State of nonspecific resistance in germfree and Escherichia coli contaminated miniature piglets

The phagocytic activity of the leukocytes and the serum complement, properdin, and lysozyme levels were studied in germfree miniature piglets and similar animals ocntaiminated withEscherichia coli 055 andE. coli 083. In the presence of autologous serum and complement phagocytosis ofE. coli 055 cells was inhibited, but it was considerably intensified under the influence of specific opsonins (antibodies againstE. coli 055). Lowered levels of complement, properdin, and lysozyme were found in the germfree animals. After peroral monocontamination withE. coli the formation of properdin and complement was stimulated the most, and that of lysozyme the least. Antibodies againstE. coli 055 were not found in the monocontaminated piglets. The highest lysozyme levels were found in the previously germfree animals, which points to the role of other contamination factors thanE. coli cells in the stimulation of lysozyme. It is concluded that microbial contamination plays an important role in the development of the cellular and humoral factors of resistance.Research Laboratory of Experimental Biological Models, Academy of Sciences of the USSR, Moscow Region. (Presented by Academician of the Academy of Medical Sciences of the USSR, N. N. Zhukov-Verezhnikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 9, pp. 341–344, September, 1978.     

A flow cytometric method for the measurement of phagocytosis by polymorphonuclear leucocytes

A new method for the measurement of phagocytosis of Candida albicans by human polymorphonuclear leucocytes (PMN) is described using a fluorescence activated cell sorter. We have used acridine orange to discriminate between PMN which have internalised yeast particles and those which have not. This method allows accurate measurement of particle phagocytosis as an event distinct from particle adherence. It also permits detailed examination of the kinetics of phagocytosis, the study of which is likely to be of value in the investigation of diseases where abnormalities of PMN function are suspected.     

Effect of Azelastine Hydrochloride on Macrophage Chemotaxis and Phagocytosis in Vitro

Azelastine, a newly synthesized anti-allergic agent, was tested for its effects on guinea pig macrophage chemotaxis and phagocytosis. As specific macrophage chemo-attractants, we used macrophage chemotactic factors a and c; separated and highly purified from inflamed skin sites. Macrophage chemotaxis induced by skin extract or chemotactic factors was significantly suppressed by a low concentration of the agent (1 microgram/ml); the effect was dose-dependent. The inhibition of chemotaxis was reversible, because chemotactic activity was restored when the agents was removed by washing cells before chemotactic assay. Inactivation of chemotactic factors was not detected by mixing azelastine and factors a and c. Azelastine may directly interact with macrophages to decrease their chemotactic responsiveness. beta-Glucuronidase activity in the medium and macrophages after phagocytosis of polystyrene latex particles was not affected by this agent at concentrations ranging from 1 to 10 micrograms/ml. The phagocytosis of latex particles or sheep red blood cells opsonized with IgG antibodies (EA) and anchoring of macrophages to substrate were not inhibited and azelastine did not damage the macrophages as determined by lactate dehydrogenase (LDH) release assay.     

伤寒杆菌耐药质粒pRST98与细菌在巨噬细胞内存活关系的研究

目的　研究伤寒杆菌的不相容性C群接合性耐药质粒 pRST98对其宿主菌在巨噬细胞内存活的影响。方法　用标准系列稀释法检测 3株同源染色体伤寒杆菌———含 pRST98的野生型伤寒菌株STpRST98、经SDS人工消除pRST98的突变体菌株ST(R-)及 pRST98重新导入突变体的接合子 pRST98/ST(R-) ,研究它们在BALB/c小鼠腹腔巨噬细胞M J774A .1内的存活情况。结果　巨噬细胞吞噬后 6 0min内大多数待检菌被杀死 ,但 pRST98宿主菌在巨噬细胞内的活菌数明显高于无此质粒的菌株 (P <0 .0 5 )。结论　pRST98能提高其宿主菌在巨噬细胞内的存活能力。     

糖尿病多形核白细胞吞噬功能的改变

目的 研究正常人及糖尿病患者多形核白细胞（polymorphonuclear leukocytes,PMNLs)吞噬功能的改变及钙通道阻滞剂对其的影响。方法 应用过氧化氢释放法测定白细胞吞噬功能,同时研究PMNLs细胞外不同浓度糖环境及钙通道阻滞剂对细胞吞噬功能的影响。结果 糖尿病患者的PMNLs吞噬功能较正常人低下。体外培养基葡萄糖浓度增高,吞噬功能越差,硝苯啶可明显改善糖尿病患者PMNLs的吞噬功能。结论 推测葡萄糖可能是通过激活L型钙通道引起细胞内游离钙浓度升高,从而影响吞噬功能,而硝苯啶可阻断该通道,恢复吞噬功能。     

眼镜蛇蛇毒因子与大鼠中性白细胞吞噬功能减退的量效研究

:研究中华眼镜蛇的蛇毒因子(CVF)与中性白细胞吞噬功能的量效关系及CVF在非协调性异种心脏移植应用中的意义。方法:①用不同剂量的CVF腹膜腔内注射(i.p.)SD大鼠,测定大鼠血液中性白细胞噬菌率;②经CVFi.p.处理后大鼠进行颈部豚鼠—大鼠异种心脏移植,然后测定移植前后大鼠不同时点的血液中性白细胞噬菌率。结果:①CVF能使大鼠血液中性白细胞噬菌功能减退,并呈量效关系;②经CVF处理的大鼠移植后无发生超急排斥反应(HAR),且血液中性白细胞噬菌率明显减退,至术后3d内仍未恢复。结论:①CVF有抑制中性白细胞噬菌功能,且呈量效关系;②CVF可抑制非协调性异种心脏移植的HAR,测定中性白细胞噬菌功能是一种简便测定CVF体内抗补体活性的方法。     

