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991.
卵巢癌细胞株冻融抗原负载的树突状细胞诱导CTL抗卵巢癌免疫应答的体外研究 总被引:6,自引:0,他引:6
目的研究卵巢癌冻融抗原负载的树突状细胞(dendriticcells,DC)诱导细胞毒性T淋巴细胞(CTL)体外杀伤卵巢癌细胞的细胞毒性效应。方法利用免疫磁珠分离法(MACS)分离纯化脐血CD34 细胞并在体外诱导分化为DC,用反复冻融法从卵巢癌细胞系SKOV3中提取的可溶性相关抗原负载DC。流式细胞学检测负载抗原后DC表面各种分化相关抗原的表达,ELISA法检测DC上清中IL12的表达,混合淋巴细胞反应(MLR)测定DC体外刺激T细胞增殖的能力,MTT法检测抗原负载DC激活的抗原特异性CTL对卵巢癌细胞的杀伤作用。结果与未经抗原负载的DC相比,经卵巢癌抗原负载的DC不仅能更高地表达各种DC分化相关抗原CD1α(73.35%±2.94%vs34.1%±2.35%)、CD83(73.9%±8.46%vs54.68%±3.26%)、CD80(91.95%±2.48%vs52.53%±3.18%)、HLADR(70.05%±2.35%vs48.7%±2.07%)以及CD54(88.9%±5.52%vs71.45%±2.29%),同时具有更强的刺激同种异体T淋巴细胞增殖和IL12分泌的能力(P均<0.05)。此外,卵巢癌细胞SKOV3冻融抗原负载DC激活的CTL在体外对SKOV3的杀伤率为77.35%,显著高于未经抗原负载的DC(P=0.0001)。结论经卵巢癌细胞冻融抗原负载DC激活的CTL在体外具有更强的增殖能力和杀伤卵巢癌细胞的作用。 相似文献
992.
氟尿嘧啶引起大鼠气管损伤修复过程中干细胞的定位 总被引:10,自引:2,他引:10
目的 气管干细胞的原位观察。方法 使用5-氟尿嘧啶(5-FU)造成大鼠离体气管环的严重损伤,采用光镜、PCNA免疫组织化学,以及Hoechst33342染色观察气管黏膜的修复过程。结果 5-Fu作用12h后,气管上皮脱落,可见少量间隔分布的类似裸核的细胞呈钉状位于基底膜上,PCNA染色阴性(Go期细胞),其中少数细胞Hoechst33342染色阴性。去除5-FU6h后,气管黏膜由扁平上皮覆盖;48h后,气管环恢复假复层纤毛柱状上皮。结论 5-Fu能杀死处于细胞周期的气管上皮细胞,对Go期细胞作用很小,残留于基底膜上的裸核样的Go期细胞中含有干细胞,其Hoechst33342染色阴性,并具有排出荧光染料的能力,正是这些细胞的增殖分化,修复了损伤的气管环。 相似文献
993.
994.
Interferon-gamma (IFN-gamma) and IL-4 expressed during mercury-induced membranous nephropathy are toxic for cultured podocytes. 下载免费PDF全文
W Coers J T Vos P H Van der Meide M L Van der Horst S Huitema J J Weening 《Clinical and experimental immunology》1995,102(2):297-307
The subepithelial immune deposits of Dorus Zadel Black (DZB) rats with mercury-induced membranous nephropathy consist of autoantibodies directed to laminin P1 and of complement. The animals develop massive proteinuria within 10-14 days which is associated with obliteration of foot processes of glomerular visceral epithelial cells (GVEC), or podocytes. Previous studies indicate that these autoantibodies are probably not the sole mediator of proteinuria and GVEC damage. In this study we investigated whether circulating or macrophage-derived cytokines can contribute to the GVEC changes as detected in vivo. In vivo at the height of the proteinuria, increased intraglomerular IFN-gamma immunoreactivity was found. In diseased rats a five-fold increase in intraglomerular macrophages was found, but we could not detect intraglomerular IFN-alpha, IFN-beta, IL-1 beta or tumour necrosis factor-alpha (TNF-alpha) by using immunohistology. Subsequently, we exposed cultured GVEC to these cytokines to investigate their cytotoxic effects on several physiological and structural parameters. IFN-gamma and IL-4 were the only cytokines that exerted toxic effects, resulting in a rapidly decreased transepithelial resistance of confluent monolayers, which was closely associated with altered immunoreactivity of the tight junction protein ZO-1. IL-4 also affected vimentin and laminin immunoreactivity. IFN-gamma and IL-4 only interfered with monolayer integrity when added to the basolateral side of the GVEC, indicating specific (receptor-mediated) effects. Only IL-4 decreased the viability of the cells, and treated monolayers demonstrated an increased passage of the 44-kD protein horseradish peroxidase. From our experiments we concluded that IFN-gamma subtly affected monolayer integrity at the level of the tight junctions, and that IL-4 additionally induced cell death. We hypothesize that the toxic effects of the cytokines IFN-gamma and IL-4 as seen with cultured podocytes are necessary together with the autoantibodies, for the ultimate induction of proteinuria in mercury nephropathy in the DZB rat. 相似文献
995.
