CD40反义RNA对系统性红斑狼疮患者B淋巴细胞功能的影响

目的　探讨瞬间表达的CD4 0反义RNA对EB病毒转染的系统性红斑狼疮 (SLE)患者B淋巴细胞CD4 0分子表达、细胞增生以及免疫球蛋白 (Ig)分泌功能的影响。 方法　构建人CD4 0反义RNA的真核表达载体CD4 0 /pcDNA3,并将其转染入EB病毒转染的SLE患者B淋巴细胞中。应用流式细胞仪 (FACS)检测观察B淋巴细胞膜上CD4 0分子表达的变化 ;应用四甲基偶氮唑盐微量酶反应比色法 (MTT)观察反义CD4RNA对B淋巴细胞增生能力的影响 ;应用酶联免疫吸附试验(ELISA)测定转染后的B淋巴细胞的Ig分泌功能。结果　与转染pcDNA3空载体组相比 ,转染CD4 0 / pcDNA3组的CD4 0分子的表达明显降低 (P <0 0 1) ;细胞的增生能力明显降低 (P <0 0 5 ) ;细胞的Ig分泌功能明显受抑制 (P <0 0 1)。结论　CD4 0反义RNA对SLE患者的B淋巴细胞有明显的免疫调控作用。     

免疫治疗联合化疗对多发性骨髓瘤患者外周血T、B淋巴细胞及调节性T细胞水平的影响

【目的】探讨免疫联合化疗对多发性骨髓瘤（MM）患者外周血T、B淋巴细胞及调节性T细胞（Tregs）水平的影响。【方法】本院收治的MM患者60例,采用数字随机对照表分为两组,每组各30例;对照组给予DC-CIK免疫治疗＋化疗,对照组仅给予化疗,评估两组临床疗效,观察两组T淋巴细胞、Tregs水平、B细胞、浆细胞比例水平变化;随访1年,比较两组复发率。【结果】观察组治疗有效率为53．33％（i6／30）显著高于对照组26．67％（8／30）,1年复发率为6．67％（2／30）显著低于对照组30．0％（9／30）,且差异有显著性（P〈0．05）。观察组治疗后CD4＋T淋巴细胞比例、CD4＋／CD8＋T淋巴比值显著高于对照组,CD8＋T淋巴细胞比例显著低于对照组,且差异有显著性（P〈0．05）。观察组治疗后CD4＋CD25＋Tregs、CD4＋CD25 high Tregs显著低于对照组,且差异有显著性（P〈0．05）。观察组治疗后B细胞比例高于对照组,浆细胞比例低于对照组,且差异有显著性（P〈0．05）。【结论】DCcIK免疫治疗＋化疗能够改善MM患者免疫功能紊乱症状,降低浆细胞比例,提高临床疗效。     

催乳素对人B淋巴细胞IgG基因表达影响的实验研究

目的:研究人催乳素(PRL)对B淋巴母细胞系IM-9细胞IgG基因表达的影响。方法:应用人PRL(10ng/ml)刺激IM-9细胞,培养48h后用试剂盒提取细胞总RNA,反转录为cDNA,RT-PCR扩增IgGcDNA,琼脂糖凝胶电泳观察扩增情况。结果:该cDNA片段的大小为98bp,LPS、PRL与LPS-PRL组cDNA条带在第24循环时出现,LPS-Br与LPS-PRL-Br组cDNA条带在第28循环时出现,并随循环数的增加亮度逐渐增强,LPS-PRL组该cDNA条带亮度最强。结论:PRL能够上调B淋巴细胞中IgG基因的表达,但该功能能够被Br拮抗。     

同种大鼠骨髓间充质干细胞诱导调节性B淋巴细胞的免疫负调节作用

目的 研究骨髓间充质干细胞(MSC)诱导调节性B淋巴细胞(Breg细胞)的免疫负调节作用.方法 取DA大鼠MSC和Lewis大鼠B淋巴细胞共培养后,酶联免疫吸附试验(ELISA)检测MSC和B淋巴细胞混合培养组上清液中白细胞介素10(IL-10)的水平和分泌因子的变化;采用流式细胞技术分析B淋巴细胞增殖、凋亡、表型的变化和对抗原特异性T淋巴细胞增殖的影响,采用激光共聚焦显微镜观察Breg细胞对免疫复合物吞噬能力的变化.结果 MSC成功诱导产生高分泌IL-10的Breg细胞产生,表型为CD19+ CD1d+ CD5+,MSC显著抑制B淋巴细胞的增殖,Breg细胞能抑制抗原特异性T淋巴细胞的增殖,并增加对免疫复合物的吞噬能力.结论 MSC诱导的Breg细胞具有明显的免疫负调节作用.     

XTT法和MTT法测定B淋巴细胞株细胞活性的实验研究

目的：比较两种常用的细胞活性检测方法MTT和XTT法间的异同。方法：以B淋巴细胞株Daudi细胞株为观察对象,分别采用MTT法和XTT法测出各自的OD值、抑制率及IC50,比较IC50敏感性及变异系数的差异。结果：MTT法和XTT法所求OD值变异系数无统计学差异;所求IC50分别为14.45μg/ml和11.24μg/ml,XTT法更敏感（P=0.031）。结论：MTT法价格便宜;XTT法价格较贵,但灵敏度更高,结果更稳定。     

