Identification of 45 kD antigen in immune complexes of patients of allergic bronchopulmonary aspergillosis

A glycoprotein antigen with an apparent mw of 45 kD was observed to be predominant in the circulating immune complexes isolated from patients of allergic bronchopulmonary aspergillosis as well as in the immune complexes prepared in vitro. Further characterisation of 45 kD antigen with trypsin showed four immunologically active peptides with mw 43, 36, 33 and 16 kD. Carbohydrate characterization using various lectins (Maackia amurensis agglutinin, Sambucus nigra agglutinin, Peanut agglutinin, Galanthus nivalis agglutinin and Datura stramonium agglutinin) showed presence of both N-linked and O-linked sugar moieties such as mannose, glucose, galactose and N-acetyl glucosamine. Predominant presence of 45 kD antigen in immune complexes, recognition of 45 kD by monoclonal antibodies raised against glycoprotein rich fraction of A. fumigatus and presence of elastinolytic protease activity indicate that 45 kD antigen is probably a potent virulence factor and may be contributing to the pathogenesis of ABPA by its biological as well as immunological activities. (Mol Cell Biochem 166: 111-116, 1997)     

HLA‐DRB1 and HLA–DQB1 genes on susceptibility to and protection from allergic bronchopulmonary aspergillosis in patients with cystic fibrosis

Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity pulmonary disease that affects both patients with cystic fibrosis (CF) and those with asthma. HLA‐DRB1 alleles have previously been associated with ABPA–CF susceptibility; however, HLA‐DQB1 allele associations have not been clearly established. The aim of the present study was to investigate HLA class II associations in patients with ABPA–CF and determine their roles in susceptibility or protection. Patients with ABPA–CF, patients with CF without ABPA, patients with asthma without ABPA (AST), and healthy controls were included in this study. DNA was extracted by automatic extractor. HLA‐DRB1 and ‐DQB1 genotyping was performed by the Luminex PCR‐SSOP method (One Lambda, Canoga Park, CA, USA). Allele specific PCR‐SSP was also performed by high‐resolution analysis (One Lambda). Statistical analysis was performed with SSPS and Arlequin software. Both HLA‐DRB1*5:01 and ‐DRB1*11:04 alleles occurred with greater frequency in patients with ABPA–CF than in those with AST and CF and control subjects, corroborating previously published data. On the other hand, analysis of haplotypes revealed that almost all patients with ABPA–CF lacking DRB1*15:01 or DRB1*11:04 carry either DRB1*04, DRB1*11:01, or DRB1*07:01 alleles. In the HLA‐DQB1 region, the HLA‐DQB1*06:02 allele occurred more frequently in patients with ABPA–CF than in those with AST and CF and healthy controls, whereas HLA‐DQB1*02:01 occurred less frequently in patients with ABPA–CF. These data confirm that there is a correlation between HLA‐DRB1*15:01, –DRB1*11:04, DRB1*11:01, –DRB1*04 and –DRB1 * 07:01 alleles and ABPA–CF susceptibility and suggest that HLA‐DQB1*02:01 is an ABPA–CF resistance allele.     

变态反应性支气管肺曲霉病临床特点探讨

目的分析变态反应性支气管肺曲霉病(ABPA)的临床特征,以期早期诊断,合理治疗及预防。方法回顾性分析浙江大学医学院附属第一医院2009年6月至2011年7月收治的13例ABPA患者的临床资料,所有患者均依照目前通用的ABPA诊断标准进行诊断,分析其临床表现,影像学特征,治疗方案及疗效等。结果 ABPA好发于中老年患者,且都好发支气管扩张及哮喘患者。13例ABPA患者均有咳嗽,咳痰,8例气喘,4例发热,9例肺部可闻及哮鸣音。13例ABPA患者中有12例CT检查发现囊状或静脉曲张样中心性支气管扩张和支气管粘液栓形成及树芽征为主要特点的小叶中心结节形成。总IgE升高12例,嗜酸粒细胞绝对值升高6例,烟曲菌特异性IgE升高10例。12例接受激素及抗真菌药物治疗,均达到临床症状好转。结论 ABPA临床表现缺乏特异性,为提高对ABPA的认识,对可疑患者早期进行外周血嗜酸性粒细胞数、血清总IgE、烟曲霉菌特异性IgE、肺部CT等检查以确诊,糖皮质激素及伊曲康唑治疗ABPA能取得良好的疗效。     

