血红蛋白β亚基:一种多效性抗病毒天然免疫调节因子

血红蛋白是动物体内最重要的呼吸蛋白之一。在脊椎动物的红细胞内含量尤其丰富,可与氧分子呈可逆性结合。随着对血红蛋白研究的不断深入,相继有研究报道血红蛋白还具有储存能量、维持渗透压、抗菌和发挥类酚氧化酶活性等多种功能,现已成为研究蛋白质多重生物学功能的理想模式分子之一。近年来的研究发现,血红蛋白在抵抗病原体入侵、调控机体天然免疫应答过程中也发挥了重要作用,但其相关机制仍有待进一步阐明。     

Identifying innate immune pathways of the chicken may lead to new antiviral therapies

Fractalkine, also known as CX(3)CL1, is a unique chemokine that mediates inflammatory responses and is involved in the pathogenesis of several inflammatory disorders, including inflammatory bowel disease (IBD) in humans. In this study, we isolated cDNAs encoding canine fractalkine and its receptor CX(3)CR1, and assessed the biological activity of these molecules. The deduced amino acid sequence of the canine fractalkine cDNA showed 66% and 57% identity to human and mouse homologs, respectively. The N-terminal chemokine domain of the canine fractalkine showed 68% and 65% identity to human and mouse counterparts, respectively. The canine CX(3)CR1 amino acid sequence showed close homology to its human (83% identity) and mouse (81% identity) counterparts. Fractalkine and CX(3)CR1 mRNA were detected in all tissues in this study. Relatively higher expression levels of fractalkine mRNA were observed in the brain, medulla spinalis, small intestine, and mesenteric lymph nodes (MLNs), whereas higher expression levels of CX(3)CR1 mRNA were observed in the medulla spinalis, brain, liver, small intestine, and MLNs. The cross-reactivities of anti-human fractalkine antibody and anti-rat CX(3)CR1 antibody to canine proteins were confirmed using recombinant canine fractalkine and a cell line overexpressing canine CX(3)CR1, respectively. A transwell chemotaxis assay showed that the recombinant canine fractalkine induced migration in canine lymphoid cells expressing CX(3)CR1. The present study will be useful in understanding the canine immune system and the immunopathogenesis of canine inflammatory diseases.     

Glandular kallikrein in the innate immune system of Atlantic salmon (Salmo salar)

Glandular Kallikrein is a serine-protease with trypsin-like activity and is able to generate bioactive peptides from inactive precursors. We have evaluated the presence of this protease in the different organs of the Atlantic salmon (Salmo salar). The results clearly indicate that GK and PRL are generated in the same pituitary cells based on a co-localization by confocal microscopy. Based on probed cross-reactivity between C. striata and C. carpio glandular anti-GK antibodies, we used a homologous antibody to detect the presence of GK in several salmon tissues. We have evaluated the GK expression in healthy and defied fish. P. salmonis and V. ordalii. The GK immunoreaction in organs such as leukocytes, gills and skin is considerably increased in defied fish compared to healthy fish. This increase was present in the cells of the excretory kidney and in the intercellular tissue, where the development of hematopoietic and lymphocytic lines in fish take place. One of the most interesting organs to study was the skin, bearing in mind that this is a primary barrier to all pathogens. The skin of the defied fish exhibited an increase in immunoreactivity for glandular kallikrein similar to the protease found in mucus. An immunoreactive tissue kallikrein-like protein was identified and partially separated by perfusion chromatography. Enzymatic activity of salmon muscle prokallikrein was determined before and after trypsin activation. Kallikrein activity was characterized with respect to their ability to cleave the chromogenic leaving group, p-nitroanilide, from the peptidyl kallikrein and trypsin substrate. These findings constitute a important contribution to reveal the role of kallikrein in the innate immune system of fish.     

