Immune expulsion of the trichostrongylid Cooperia oncophora is associated with increased eosinophilia and mucosal IgA

Previous experiments have shown that a primary infection with 100000 infective larvae of the trichostrongylid Cooperia oncophora allows discrimination between different type of responder animals based on the speed by which the parasite is expelled from the host. In most of the animals (intermediate responders) the expulsion occurs 35-42 days after infection. This experiment was carried out to investigate which mechanisms contribute to the clearance of the parasite from the intestine. Sequential necropsy of the animals 14, 28 and 42 days after infection together with a segmental division of the small intestine, allowed us to characterise essential components associated with development of immunity and expulsion of the parasite from its niche. The results show that during the patent phase of the infection the parasite preferentially resides in the proximal gut. Forty-two days after infection ongoing expulsion is characterised by a migration of the worms to the more distal part of the intestine. Expulsion of the adult worm population appears to be mast-cell independent and is associated with a significant increase in parasite-specific mucous IgA and IgG1 as well as with an influx of eosinophils in the intestinal lamina propria. Although we did not observe a specific lymphocyte recruitment into the intestinal mucosa, the accumulation of eosinophils seems to be mediated by CD4+ cells. We measured significant negative correlations between the number of eosinophils and the expulsion rate of the parasite expressed by sex ratio and ratio eggs per gram faeces. Parasite-specific mucosal IgA levels were negatively correlated to the fecundity of the worms, expressed as number of eggs per female worm. Our results describe the involvement of both eosinophils and mucosal IgA in the regulation of C. oncophora expulsion and suggest the development of a Th2 effector immune response.     

T-cell mediated immune responses in calves primary-infected or re-infected with Cooperia oncophora: similar effector cells but different timing

Cooperia oncophora is the most prevalent intestinal nematode of cattle occurring in Western Europe. Primary infection with 100000 third stage infective larvae (L3) induces acquired immunity in a high proportion of the animals but there is little information on immunity against re-infection. In the current experiment, the contribution of the T-cell mediated immunity in protection against re-infection with C. oncophora was investigated in detail. Priming elicited long-lasting protective immunity that was evidenced by a significantly decreased worm burden and egg excretion in primed animals compared to challenge control animals. Lymphocyte proliferation tests with excretory/secretory products (ESP) of C. oncophora and with three distinct ESP fractions indicated an enhanced reactivity in primed animals and suggested that by fractionating of ESP we selected for proteins involved in protective immunity against re-infection with C. oncophora. Phenotypic analysis of T cell subsets at diverse anatomical locations revealed that the enhanced reactivity of lymphocytes from peripheral blood and lymph nodes of the infected animals coincided with a significantly increased frequency of CD4(+) cells at these locations but a deceased frequency of CD4(+) cells in the lamina propria. These findings were independent of the immune status of the animals but more pronounced in the primed animals than in the challenge control animals. In addition we demonstrated that primary and secondary infections with C. oncophora were associated with two waves of eosinophils and that the kinetics of this cell population differed as a result of priming. Based on the observed correlations we propose that the early increase of eosinophils is T cell independent and merely a consequence of inflammation in the parasitised gut. In contrast, the second wave of eosinophils depends upon CD4(+) cells and correlations with parasitological parameters at this time point support a role of eosinophils as effector cells against adult stages of C. oncophora.     

Positive coupling of atypical adenosine A3 receptors on human eosinophils to adenylyl cyclase

Adenosine A(3) receptors are reported to couple negatively to adenylyl cyclase (AC) but their mediation of anti-inflammatory effects in human eosinophils prompted us to investigate their coupling to AC. The A(3)-selective agonists IB-MECA and Cl-IB-MECA evoked a concentration-dependent generation of cAMP (EC(50), 3.2 and 1.8 microM, respectively) and were more potent than the A(2A) agonist CGS 21680 (EC(50)=15.4 microM) and adenosine (EC(50)=19.2 microM). The cAMP response was additive to that produced by forskolin (10 microM). The effect of IB-MECA was insensitive to A(1) and A(2A) receptor antagonists, but was antagonized by the A(3)-selective antagonist MRS 1220 (0.1-2.5 microM) in a competitive manner. The estimated K(B) of 190 nM was, however, atypical. The cyclo-oxygenase inhibitor, indomethacin, had no effect on the cAMP response. A general inverse relationship between cAMP generation and inhibition of degranulation was seen. We conclude that in human eosinophils, an atypical form of A(3) receptors positively coupled to AC may exist. The resulting cAMP generation may underlie the anti-inflammatory actions of A(3) agonists in eosinophils.     

嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的诊断及预后预测价值分析

摘要 目的：探讨嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的诊断及预后预测价值。方法：选取我院2020年3月到2023年3月收治的100例慢性鼻窦炎伴鼻息肉患者作为研究对象,将其分为慢性鼻窦炎伴鼻息肉组,选取同期来我院治疗的100例单纯慢性鼻窦炎患者作为慢性鼻窦炎组,另选取同期来我院体检的100名健康志愿者作为对照组。对比三组受检者嗜酸性粒细胞、调节性T细胞表达水平,并建立受试者工作特征（ROC）曲线,分析嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的诊断效能。随后对100例性鼻窦炎伴鼻息肉患者手术治疗后进行1年随访,依照患者的复发情况评价其预后情况,并分为两个亚组,将术后1年内复发的21例患者分为预后不良组,两未复发的79例患者分为预后良好组,对比两组患者一般临床情况,嗜酸性粒细胞、调节性T细胞表达水平,并分析嗜酸性粒细胞、调节性T细胞对慢性鼻窦炎伴鼻息肉的预后预测价值。结果：三组受检者嗜酸性粒细胞、调节性T细胞表达水平对比差异显著,慢性鼻窦炎伴鼻息肉组患者嗜酸性粒细胞（EOS）个数高于慢性鼻窦炎组和对照组,调节性T细胞（Tregs）水平低于慢性鼻窦炎组和对照组（P＜0.05）;通过绘制ROC曲线,确定EOS、Tregs其对慢性鼻窦炎伴鼻息肉的诊断效能,结果显示,EOS、Tregs两者联合对慢性鼻窦炎伴鼻息肉的诊断效能优于单一检测（P＜0.05）;预后良好组与预后不良组患者性别、年龄、BMI、吸烟史、饮酒史、白细胞介素-35（IL-35）、转化生长因子-β（TGF-β）、白细胞介素-1β（IL-1β）表达水平对比无明显差异（P＞0.05）,预后良好组与预后不良组患者病程、合并哮喘、合并变应性鼻炎、组织淋巴细胞占比、白细胞介素-10（IL-10）、肿瘤坏死因子-α（TNF-α）、EOS个数以及Tregs表达水平对比差异显著（P＜0.05）;logistic回归分析结果表明：合并哮喘、组织淋巴细胞占比、EOS个数以及Tregs为慢性鼻窦炎伴鼻息肉的预后独立影响因素（P＜0.05）。结论：嗜酸性粒细胞、调节性T细胞不仅对慢性鼻窦炎伴鼻息肉的临床诊断具有重要价值,而且能够预测慢性鼻窦炎伴鼻息肉患者的预后情况,因此临床上对于EOS个数增加,Tregs降低的患者要及时改善治疗措施,预防患者预后不良的发生。     

Zur Feinstruktur der eosinophilen Granulozyten der Hausgans (Anser anser dom.)

Zusammenfassung Die Feinstruktur der Blut-Eosinophilen wurde bei 7 klinisch gesunden Toulouser Hausgänsen (Anser anser dom.) untersucht. Sie unterscheidet sich deutlich von der der Heterophilen.Die Form der Eosinophilen ist oval bis rundlich. Das Chromatinmaterial liegt hauptsächlich in dichten Chromocentren. Unter den Zellorganellen treten Mitochondrien, die sich in osmiophile Granula umwandeln, Vakuolen, multivesikuläre Körper und Mikrofilamente hervor. Für die Eosinophilen bezeichnend sind zahlreiche ellipsoide, ovoide oder rundliche osmiophile Granula mit einem häufig band- oder stabförmigen Internum. An dem Internum läßt sich eine Substruktur aus parallel geordneten, massendichten Banden nachweisen, die bisher nur bei Säugetieren sichtbar gemacht wurde. Die ultrastrukturellen und biochemischen Merkmale der internumhaltigen Granula der Eosinophilen sprechen eher für das Vorliegen eines ionotropen mikrohetercgenen Gels als eines kristallähnlichen Körpers. Im Internum einiger Granula zeigen sich membranöse, vesikuläre und tubulöse Strukturen, die Ausdruck einer bestimmten Entwicklungsphase sein können.

