MELATONIN AND IMMUNOMODULATION IN AGED AND IMMUNODEFICIENT MICE

Melatoninisaindolehormonesecretedbythepinealglandinresponsetophotoperiodsandinflu encesmanyimportantbiological processes,recentstudiesrevealedthatMelatonincouldincreasetheweightofimmunorgans,raisethelymphocyteper centageinperipheralblood ,acceleratetheantibody’ssynthesisandcell’sproliferation ,stirupcytokine’sdevelopment,helptheregulationinneuroen docrineimmunologicalnetwork ,andalsoplayanimportantroleininducingtheimmuno circadianrhythm[1 ] .HelperandsuppressionTlymphocyteswerecapitalcellsi…     

IL-17A在博来霉素致小鼠肺纤维化形成中的表达

目的探讨白细胞介素17A(IL-17A)在肺纤维化形成中的表达。方法 40只C57BL/6的雄性小鼠随机分为正常对照组(N组)、博来霉素组(BLM组),每组20只。BLM组采用气管内注入博来霉素5mg/kg,N组经气管注入等体积的9g/L氯化钠注射液,分别于3、7、14d和28d处死小鼠。免疫组化法和半定量RT-PCR观察IL-17A在肺纤维化形成中的定位和表达。流式细胞术检测脾淋巴细胞CD4+IL-17+T细胞量。结果①IL-17A定位在终末支气管、呼吸性细支气管和肺泡上皮。②IL-17A在第7天表达开始增加,14d达到高峰,28d有所下降,但均高于正常组。③脾淋巴细胞CD4+IL-17+T/CD4+T量变化规律与IL-17A表达量相同。结论 IL-17A在博来霉素致小鼠肺纤维化形成中表达增加,可能参与肺纤维化的发生发展过程。     

THE HUMORAL AND CELLULAR IMMUNE RESPONSES INDUCED BY HPV18L1-E6/E7 DNA VACCINES IN MICE

Objective To construct eukaryutic expression vector of HPV18 L1- E6, E7 chimeric gene and examine the humorul and cellular immune responses induced by this DNA vaccines in mice. Methods The C-terminal of major rapsid protein L1 gene and mutant zinc finger domains of early E6/7 oncogenes in HPV18 were integrated and inserted into eukaryotic expression vector pVAX1 to generate vaccines pVAX1-L1E6Mxx, ETMxx. CHO cells were transiently transfected with the individual construct. Target protein expressions in the lysate of the transfected cells were measured by ELISA and immunocytochemistry After BALB/c mice were vaccinated with various recombinant plusmids（pVAX1- L1-E6M3 or pVAX1-L1-E7M3 ） and immunie adjuvants （pLXHDmB7-2 or LTB） through different administration routes （intramuscular or intranasal） , the great cellular immune responses were produced us revealed by delayed-type hypersensitivity （DTH） and lymphocyte proliferation, and the expression of IL-4 and IFN- 7 cells in CD4^＋ and CD8^＋ subpopulations. Results The highly efficient expression of pVAX1-L1E6Mxx, E7Mxx vector in host eukaryotic cells were demonstrated both by ELISA and immunocytochemistry. The level of specific serum IgG against HPV in experiment groups mice was much higher than that of control group, and intranuscular immunization group had the highest antibody level. Intramuscular immunization groups were superior to intranasal immunization groups in DTH response, splenocyte proliferation and CD8^＋ IFN-γ^＋ cells number, but CD4^＋ IL4^＋ cell number was higher in intranasul immunization groups. The immunization groups using pLXHDmB7.-2 as adjuvant were superior to other groups in immunorespouse. Conclusion These DNA vaccines produce remarkable cellular and humorul immune responses in the mouse and may provide us prophylatic and therapeutic candidates for HPV induced cancer treatment.     

