结果:SDS-PAGE和Western blot结果显示原核表达的重组蛋白与预期大小一致,mPD-L1和mPD-1分别为25 kDa和40 kDa。经mPD-L1蛋白免疫后的日本大耳白兔能够产生特异性抗体,抗体效价在免疫后第6周达到高峰,Western blot检测结果显示多克隆血清可特异性识别mPD-1。ELISA结果表明,原核表达的mPD-L1蛋白能够和mPD-1蛋白结合,实验组OD450值明显高于对照组。免疫荧光结果显示,在小鼠PD-L1阳性细胞B16(小鼠黑色素瘤细胞株)的胞膜区出现绿色荧光团块。结论:利用原核细胞表达并纯化后获得mPD-L1和mPD-1蛋白,并在体外成功验证了mPD-L1和mPD-1之间的结合特性,为后期利用mPD-1和PD-L1的生物学特性研究提供了基础。

关键词： mPD-1 mPD-L1 原核表达干扰素 ELISA 免疫荧光小鼠

新型冠状病毒E蛋白结构与免疫优势表位的信息学预测分析

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温州医科大学学报 2021年第3期51卷 209-214,219页

作者：李明洋 saidu kamara 汪琪郭艳茹李青峰朱珊丽陈俊蒋朋飞张丽芳温州医科大学分子病毒与免疫研究所病原生物与免疫学系浙江温州325035

目的:应用生物信息学方法预测和分析新型冠状病毒(SARS-CoV-2)包膜蛋白(E蛋白)的结构及免疫优势表位。方法:从NCBI GenBank数据库中检索SARS-CoV-2 E蛋白的氨基酸序列,通过Expas y服务器上的软件分析预测其理化性质、二级结构、跨膜区域... 详细信息

目的:应用生物信息学方法预测和分析新型冠状病毒(SARS-CoV-2)包膜蛋白(E蛋白)的结构及免疫优势表位。方法:从NCBI GenBank数据库中检索SARS-CoV-2 E蛋白的氨基酸序列,通过Expas y服务器上的软件分析预测其理化性质、二级结构、跨膜区域;并结合其亲水性、表面可及性、极性、抗原性和柔韧性等综合分析,预测其可能的B细胞表位;利用Net CTL1.2 server预测CTL表位;通过Vector NTI对冠状病毒属E蛋白进行同源性分析;同时应用Swiss Model对E蛋白进行三维结构同源建模及功能结构域预测分析。结果:SARS-CoV-2 E蛋白由75个氨基酸残基组成,为跨膜蛋白,跨膜区域以α螺旋为主;E蛋白可能含有2个B细胞表位和6个CTL表位,其免疫优势表位区域分别位于其N端16-34aa和C端50-70aa区段;SARS-CoV-2与SARS-CoV和Bat-SARS-like-CoV E蛋白的免疫优势表位存在极高的同源性;其疏水性跨膜结构和PDZ结合基序分别位于N端12-34aa和C端72-75aa区段。结论:E蛋白为跨膜蛋白,其氨基酸序列的16-34和50-70区段可能为免疫优势表位。

关键词：新型冠状病毒包膜蛋白生物信息学免疫优势表位同源性分析

EB病毒LMP1 C端重组蛋白多克隆抗体的制备和鉴定

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温州医科大学学报 2018年第7期48卷 469-473页

作者：毛珊珊王路得周萌吕开绩朱进顺 kamara saidu 叶晓鲜张丽芳温州医科大学分子病毒与免疫研究所病原生物与免疫学系浙江温州325035

目的:制备EB病毒(EBV)潜伏膜蛋白1(LMP1)C端(187～386)的原核表达蛋白及其多克隆抗体并检验其生物学特性。方法:选择LMP1C端的基因并进行原核密码子优化,全基因合成pET21a(+)/EBVLMP1C端重组质粒,并转入***21(DE3)感受态细菌进行诱导表... 详细信息

目的:制备EB病毒(EBV)潜伏膜蛋白1(LMP1)C端(187～386)的原核表达蛋白及其多克隆抗体并检验其生物学特性。方法:选择LMP1C端的基因并进行原核密码子优化,全基因合成pET21a(+)/EBVLMP1C端重组质粒,并转入***21(DE3)感受态细菌进行诱导表达重组蛋白,利用His标签提取纯化重组蛋白并经SDS-PAGE及Westernblot分析鉴定。将纯化的LMP1C端重组蛋白免疫日本大耳白兔,获得多克隆抗体,分别采用ELISA、Westernblot及免疫荧光方法对兔多克隆抗体进行分析。结果:EBVLMP1C端(187～386)重组蛋白可经原核表达系统表达;通过免疫兔获得了特异性的多克隆Ig G抗体;制备的兔多克隆抗体可特异性结合并识别LMP1C端蛋白。结论:EBVLMP1C端重组蛋白免疫原性较强,制备的兔多克隆抗体效价高、特异性强。

