昆虫天然免疫反应分子机制研究进展

昆虫体内缺乏高等脊椎动物所具有的获得性免疫系统, 只能依赖发达的天然免疫系统抵抗细菌、 真菌、 病毒等外源病原物的侵染。本文概括了昆虫天然免疫反应发生和作用的分子机制相关进展, 重点阐述了重要免疫相关因子在昆虫天然免疫反应中的功能和作用机制。昆虫天然免疫反应分为体液免疫和细胞免疫两种, 二者共同作用完成对病原物的吞噬 (phagocytosis)、 集结 (nodulation)、 包囊 (encapsulation)、 凝结 (coagulation)和黑化（melanization）等。当昆虫受到外界病原物的侵染时, 首先通过体内的模式识别蛋白（pattern recognition proteins/receptor, PRPs）识别并结合病原物表面特有的模式分子（pathogen-associated molecular pattern, PAMPs）, 继而一系列包括丝氨酸蛋白酶和丝氨酸蛋白酶抑制剂在内的级联激活反应被激活和调控, 产生抗菌肽、 黑色素等免疫效应分子, 清除或杀灭外源物。抗菌肽是一类小分子量的阳离子肽, 具有广谱抗菌活性, 针对不同类型的病原物, 抗菌肽的产生机制也不尽相同。昆虫体内存在着两种信号转导途径调节抗菌肽的产生： 一是由真菌和大部分革兰氏阳性菌激活的Toll途径; 二是由革兰氏阴性菌激活的Imd途径（immune deficiency pathway）。这两个途径通过激活不同转录因子调控不同抗菌肽基因的表达参与昆虫体内的天然免疫反应。     

小菜蛾Gloverin like抗菌肽对蝉拟青霉侵染的响应研究

抗菌肽是昆虫天然免疫的重要成分,在抵御病原物的侵染中发挥着重要的作用。昆虫Gloverin抗菌肽是一类仅在鳞翅目昆虫中存在的富含甘氨酸抗菌肽。本研究在转录组测序的基础上对小菜蛾抗菌肽gloverin like基因进行了克隆,其开放阅读框序列全长519 bp,编码172 aa,信号肽为1-17 aa。序列分析表明其序列中甘氨酸的比例为15%,与鳞翅目昆虫Gloverin抗菌肽具有较高的同源性。qRT-PCR结果表明小菜蛾抗菌肽gloverin like基因在脂肪体和血细胞组织中表达量较高。蝉拟青霉、金黄色葡萄球菌及大肠杆菌均可诱导小菜蛾抗菌肽gloverin like基因上调表达,蝉拟青霉诱导12 h后表达量达到最高。利用LC-MS方法在蝉拟青霉侵染的小菜蛾幼虫血淋巴中鉴定出小菜蛾Gloverin like、Gloverin抗菌肽以及Lysozyme II、Transferrin、Prophenoloxidase等抗菌效应蛋白。与对照相比,蝉拟青霉侵染后小菜蛾Toll通路4个免疫基因(βGBP、Toll、Cactus和Dorsal)表达量上升。本研究在抗菌肽方面为进一步研究小菜蛾抵御病原真菌入侵的分子机制提供基础信息,也为新的害虫防治方法提供了思路和靶标。     

昆虫病原微生物对其寄主行为的调控作用研究进展

昆虫病原微生物是调控昆虫种群数量动态的重要因子,其作为生物防治害虫的重要手段被广泛应用。昆虫病原物往往通过调控其寄主行为来提高自身的适应性,而有些寄主行为的改变却是其应对病原物侵染的免疫反应。发烧行为被证明可抑制病原增殖并延长寄主死亡;取食行为变化影响病原物或寄主的适应性;繁殖行为主要表现在产卵力、交配行为和性信息素等方面的变化;社会性行为改变对整个社会群体的适应性或病原物传播有影响;病原物所引起的寄主防卫和群集能力下降被认为对病原传播不利;病虫趋光和趋地行为、顶峰行为和活体寄主传病行为等被认为是病原物操纵的有利于病原微生物扩散和传播的行为。明确昆虫病原物调控其寄主行为的策略和机制对于寻找到新的害虫防治方法有指导意义。     