THE STUDY ON RELATION BETWEEN CD1a^＋ CELLS AND INFILTRATIVE CD3^＋ AND CD8^＋ CELLS IN RENAL TISSUE OF GLOMERULONEPHRITIS

Objective To discuss the role of dendritic cells (DCs) in cellular immunity pathogenesis of glomerulonephritls (GN). Methods 114 patients with GN were selected randomly and divided into two groups,primary GN (pGN) and secondry GN (sGN). CDIa^ , CD3^ and CD8^ cells in bioptic renal tissues were examine dy immunohistochemically. The distribution of CDla cells and the infiltration of CD3^ and CD8^ cells in renal tissues were observed. Results There was no significant difference of CDIa^ , CD3^ , and CD8^ cells between pGN and sGN group (P＞0. 05). CDIa^ cells had significant positive correlation with the infiltrative CD3^ and CD8 cells,respectively (P＜0.01). The infiltrative CD3^ cells had significant positive correlation with the CD8 cells in the same area, respectively (P＜0.01). CD1a^ cells, CD3^ cells infiltrating in both glomeruli and renal interstitial tissues, and CD8^ cells only infiltrating in renal interstitial tissues, all of them had significant positive correlation with the degree of glomerular proliferation, respectively (P＜0.05). The infiltrative CD3^ and CD8^ cells in renal interstitial tissues had significant positive correlation with the degree of glomerular sclerosis and the lesion degree of renal tubule and interstitial, respectively (P~ 0. 05). There were significant positive correlation between CDla cells and the lesion degree of renal interstitial (P＜0.05). Collusion DCs could activate T lymphocyte by presenting antigen, then the activated T lymphocyte participate in the pathogenesls of GN through releasing cytokine and/or directly damaging the renal tubule and interstitial, which produce more serious glomeralar lesion.     

EXPRESSION OF PRE S ROTEIN OF HBV AS A ALKALINE PHOSPHATASE FUSION PROTEINS IN NIH 3T3 CELLS

ＥＸＰＲＥＳＳＩＯＮＯＦＰＲＥＳＲＯＴＥＩＮＯＦＨＢＶＡＳＡＡＬＫＡＬＩＮＥＰＨＯＳＰＨＡＴＡＳＥＦＵＳＩＯＮＰＲＯＴＥＩＮＳＩＮＮＩＨ３Ｔ３ＣＥＬＬＳＸｉａｏＳｈｅｎｇｘｉａｎｇ；Ｔ．Ｓ．Ｂ．Ｙｅｎ（ＬａｂｏｒａｔｏｒｙｏｆＭｏｌｅｃｕｌａｒＰａｔ...     

褪黑素与免疫调节细胞的关系

目的　探讨褪黑素与免疫调节细胞之间的关系。方法　采用流式细胞计数测定外周血T淋巴细胞亚群CD4+和CD8+T细胞的百分含量、比值。结果　褪黑素可提高外周血中CD4+和CD8+T细胞的百分含量 ,调整CD4+/CD8+比值。结论　褪黑素可通过调整免疫调节细胞TH/TS 的比例发挥免疫调节作用。     

STUDY ON B7-1 PROTEIN‘S EXPRESSION IN BIOPSIES IN RENAL TRANSPLANTATION

Ａｌｌｏｇｒａｆｔｒｅｊｅｃｔｉｏｎｉｓａｎｉｍｐｏｒｔａｎｔｆａｃｔｏｒｏｆｉｎ ｆｌｕｅｎｃｉｎｇｔｈｅｌｏｎｇｔｅｒｍｓｕｒｖｉｖａｌｏｆａｌｌｏｇｒａｆｔｉｎｈｕ ｍａｎｏｒｇａｎｔｒａｎｓｐｌａｎｔａｔｉｏｎ .Ｉｍｍｕｎｏｌｏｇｉｃａｌｔｏｌｅｒａｎｃｅｉｎｄｕｃｔｉｏｎｉｓａｍａｉｎ ｐａｒｔｉｎｔｈｅｒｅｓｅａｒｃｈｏｆｏｒｇａｎｔｒａｎｓｐｌａｎｔａｔｉｏｎ .Ｔｏｓｔｕｄｙｔｈｅｉｍｍｕｎｏｌｏｇｉｃａｌａｎｄｍｏｌｅｃｕｌａｒｍｅｃｈａｎｉｓｍｏｆｒｅｊｅｃｔｉｏｎａｎｄｓｅｅｋ…     