关键词： EB病毒 LMP1蛋白多克隆抗体

Extent of Reduction of the Fallow Period and Its Impact on Upland Rice Production in the Nongowa Chiefdom of Kenema District in Eastern Sierra Leone

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Agricultural Sciences 2016年第11期7卷 805-812页

作者： Alie kamara Osman Sidie Vonu John Lansana John Lansana Foday saidu Sesay Department of Soil Science School of Agriculture Njala Campus Njala University Njala Sierra Leone Department of Agriculture Eastern Polytechnique Kenema Sierra Leone

Slash-and-burn agriculture is the common practice for upland rice intercropping system in Sierra Leone and it has been blamed for the loss of forest vegetation across the country. Shortening of the fallow period in the Eastern region of Sierra Leone has implications on agricultural food production and the persistence of the remaining tropical rainforest. This study was therefore undertaken to assess: 1) the extent of reduction of the fallow period in the Nongowa Chiefdom of Kenema District;2) the causes of reduction in the fallow period;3) farmers’ perception of the relevance of fallow period and the implications of reduction of fallow period on crop production. The study revealed a drastic reduction of the fallow period in the chiefdom with a mean of 3.8 years which falls far below the country mean fallow period of 8.8 years as estimated by FAO. Farmers pointed out that the reduction of the fallow period was as a result of land scarcity due to competing land uses such as large scale commercial agriculture, logging, mining, charcoal burning and expansion of settlements. Farmers agreed that a fallow period of at least 10 years is necessary for upland rice production. Farmers were aware of the impact of reduction of the fallow period on upland rice production and were able to estimate rice yields based on the age of the fallow. Also, farmers have attempted to adapt to the constraints posed by shorter fallow periods by selection of rice varieties to suit the length of the fallow periods.

关键词： Shifting Cultivation Slash-and-Burn Fallow Period Upland Rice Farming Nongowa Chiefdom Eastern Sierra Leone

Effect of Land Uses on Soil Erodibility in the Njala Area of Southern Sierra Leone

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Open Journal of Soil Science 2022年第10期12卷 475-489页

作者： Denis Magnus Ken Amara Ibrahim Benya Sheku Alfred Kanu Daniel Hindogbe saidu Raymond Morie Musa Osman Sidie Vonu Fayia Brima Joseph Christian Adamu Mboma Michael Jusu Foday Turay Abdulai kamara Department of Soil Science School of Agriculture & Food Sciences Njala University Njala Campus Njala Sierra Leone Department of Soil Science Faculty of Development Agriculture & Natural Resources Management Eastern Technical University of Sierra Leone Private Mail Bag Kenema Sierra Leone Project Management Office Mamour Agricultural Foundation Freetown Sierra Leone Department of Crop Science School of Agriculture & Food Sciences Njala University Njala Campus Sierra Leone Department of Agriculture Food Security & Livelihood Sustainable Cassava Commercialization through Entrepreneurs Strengthening & Support (SUCCESS) Project Bangladeshi Rural Agro Committee (BRAC) Bo Sierra Leone Department of Agricultural Education Faculty of Education Eastern Technical University of Sierra Leone Kenema Sierra Leone Department of Social Studies Faculty of Education Eastern Technical University of Sierra Leone Kenema Sierra Leone Department of Agriculture & Forestry Faculty of Development Agriculture & Natural Resources Management Eastern Technical University of Sierra Leone Kenema Sierra Leone Department of Agricultural Engineering Faculty of Development Agriculture & Natural Resources Management Eastern Technical University of Sierra Leone Kenema Sierra Leone

Soil loss through erosion continues to pose serious challenges to increasing the smallholder agricultural productivity in Sierra Leone. While emphasis on sustainable land use practices continue to gain attention among land users, however, the rate of adoption among smallholder farmers is still very low and hence, in most part of the country soil fertility has been declining at alarming rates. In the Njala area, studies have shown that soil loss ranges from moderate to high. Though soil erosion has been identified as a major soil fertility declining factor, however, the effect of land use practices on the inherent resistance of soil materials to erosion is lacking. This study was therefore conducted to assess the effect of land uses on soil erodibility. The results showed that the soils are coarse sandy loam with high sand content. The dispersion ratios and erosion indices of soils under cassava, plantain, maize and guava were above the minimum thresholds of 15.0 and 10.0 respectively, thus indicating that these soils are highly erodible compared to soils under fallow and sweet potato which have dispersion ratios and erosion indices below the minimum thresholds. Clay content was inversely proportional to and significantly correlated with dispersion ratio and erosion index while the correlations between silt + clay, dispersion ration and erosion index, and silt, silt + clay and dispersion ratio were negative and non-significant. Considering the coarse nature of soils, landscape features and high erodibility indices, these soils would need special soil and water conservation practices to counter further degradation. These may include following, agroforestry, silvi-pasture, controlled and rotational grazing. In addition, awareness of sense of self-responsibility and forest policies and regulations are needed.

关键词： Soil Erosion Land Use Agriculture Smallholders Soil Texture Erodibility Soil Fertility Deforestation