儿童癫痫患者外周血B淋巴细胞CD20的表达

目的 探讨癫痫患儿外周血B淋巴细胞CD20表达的变化与癫痫发作的关系.方法 应用流式细胞仪测定本院2008年1月至2010年12月就诊的620例儿童癫痫患者外周血B淋巴细胞CD20的表达,并与健康对照组(n=36)进行比较.同时分析96例儿童癫痫患者(症状性癫痫患者37例,特发性癫痫患者59例)应用静脉注射免疫球蛋白(IVIG)治疗前后的B淋巴细胞CD20表达的改变,并评价IVIG治疗各型癫痫的有效率.结果 癫痫儿童外周血B淋巴细胞CD20+比例较健康对照组明显升高[(21.50±8.41)％比(16.13±4.19)％,P＜0.05].96例儿童癫痫患者经IVIG治疗6个月后外周血B淋巴细胞CD20+细胞比例较治疗前下降[(16.74±5.12)％比(21.61±8.03)％,P＜0.05].IVIG治疗癫痫总体有效率为76.04％(73/96),症状性癫痫治疗有效率为86.49％ (32/37),特发性癫痫治疗有效率为69.49％(41/59).结论 儿童癫痫患者存在B淋巴细胞功能异常,IVIG治疗能改善癫痫患儿的细胞免疫功能.     

B cells in murine cervical lymph nodes are conventional B-2 cells

We investigated the characteristic features of cervical lymph node B cells to determine whether their behavior differs from that of B cells located elsewhere, because cervical lymph nodes may be exposed to continual antigenic stimulation from the naso- and/or oropharynx. B cells were isolated from cervical lymph nodes, spleen and peritoneal fluid of mice, cultured in medium, and exposed to various stimuli. The expression of various surface molecules characteristic of lymphoid B cells was assayed by flow cytometry, and immunoglobulin secreted into the culture supernatants was evaluated by enzyme-linked immunosorbent assay. B220+ cells were cultured in medium alone or with lipopolysaccharide, and their entrance into S phase in response to stimuli was measured by proliferative assays. Phenotypic characteristics of cervical lymph node B cells included CD5 low, CD23high, CD43low, B7.1low, B7.2low, and Syndecan-1low. Unstimulated lymphoid B cells did not secrete immunoglobulin, but, upon stimulation, secretion of IgM was increased more than secretion of IgA and IgG. B cells actively entered S phase after 48 hr stimulation. These results show that B cells in cervical lymph nodes are conventional B2 cells, like splenic B cells.     

免疫相关性血细胞减少症23例临床分析

目的探讨免疫相关性血细胞减少症（IRP/IRH）的发病机制及预后。方法对我院收治的23例患者病例进行分析。结果 23例中并发甲状腺功能亢进、干燥综合征各2例,桥本甲状腺炎、系统性红斑狼疮、糖尿病各1例;网织红细胞正常或增高20例,减低3例;乳酸脱氢酶活性增高14例;外周血单核细胞百分比增高19例,形态异常5例;骨髓单核细胞百分比增高9例,形态异常8例;骨髓增生活跃22例,增生减低1例;白细胞碱性磷酸酶阳性率减低16例,增高3例;单用或联合糖皮质激素、环孢素A及丙种球蛋白为该疾病的主要治疗方法,总有效率为95.6%。结论单核细胞百分比增高及形态异常是IRP/IRH特征的新发现。     

Nonselective cationic currents elicited by extracellular ATP in human B-lymphocytes

Adenosine 5-triphosphate-(ATP)-induced whole-cell currents were studied in human B-lymphocytes, transformed by the Epstein-Barr virus, by means of the tight-seal voltage-clamp technique. During bath application of ATP, the membrane conductance was increased. The change of membrane conductance occurred within milliseconds. The dose response relationship for the ATP^4–-elicited membrane current (I _p) was fitted by the Hill function with a Hill coefficient of 1 and a K _D value of 0.2 mmol/l. Adenosine, as well as the Mg²⁺-bound form of ATP, did not effect the membrane conductance. I _p did not desensitize within 1 min and could be evoked repeatedly up to 100 times in 1 cell in the presence of the G-protein blocker Guanosine 5-o-(2-thiodiphosphate) (GDP [S]). Therefore, it seems that ion channels in form of P _2Z purinoceptors are involved in the observed effects. The permeability (P) sequence for cations carrying I _p was P _Ca:P _K:P _Cs:P _Na:P _TRIS= 3521.210.1. The reversal potential of I _p was not changed by substitution of intracellular Cl^– for aspartate, indicating that anions are not involved in the purinoceptor-dependent conductance. A single-channel conductance of P _2Z-receptor-dependent ion channels of about 3 pS was determined by noise analysis of I _p.     

转人B7-H3基因鳞癌细胞株的建立及其检测

目的探讨转入B7-H3基因鳞癌细胞株的建立及其检测。方法采用脂质体介导法将已建好的真核表达质粒pEGFP-C1-B7-H3导入人鳞癌细胞Tca8113中(Tca8113/B7-H3),RT-PCR检测B7-H3在该细胞中的表达,Western blot检测其蛋白的表达。经G418筛选后,用有限稀释法建立稳定高表达的带有B7-H3基因的Tca8113鳞癌细胞株。结果RT-PCR方法检测到目的基因B7-H3的一个215bp大小的cDNA扩增产物;DAB显色,裂解的Tca8113/B7-H3基因转染细胞膜蛋白中相对分子质量约65～86ku大小的特异性条带。转染B7-H3的Tca8113细胞,经过细胞传代培养后仍然能够检测到高表达的B7-H3基因,经过有限稀释法建立了Tca8113/B7-H3细胞株。结论成功建立了Tca8113/B7-H3细胞株,为以后的体内外试验提供了很好的平台,为近一步研究其抗肿瘤作用奠定了基础。