灵芝对小鼠B淋巴细胞分泌特异性抗体及T淋巴细胞产生γ－干扰素的调节

报道了灵芝对Ｂａｌｂ／ｃ小鼠产生抗ＳＲＢＣ抗体能力的调节及产生γ－干扰素的影响。实验组小鼠分成４组，分别经胃灌注不同灵芝制剂：１号、２号、３号及４号制剂，每天一次，连续９天，对照组小鼠则以等量蒸馏水代替。在实验第７天用ＳＲＢＣ免疫小鼠，于免疫后第４天进行溶血空斑试验和血清抗ＳＲｌＢＣ抗体凝集效价测定，并取小鼠脾细胞加ＰＨＡ刺激、诱导，于３７℃、５％ＣＯ＿２培养７２小时，测定培养液中γ－干扰素活性。结果表明，１号、２号制剂能明显提高实验组小鼠的溶血空斑数目及血清抗体凝集效价，与对照组相比，Ｐ＜０．０５，２号、４号制剂则能明显提高实验组小鼠产生的γ－干扰素活性，与对照组相比，Ｐ＜０．０５。提示灵芝能增强小鼠的免疫功能─—Ｂ细胞产生特异性抗体的能力及Ｔ细胞产生（经ＰＨＡ诱导）的γ－干扰素的能力。     

羧甲基淀粉对小鼠产生抗体和免疫器官的影响

本实验研究了国产羧甲基淀粉(CMS)对小鼠产生 IgE及 IgM抗体和对小鼠胸腺及脾脏重量的影响,结果证明,无论给予或不给予小剂量环磷酰胺(Cy),CMS均能显著或非常显著地抑制小鼠体内诱生抗绵羊红细胞(SRBC)的 IgM直接空斑形成细胞(PFC)数及显著降低小鼠产生抗 SRBC的凝集抗体效价;卵清蛋白致敏的小鼠如加注 Cy能促进IgE产生,但若使用CMS,即使给 Cy仍能非常显著地抑制抗卵清蛋白的 IgE产生。单用CMS能显著增高胸腺的重量,但对脾脏重量无明显影响。机理可能与该药能促进胸腺发育而产生或激活较多的T抑制细胞有关,提示 CMS可能具有免疫调节效应。     

一种根瘤菌胞外多糖对小鼠免疫功能的影响

目的研究一种根瘤菌胞外多糖(REPS)对正常小鼠免疫功能的影响。方法将该多糖以大、中、小3个剂量[(40、20、10 mg/(Kg.d)]给小鼠连续腹腔注射(ip)10 d,测定胸腺质量、脾脏质量、碳粒廓清指数、血清抗绵羊红细胞(SRBC)抗体凝集效价和淋巴细胞转化值。结果与对照组比较,该多糖使小鼠脾脏质量明显增加,单核吞噬细胞的吞噬能力显著增强,提高了血清抗SRBC抗体凝集效价,增强了小鼠脾淋巴细胞转化功能。结论此根瘤菌胞外多糖对小鼠的非特异性和特异性免疫功能均有明显的增强作用。     

空斑形成细胞测定技术的进展及临床免疫应用

一、简介空斑形成细胞(Plaque-forming cell.PFG)测定技术又称溶血空斑技术,为Jerne等在1963年建立,主要用于体外单个抗体分泌细胞的检测。PFC技术具有敏感、特异、简便、快速的优点,因而在免疫学各领域得到广泛应用,成为免疫学研究不可缺少的工具之一。Jerne等建立的方法为平皿法,即用羊红细胞(SRBC)免疫小鼠,经一段时间取脾制成细胞悬液,加入含SRBC的琼脂介质中,倾到平皿,孵育后加入补体。由于抗体分泌细胞分泌抗SRBC抗体,致敏了周围的SRBC,经补体作用使SRBG溶解,形成溶血空斑。     

黄芪多糖对去T细胞小鼠促进抗体产生机理的探讨

黄芪多糖(APS)100mg/kgx6天 ip 可以增加正常小鼠脾脏重量,而对去胸腺与抗胸腺细胞血清处理过的小鼠(T_x-ATS)的作用明显降低。APS 能够显著提高正常小鼠 SRBC 免疫后脾细胞直接溶血空斑(d-PFC)和间接溶血空斑(i-PFC)数量,但对 T_x-ATS 小鼠不能促进d-PFC 数升高。APS 可以提高 T_x-ATS 小鼠在 LPS 免疫后 d-PFC 数,这个作用同 APS 对正常小鼠 SRBC 免疫后的效应相比显著降低。这些结果提示,APS 促进抗体形成作用机理可能是通过 T 细胞介导的。     

