BXSB小鼠血清抗DNA抗体水平及其对巴细胞转化的影响

用酶联免疫吸附剂测定法(ELISA)和3H-胸腺嘧啶核苷掺入法,观察一种自发性系统性红斑狼疮(SLE)好发小鼠(BXSB小鼠)在不同月龄时,血中抗DNA抗体水平的变化,并观察该血清对正常LACA小鼠淋巴细胞转化的影响.结果显示,自3月龄起,BXSB雄性鼠血中的抗DNA抗体水平开始升高,4月龄上升至高峰,5月龄仍保持较高水平,各月龄BXSB血清与对照血清相比,P<0.05(ANOVA).同时,该血清对正常LACA小鼠淋巴细胞转化也有抑制作用,以4月龄血清的抑制作用最大.如果将同一只小鼠血清抗DNA抗体水平的变化与该血清对淋巴细胞转化的影响做回归和相关分析,发现两者有明显的正相关关系(r=0.232,n=53, P<0.05).提示雄性BXSB小鼠可能从3月龄开始表现SLE.发病后,血清中产生某种能够抑制淋巴细胞转化的物质,它的产生与抗DNA抗体有平行关系.     

胃癌血清中免疫抑制因子的初步探讨

带瘤动物血清能抑制小鼠脾细胞抗绵羊红细胞的空斑形成细胞反应(PFC),说明其血清中存在免疫抑制因子。1980年Takateru等报道用肺癌血清作PFC免疫抑制试验,阳性率达78％,非癌肺部疾病者为6％,正常人为7％,并提出有可能将此法用于肺癌的免疫诊断。由于此免疫抑制试验的结果与肺癌的组织分型无显著关系,提示可用于其他肿瘤的测定。我们尝试用胃癌血清作PGC免疫抑制试验,以了解,胃癌血清的免疫抑制活性,现将结果简要报道如下。 材料与方法 取C57BL/6J小鼠,每组3只。将0.4ml经56℃     

几种阿片肽对小鼠腹腔巨噬细胞吞噬功能的影响

本实验观察了几种阿片肽及氢化考的松对小鼠腹腔巨噬细胞吞噬染料中性红能力的影响。 18-20克雄性LACA小鼠断颈髓处死后用Hanks＇液洗出腹腔巨噬细胞。用含10％小牛血清的RpMI_1,640液将细胞制成悬液(2×10~6细胞/ml)。在40孔细胞培养板中,每孔加入100μl细胞悬液。药物用RpMI_1,640液稀释成10~(-5)、10~(-7)、10~(-9)、10~(-11)M,分别加入细胞悬液中(每孔容量10μl),药物最终浓度分别为10~(-6)、10~(-8)、10~(-10)、10~(-12)M,同一浓度加入10孔中,即为10复管。细胞经37℃、24小时孵育后,弃去培养液,每孔加入     

同种特异T细胞疫苗诱导移植物存活期延长的研究——Ⅲ.抗T细胞抗体的活性分析

本文观察了TCV免疫小鼠血清对同种抗原和非特异抗原刺激的淋巴细胞反应性的影响,结果显示,TCV免疫小鼠血清能显著抑制ConA诱导的淋巴细胞增殖和同种MLR,表明该血清中存在有抑制T细胞功能的“抑制因子”。实验还发现TCV免疫小鼠血清中存在针对多种不同抗原决定簇的抗体,包括有TCV特异的、活化T细胞的和正常淋巴细胞的。TCV吸收实验证明TCV免疫小鼠血清中能特异地与TCV结合的抗体不能与其它活化T细胞结合。经细胞膜分子SDS-PAGE和免疫印迹分析,TCV免疫小鼠血清中的抗体所识别的细胞膜抗原分子量大约为15kD,35/38kD左右。     

试管内诱生抗体的研究——Ⅱ.对实验条件的进一步探讨

本文对体外诱生溶血空斑形成细胞(PFC)的实验条件进行了进一步研究,包括筛选新生小牛血清、挑选绵羊红细胞(SRBC)的供羊;和诱生抗体反应的最适SRBC剂量,以及选择适合于本实验的小鼠品系。结果证明不同批的小牛血清支持PFC反应的能力及不同个体的SRBC诱生PFC反应的能力存在着差异;每毫升脾细胞培养物中加入50微升1％SRBC悬液及用615小鼠作为脾脏细胞的供体较为合适。还对贮存不同时间的SRBC和用SRBC吸收过的小牛血清进行了比较。通过选用合适的条件,能够获得稳定的实验结果和诱生出较高的PFC反应。文内分析和讨论了可能引起实验失败的原因。     