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On the finestructure of the goose eosinophils

Summary Investigations of the fine structure of blood eosinophils of seven clinically healthy Toulouse-geese (Anser anser dom.) reveal striking differences between these cells and the heterophils.The eosinophils are characterized by an oval or nearly round profile. The nucleus contains chromatin mainly concentrated in dense chromocentres. Mitochondria turning into osmiophilic granules, vacuoles, multivesicular bodies, and microfilaments are prominent cytoplasmatic elements. Typical cytoplasmic organelles are numerous elliptic, oval, or round osmiophilic granules, frequently containing a nearly bacilliform internum. The internum possesses a substructure of parallel arranged dense bands, until now observed only in mammalian eosinophils. The ultrastructural and biochemical characteristics imply that the internum-containing granules of the eosinophils represent a state of an ionotropic, microheterogeneous gel and not that of a cristalloid body. The appearence of membraneous, vesicular, and tubular structures in the internum of several granules may be an equivalent of a certain phase of development.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Association of immune cells with neuroepithelial bodies in the lungs of neonatal dogs,cats and hamsters

A close topographical association between neuroepithelial bodies and immune cells is occasionally observed in the lungs of various neonatal mammalian species. The immune cells concerned are mast cells and neutrophil or eosinophil granulocytes. In the lungs of newborn puppies having undergone left lung autotransplantation, mast cells are particularly numerous in the airway mucosa of both right and left lungs and their association with neuroepithelial bodies is highly significant. Several of the substances known to be synthesized by the neuroepithelial bodies have a chemoattractive effect on immune cells. Thus, our observations indicate that intrapulmonary neuroepithelial bodies contribute to the local immune response.This investigation was supported by a grant from the Fonds voor Geneeskundig Wetenschappelijk Onderzoek (FGWO-NFWO), Belgium     

中华蟾蜍足垫部位对植物血凝素的反应及其组织学和血液学变化

为探究中华蟾蜍(Bufogargarizans)对植物血凝素(PHA-P)反应的最适浓度及足垫部位皮肤组织学和血液学的变化,比较了在足垫部位注射1.0、2.5和5.0g/L植物血凝素条件下的PHA-P反应,以及最适注射浓度条件下PHA-P反应高峰值时足垫部位皮肤组织和循环血中各型白细胞的百分比,血液中血清杀菌能力的变化。结果表明,中华蟾蜍对PHA-P的反应无性别差异(P 0.05),最适注射浓度为2.5 g/L,注射后12 h达反应峰值;注射PHA-P或生理盐水的左足与注射生理盐水或不注射任何物质的右足皮肤组织相比,各型白细胞的百分比均无显著差异(P0.05),但循环血中淋巴细胞的百分比升高(P 0.05),嗜酸性粒细胞的百分比下降(P 0.05),血清杀菌能力无明显变化(P 0.05)。2.5 g/L的注射浓度适用于中华蟾蜍,反应高峰值与足垫部位皮肤组织中各型白细胞的动态无关,但循环血中淋巴细胞被激活、嗜酸性粒细胞功能弱化,这可能是导致中华蟾蜍PHA-P反应的主要原因。     

Periostin: its role in asthma and its potential as a diagnostic or therapeutic target