日本血吸虫感染宿主的免疫抑制现象

本文报告用巨噬细胞移动抑制试验,溶血空斑试验及T细胞辅助活力测定观察小鼠感染日本血吸虫后对植物血凝素、血吸虫成虫抗原、羊红细胞及半抗原-载体(TNP-童虫)免疫应答的抑制现象。MMIT的结果表明:对PHA的应答于感染后28天出现抑制;对血吸虫成虫抗原的应答于感染后2周内正常,第3周开始出现抑制,14周后又基本恢复正常。溶血空斑试验结果表明,感染后头3周对SRBC的应答水平递增,第3周达最高水平,第4周开始应答逐渐下降。不同H-2的纯系小鼠感染日本血吸虫后对SRBC的应答规律基本相似,但于感染后同一时间内不同H-2的小鼠应答水平随小鼠品系的不同而不同。T细胞辅助活力测定的结果表明:小鼠感染后3周内对TNP-童虫的应答逐渐增加,第3周最高,之后逐渐下降。上述结果提示感染宿主存在免疫应答的抑制现象。这种抑制现象不仅表现在T细胞对促有丝分裂因子PHA和血吸虫成虫抗原的应答上;也表现在B细胞对胸腺依赖抗原SRBC的应答上,以及在T-B细胞协作产生抗半抗原抗体时,T细胞辅助应答也受到抑制的影响。     

精制厌氧棒状杆菌细胞壁的佐剂作用及其对巨噬细胞的激活性能

本文主要观察经冷冻高压破碎菌体、SDS处理以及蛋白酶消化提纯的精制厌氧棒菌细胞壁的佐剂作用;用溶血空斑法测定抗体形成细胞数以及间接血凝法测定免疫小鼠血清抗体,证明其对抗原SRBC及BSA具有明显佐剂作用;此外亦能恢复带瘤小鼠(艾氏腹水癌)的吞噬功能。     

MIMF—1对B细胞功能的抑制效应

人精浆含有男性抑制物质(MIM)及其他抑制因子,它们可防止对精子及精浆产生免疫应答。本文报告,用Sephadex G-100过滤法从液化的正常人精液中制备MIM。犹如以往报道,从凝胶过滤中可分离出五个峰;而第一峰(称为MIMF-1)在SDS-PAGE上呈现一个区带。MIMF-1不仅能抑制SRBC进入小鼠体内所引起的免疫应答(以溶血空斑多少表示);亦能降低小鼠血清中SRBC抗体滴度(表1)。MIMF-1所致的免疫抑制作用或许与精液过敏或不育有关。     

绵羊红细胞溶血素制备方法的改进及其在教学实验中的应用

目的:制备高效价、具有较高应用价值的绵羊红细胞(SRBC)溶血素。方法:采用两种免疫方法制备绵羊红细胞溶血素:A组采用传统SRBC悬液兔耳缘静脉注射的免疫方法,B组采用SRBC悬液加弗氏佐剂作为免疫原皮下注射的免疫方案。结果:用SRBC溶血实验及酶联免疫吸附实验(ELISA)检测抗体效价。结果表明,B组抗体效价均比A组高4倍,在溶血实验中获得了良好的教学效果。结论:SRBC悬液加弗氏佐剂的免疫方案对制备高效价的SRBC溶血素具有较高的应用价值。     