酶联免疫斑点方法改进及应用

建立了ELISPOT方法并通过改进 ,应用到抗人CD8杂交瘤细胞及LACA小鼠体外脾细胞分泌IgG量的检测 ,同时观察六味地黄多糖CA4 3在体外对LACA小鼠脾细胞分泌IgG的影响。结果表明 10 μg/mlLPS作用下可明显提高小鼠脾细胞体外产生抗体的量和提高分泌抗体的细胞数。 5 0 μg/mlCA4 3在体外与LPS具有相似的作用 ,可以刺激小鼠脾细胞分泌IgG的量和分泌抗体的细胞数量增多。     

含活性炭纤维敷料类产品的体外细胞毒性研究

活性炭纤维因其高效吸附特性而广泛应用于医疗卫生领域。我们设计不同的浸提液制备方法,通过MTT法评价该浸提液对小鼠成纤维细胞L929活性及增殖的影响,以寻找适合评价含活性炭产品体外细胞毒性的浸提方法。结果显示,由于活性碳纤维的强吸附性,相对于传统的含血清细胞培养基直接作为浸提介质,采用无血清培养基浸提,试验前在浸提液中加入10%血清更适合评价含活性炭产品的体外细胞毒性。     

创伤对小鼠抗体分泌细胞的抑制作用及其纠正实验观察

<正> 本实验利用空斑形成细胞(简称PFC)测定法检测SRBC免疫小鼠脾细胞的抗体分泌细胞数,观察创伤对B细胞系统的抑制作用及抗体生成素对该抑制作用的纠正效应。 重18～22g的雄性昆明小鼠随机分为创伤组、创伤治疗(简称洽疗)组和对照组,每组10只,在实     

对IgA缺乏和有抗IgA抗体母亲所生两个孩子的免疫学研究

1968年首次确认抗人IgA自身抗体,在选择性IgA缺乏病人其检出率可达40％。在小鼠实验中亦证明抗IgA抗体能抑制IgA的合成。新生小鼠于Ig开始产生前、应用抗IgA亦看到抑制IgA合成,停止用抗IgA时,则小鼠再次产生TgA,血清中IgM和IgG正常     

低剂量电离辐射对免疫器官局部微环境的免疫调节作用

目的和方法：观察７５ｍＧｙＸ射线全身照射昆明系雄性小鼠后２４ｈ，其血清及胸腺和脾细胞外液对免疫功能的影响。结果：小鼠经７５ｍＧｙ照射后，其血清抑制脾细胞对ＣｏｎＡ反应性低于正常血清，其胸腺（脾）细胞外液抑制胸腺细胞３Ｈ－ＴｄＲ自发掺入能力（脾细胞对ＣｏｎＡ反应性）也明显低于正常对照组，尤以ＭＷ＜１０ｋＤ组份为著；胸腺和脾细胞外液经ＳｅｐｈａｄｅｘＧ１００层析获得的第一峰较正常对照的高，而第二峰较之为低，其中第二峰对免疫功能的抑制作用低于正常对照的第二峰更为明显。结论：低剂量辐射能够改变免疫器官局部微环境，使免疫功能增强，可能与其神经体液因子等因素的变化有关。     

IgE-mediated sensitization to English plantain pollen in seasonal respiratory allergy: identification and partial characterisation of its allergenic components.

Characterisation by SDS-PAGE immunoblotting of plantain pollen extract showed that components of 16,000-20,000 M(r) were frequently reactive with IgE antibody in the sera of subjects with seasonal respiratory allergy. Other, more weakly IgE-binding allergens were seen in the range of 40,000-60,000 M(r). HPLC followed by RAST inhibition demonstrated that components of approximately 17,000 M(r) were also responsible for much of the IgE-binding activity of the extract. These components appeared to have pI values between 4.5 and 5.2. RAST inhibition showed that there were no common IgE-binding epitopes in grass pollen and plantain pollen extracts, indicating that skin test responses should not necessarily be interpreted in terms of cross-reaction. 82 subjects with a clinical history of seasonal, respiratory allergy were screened in a skin prick test survey. 28% were skin test positive to plantain pollen extract. The frequency of positive skin test reactions to plantain pollen extract was greater than that to Betula (23%) and Artemisa (16%), both which are considered to be important allergens. In a larger survey positive RAST scores to plantain pollen were given by 34% of sera from subjects with respiratory allergy. Plantain pollen sensitivity should therefore be considered during diagnosis of seasonal allergy.     

