【正文】 nspecific Memory CD8 T Cells. OT1 lymph nodes cells were transferred to RAG/ mice through tail vein injection, and mice were immunized with 50ug of SIINFEKL peptide in CFA 1 day later. At day 7 , 14 , 21, mice were sacrificed, and splenocytes or lymph nodes cells were isolated and analyzed. The percentage of CD44high, SIINFEKLspecific CD8 T cells was assessed by threecolor FACS staining with SIINFEKL MHC class I tetramer and antibodies to CD8 and CD44. Plots shown are gated on the SIINFEKL tetramer positive lymphocytes。 ? 棄細(xì)胞懸液，重新解離細(xì)胞與磁珠。在激發(fā)光的作用下，可直接發(fā)射熒光，前者發(fā)黃綠色熒光，后者發(fā)紅色熒光。 ? 抗體含量固定并處于抗體過剩時(shí)，免疫復(fù)合物的多少直接取決于抗原的濃度，抗原的終濃度通過標(biāo)準(zhǔn)品繪出的標(biāo)準(zhǔn)曲線查出。 常用的抗原抗體反應(yīng) ? 凝集反應(yīng) ? 沉淀反應(yīng) ? 免疫標(biāo)記技術(shù) 凝集反應(yīng) ? 直接凝集反應(yīng) ? 血型鑒定 ? 肥大反應(yīng) ? 間接凝集反應(yīng) ? 間接凝集抑制試驗(yàn) ? 微粒捕獲酶免疫分析技術(shù) 間接凝集反應(yīng) ? 將可溶性抗原或抗體吸附在與免疫無關(guān)的顆粒載體上，形成致敏顆粒，再與相應(yīng)抗體或抗原進(jìn)行反應(yīng)產(chǎn)生的凝集反應(yīng)，稱為間接凝集反應(yīng)。 ? 本法靈敏度高，檢測(cè)可溶性抗原或抗體、組織或細(xì)胞表面特異性抗原。 ? 膠體金顏色隨顆粒大小而變化，大于 20 nm的金顆粒在光鏡下呈現(xiàn)磚紅色，可在光鏡水平行免疫分析，也可用銀顯影劑增強(qiáng)，進(jìn)一步提高靈敏度。 values are mean percentages of CD44 cells within the SIINFEKL tetramerpositive population. CD8+ Memory T Cells 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 longliv