免疫學(xué)技術(shù)在科研中的應(yīng)用(存儲(chǔ)版)
【正文】 s Need Antigenspecific CD4+Thelper Cells to Achieve Tumor Protection Ovalbumin CTL epitope (SIINFEKL)specific T cells were parked in Rag/ mice for 42 days to generate mCTL and then adoptively transferred to C57BL/6 mice . Two groups of recipient C57BL/6J mice were immunized 8 days prior with 50 ug of OVT in inplete Freund’s adjuvant or 20 ug keyhole limpet hemocyanin (KLH) protein as control in inplete Freund’s adjuvant . Mice were challenged with ovalbuminexpressing tumor cells (EG7) in the scruff of the neck 1 day after adoptive transfer. Normal C57BL/6 control mice were challenged with EG7 without any treatment. CTL+ThCTL+KLHCTLThKLHControl0. 00. 20. 40. 60. 81. 0CT L + T hCT L + K L HC T LThK L HCo n tr o lTumour weight (g)Absence of tumor protection in mice without antigenspecific Thelper cells is not because of lower levels of tumor antigen (SIINFEKL)specific CD8+ T cells. mCTLs were transferred to groups of C57BL/6 mice with or without immunization 8 days prior with 50 ug of OVT or with 20 ug of KLH protein control. C57BL/6 control group was given no treatment. IFN Elispot assays were performed using the splenocytes of these mice, which were challenged in vitro with (A) 1 ug/ml SIINFEKL peptide or (B) 8ug/ml OVA Thelper peptide to determine the presence of ovalbuminspecific CD8+ CTLs and CD4+ Thelper cells. In contrast to mCTLs, eCTLs do not need T help to kill tumor. eCTLs generated from RAG/ mice were transferred at (A) day 7 or (B) day 14 to C57BL/6 mice that had been immunized 8 days earlier with 50 ug of OVT or as control 20 ug of KLH. C57BL/6 mice without any treatment were used as controls (EG7 and EL4 controls). Ovalbuminexpressing tumor cells (EG7 ) were injected under scruff of the neck of some groups of mice at (A) day 7 or (B) day 14. The parent tumor cell line EL4 that does not contain ovalbumin gene was injected into other groups of mice at day7 (A) as nonspecific tumor control. Study of the longlived memory CD8+ T cells generated in C57BL/6J mice. Ovalbumin CTL epitope (SIINFEKL)specific OT1 cells were adoptively transferred to syngeneic C57BL/6 mice 。 ? 轉(zhuǎn)基因動(dòng)物可以作為疾病模型。 ? 反應(yīng)管置于磁場(chǎng)中，鐵顆粒受磁場(chǎng)的吸引，攜帶有相應(yīng)細(xì)胞的磁球吸附于靠近磁鐵的管壁上。 ? 膠體金可標(biāo)記白蛋白、免疫球蛋白、糖蛋白、激素、脂蛋白、植物血凝素、卵白素等。 ? 常用 的熒光素有異硫氰酸熒光素(nuorescein isothiocyanate， FITC)、藻紅蛋白 (phycoerythrin， PE)等。 ? 將 SDSPAGE分離到的蛋白條帶轉(zhuǎn)移至固相的硝酸纖維素膜上。 沉淀反應(yīng) ? 速率散射比濁法／免疫比濁法 ? 瓊脂擴(kuò)散法 ? 單向瓊脂擴(kuò)散 ? 雙向瓊脂擴(kuò)散 ? 火箭電泳 ? 對(duì)流免疫電泳 ? 免疫印跡技術(shù) 速率散射比濁法／免疫比濁法 ? 將已知抗體與相應(yīng)抗原在液相中按一定比例混合形成可溶性免疫復(fù)合物，這些復(fù)合物微小粒子對(duì)一定波長(zhǎng)的光照射發(fā)生散射，可通過三個(gè)光路系統(tǒng)測(cè)定光散射(OD值 )。這種非共價(jià)鍵不如共價(jià)鍵結(jié)合穩(wěn)定，極易受溫度、酸堿度和離子強(qiáng)度的影響而解離。 ? 顆粒載體有紅細(xì)胞、聚苯乙烯乳膠顆粒、活性炭顆粒，而相應(yīng)的凝集現(xiàn)象分別稱為間接血球凝集、間接乳膠凝集、間接炭粒凝集反應(yīng)。 ? 檢測(cè)可溶性抗原、細(xì)胞成分的鑒定與分析，檢測(cè)與自身變性細(xì)胞核成分結(jié)合的抗體 (抗核抗體 )， HIV的明確診斷。 ELISA ? 雙抗體夾心法 (sandwich assay) ? 檢測(cè)血清、腦脊液、胸、腹水等各種液相中的可溶性抗原 ? 間接法 ? 測(cè)定細(xì)胞及組織表面抗原 酶聯(lián)免疫斑點(diǎn)試驗(yàn) (enzyme—linked immunospot assay， ELISPOT) ? 用已知細(xì)胞因子的抗體包被固相載體 ， 加入待檢效應(yīng)細(xì)胞 ， 溫育一定時(shí)間后洗去細(xì)胞 ， 如待檢效應(yīng)細(xì)胞產(chǎn)生相應(yīng)細(xì)胞因子 ， 則與已包被的抗體結(jié)合 ， 再加入酶標(biāo)記抗該細(xì)胞因子抗體 ， 加底物顯色 。 ? 廣泛應(yīng)用于激素、藥物等微量物質(zhì)的檢測(cè)。 ? 當(dāng)膠體金的粒徑較大、濃度密集時(shí)肉眼水平即可觀察，即膠體金斑點(diǎn)滲濾試驗(yàn)和膠體金斑點(diǎn)免疫層
醫(yī)療健康相關(guān)推薦
鄂ICP備17016276號(hào)-1