免疫应答过程对小鼠习得性行为的影响及其相关分子机制的研究

目的探讨免疫应答和免疫记忆的建立对大脑学习记忆能力的影响及分子机制。方法以绵羊红细胞作为外源性抗原激发小鼠免疫应答,建立免疫记忆;同时注射生理盐水作为对照组。应用跳台法对两组小鼠进行被动回避行为训练,以步下潜伏期为学习记忆成绩指标比较分析两组的差异。再取行为训练后小鼠脑做石蜡切片采用免疫组化法检测其海马结构内cFos蛋白的表达。结果小鼠经被动行为训练后学习记忆成绩免疫组优于对照组,有统计学显著性差异(P<0.05);小鼠海马内cFos蛋白表达免疫组显著高于对照组(P<0.05);相关性分析发现实验组小鼠抗SRBC抗体效价与海马cFos蛋白表达的免疫组化灰度值之间,实验组小鼠抗SRBC抗体效价与行为训练后学习记忆成绩之间不呈统计学相关(P>0.05)。结论免疫应答和免疫记忆的建立可以提高大脑的学习记忆能力,免疫应答过程增强小鼠脑海马内的cFos蛋白的表达可能是免疫系统对大脑高级活动产生影响的重要因素。     

Lack of age-associated auto-anti-idiotypic antibody regulation in mucosal-associated lymph nodes

It has previously been shown that the loss of immune competence in the splenic B cell population with age may be due to auto-anti-idiotypic antibody regulation (M. R. Szewczuk and R. J. Campbell, Nature 1980. 286: 164). In the present study we have investigated the appearance of auto-anti-idiotypic antibody on immune B cells from the mucosal-associated lymph nodes of old and young mice of the same strain. Various aged C57BL/6J male mice were immunized with 500 μg trinitrophenylated bovine gamma globulin (TNP-BGG) in complete Freund's adjuvant (CFA) intraperitoneally. IgM, IgG and IgA anti-TNP plaque-forming cell (PFC) responses in the spleen, mediastinal and mesenteric lymph nodes were assayed for anti-idiotype-blocked, hapten-augmentable PFC, 14 days later. It was found that 8 months or older C57BL/6J male mice produced a significantly high percentage of hapten-augmentable IgM, IgG and IgA anti-TNP PFC in the spleen. In contrast, there was a lack of hapten-augmentable anti-TNP PFC in the mesenteric and mediastinal lymph nodes with increasing age of the animal. Mice receiving antigen in the footpads and base of the tail also produced a significantly high percentage of hapten-augmentable IgG PFC in the draining peripheral lymph nodes. TNP-ε-amino-n-caproic acid (EACA) as hapten was shown to specifically augment anti-TNP PFC. Immune sera from 15-month-old mice caused a specific inhibition of anti-TNP PFC in vitro. The inhibition of plaque formation was completely reversible by addition of TNP-EACA as hapten. This PFC inhibitory activity in immune sera lacked anti-TNP antibody activity, but reacted with anti-TNP antibody of C57BL/6J origin. Immune sera from 8 month or older mice also revealed anti-(anti-TNP F(ab')₂ IgG) titer as assayed by passive hemagglutination. Thus, the results of this study indicate a division of the immune system into regulatory compartments. Auto-anti-idiotypic antibody may be involved in a down-regulation of systemic responses but with no apparent effect in the mucosal-associated lymph nodes.     

Alterations of Immune Reactivity by Haloperidol and Delta-9-Tetrahydrocannabinol

Studies were performed to determine whether Δ⁹-tetrahydrocannabinol (THC) or haloperidol suppress or ablate humoral or cellular immune responses against sheep erythrocytes. Both agents produced dose-dependent reductions in henolytic plaque-forming cell (PFC) numbers at the time of peak reactivity (Day 4) in vehicle-treated, control mice. However, both Δ⁹-THC and haloperidol only delayed the time of peak PFC formation by 24-48 hours. These changes in kinetics of humoral immune responsiveness took place at doses of Δ⁹-THC and haloperidol that produced signs of gross behavioral toxicity. Neither Delta-9-THC, cannabinol (CBN) or cannabidiol (CBD) had an effect on the titer of serum hemagglutinat-ing antibody measured seven days after immunization. Further, haloperidol did not alter the delayed-type hypersensitivity response to dinitroflorobenzene (DNFB).     