变应性支气管肺曲霉病的CT表现——附17例分析

目的探讨变应性支气管肺曲霉病(ABPA)的CT特点,进一步提高对该病的认识。方法回顾性分析17例AB-PA的CT表现,所有的患者均按目前的ABPA诊断标准进行诊断。结果 17例患者均有中心性支气管扩张征象,13例并支气管黏液栓形成,为分支状或挤牙膏状或长管状"Y"形,7例以树芽征为主要表现的小叶中心结节形成,9例有斑片状浸润影,2例随访见中心性支气管扩张有游走性。结论变应性支气管肺曲霉病(ABPA)的CT表现相对有特征性,以中心性支气管扩张为主,常伴有较高密度的支气管腔内黏液栓形成,结合临床一般能做出诊断。     

Animal models of skin regeneration

Cutaneous injury in the majority of vertebrate animals results in the formation of a scar in the post-injured area. Scar tissues, although beneficial for maintaining integrity of the post-wounded region often interferes with full recovery of injured tissues. The goal of wound-healing studies is to identify mechanisms to redirect reparative pathways from debilitating scar formation to regenerative pathways that lead to normal functionality. To perform such studies models of regeneration, which are rare in mammals, are required. In this review we discussed skin regenerative capabilities present in lower vertebrates and in models of skin scar-free healing in mammals, e.g. mammalian fetuses. However, we especially focused on the attributes of two unusual models of skin scar-free healing capabilities that occur in adult mammals, that is, those associated with nude, FOXN1-deficient mice and in wild-type African spiny mice.     

Diminished levels of nasal S100A7 (psoriasin) in seasonal allergic rhinitis: an effect mediated by Th2 cytokines

Background

S100A7 is an antimicrobial peptide involved in several inflammatory diseases. The aim of the present study was to explore the expression and regulation of S100A7 in seasonal allergic rhinitis (SAR).

Methods

Nasal lavage (NAL) fluid was obtained from healthy controls before and after lipopolysaccharide (LPS) provocation, from SAR patients before and after allergen challenge, and from SAR patients having completed allergen-specific immunotherapy (ASIT). Nasal biopsies, nasal epithelial cells and blood were acquired from healthy donors. The airway epithelial cell line FaDu was used for in vitro experiments. Real-time RT-PCR and immunohistochemistry were used to determine S100A7 expression in nasal tissue and cells. Release of S100A7 in NAL and culture supernatants was measured by ELISA. The function of recombinant S100A7 was explored in epithelial cells, neutrophils and peripheral blood mononuclear cells (PBMC).

Results

Nasal administration of LPS induced S100A7 release in healthy non-allergic subjects. The level of S100A7 was lower in NAL from SAR patients than from healthy controls, and it was further reduced in the SAR group 6 h post allergen provocation. In contrast, ASIT patients displayed higher levels after completed treatment. S100A7 was expressed in the nasal epithelium and in glands, and it was secreted by cultured epithelial cells. Stimulation with IL-4 and histamine repressed the epithelial S100A7 release. Further, recombinant S100A7 induced activation of neutrophils and PBMC.

Conclusions

The present study shows an epithelial expression and excretion of S100A7 in the nose after microbial stimulation. The levels are diminished in rhinitis patients and in the presence of an allergic cytokine milieu, suggesting that the antimicrobial defense is compromised in patients with SAR.     

Animal models for respiratory chain disease

Elucidation of the pathogenesis in respiratory chain diseases is of great importance for developing specific treatments. The limitations inherent to the use of patient material make studies of human tissues often difficult and the mouse has therefore emerged as a suitable model organism for studies of respiratory chain diseases. In this review, we present an overview of the field and discuss in depth a few examples of animal models reproducing pathology of human disease with primary and secondary respiratory chain involvement.     