鼻咽癌组织中BCRF-1基因表达及其与免疫功能的相关性分析

目的研究鼻咽癌组织中BCRF-1mRNA及病毒白介素-10(VIL-10)蛋白质的表达,明确VIL-10对鼻咽癌患者免疫细胞的影响。方法在鼻咽癌组织和鼻咽炎症组织中,分别应用RT-PCR及Western Blot检测BCRF-1mRNA和蛋白产物VIL-10的表达;流式细胞术检测两组患者外周血CD4+T、CD8+T细胞的比例。结果①BCRF-1mR-NA在34/40例(85%)鼻咽癌组织和16/20例(80%)鼻咽炎症组织中表达阳性,两者差异无统计学意义(P>0.05)。②30/40例(75%)鼻咽癌组织VIL-10呈阳性,10/20例(50%)鼻咽炎症组织VIL-10呈阳性,两者差异具有统计学意义(P<0.05)。③40例鼻咽癌患者分为VIL-10阳性组和VIL-10阴性组,VIL-10阳性鼻咽癌患者组CD4+T细胞及CD4+T/CD8+T比值明显低于VIL-10阴性鼻咽癌患者组(24.99%±3.88%vs.35.92%±6.31%,0.86 vs.1.99,均P<0.05),而VIL-10阳性鼻咽癌患者组CD8+T细胞明显高于VIL-10阴性鼻咽癌患者组(30.61%±3.34%vs.20.10%±7.68%,P<0.05)。结论鼻咽癌组织中高表达BCRF-1基因及其蛋白产物VIL-10;VIL-10使鼻咽癌患者的免疫细胞发生偏移,发挥免疫抑制作用。     

Study on T lymphocyte subsets and NK cells in patients with Graves＇ disease combined with type 2 diabetes

Objective To investigate changes in T lymphocyte subsets and NK cells in patients with simple Graves' disease (GD)and Graves' disease combined with type 2 diabetes mellitus (GD/T2DM). Methods Fifteen cases of GD/ T2DM were selected from our hospital from November 2001 to November 2004. Before and after therapy thyroid function, thyroglobulin antibody (TGA), thyroid microsomal antibody (TMA) and blood glucose level were measured, and T lymphocyte subsets (CD3, CD4, CD8, CD4/CD8) and NK cells (CD56) were measured by immunofluorescence double labeling monoclonal antibody and flow cytometry, respectively. At the same time, comparison was made with simple GD (15 cases), T2DM (15 cases) and healthy control (20 cases). Results Before therapy, CD4/CD8, CD4 and NK cells in GD/T2DM were less than normal, and there was no significant difference in comparison with simple GD (P<0.05). In T2DM group, only CD4/CD8 and CD4 were less than those of healthy controls (P<0.05). When thyroid function recovered after 1 to 3 months of methimazole treatment in both GD/T2DM and simple GD groups, various indexes recovered, which were more obvious in simple GD. Conclusion Immune hypofunction of GD may be the key to the immune abnormality of GD/T2DM, which is more significant than that of simple GD or T2DM. The recovery of thyroid function and immune abnormality is not consistent, and the recovery of GD is more significant than that of GD/T2DM.     

B－CELL APOPTOSIS INDUCED BY ANTIGEN RECEPTOR CROSSLINKING IS BLOCKED BY A T－CELL SIGNAL THROUGH CD40

Ｂ－ＣＥＬＬＡＰＯＰＴＯＳＩＳＩＮＤＵＣＥＤＢＹＡＮＴＩＧＥＮＲＥＣＥＰＴＯＲ　ＣＲＯＳＳＬＩＮＫＩＮＧＩＳＢＬＯＣＫＥＤＢＹＡＴ－ＣＥＬＬＳＩＧＮＡＬＴＨＲＯＵＧＨＣＤ４０ＴａｋｅｓｈｉＴｓｕｂａｔａ，ＷｕＪｉｎｇ；ＴａｓｕｋｕＨｏｎｊｏ（Ｄｅｐａ...     

流式细胞术CD45/SSC设门在急性白血病免疫分型中的应用

目的探讨流式细胞术CD45／SSC设门在急性白血病免疫分型中的应用。方法选用三色荧光抗体直接标记初诊急性白血病患者骨髓，应用流式细胞术CD45／SSC设门进行免疫分型。结果49例急性白血病患者分为三型，其中系列专一型25例，占51．2％；系列非专一型21例，占42．8％；裸型3例，占6．0％。结论流式细胞术CD45／SSC设门，容易将原始／幼稚细胞与正常成熟细胞群区分开，使白血病分型结果更准确、可靠。     