The innate immune response of the bovine mammary gland to bacterial infection

Intra-mammary (IM) bacterial infection in cattle can result in clinical outcomes that range from being acute and life-threatening to those that are chronic and sub-clinical. The typical bacteria involved in IM bacterial infections activate the mammary immune system in different ways which can influence the severity of the outcome. A clear understanding of the mechanisms that activate and regulate this response is central to the development of effective preventative and treatment regimes. This review focuses on the different immune responses of the bovine mammary gland to common mastitis-causing pathogens. There is special emphasis on comparing the responses to Escherichia coli and Staphylococcus aureus infections, as these are typically associated, respectively, with acute/severe and chronic/sub-clinical forms of the disease.     

Prolonged suppression of the innate immune system in the horse following an 80 km endurance race

REASONS FOR PERFORMING STUDY: An increased susceptibility to bacterial and viral infections of the respiratory tract, which results in a loss of performance, has been reported in racehorses. Much research has focused on the influence of high-intensity exercise of a short duration on immune system function in horses, but scant attention has been given to prolonged endurance exercise as an immune modulator. OBJECTIVES: The objective of this study was to evaluate the effect of an 80 km endurance race on the monocyte and neutrophil oxidative burst, serum cortisol, glutamine and plasma glucose concentrations in 8 endurance-trained horses (mean +/- s.d. age 9.4 +/- 2.2 years). METHODS: Blood samples were drawn from the horses prior to and following an 80 km ride. RESULTS: Mean time for completion of the 80 km race was 306 +/- 40 mins. Immediately post race mean serum cortisol concentration, blood monocyte and neutrophil counts were higher and blood lymphocyte counts and plasma glucose concentration were lower compared with prerace values (P < 0.05). Neutrophil and monocyte oxidative burst activity decreased following the race and had not regained prerace values after 3 days of rest (P < 0.05). CONCLUSIONS: The present study indicates that long duration exercise in horses has a negative impact on the function of the innate immune system that lasts several days post race. Precise mechanisms instigating the fall in innate immune system function are unclear and multifactorial, but may be attributed, at least in part, to a high serum cortisol response during very prolonged exercise. POTENTIAL CLINICAL RELEVANCE: A prolonged bout of exercise results in a long-term suppression of the innate immune system function in horses which may, in part, account for the observed increase of infectious episodes in horses during training.     

抗病毒药物作用机理的分子水平研究

真核细胞内病毒复制周期包括吸附、穿入、脱蛋白衣壳、生物合成以及装配释放等阶段。抗病毒药物干扰病毒生长周期的少数程序，从而阻断了病毒核酸的复制，起到了抗病毒的作用。本文将在分子水平上对抗病毒药物如何阻断或干扰不同病毒复制周期作一综述。     

Effects of tramadol and o‐desmethyltramadol on canine innate immune system function

ObjectiveTramadol is a commonly used opioid analgesic in dogs, particularly in dogs with a compromised immune system. An opioid may be selected for its immunomodulatory effects. Consequently, the objective of this study was to investigate the effects of tramadol on immune system function by evaluating the effect of tramadol and o-desmethyltramadol (M1) on the function of canine leukocytes in vitro. The hypothesis was that tramadol and M1 would not alter polymorphonuclear leukocyte (PMN) phagocytosis, PMN oxidative burst, or stimulated leukocyte cytokine production capacity of tumor necrosis factor (TNF)-a, interleukin (IL)-6, and IL-10.Study designIn vitro pharmacodynamic study.AnimalsSix healthy dogs.MethodsBlood from six dogs was obtained and incubated with various concentrations of tramadol and M1. Phagocytosis and oxidative burst were assessed using flow cytometry, and lipopolysaccharide (LPS), lipoteichoic acid (LTA) and peptidoglycan (PG)-stimulated leukocyte production of TNF, IL-6, and IL-10 were measured using a canine specific multiplex assay.ResultsNo differences were detected in phagocytosis or oxidative burst with any drug concentration. Tramadol did not alter leukocyte cytokine production, however, M1 significantly blunted IL-10 production.ConclusionsTramadol and its metabolite M1 were sparing to PMN phagocytosis and oxidative burst in dogs in vitro. Tramadol did not alter leukocyte cytokine production, however, M1 blunted IL-10 production at clinically achievable concentrations suggesting that M1 may promote a proinflammatory shift.Clinical relevanceThese data suggest that tramadol has minimal effect on phagocytosis and oxidative burst, and may promote a proinflammatory shift. Therefore, tramadol may be an ideal opioid analgesic in dogs at high risk of infection. Further investigation in vivo is warranted.     