Animal disease models generated by genetic engineering of polyamine metabolism

The polyamines putrescine, spermidine and spermine are natural components of all living cells. Although their exact cellular functions are still largely unknown, a constant supply of these compounds is required for mammalian cell proliferation to occur. Studies with animals displaying genetically altered polyamine metabolism have shown that polyamines are intimately involved in the development of diverse tumors, putrescine apparently has specific role in skin physiology and neuroprotection and the higher polyamines spermidine and spermine are required for the maintenance of pancreatic integrity and liver regeneration. In the absence of ongoing polyamine biosynthesis, murine embryogenesis does not proceed beyond the blastocyst stage. The last years have also witnessed the appearance of the first reports linking genetically altered polyamine metabolism to human diseases.     

TNF-alpha promotes LPA1- and LPA3-mediated recruitment of leukocytes in vivo through CXCR2 ligand chemokines

Lysophosphatidic acid (LPA) is a bioactive lysophospholipid present in low concentrations in serum and biological fluids but in high concentrations at sites of inflammation. LPA evokes a variety of cellular responses via binding to and activation of its specific G protein-coupled receptors (GPCR), namely LPA(1-6). Even though LPA is a chemoattractant for inflammatory cells in vitro, such a role for LPA in vivo remains largely unexplored. In the present study, we used the murine air pouch model to study LPA-mediated leukocyte recruitment in vivo using selective LPA receptor agonist/antagonist and LPA(3)-deficient mice. We report that 1) LPA injection into the air pouch induced leukocyte recruitment and that both LPA(1) and LPA(3) were involved in this process; 2) LPA stimulated the release of the pro-inflammatory chemokines keratinocyte-derived chemokine (KC) and interferon-inducible protein-10 (IP-10) in the air pouch; 3) tumor necrosis factor-α (TNF-α) injected into the air pouch prior to LPA strongly potentiated LPA-mediated secretion of cytokines/chemokines, including KC, IL-6, and IP-10, which preceded the enhanced leukocyte influx; and 4) blocking CXCR2 significantly reduced leukocyte infiltration. We suggest that LPA, via LPA(1) and LPA(3) receptors, may play a significant role in inducing and/or sustaining the massive infiltration of leukocytes during inflammation.     

Mechanisms of drug-induced allergic contact dermatitis

Allergic contact dermatitis is induced by a wide variety of drugs that trigger specific immune responses following topical exposure. Identified chemical stuctures involved in such reactions include the mercuric and thiosalicylic acid groups of thimerosal, the diphenylketone group of the anti-inflammatory drug ketoprofen, the amide or ester structure of local anesthetics, and the side-chain and thiazolidine ring of -lactams. The T cell responses to such compounds involve CD4+ and CD8+ + T lymphocytes and also CD4–/CD8– + T cells. Although "T helper 2" cytokine production by drug-specific human T cells from patients with allergic contact dermatitis has been described, T helper 1-like and T cytotoxic 1-like responses clearly play key roles in this cutaneous reaction.     

Tryptophan residue is essential for immunoreactivity of a diagnostically relevant peptide epitope of A. fumigatus

The role of tryptophan (Trp¹⁷) in immunoreactivity of P1, the diagnostically relevant peptide from a major allergen/antigen of Aspergillus fumigatus, was evaluated by chemically modifying tryptophanyl residue of P1. In BIAcore kinetic studies, unmodified P1 showed a 100-fold higher binding with ABPA (Allergic Bronchopulmonary Aspergillosis) patients’ IgG [K_D (equilibrium dissociation constant) = 2.74 e⁻⁸ ± 0.13 M] than the controls’ IgG (K_D = 2.97 e⁻⁶± 0.14 M), whereas chemically-modified P1 showed similar binding [K_D patients’ IgG = 3.25 e⁻⁷± 0.16 M, K_D controls’ IgG = 3.86 e⁻⁷± 0.19 M] indicating loss of specific immunoreactivity of P1 on tryptophan modification. Modified P1 showed loss of specific binding to IgE and IgG antibodies of ABPA patients in ELISA (Enzyme-Linked Immunosorbent Assay). The study infers that tryptophan residue (Trp¹⁷) is essential for immunoreactivity of P1.     