狼疮性肾炎肾活检组织中PCNA与Bcl-2的表达及意义

目的　观察狼疮性肾炎 (lupusnephritis ,LN)肾组织内凋亡调控基因Bcl 2及增殖细胞核抗原 (PCNA)的表达 ,探讨凋亡在LN发病中的作用。方法　应用ELPS(enhancedlabelled polymersystem)检测 2 9例LN肾穿刺活检组织PCNA和Bcl 2的表达 ,并与正常组进行对比分析。结果　LN组肾小球内PCNA阳性细胞数与正常对照组肾组织比较 ,有显著性差异 (P <0 .0 0 1)。LN组肾小球内Bcl 2阳性表达细胞明显高于正常对照组 (P <0 .0 0 1) ,阳性表达细胞见于系膜细胞、毛细血管内皮细胞和细胞性新月体内。LN组肾小球Bcl 2阳性细胞数与PCNA阳性细胞数、肾小球细胞总数有显著正性线性相关 (r =0 .70 1.P <0 .0 1;r =0 .5 5 0 ,P <0 .0 1)。结论　LN肾组织肾小球内细胞高增殖和Bcl 2高表达 ,提示细胞增殖与凋亡调控异常均参与了LN肾小球的增生性病变     

ROLE OF PERIPHERAL LYMPHOCYTES SIALYL LEWIS（X） （CD15s） ANTIGEN BEFORE AND AFTER KIDNEY TRANSPLANTATION

Objective To investigate the role of peripheral lymphocytes Sialyl Lewis（x） （CD15s） antigen before and after kidney transplantation. Methods Flow cytometry technique was applied to examine the expression of peripheral lymphoid cell surface CD15s antigen after renal transplantation, and to evaluate various therapeutic regimen. Results The statistic analysis results of peripheral lymphoid cell surface CD15s antigen expression level showed that there was significant difference among the patients with acute rejection, long-term dialysis and with normal renal function post-transplant; significant difference of CD15s expression level between group of rejection and infection; no significant difference of CD15s expression among the different groups treated by various therapeutic regimens. Conclusion The different therapeutic regimen has no influence to CD15s expression; Detection of peripheral lymphoid cell surface CD15s antigen expression periodically, intelligently make convenience to understand suitable status of immunosuppression.     

慢性粒细胞白血病树突状细胞的体外无血清培养及杀瘤效应

目的　建立慢性粒细胞白血病 (CML)树突状细胞 (DCs)体外无血清培养体系。方法　小牛血清 (FCS)、无血清 (SFM)及自体血清分别培养CML病人CD34 + 细胞或单个核细胞 (MNCs) ,以SCF、GM CSF、TNF α、IL 4 (A)与GM CSF、TNF α、IL 4 (B)两组不同的细胞因子作对照。结果　CML DCsHLA DR高表达 ,CD80、CD83和CD86表达比例不高 ,异体混合淋巴细胞反应能力不强。SFM同FCS体系相比无显著差异。 5例CML DCsPh染色体比例与培养前MNCsPh染色体比例基本吻合 ;同时以CML DCs同自身T细胞、自体Ph+ MNCs共孵育 ,测其杀伤率为 (38.5± 6 .5 ) %。结论　CML DCs携带Ph染色体 ,能刺激自体T细胞杀伤自身白血病细胞 ,但需进一步改善其功能。     

胃癌患者外周血T淋巴细胞亚群变化的临床意义

目的　探讨胃癌患者细胞免疫功能改变的临床意义。方法　采用流式细胞术检测胃癌患者外周血T淋巴细胞亚群。结果　 5 0例胃癌患者与正常对照组比较 ,CD4降低 ,CD8升高 (P<0 .0 5 ) ,CD4/CD8比值下降 (P <0 .0 5 )。胃癌患者中 ,2 0例有转移单纯化疗组与 2 1例无转移手术联合化疗组和对照组相比 ,CD4下降 ,CD8升高 ,CD4/CD8有下降趋势 ;但CD4、CD8、CD4/CD8均无显著性差异 (P >0 .0 5 )。结论　胃癌的发生发展与机体免疫系统功能状态密切相关。     

慢性肾衰血液透析患者外周血淋巴细胞CD15s抗原的表达

目的了解血液透析患者的外周血淋巴细胞表面CD15s抗原的表达。方法应用流式细胞技术检测慢性肾衰(CRF)血液透析患者的外周血淋巴细胞表面CD15s抗原的表达,并进行临床分析。结果长期透析患者外周血淋巴细胞CD15s表达水平为1.81±0.79,与正常对照(1.05±0.13)相比有所升高(P<0.05)。结论长期行血液透析的慢性肾衰患者外周血淋巴细胞表面CD15s表达升高。     