IMMUNOLOGICAL AND HISTOLOGICAL STUDIES OF THE PROLONGED ADMINISTRATION OF γG FRACTION OF ANTILYMPHOCYTE SERUM

Rabbits were treated with the prolonged administration of γG fraction of goat anti-rabbit lymphocyte serum (ALγG) over 180 days. The effect of ALγG on various kinds of immune responses was examined at different stages of the experiment.
The antibody against ALγG appeared at an early stage. Especially the formation of γM antibody was rather enhanced as compared to the control animals which received normal goat γG (NγG). The antibody titer increased, and then maintained its level through the course of experiment. However, a slight decline of the titer was noted in the last stage. The Immune responses against sheep red blood cells, bovine serum albumin and skin homograft were greatly suppressed by the short-term treatment of ALγG. On the other hand, no suppression of the immune responses was demonstrated in the group treated with ALγG for 180 days as compared with the control group.
The results indicate that ALγG used is effective to suppress the immune responses in the short-term treatment, but it is probably neutralized by the antibody and causes no further immunosuppression. In addition, it has also been confirmed that an exhaustion of the lymphoid tissues accompanying occasional amyloidosis might be induced by the long-term administration of ALγG.     

Detection of cytomegalovirus antibody with latex agglutination.

Transfusion-acquired cytomegalovirus (CMV) infections should be prevented in seronegative immunocompromised patients by providing blood products from donors who are also seronegative. Latex agglutination was investigated as a simple and rapid method for detecting antibody against CMV. Latex beads were coated with CMV antigen, incubated for 8 min at room temperature with 25 microliter of sera, and examined for agglutination. The sensitivity and specificity of latex agglutination was compared with that of indirect hemagglutination (IHA, Cetus Corp., Emeryville, Calif.) and enzyme immunoassay (EIA) with sera from 604 random blood donors or patients. Of 327 serum samples shown to be seronegative by EIA and IHA, 327 had a latex agglutination titer of less than 1:4 (specificity, 100%). Of 236 serum samples with detectable antibody by EIA and IHA, 228 had a latex agglutination titer of 1:4 or greater (sensitivity, 97%). Plasma collected with EDTA, heparin, or citrate was satisfactory for latex agglutination. Latex agglutination results correlated quantitatively with those of EIA, and the test also detected fourfold or greater rises in antibody with paired sera from six patients with posttransfusion CMV infections. Latex agglutination is a sensitive and specific assay that is rapid and simple to perform and should be effective in selecting seronegative blood donors to prevent posttransfusion CMV infections in seronegative recipients.     

脆弱类杆菌NCTC9343脂多糖抗体的产生

本实验用间接血凝方法检测了兔抗血清中抗脆弱类杆菌NCTC 9343株脂多糖(LPS)抗体的变化情况。在初次免疫应答中,特异性抗体产生得很快,免疫局第2天即可测出,持续3～4周,高峰出现在第6～8天,最高效价为1:80;再次免疫应答,抗体高峰出现在再次免疫后的2～4天,效价为1:320,抗体效价维持6～8周。特异性抗体的类型主要是IgM。     

Effect of chinese softshell turtle egg powder on immune functions in mice

The effects of whole Chinese soft-shell turtle egg powder (WTE) on immune functions is unknown. Female ICR 6-week-old mice (n=80) were administered for 30 days with 1.125 g·kg^?1 WTE. Spleen and thymus were weighed and spleen lymphocyte proliferation response to concanavalin A (Con A), plaque forming cell(PFC) assay, phagocytosis assay and NK cell activity detection were performed on day 30. Compared to vehicle control, WTE increased the relative weight of spleen (p<0.05). Mice treated with WTE showed increased Con A stimulated lymphocyte proliferation (p<0.05) and PFC (p<0.01). Carbon clearance capacity (p<0.05) and NK cell activity (p<0.01) were also enhanced while a mild improvement was seen in phagocytosis capacity of peritoneal cavity macrophages in WTE treated mice. In conclusion, our data provides evidence of the enhanced effects of WTE in cell-mediated immunity, humoral immunity and NK cell activity in mice.     

Effect of Kalpaamruthaa on immunosuppression of both humoral and cell-mediated immunity in DMBA-induced mammary tumours of rats