Accumulating evidence shows that periostin, a matricellular protein, is involved in many fundamental biological processes such as cell proliferation, cell invasion, and angiogenesis. Changes in periostin expression are commonly detected in various cancers and pre-cancerous conditions, and periostin may be involved in regulating a diverse set of cancer cell activities that contribute to tumorigenesis, cancer progression, and metastasis. Periostin has also been shown to be involved in many aspects of allergic inflammation, such as eosinophil recruitment, airway remodeling, development of a Th2 phenotype, and increased expression of inflammatory mediators. In an in vivo model, bronchoalveolar lavage (BAL) fluid obtained from ovalbumin-challenged mice was found to contain significantly higher levels of periostin compared to BAL samples from control mice. To date, the molecular mechanisms involving periostin in relation to asthma in humans have not been fully elucidated. This review will focus on what is known about periostin and its role in the pathophysiological mechanisms that mediate asthma in order to evaluate the potential for periostin to serve as a biomarker and therapeutic target for the detection and treatment of asthma, respectively.     

Immunohistochemical Artifact for Nitrotyrosine in Eosinophils or Eosinophil Containing Tissue

Immunohistochemical artifacts for nitrotyrosine were investigated in eosinophils with regard to fixatives. Immunoreactivity for nitrotyrosine was revealed in separated eosinophils and in gastric mucosa fixed with periodate, lysine-paraformaldehyde (PLP). The increase in immunoreactivity by PLP was due to periodate itself, a component of PLP. Nitrotyrosine formed by peroxidase using NO 2 &#109 and H 2 O 2 or by peroxynitrite was not completely inhibited by 100 mM dithionite but the immunoreactivity for nitrotyrosine antibodies by PLP was completely inhibited by 5.7 mM dithionite. Although untreated eosinophils or ovalbumin (OVA) did not show protein tyrosine nitration in a standard Western blot, the treatment of the blotted membrane with PLP increased the reactivities of proteins from eosinophils with anti-nitrotyrosine antibodies. The increase in immunoreactivity of OVA with anti-nitrotyrosine antibodies by PLP did not change with pre-treatment with dithionite but was abolished by treatment with dithionite after PLP fixation. In HPLC assays, periodate did not generate nitrotyrosine from l -tyrosine and aminotyrosine. These results suggest that the treatment of eosinophils or eosinophil-containing tissues with PLP fixative augments the immunoreactivity of nitrotyrosine antibodies with eosinophils due to the formation of epitopes similar to nitrotyrosine by an oxidation reaction of periodate, which evokes an artifact in nitrotyrosine immunohistochemistry.     

稳定期慢性阻塞性肺病患者血嗜酸性粒细胞水平与炎症表型相关性研究

摘要 目的：探究稳定期慢性阻塞性肺病患者血嗜酸性粒细胞水平与炎症表型相关性研究。方法：选择淮安市第一人民医院（我院）2017年5月至2020年3月诊治的稳定期慢性阻塞性肺病患者280例,按照外周血嗜酸性粒细胞计数分为3组,嗜酸性粒细胞＜100为A组86例,嗜酸性粒细胞100~300为B组102例,嗜酸性粒细胞＞300为C组92例。同时纳入同期进行体检的健康者100例为对照组。结果：A组、B组和C组的FeNO显著高于对照组,且C组>B组>A组（P<0.05）;FEV₁占预测值和FEV₁/FVC 显著低于对照组（P<0.05）;A组、B组和C组的FEV₁占预测值和FEV1/FVC对比无差异（P>0.05）。A组、B组和C组的CRP、IL-6、IL-8和TNF-α显著高于对照组（P<0.05）,且C组>B组>A组（P>0.05）。经相关性分析发现,A组和B组患者的嗜酸性粒细胞计数与FeNO、CRP、IL-6、IL-8和TNF-α、FEV₁占预测值和FEV₁/FVC无明显相关性（P>0.05）。C组的嗜酸性粒细胞计数与FeNO、CRP、IL-6、IL-8和TNF-α呈正相关（P<0.05）,与FEV₁/FVC%呈负相关（P<0.05）;与FEV₁占预测值无显著相关性（P>0.05）。结论：稳定期慢性阻塞性肺病患者外周血嗜酸性粒细胞计数与FeNO水平呈正相关,二者能够反映稳定期慢性阻塞性肺病患者气道的炎症反应,对于稳定期慢性阻塞性肺病患者炎症的评估、病情的程度及后期的治疗具有一定的临床指导意义。