Immunoglobulin E (IgE)-mediated cross-reactivity between mesquite pollen proteins and lima bean, an edible legume

Immunoglobulin E (IgE)-mediated food allergy often develops as a consequence of allergic sensitization to pollen proteins. Mesquite (Prosopis juliflora) tree pollen is reported to be cross-reactive with other pollen species, but little has been reported on its cross-reactivity with plant-derived foods belonging to the same/different families. The present study investigates the in vitro cross-reactivity of mesquite pollen and lima bean (Phaseolus lunatus), an edible seed belonging to the Leguminosae family. Of 110 patients (asthma, rhinitis or both) tested intradermally, 20 showed marked positive reactions with Prosopis pollen extract. Of these, 12 patients showed elevated specific IgE to Prosopis pollen extract alone and four to both Phaseolus and pollen extract. In vitro cross-reactivity was investigated using inhibition assays [enzyme-linked immunosorbent assay (ELISA) inhibition, immunoblot inhibition], histamine release and lymphoproliferation. P. lunatus extract could inhibit IgE binding to P. juliflora in a dose-dependent manner, requiring 400 ng of protein for 50% inhibition in ELISA assay. Immunoblot and immunoblot inhibition demonstrated the presence of 20, 26, 35, 66 and 72 kDa as shared IgE binding components between the two extracts. Histamine release, peripheral blood mononuclear cells proliferation and interleukin (IL)-4 levels also suggested allergenic cross-reactivity. In conclusion, there is humoral and cellular cross-reactivity between Prosopis pollen and Phaseolus seed allergens.     

Clinicoimmunologic studies on Phoenix sylvestris Roxb. pollen: an aeroallergen from Calcutta, India

BACKGROUND: This study highlights the allergenicity and allergenic components of the pollen of Phoenix sylvestris Roxb. (PS), or date sugar palm, which is predominantly airborne in the air of Greater Calcutta. METHODS: A 2-year aerobiologic survey was performed by Burkard sampler. PS pollen extract was used in skin tests of allergic patients, fractionated by (NH4)2SO4 and the Sephacryl S-200 column. The allergenicity of each fraction was checked by skin test and IgE ELISA inhibition. The principal allergenic fraction, Fr.lla, was separated in 11% SDS-PAGE, and its allergenicity was confirmed by IgE ELISA inhibition and immunoblotting. RESULTS: PS pollen grains were found to be prevalent in the air of the suburban zone of Calcutta from January to March with a peak in February. The pollen extract showed high (44.07%) positive skin reaction on 540 respiratory allergic patients. Among the (NH4)2SO4 cut fractions, Fr.II was the most active one, and it was resolved into four subfractions in the Sephacryl S-200 column. Fr.lla was the principal allergenic fraction, showing the presence of two components of 33 and 66 kDa in SDS-PAGE. In IgE immunoblotting, both of the components were found to be allergenic. CONCLUSIONS: The PS pollen grain is an important aeroallergen from Calcutta, India. The 33- and 66-kDa components are the major allergens present in the relevant pollen extract.     

Allergenic cross-reactivity of olive pollen

BACKGROUND: Sera of patients allergic to olive (Olea europaea) pollen were used to analyze the IgE cross-reactivity between olive-pollen extract and other pollens obtained from phylogenetically unrelated species. METHODS: We used IgE immunostaining of pollen extracts blotted to nitrocellulose membranes after SDS-PAGE and inhibition analysis of this binding. RESULTS: A high inhibition of the IgE binding on olive-pollen extract was exhibited by birch, mugwort, pine, and cypress pollens, suggesting that these extracts contain proteins which share common epitopes and thus can be recognized by olive-allergic sera. IgE binding to Gramineae pollen extracts was not inhibited by olive-pollen extract, indicating a primary sensitization of the patients to these species. From the inhibition assays, the presence of an allergen of 45 kDa in the olive pollen, which has no homologous counterparts in other allergenic species, has been inferred. CONCLUSIONS: Olive pollen contains allergens which cross-react with pollens from unrelated species, a fact that could simplify the diagnosis and treatment of pollinosis.     