Local injections of OK432 can help the infiltration of adoptively transferred CD8+ T cells into the tumor sites and synergistically induce the local production of Th1-type cytokines and CXC3 chemokines

The effect of local injections with streptococcal preparation OK432 on the antitumor effect induced by adoptive immunotherapy (AIT) was investigated. Draining lymph node cells on day 14 after B7-P815 inoculation were used for AIT after in vitro stimulation. AIT on days 7 and 10 showed no effect on the growth of s.c. established P815 mastocytoma, but local injections with OK432 into the tumor sites on days 3, 6 and 9 resulted in a moderate antitumor effect. On the other hand, the combination therapy significantly suppressed tumor growth, and the tumor-bearing mice survived longer than those receiving only one of the treatment modalities. The significant infiltration of CD4⁺ or CD8⁺ T cells and multiple necrosis in the tumor sites were observed only when the tumor-bearing mice were treated with the combination therapy. In addition, a transfer experiment using labeled effector cells revealed these infiltrated CD8⁺ T cells and CD4⁺ T cells to be derived from the donor and the host respectively. More importantly, the combination therapy clearly led to higher expression of the mRNA for Th1-type cytokines and CXC3 chemokines in the tumor sites than resulted from each of the treatment modalities alone. Collectively, these results indicate that local injections with OK432 can help the infiltration of adoptively transferred CD8⁺ T cells into the tumor sites and synergistically induce the local production of the Th1-type cytokines and CXC3 chemokines. Received: 3 April 2000 / Accepted: 5 May 2000     

Stepwise oral immunotherapy for 10 days in an egg-white allergy mouse model did not ameliorate the severity of allergy but induced the production of allergen-specific IgA

ABSTRACT

We examined whether the stepwise oral immunotherapy (OIT) for 10 days ameliorates the severity of allergy and the biomarkers in an allergy mouse model. The OIT could not protect anaphylaxis symptoms after allergen challenges but promote the production of antibodies, especially allergen-specific IgA. It was suggested that this OIT influenced the function of immuno response against the allergen.

Abbreviations: EW: egg white; IFC: intraperitoneal food challenge; IFN-γ: interferon-γ; IL: interleukin; OVA: ovalbumin; OM: ovomucoid; OFC: oral food challenge; OIT: oral immunotherapy.     

Animal models of mucosal Candida infection

Rodent models of oral, vaginal and gastrointestinal Candida infection are described and discussed in terms of their scientific merits. The common feature of all experimental mucosal Candida infections is the need for some level of host immunocompromise or exogenous treatment to ensure quantitatively reproducible disease. A growing literature describes the contributions of such candidiasis models to our understanding of certain aspects of fungal virulence and host response to mucosal Candida albicans challenge. Evidence to date shows that T-lymphocyte responses dominate host immune defences to oral and gastrointestinal challenge, while other, highly compartmentalized responses defend vaginal surfaces. By contrast the study of C. albicans virulence factors in mucosal infection models has only begun to unravel the complex of attributes required to define the difference between strongly and weakly muco-invasive strains.     

Distinct inflammatory phenotypes of microglia and monocyte‐derived macrophages in Alzheimer's disease models: effects of aging and amyloid pathology