脂多糖诱导奶牛乳腺上皮细胞先天性免疫反应

采取荷斯坦奶牛乳腺,进行体外分离培养,并纯化细胞。用不同质量浓度(0、1、10、100mg/L)的脂多糖刺激乳腺上皮细胞,采用MTT法检测脂多糖对细胞增殖的影响,半定量PCR检测10mg/L的LPS对乳腺上皮细胞TLR4、TLR2、CD14、MD-2四个基因在不同时间(0、2、6h)mRNA表达水平的差异。结果表明,高剂量(100mg/L)的LPS对乳腺上皮细胞的增殖产生明显影响;LPS刺激乳腺上皮细胞后,导致TLR4、CD14、MD-2mRNA表达迅速升高,而TLR2mRNA弱表达。说明TLR4、CD14、MD-2参与LPS的识别,同时也说明脂多糖刺激乳腺上皮细胞后,乳腺上皮细胞能够产生先天性免疫反应。     

The molecular basis for bidirectional communication between the immune and neuroendocrine systems

IL-2 has analgesic effects in both central and peripheral nervous systems. There are two distinct domains in IL-2 molecule mediating immunologic and analgesic activity, respectively. The analgesic domain of IL-2 may be composed of the 44th Phe, 45th Tyr, 107th Tyr, and 117th Phe residues that are located closely in the tertiary structure of IL-2. The analgesic activity may be mediated through the analgesic domain interaction with opioid receptor.In addition to peptides, cytokines may directly bind to peptide receptors, other than their specific cytokine receptors themselves. Conversely, peptides may also interact with cytokine receptors. Thus, peptide neurotransmitters and hormones may serve as endogenous regulators of the immune system, and cytokines may also serve as neurotransmitters. Multiple actions might be mediated by interactions between distinct domains of bioactive molecules with different receptors.     

BCG vaccination of neonatal calves: potential roles for innate immune cells in the induction of protective immunity

Bovine tuberculosis is a disease of increasing incidence in the UK causing major economic losses and with significant impact on bovine and, potentially human health: the causative agent Mycobacterium bovis is a zoonotic pathogen. Neonatal vaccination with the attenuated M. bovis Bacille Calmette Guerin (BCG) vaccine confers a significant degree of protection in cattle, and is a widely used control strategy for human TB. The adaptive immune system is relatively immature in neonates and increased numbers of innate effector cells present in young animals and human infants may compensate for this, enabling effective immune responses to vaccination. Natural killer cells and subsets of γδ TCR+ T lymphocytes secrete high levels of interferon gamma and can interact with antigen presenting cells to promote both innate and adaptive immune responses. These cell populations may be pivotal in determining immune bias following neonatal vaccination with BCG.     