脐带血清对骨髓粒—巨噬细胞集落形成的影响 总被引:1,自引:0,他引:1
在骨髓细胞半固体琼脂培养体系中,研究脐带血清(CBs)对骨髓粒—巨噬细胞集落形成单位(CFU-GM)的影响,单纯应用CBs培养可使骨髓CFU-GM(x±s为26.0375±10.2327)显著的高于正常成人外周血清(PBs)组(x±s为11.5917±3.6047;P<0.001)。CBs加粒-巨噬细胞刺激因子,GM-CSF组CFU-GM(x±s为119.0792±35.0991)是单纯CBs组的4.57倍;而PBs加GMC-SF组(x±s为97.125±26.7559)是单纯PBs组的8.38倍。有白细胞介素-6(IL-6)存在的CBs组,CFU-GM(x±s为31.9333±10.9144)明显的高于单纯CBs组(P<0.02)。提示CBs中含有较高水平的内源性GM-CSF。 相似文献
996.
兔急性脑缺氧时脑及脑脊液内腺嘌呤核苷含量的变化 总被引:2,自引:0,他引:2
本文报告了由低张性低氧血症所致兔急笥脑缺氧时脑组织内腺苷、次黄嘌呤核苷及次黄嘌呤水平分别从正常对照的53.3±2.9、115.6±11.8及186.5±10.3增至816.4±59.0、1049.7±37.5及704.4±55.3μM/g(X±SD),各组间P值均<0.01至0.05。同步测定的脑脊液中三种腺嘌呤核苷水平分别从正常对照的1.6±0.8、5.1±1.0及133.9±50.8增至7.0 相似文献
997.
Mark M. Kockx Floris L. Wuyts Norbert Buyssens Rita M. Van Den Bossche Guido R. De Meyer Hidde Bult Arnold G. Herman 《Virchows Archiv : an international journal of pathology》1993,422(4):293-299
Intima formation in vessels, spontaneous or experimentally induced, is generally characterized by the presence of longitudinally orientated smooth muscle cells (LSMC). During an experiment of neo-intima induction in carotid arteries in rabbits, by application of a nonconstrictive silastic cuff, a study was performed to investigate the presence of LSMC in the systemic and pulmonary circulations, in both elastic and muscular arteries. Three patterns could be distinguished: intimai cushions in muscular arteries, single or small groups of LSMC in the intima in elastic and larger muscular arteries, and intra-medially located layers or columns of LSMC in the aorta, the pulmonary artery, at the bifurcation of the aorta and around orifices of branches. In order to understand this peculiar orientation a biomechanical approach was used: this showed that near the lumen the circumferential stress is 4.5 times higher than the longitudinal. Because the cell surface of the smooth muscle cells exposed to this stress per unit vessel length is much less in the longitudinal than in the circular direction we conclude that the LSMC align in the direction which allows them to cope most effectively with the mechanical stresses. 相似文献
998.
Sj?gren Anita; Hillensj? Torbj?rn; Hamberger Lars 《Human reproduction (Oxford, England)》1987,2(6):469-473
Granulosa cells were aspirated from human pre-ovulatory folliclesfollowing a combined clomiphene-gonadotrophin stimulation inan in-vitro fertilization (IVF) programme. The cells were culturedfor 8 days in medium M199 containing 10% bovine fetal calf serumunder 5% CO2 in air. Pure human FSH and human LH were addedalone or in combination to the culture in various concentrationsand the progesterone (P) and oestradiol-17 (E2) levels in themedium were measured every second day by a conventional RIAtechnique. In the presence of FSH or LH the formation of P increased2- to 3-fold with the pronounced effect after 4 to 6 days inculture. Addition of testosterone (T) (3 ? 107 M) tothe culture medium affected neither basal nor gonadotrophinstimulated P formation. In this system, only minute amountsof E2 were formed and neither FSH nor LH stimulated its formation.When the medium was fortified with T, basal E2 formation increased50- to 100-fold. FSH further stimulated this conversion significantlyafter 6 and 8 days of culture, while LH had no significant influence. 相似文献
999.
Theresa L. Whiteside Sylvia Miescher H. R. MacDonald V. Von Fliedner 《Journal of immunological methods》1986,90(2):221-233
The separation of viable tumor-infiltrating lymphocytes (TIL) from surgical biopsies of human solid tumors was achieved by velocity sedimentation at unit gravity or by discontinuous density gradients. The two methods were adapted to small volumes and cell numbers not exceeding 1 × 108. The recovery, purity and composition of the TIL-enriched fractions were comparable in the two methods. Density gradients were more rapid, simpler and more practical for preparation under sterile conditions of TIL from clinical material than velocity sedimentation. Lymphocytes in the TIL-enriched fractions obtained by either of the methods were poorly responsive to mitogens. This poor responsiveness is a characteristic of the human TIL and seems to be related to effects exerted by tumor cells. 相似文献
1000.
Perisinusoidal cells (PSCs) are currently regarded as the major source of extracellular matrix proteins during hepatic fibrogenesis in response to liver injury. However, the cellular mechanisms underlying their response to injury are not fully understood. One hypothesis is that the PSCs are stimulated by peptide growth factors produced by hepatic macrophages (Kupffer cells) in response to parenchymal cell damage. In this study we have investigated the kinetics of the PSC and macrophage populations in acute carbon tetrachloride-induced hepatic injury in rats. PSCs were identified immunohistochemically by detection of cytoplasmic desmin; monocytes and macrophages were detected using the monoclonal antibodies ED1 and ED2; cells in S phase were identified by immunohistochemical detection of nuclear-incorporated bromodeoxyuridine. The results showed an expansion of the desmin-positive PSC population, predominantly within the damaged perivenular zones, which reached a peak on days 3 and 4 following administration of carbon tetrachloride; this was contributed to by local PSC proliferation. The PSC response was preceded by an expansion of the macrophage population resulting from both local macrophage proliferation and influx of blood monocytes. These results are in keeping with the hypothesis that the PSC response to acute liver injury is mediated, at least in part, by hepatic macrophages. 相似文献