异丹叶大黄素在体外对Cu2+介导的人低密度脂蛋白过氧化的抑制作用

目的研究异丹叶大黄素(Iso)对Cu2 介导的人低密度脂蛋白(LDL)过氧化及氧化LDL(ox-LDL)对小鼠巨噬细胞毒性的抑制作用.方法用序贯超离心法分离血脂正常的供血者血清LDL,分离的LDL 1 mg·mL-1 磷酸缓冲液(pH 7.4), 与10 μmol·L-1 CuSO4于37℃水浴温育10 h 以引起LDL过氧化,给药组预先加入不同浓度Iso,对照组加等量生理盐水,测定丙二醛(MDA)的生成量、维生素E的耗竭以及LDL电泳迁移率.另外测定用ox-LDL处理小鼠腹腔巨噬细胞线粒体的膜电位、吞噬刚果红及释放NO的量.结果 1-100 μmol·L-1 Iso 剂量依赖性地抑制Cu2 引起的人LDL氧化时MDA的生成量、维生素E的耗竭及电泳迁移率的升高.10 μmol·L-1 Iso能防止0.1 mg·mL-1 ox-LDL与小鼠腹腔巨噬细胞温育4 h 后线粒体膜电位的损伤、吞噬刚果红功能以及NO释放量的降低.结论 Iso在体外对Cu2 介导的LDL过氧化以及ox-LDL对小鼠巨噬细胞的毒性有保护作用.     

静脉注射免疫球蛋白对急性川崎病患儿淋巴细胞凋亡的影响

通过观察静脉注射免疫球蛋白（ＩＶＩＧ）对川崎病（ＫＤ）患儿淋巴细胞凋亡（ＡＰＯ）的影响，进一步探讨ＩＶＩＧ对免疫性疾病的作用机理。对２６例川崎病患儿和２０名健康儿童外周血单个核细胞（ＰＢＭＣ）经抗－ＣＤ３单克隆抗体刺激培养不同时间（０，１２，２４，４８，７２小时）ＡＰＯ百分率和ＤＮＡ片断化分析，２６例患儿随机分为两组，阿司匹林＋ＩＶＩＧ治疗组（ｎ＝１６）和阿司匹林治疗组（ｎ＝１０），并对ＰＢＭＣ经植物血凝素（ＰＨＡ）刺激淋巴细胞增殖反应进行了观察。结果：ＫＤ患儿ＡＰＯ百分率和ＤＮＡ片断化较正常儿童明显降低（Ｐ＜０．００１）和延迟；ＩＶＩＧ治疗后，降低的ＡＰＯ百分率和延迟的ＤＮＡ片断化被逆转，同时与单用阿司匹林组比较，临床症状明显改善。淋巴细胞增殖反应下调（Ｐ＜０．００１）。外周血淋巴细胞ＡＰＯ下调可能参与了ＫＤ的发病。ＩＶＩＧ治疗ＫＤ的机理可能部分归于对被抑制的淋巴细胞ＡＰＯ的逆转。ＩＶＩＧ对其它淋巴细胞凋亡不足的自身免疫性疾病治疗可能存在同样机理。     

IL—6抗化疗药所致血细胞和巨噬细胞功能减低效应的观察

将5-氟脲嘧啶（5－Fu125mg／kg）为化疗组；注入小鼠后5h后再给IL－6（10000U／0．5ml每日1次）为IL－6组；给生理盐水为阴性对照组。第1、3、5、7天后，阴性对照组血白细胞、血小板数值无明显变化，化疗组则呈明显下降趋势，应用IL－6后在第3天以后白细胞下降趋势略小于化疗组，血小板下降明显小于化疗组。化疗组巨噬细胞吞噬率（％）和指数明显下降，应用IL－6后则升高，并发现脾组织红髓中大量巨噬细胞增生，白髓中大量淋巴细胞及母细胞聚集。说明IL－6可促进受抑制造血和免疫机能恢复。     

地塞米松对小梁细胞粘附及吞噬功能的影响

目的：探讨地塞米松对小梁细胞的粘附及吞噬功能的影响。方法：不同浓度地塞米松(10^-7M，10^-6M，10^-5M)处理体外培养的牛眼小梁细胞3d，消化、漂洗后加入用不同的细胞外基质(Extracellular matrix，ECM)包被的培养板中，37℃孵育90min后．采用甲基噻唑基四唑(Methyl thiazolyl tetrazolium，MW)法测定各组吸光度值反应粘附细胞的量；另外，以乳胶微球为标记，观察不同浓度的地塞米松对小梁细胞吞噬功能的影响。结果：与对照组比较，三种浓度的地塞米松均对小梁细胞与ECM粘附有抑制作用．且呈浓度依赖性；对小梁细胞的吞噬功能亦呈现浓度依赖性抑制作用。结论：地塞米松对小梁细胞的粘附及吞噬功能有抑制作用，抑制小梁细胞的粘附及吞噬功能可能是皮质类固醇性青光眼的发病原因之一。