Fenbendazole and its effect on the immune system of the sheep

Ten parasite-free 6-month-old lambs were drenched on days 0 and 28 with fenbendazole and 1 day after each drench were injected with human erythrocytes and ovalbumin. Ten other lambs injected with the antigens were not drenched with anthelmintic and served as controls. Lymphocytes from the fenbendazole-drenched lambs collected 3 days after the first antigen injections and cultured in vitro in RPM11640 plus 5% foetal calf serum, and lymphocytes collected at 3 and 7 days and cultured in RPM1 plus 50% autologous serum, had decreased blastogenic activity compared with lymphocytes from control lambs. Similiarly, decreased blastogenesis was observed with lymphocytes collected 7 days after the second antigen injections from drenched lambs and cultured in 50% autologous serum containing concanavalin A. In contrast, increased blastogenesis was seen with lymphocytes collected 14 days after the second antigen injections from the drenched lambs and cultured in 50% autologous serum containing phytohaemagglutinin. Similar antibody responses were seen for the drenched and control lambs in response to the injections of both antigens except that, after the second injection, there was a significant reduction in antibody response to human erythrocytes in the fenbendazole-treated lambs. Decreased serum complement levels were seen particularly 3 and 7 days after the second antigen injections in drenched lambs. These serum samples had increased conglutinin activity. At the end of the experiment, the fenbendazole-drenched lambs were significantly heavier than the control lambs. However, this did not appear to be related to any effects of fenbendazole on levels of growth promoting hormones.     

Fenbendazole and its effect on the immune system of the sheep

Ten parasite-free 6-month-old lambs were drenched on days 0 and 28 with fenbendazole and 1 day after each drench were injected with human erythrocytes and ovalbumin. Ten other lambs injected with the antigens were not drenched with anthelmintic and served as controls. Lymphocytes from the fenbendazole-drenched lambs collected 3 days after the first antigen injections and cultured in vitro in RPM1 1640 plus 5% foetal calf serum, and lymphocytes collected at 3 and 7 days and cultured in RPM1 plus 50% autologous serum, had decreased blastogenic activity compared with lymphocytes from control lambs. Similarly, decreased blastogenesis was observed with lymphocytes collected 7 days after the second antigen injections from drenched lambs and cultured in 50% autologous serum containing concanavalin A. In contrast, increased blastogenesis was seen with lymphocytes collected 14 days after the second antigen injections from the drenched lambs and cultured in 50% autologous serum containing phytohaemagglutinin. Similar antibody responses were seen for the drenched and control lambs in response to the injections of both antigens except that, after the second injection, there was a significant reduction in antibody response to human erythrocytes in the fenbendazole-treated lambs. Decreased serum complement levels were seen particularly 3 and 7 days after the second antigen injections in drenched lambs. These serum samples had increased conglutinin activity. At the end of the experiment, the fenbendazole-drenched lambs were significantly heavier than the control lambs. However, this did not appear to be related to any effects of fenbendazole on levels of growth promoting hormones.     

The innate immune response of equine bronchial epithelial cells is altered by training

Respiratory diseases, including inflammatory airway disease (IAD), viral and bacterial infections, are common problems in exercising horses. The airway epithelium constitutes a major physical barrier against airborne infections and plays an essential role in the lung innate immune response mainly through toll-like receptor (TLR) activation. The aim of this study was to develop a model for the culture of equine bronchial epithelial cells (EBEC) in vitro and to explore EBEC innate immune responses in trained horses. Bronchial epithelial biopsies were taken from 6 adult horses during lower airway endoscopy. EBEC were grown in vitro by an explant method. The innate immune response of EBEC was evaluated in vitro by treatment with TLR ligands. TLR3 is the most strongly expressed TLR at the mRNA level in EBEC and stimulation of EBEC with Poly(I:C), an analog of viral dsRNA, triggers a strong secretion of IFN-β, TNF-α, IL-6 and CXCL8. We further evaluated the EBEC innate immune response in horses that underwent a 4-month-training program. While training had no effect on TLR mRNA expression in EBEC as well as in bronchial biopsies, it increased the production of IFN-β after stimulation with a TLR3 ligand and decreased the secretion of TNF-α and IL-6 after stimulation with a TLR2 and TLR3 ligand. These findings may be implicated in the increased risk for viral and bacterial infections observed in sport horses. Altogether, we report a successful model for the culture of EBEC that can be applied to the investigation of pathophysiologic conditions in longitudinal studies.     