Alzheimer's disease (AD) is a neurodegenerative disease characterized by formation of amyloid‐β (Aβ) plaques, activated microglia, and neuronal cell death leading to progressive dementia. Recent data indicate that microglia and monocyte‐derived macrophages (MDM) are key players in the initiation and progression of AD, yet their respective roles remain to be clarified. As AD occurs mostly in the elderly and aging impairs myeloid functions, we addressed the inflammatory profile of microglia and MDM during aging in TgAPP/PS1 and TgAPP/PS1dE9, two transgenic AD mouse models, compared to WT littermates. We only found MDM infiltration in very aged mice. We determined that MDM highly expressed activation markers at basal state. In contrast, microglia exhibited an activated phenotype only with normal aging and Aβ pathology. Our study showed that CD14 and CD36, two receptors involved in phagocytosis, were upregulated during Aβ pathogenesis. Moreover, we observed, at the protein levels in AD models, higher production of pro‐inflammatory mediators: IL‐1β, p40, iNOS, CCL‐3, CCL‐4, and CXCL‐1. Taken together, our data indicate that microglia and MDM display distinct phenotypes in AD models and highlight the specific effects of normal aging vs Aβ peptides on inflammatory processes that occur during the disease progression. These precise phenotypes of different subpopulations of myeloid cells in normal and pathologic conditions may allow the design of pertinent therapeutic strategy for AD.     

In vitro models for analysis of the hepatitis C virus life cycle

Chronic hepatitis C virus (HCV) infection affects approximately 170 million people worldwide. HCV infection is a major global health problem as it can be complicated with liver cirrhosis and hepatocellular carcinoma. So far, there is no vaccine available and the non-specific, interferon (IFN)-based treatments now in use have significant side-effects and are frequently ineffective, as only approximately 50% of treated patients with genotypes 1 and 4 demonstrate HCV clearance. The lack of suitable in vitro and in vivo models for the analysis of HCV infection has hampered elucidation of the HCV life cycle and the development of both protective and therapeutic strategies against HCV infection. The present review focuses on the progress made towards the establishment of such models.     

裸鼠呼吸道合胞病毒感染的动物模型

呼吸道合胞病毒(RSV)是全世界婴幼儿下呼吸道感染的首位病毒病原体,免疫缺陷个体容易发生严重感染,目前尚无理想RSV感染动物模型用于研究.我们用细胞免疫缺陷裸鼠感染RSV,旨在建立理想的动物模型,为RSV感染的防治研究奠定基础.裸鼠滴鼻感染RSV后肺组织分离到病毒,直接免疫荧光检测到支气管肺泡灌洗液RSV抗原阳性,空斑形成实验检测肺组织病毒滴度在感染后第3天达高峰,并持续到第9天仍能检测到病毒.免疫组化检测RSV抗原主要分布在细支气管、毛细支气管和肺泡上皮细胞胞浆内.肺组织病理学显示RSV感染导致裸鼠淋巴细胞浸润为主的肺间质性炎症,电镜分析超微结构可见到细胞内病毒颗粒和气血屏障的破坏.支气管肺泡灌洗液白细胞计数显示裸鼠RSV感染炎症高峰在感染后第9天.裸鼠RSV感染的病毒复制和病理改变特点与人相似,病毒持续高水平复制,是客观而实用的评价抗RSV制剂效果的小鼠模型.     

裸鼠呼吸道合胞病毒感染的动物模型

呼吸道合胞病毒(RSV)是全世界婴幼儿下呼吸道感染的首位病毒病原体,免疫缺陷个体容易发生严重感染,目前尚无理想RSV感染动物模型用于研究。我们用细胞免疫缺陷裸鼠感染RSV,旨在建立理想的动物模型,为RSV感染的防治研究奠定基础。裸鼠滴鼻感染RSV后肺组织分离到病毒,直接免疫荧光检测到支气管肺泡灌洗液RSV抗原阳性,空斑形成实验检测肺组织病毒滴度在感染后第3天达高峰,并持续到第9天仍能检测到病毒。免疫组化检测RSV抗原主要分布在细支气管、毛细支气管和肺泡上皮细胞胞浆内。肺组织病理学显示RSV感染导致裸鼠淋巴细胞浸润为主的肺间质性炎症,电镜分析超微结构可见到细胞内病毒颗粒和气血屏障的破坏。支气管肺泡灌洗液白细胞计数显示裸鼠RSV感染炎症高峰在感染后第9天。裸鼠RSV感染的病毒复制和病理改变特点与人相似,病毒持续高水平复制,是客观而实用的评价抗RSV制剂效果的小鼠模型。