The present study was carried out to elucidate the protective effect of Kalpaamruthaa on improving 7,12-dimethylbenz(a)anthracene (DMBA)-induced immunosuppression of both humoral and cell-mediated immunity in mammary carcinoma-induced rats. Breast cancer was induced in rats by administering DMBA orally (25 mg/rat) as a single dose. After 90 days of induction, SA (200 mg/kg body weight) and KA (300 mg/kg body weight) were administered for 14 days, by gastric intubation. Several immunotoxicological assays such as T cell rosette delayed type hypersensitivity (DTH) response, migration inhibition factor (MIF) assay, lymphocyte proliferation assay, plaque forming cell (PFC) assay and haemagglutination assay, plaque forming cell (PFC) assay, serum soluble immune complex and cytokine production, T and B cell mitogenesis induced by Con A and nonspecific cell-mediated immunity were evaluated using phagocytosis activity and NBT reduction. In cancer-induced animals (group II), the leukocyte migration inhibition declined markedly (p?<?0.001), the levels of cytokines IFN-γ and IL-2 were significantly decreased (p?<?0.001) and also the antibody titre level (p?<?0.001) was significantly reduced when compared with control rats. A marked decline in PFC (p?<?0.001) and serum soluble immune complex (PEG) formation (p?<?0.001) was also observed. Hence, the present study clearly demonstrates the immunoprotective effect of KA.     

The relationship of antibody-forming cells to rosette-forming cells

Rosette-forming cells (RFC) increased in numbers in the spleens of mice following injection of sheep red blood cells (SRBC). Very sensitive assay system for detecting plaque-forming cells (PFC) showed that RFC and PFC were present in approximately equal numbers at the height of the immune response. Thereafter PFC numbers declined much more rapidly than RFC.

Two techniques were used to study the contribution of PFC to RFCs (a) velocity sedimentation through foetal calf serum gradients and (b) transfer of individual RFC by micromanipulator into the PFC assay gel containing complement and rabbit antimouse IgG antiserum to identify what proportion of RFC were secreting antibody.

It was shown that, when rosettes were prepared at 4°, <10 per cent were formed by PFC at 4 days after immunization, and <2 per cent at 6 days. Rosettes prepared at 37° contained up to 16 per cent PFC.

It was concluded that PFC had either no cell bound antigen-binding receptors or that the receptors were not demonstrable at 4° by the particular rosette preparation used in the study.

Rosettes prepared late in the immune response were more resistant to mechanical agitation than those prepared early in the response.

     

IMMUNOHISTOLOGICAL ANALYSIS OF INTRAVASCULAR AGGLUTINATION IN A CASE OF ABO INCOMPATIBLE BLOOD TRANSFUSION BY MEANS OF MIXED CELL AGGLUTINATION REACTION

Applicability of the mixed cell agglutination reaction (MGAR of Davidsohn) in histological examination was tested in a case of ABO incompatible blood transfusion, who had died of severe head injury two days after an accident. Massively disseminated intravascular aggregation of erythrocytes was identified as immune agglutination by the modified method of MGAR (I shiyama & O kada ) with precise knowledge of topographic isoantigen localization in tissues. The immune specific agglutination mimics intravascular changes such as hyperemia, stagnation and hemorrhage derived from various circulatory disturbances obtained in routine autopsy materials. ACTA PATH. JAP. 27: 729 ˜ 738, 1977.     

Modulation of Primary Antibody Response by Protein a in Tumor Bearing Mice

Abstract

Protein A (PA) is a cell wall glycoprotein of Staphylococcus aureus Cowan I, which possess a number of immunomodulatory and antitumor properties. We have previously shown that PA suppresses the anti-sheep erythrocyte primary antibody response in normal mice. The present investigation evaluates the effect of protein A on the anti-sheep erythrocyte primary antibody response in tumor-bearing mice. The primary antibody response in tumor-bearing mice immunized with sheep red blood cells (SRBC) was suppressed by the intraperitoneal administration of PA in a dose-dependent fashion. The plaque forming cell (PFC) assay was used to assess this response. Maximum suppression of the PFC response was observed at 12 μg PA/animal (p<0.001) and could be observed at doses as low as 1 μg PA/animal (p<0.01). The amount of suppression was proportional to the number of PA doses administered. In addition this effect was critically dependent on the timing of PA administration. PA showed no significant effect on PFC when injected after immunization, but it produced pronounced suppression when injected prior to the immunization with SRBC. Maximum suppression of the PFC response was observed when PA was administered one day before the antigen challenge. PA also reduced splenic localization of ⁵¹Cr labeled SRBC to 42% (p<0.01). The altered localization of antigen in spleen may be responsible for reduced PFC response in tumor-bearing mice. Depletion of B-lymphocyte is reported to exhibit tumor inhibition. Therefore, we propose that the suppression of the primary antibody response by PA helps in tumor regression by reducing the soluble immunosuppressive immune complexes.