不同品种绵羊和山羊主要先天性免疫指标的比较

对4个山羊品种和4个绵羊品种的7项主要先天性免疫学指标进行了检测并比较。对动物进行颈静脉采血,制备血清,用ELISA试剂盒测定IL-1、IL-2、IL-6、IL-18、IFN-γ、NK细胞、血清溶菌酶。利用生物统计学的方法和原理,以品种和品种来源地为应变量对检测结果进行比较分析。结果表明,山羊当地品种的NK和LYS高于引进品种（P〈0.05）;绵羊引进品种的IL-1、IL-2、IL-6、IL-18、IFN-γ、NK、LYS均明显高于地方品种（P〈0.01）。在不同品种之间,莱芜黑山羊NK和LYS水平高于崂山奶山羊、波尔山羊和鲁波山羊,萨福克羊的IL-1、IL-6、IL-18、IFN-γ、LYS水平最高且差异极显著（P〈0.01）。小尾寒羊IL-18、IFN-γ水平最低且差异极显著（P〈0.01）。说明不同种群绵羊、山羊的先天性免疫水平存在显著差异,为以后抗病育种提供合理的试验数据。     

The avian lung-associated immune system: a review

The lung is a major target organ for numerous viral and bacterial diseases of poultry. To control this constant threat birds have developed a highly organized lung-associated immune system. In this review the basic features of this system are described and their functional properties discussed. Most prominent in the avian lung is the bronchus-associated lymphoid tissue (BALT) which is located at the junctions between the primary bronchus and the caudal secondary bronchi. BALT nodules are absent in newly hatched birds, but gradually developed into the mature structures found from 6-8 weeks onwards. They are organized into distinct B and T cell areas, frequently comprise germinal centres and are covered by a characteristic follicle-associated epithelium. The interstitial tissue of the parabronchial walls harbours large numbers of tissue macrophages and lymphocytes which are scattered throughout tissue. A striking feature of the avian lung is the low number of macrophages on the respiratory surface under non-inflammatory conditions. Stimulation of the lung by live bacteria but not by a variety of bacterial products elicits a significant efflux of activated macrophages and, depending on the pathogen, of heterophils. In addition to the cellular components humoral defence mechanisms are found on the lung surface including secretory IgA. The compartmentalisation of the immune system in the avian lung into BALT and non BALT-regions should be taken into account in studies on the host-pathogen interaction since these structures may have distinct functional properties during an immune response.     

家禽神经内分泌系统与免疫系统之间的相互作用

神经内分泌系统是机体最主要的调节系统，机体内部各系统之间的协调平衡以及机体与外界环境之间的适应与统一，无不受到神经内分泌的整合与调节。免疫是机体的正常生理机能，免疫系统能识别异物，并将其代谢、中和或清除以保卫自身。现已知免疫系统不仅是机体的防卫系统，同时也是机体的一个重要感受和调节系统，可感受肿瘤、病毒、细菌和毒素的刺激，因而免疫反应具有维持体内平衡 (Homeostasis)的功能。由于免疫细胞可随血液循环流动，因而有人提出免疫系统可起一种“移动大脑” (mobile brain)的作用。近十多年研究证明，哺乳动物的神经…     