Primary sensitization to sweet bell pepper pollen in greenhouse workers with occupational allergy

BACKGROUND: In a previous investigation, a high prevalence of allergy to sweet bell pepper pollen was found among exposed horticulture workers. Allergy to plant-derived food is often the consequence of primary sensitization to common pollen allergens. OBJECTIVE: We therefore investigated the cross-reactivity between sweet bell pepper pollen and pollen from grass, birch or mugwort. METHOD: We selected 10 sera from greenhouse workers who had, besides specific IgE against sweet bell pepper pollen, also IgE to grass, birch or mugwort pollen. Cross-reactivity was tested by the inhibition of IgE binding to solid-phase coupled sweet bell pepper pollen extract. The 10 sera were also analysed for IgE binding to sweet bell pepper pollen by immunoblotting. RESULTS: With these sera, no or small inhibition of IgE binding to sweet bell pepper pollen extract was observed with grass, birch and mugwort pollen. With immunoblotting, major IgE-binding structures were seen at 14, 29 and 69 kDa in sweet bell pepper pollen extract. CONCLUSION: The results of our study demonstrate that sweet bell pepper pollen contains allergens that have no or limited cross-reactivity with common pollen allergens. With sera from the 10 patients tested, sensitization to sweet bell pepper pollen was not the consequence of primary sensitization to common pollen allergens.     

Occupational Allergy to Flowers: Immunoblot Analysis of Allergens in Freesia,Gerbera and Chrysanthemum Pollen

High exposure to pollen from ornamental flowers can induce an IgE‐mediated occupational allergy in florists and horticulture workers. We investigated IgE‐binding antigens in chrysanthemum, freesia and gerbera pollen by immunoblot analysis and analysed the cross‐reactivity of these pollen with birch, grass and mugwort pollen. In immunoblots with chrysanthemum pollen, major IgE‐binding structures were seen with a molecular weight (MW) of approximately 25, 45 and 65 kD. In the immunoblots with freesia pollen, IgE from freesia pollen was directed against two proteins with an MW of approximately 15 kD. Most sera showed IgE binding to an approximately 15 kD band in gerbera pollen; with some sera additional bands were seen in the range of 30–50 kD. IgE binding to chrysanthemum pollen was inhibited by mugwort pollen only, whereas IgE binding to freesia pollen was suppressed by birch, grass and mugwort pollen. The inhibitory activity of birch and grass pollen extract on IgE binding to gerbera pollen extract was serum dependent and ranged from no inhibition to complete inhibition. Occupational exposure to many different flowers induced IgE against all three types of pollen. Exposure in greenhouses to gerbera flowers elicited mainly IgE against gerbera pollen. Mugwort pollen extract inhibited IgE binding to pollen from all three flowers.     

Platanus acerifolia pollinosis and food allergy

BACKGROUND: In Mediterranean areas, oral allergy syndrome (OAS) occurs independently of an associated birch pollinosis; moreover, on occasions it presents with no other associated pollinosis. The aim of this study was to assess the possible association of OAS with Platanus acerifolia pollinosis. METHODS: We evaluated consecutive patients seen for pollinosis in an allergy department. Seven hundred and twenty patients were selected on the basis of seasonal or perennial rhinitis, or asthma, or both. Respiratory and food allergies were studied in all patients. Clinical history was recorded and examinations and skin prick tests were performed with a battery of available common inhalant allergens and plant-derived food allergens. Specific IgE levels to P. acerifolia pollen extract and food allergens tested were measured. Molecular masses of the IgE-binding proteins and cross-reactivity among the P. acerifolia pollen and different food extracts were also determined. RESULTS: Of the 720 patients evaluated, 61 (8.48%) were sensitized to P. acerifolia pollen. Food allergy was observed in 32 (52.45%) of the 61 patients sensitized to P. acerifolia pollen. Food allergens most frequently implicated were hazelnuts, peach, apple, peanuts, maize, chickpea and lettuce. Enzyme allergosorbent (EAST)-inhibition showed high inhibition values when P. acerifolia pollen extract was used as free phase. On the contrary low inhibition was observed when plant-derived food allergens were used as free phase and P. acerifolia pollen extract as solid phase. CONCLUSIONS: Cross-reactivity was observed among P. acerifolia pollen and plant-derived foods. OAS in these patients may have been caused by primary respiratory sensitization.     

Antibody responses of mice to intragastric and parenterally administered aeroallergens

Intragastric administration of aeroallergens (pollen extract)-primed mice to produce transient serum IgE antibody responses following subsequent parenteral stimulation while the same initial dose of extract, given parenterally, did not have this effect. In previously immunized animals, intragastric administration of pollen extract was found to enhance systemic antibody production. These observations indicate that exposure of gut-associated lymphoid tissue to aeroallergens can have a profound effect on subsequent reaginic antibody production. This procedure provides a useful model for studying IgE responses to allergens without the complication of an initial injection with adjuvant. A combination of parenteral immunization with oral administration may therefore offer a convenient immunotherapeutic manoeuvre for patients with seasonal rhinitis/asthma.     