流式细胞仪检测支原体肺炎及呼吸道合胞病毒肺炎免疫状态的研究

使用流式细胞位（FCM）双色分析法，对1名支原体肺炎（MP）及1名呼吸道合胞病毒（RSV）肺炎患性期及恢复期14项淋巴细胞亚群进行了检测，结果显示：急性期两者均有CD4+CD45RO+细胞、CD8+细胞、CD8+CD45RO+细胞、CD56+细胞、CD57+细胞明显降低，CD19+细胞明显升高；恢复期MP上述淋巴细胞亚群均能迅速恢复，而RSV肺炎各项淋巴细胞亚群基本上没有好转，其中CD3+细胞、CD4+细胞、CD4+CD45RA+细胞、CD4+CD45SRO+细胞、CD8+CD45RO+细胞、CD57+细胞比急性期更低，CD19+细胞比急性期更加升高。提示：RSV肺炎患儿免疫功能受抑制比MP更重，抑制时间持续长，推测这一特点可能正是导致RSV肺炎易反复咳喘的重要原因之一。     

人胃肠癌组织中浸润淋巴细胞活性的原位分析

利用一组抗淋巴细胞亚群的单克隆抗体和免疫组化方法对63例进展期胃癌和肠癌组织中浸润单个核细胞,尤其是散在浸润淋巴细胞(主要是T淋巴细胞)的I_2(HLA—DR)抗原和OKT_9抗原的表达进行了分析。同时,无选择的选取了18例慢性胃炎和慢性肠炎活检标本做为对照。在癌肿标本中,散在浸润淋巴细胞的I_2抗原表达显著变异。少数标本中很难查见I_2~+T细胞,大部分病例只能查见少数细胞呈I_2~+。63例中仅有10例其I_2~+细胞占全部散在浸润淋巴细胞的10—30%,所有瘤组织中几乎无OKT_9~+浸润T细胞。这种表现完全不同于正常粘膜及炎症粘膜中的I_2~+浸润单个核细胞分布。这说明就大多数病例来说,肿瘤组织中浸润的T细胞多数是非活化的。     

头孢地嗪对免疫性肝损伤小鼠外周血T细胞亚群的影响

目的观察头孢地嗪对免疫性肝损伤小鼠外周血T细胞亚群的调节作用。方法采用卡介苗联合脂多糖诱导法建立免疫性肝损伤的小鼠模型;将造模成功的免疫性肝损伤小鼠随机分为胸腺肽组、头孢地嗪大、中、小剂量组、头孢曲松组和生理盐水组,连续给药7d,同时以正常小鼠作为对照组;采用免疫荧光染色技术,经流式细胞仪对小鼠的外周血T细胞亚群进行检测。结果免疫性肝损伤合并败血症小鼠的CD3 (%)、CD4 (%)和CD8 (%)均明显低于正常小鼠,而头孢地嗪可有效地提高免疫性肝损伤合并败血症小鼠的CD3 (%)、CD4 (%)和CD4 /CD8 。结论头孢地嗪可通过调整CD4 和CD8 之间的平衡,有效地提高机体的免疫功能。     

鼻咽癌患者中CD4~+CD25~+T细胞与CCR4的相关性

目的 探讨鼻咽癌患者全身及肿瘤局部的免疫状态及其与CC类趋化因子受体-4(CCR4)的关系.方法 采用流式细胞术检测初诊鼻咽癌患者组织(25例)及外周血(35例)中CD4~+CD25~+T细胞与CCR4阳性细胞在淋巴细胞中的比例,并进行统计学分析.结果 鼻咽癌组织及外周血中CD4~+CD25~+T细胞及CCR4阳性细胞的比例高于对照组(P<0.05);两种细胞在鼻咽癌组织中的比例高于外周血(P<0.05),而对照组外周血与组织中却无明显差别(P>0.05);鼻咽癌Ⅲ+Ⅳ期组织中的 CD4~+CD25~+T细胞比例高于Ⅰ+Ⅱ期(P<0.05),外周血中两组无明显差别(P>0.05);鼻咽癌组织及外周血中的CD4~+CD25~+T细胞与CCR4阳性细胞存在正相关关系.结论 鼻咽癌患者存在不同程度的免疫抑制,肿瘤局部免疫抑制更为严重;CD4~+CD25~+T细胞的比例与鼻咽癌分期相关,分期越晚,该细胞比例越高;CCR4在介导CD4~+CD25~+T 细胞趋化到肿瘤局部的过程中起了一定作用.