Effects of midazolam on equine innate immune response: a flow cytometric study

Benzodiazepines (BDZ) are among the most frequently used class of psychotropic drugs employed in veterinary medicine in Brazil and worldwide due to their anxiolytic, muscle relaxant and anticonvulsant effects [J. Clin. Pharmacol. 33 (1993) 124]. Peripheral benzodiazepine receptor (PBR) sites were described in peripheral organs, endocrine steroidogenic tissues and immune organs and cells. Midazolam is a mixed-type agonist of PBRs. The present study is focused on the effects of midazolam on equine peripheral blood neutrophils, peritoneal macrophages and cortisol levels in plasma. Adult horses were treated with a single dose of midazolam (0.06 or 0.1 mg/kg) or with 0.9% NaCl. Immune cells were collected 24 h after treatment for flow cytometry analysis of Staphylococcus aureus-induced phagocytosis and oxidative burst. Plasma cortisol concentration was measured 30, 90, 180 and 360 min after midazolam treatment. Midazolam induced a dose-dependent reduction on: (1) peripheral blood neutrophil and peritoneal macrophage oxidative burst; (2) the capacity of both peripheral blood neutrophils and peritoneal macrophages to phagocyte S. aureus. Increments on plasma cortisol concentration were not found after 0.06 and 0.1 mg/kg of midazolam. The effects on oxidative burst of neutrophils and macrophages from horses treated with midazolam were interpreted as a consequence of an impairment of S. aureus-induced phagocytosis. The present data suggest that midazolam, most probably acting on PBRs present on equine macrophage and neutrophil membranes, might have changed some mechanisms related to both phagocytosis and oxidative burst. These results support the use of flow cytometry to identify functional properties and dysfunction of equine immune cells. They also confirm the notion that changes in the functional capacity of the immune system may represent an important hazard of exposure to drugs or chemicals.     

In utero heat stress alters the postnatal innate immune response of pigs

The effects of in utero heat stress (IUHS) range from decreased growth performance to altered behavior, but the long-term impact of IUHS on postnatal innate immune function in pigs is unknown. Therefore, the study objective was to determine the effects of early gestation IUHS on the immune, metabolic, and stress response of pigs subjected to an 8 hr lipopolysaccharide (LPS) challenge during postnatal life. Twenty-four pregnant gilts were exposed to thermoneutral (TN; n = 12; 17.5 ± 2.1 °C) or heat stress (HS; n = 12; cyclic 26 to 36 °C) conditions from days 6 to 59 of gestation, and then TN conditions (20.9 ± 2.3 °C) from day 60 of gestation to farrowing. At 12 wk of age, 16 IUHS and 16 in utero thermoneutral (IUTN) pigs were selected, balanced by sex and given an intravenous injection of LPS (2 µg/kg BW mixed with sterile saline [SAL] and injected at 2 µL/kg BW) or SAL (2 µL/kg BW). Body temperature was monitored every 30 min, and blood was obtained at 0, 1, 2, 3, 4, 6, and 8 hr following the LPS challenge. Blood samples were analyzed for glucose, insulin, non-esterified fatty acids (NEFA), cortisol, and cytokine concentrations. In addition, white blood cell counts were determined at 0 and 4 hr. Hour 0 data were used as covariates. Body temperature was increased (P < 0.01) in LPS (40.88 ± 0.08 °C) vs. SAL (39.83 ± 0.08 °C) pigs. Eosinophils tended to be decreased overall (P = 0.09; 43.9%) in IUHS vs. IUTN pigs. Glucose concentrations were reduced overall (P = 0.05; 5.9%) in IUHS vs. IUTN pigs. The NEFA concentrations tended to be greater (P = 0.07; 143.4%) in IUHS-LPS pigs compared with all other treatments, and IUTN-LPS pigs tended to have greater (127.4%) circulating NEFA concentrations compared with IUTN-SAL and IUHS-SAL pigs. Cortisol was increased (P = 0.04) in IUHS-LPS compared with IUTN-LPS pigs at 3 hr (21.5%) and 4 hr (64.3%). At 1 hr, tumor necrosis factor α was increased (P = 0.01; 115.1%) in IUHS-LPS compared with IUTN-LPS pigs. Overall, interleukin-1β (IL-1β) and interleukin-6 (IL-6) were greater (P < 0.04; 281.3% and 297.8%, respectively) in IUHS-LPS pigs compared with all other treatments, and IUTN-LPS pigs had increased IL-1β and IL-6 concentrations compared with IUTN-SAL and IUHS-SAL pigs. In summary, IUHS altered the postnatal cytokine, metabolic, and physiological stress response of pigs during postnatal life, which may have negative implications toward the innate immune response of IUHS pigs to pathogens.