Occupational rhinitis and bronchial asthma due to artichoke (Cynara scolymus).

BACKGROUND: The artichoke is a perennial horticultural plant that belongs to the Compositae family. OBJECTIVE: To present case studies of 2 vegetable warehouse workers who developed occupational rhinitis and bronchial asthma by sensitization to artichoke. METHODS: Skin prick tests with common inhalants and foods were performed. Specific IgE to artichoke, Parietaria judaica pollen, and Olea europaea pollen extracts was measured by a specific IgE enzyme immunosorbent assay kit. Molecular mass of the allergens was studied by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) immunoblotting technique. Patients underwent a nasal challenge test, and one patient provided peak expiratory flow rate (PEFR) measurements in her workplace. RESULTS: In both patients, results of skin prick tests to artichoke were positive. Levels of specific IgE for artichoke were 0.68 kU/L in patient 1 and 2.14 kU/L in patient 2. The protein composition of the artichoke extract, studied by SDS-PAGE, showed that most bands ranged from 30 to 14 kDa. The IgE-binding bands with the serum samples of patient 1 showed apparent molecular masses of 56, 48, 38, 31, 27, 25, 16, and 15 kDa; however, the serum samples of patient 2 showed IgE bands of 21 and 19 kDa. Western blotting of artichoke extract showed a complete inhibition of IgE-binding bands when serum samples were preincubated with P. judaica pollen extract. Nasal challenge with artichoke extract triggered a peak nasal inspiratory flow decrease of 81% and 85% in patient 1 and patient 2, respectively. Finally, patient 1 recorded a PEFR decrease of up to 36% after exposure to artichoke in her workplace. CONCLUSIONS: SDS-PAGE immunoblotting inhibition performed for the artichoke extract showed a total disappearance of the specific IgE binding bands when serum samples were previously incubated with P. judaica pollen extract, thus establishing the existence of a serologic cross-reactivity between artichoke and P. judaica pollen.     

Aerobiological and immunochemical studies on Carica papaya L. pollen: an aeroallergen from India

BACKGROUND: Carica papaya L. is a fruit yielding tree, wildly grown or cultivated in the tropics and subtropics. Its pollen grain has been reported to be airborne and cause immunoglobulin E (IgE)-mediated hypersensitivity. OBJECTIVE: To conduct long-term aerobiological study on Carica pollen, along with aeroallergenic particles originating from it and to identify vis-a-vis characterize an important IgE-reactive component present in this pollen. METHODS: The seasonal and diurnal periodicities of airborne C. papaya pollen were recorded in a 5-year survey using a Burkard volumetric sampler. The allergenic potential was studied by skin prick tests, IgE-enzyme-linked immunosorbent assay (ELISA) and also by aeroallergen immunoblotting. The total pollen extract was fractionated by Sephacryl S-200 column, and out of the eluted five fractions, the maximum IgE-reactive fraction (as found in ELISA inhibition) was resolved into five major subfractions in reverse-phase high-performance liquid chromatography (RP-HPLC). The subfraction with optimum IgE reactivity was studied by activity gel, native and nonreducing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The homogeneity of the isolated protein fraction was checked by crossed immunoelectrophoresis with rabbit antisera and IgE reactivity was confirmed by ELISA inhibition and immunoblotting using individual patient sera. RESULTS: The Carica pollen occurred in the air round the year with peaks during January and September-October. Among a patient population of 1000, skin-test results showed 27.8% +1 level and 5.6% +2/+3 level reactions. In aeroallergen immunoblotting of exposed Burkard tape segments, the detected allergen spots showed a significant correlation with airborne pollen count recorded. The pollen extract elicited loss of IgE reactivity when treated with reducing agent-like beta-mercaptoethanol and heat, but showed six IgE-reactive components in nonreducing IgE-immunoblot. The fraction 1 eluted from Sephacryl S-200 column showed highest IgE reactivity and resolved into five major components in RP-HPLC. Out of these, the fraction showing optimum IgE reactivity in IgE-ELISA inhibition and immunoblotting with patient antisera, elicited esterase activity and found to be a homogenous protein of 100 kDa. CONCLUSION: Carica papaya tree contributes significantly to the aeropollen and aeroallergen load of the suburban outskirts of Calcutta metropolis, India. The pollen extract contains an important IgE-reactive protein component of 100 kDa molecular weight with esterase activity.