某些抗菌肽的研究进展

抗菌肽是各种生物防御系统的一个组成部分,是由于微生物等因素的侵染而产生的免疫应答反应产物,具有分子量低,热稳定,强碱性和广谱抗菌等特点,抗菌肽的正电荷与细菌细胞的磷脂头负电荷之间的相互作用是重要,肽分子氮端的两性螺旋是裂解细菌的主要部分,碳末端酰胺化与抗菌肽的广谱抗菌有关,抗菌肽具有独特的抗菌机理,能在细菌质膜上形成离子通道,破坏膜势,引起胞内物质泄漏,从而杀灭细菌。     

多种病原物混合侵染的昆虫流行病模型

两种或两种以上的病原物同时侵染昆虫寄主时,病原物之间的相互作用表现为偏利、偏害、中性及竞争等类型,寄生群体的病症可呈多种形式．根据单种病菌的重叠侵染原理,建立了多种病原物混合侵染时以温度、病原接种量、虫龄及湿度为因子的昆虫流行病模型．由模型可计算寄生群体中不同病原物的致病比率,及寄主群体的总发病率,给出了模型的参数求解算法,以及病原物相互作用类型的判定准则．这类模型可用于多种病原物混合侵染的昆虫流行病预测,也可作为多种病原物混合施用防治害虫的最优化模型．     

昆虫免疫识别与病原物免疫逃避机理研究进展

昆虫在长期进化过程中形成复杂的天然免疫系统,病原识别是启动下游免疫反应的第一步,这一过程主要是由不同的模式识别蛋白来完成的。目前发现并鉴定的昆虫模式识别蛋白主要包括肽聚糖识别蛋白、类免疫球蛋白、β-1,3-葡聚糖结合蛋白、C型凝集素及具多功能的载脂蛋白等,不同的蛋白种类具有不同的结构、功能及识别对象。与昆虫免疫识别相对应的是,不同昆虫病原物在进化过程中发展出不同策略的免疫逃避能力,以战胜宿主免疫而致病或最终杀死昆虫。本文就昆虫免疫过程中不同模式识别蛋白的结合对象、结构与功能,以及逐渐兴起的病原物通过分子伪装等进行免疫逃避的研究进展进行了综述。并在此基础上,作者就昆虫免疫与昆虫病理研究的发展方向进行了展望,认为只有当两方面研究相结合时,才能更好地揭示昆虫宿主与病原物之间免疫与抗免疫的动态相互作用过程。     

哺乳动物抗菌肽研究进展

自昆虫中发现抗菌肽以来,迄今已有约2000多种抗菌肽在生物体中被分离,仅在哺乳动物中就发现了上百种抗菌肽,哺乳动物抗菌肽是哺乳动物抵抗外界病原物质侵染而产生的一系列非专一性免疫应答物,它具有分子量低,热稳定,广播谱抗菌等特点,本文就近年来发现的哺乳动物的抗菌从分类,结构、生物学特性,表达控制,作用机制及应用前景方面作一介绍。     

植物NLR免疫受体的识别、免疫激活与信号调控

高等植物进化出大量膜表面和胞内免疫受体以感知各种病原信号,抵御病原物入侵。其中,细胞表面的模式识别受体感知模式分子后激活基础免疫反应,核苷酸结合和富亮氨酸重复蛋白(NLRs)则通过感知病原微生物分泌的效应蛋白激活特异免疫反应,导致超敏反应与细胞死亡。该文主要综述了NLRs对效应蛋白的识别、植物免疫激活及下游信号调控的最新研究进展。     

RXLR效应分子抑制植物免疫机制研究进展

植物病原卵菌是一类农业生产上为害巨大的病原物,其分泌大量的RXLR效应分子进入寄主植物细胞并干扰植物免疫系统,以协助病原菌成功侵染。尽管有一小部分RXLR效应分子会被植物识别成为无毒蛋白,但大部分RXLR效应分子则会逃避识别和抑制植物免疫。随着高通量测序和蛋白互作技术的广泛应用,大量RXLR效应分子干扰植物免疫的分子机制已经被揭示。本文综述了RXLR效应分子操纵植物免疫系统的分子策略,探讨了RXLR效应分子与植物免疫互作的研究方向和应用前景。     

次生代谢产物与植物抗病防御反应

次生代谢产物是由植物次生代谢产生的许多结构不同的小分子有机化合物,它们广泛参与植物的生长、发育、防御等生理过程。次生代谢产物在植物的抗病防御反应中发挥着重要作用,可以作为生化壁垒防御病原物侵染,还可以作为信号物质参与植物的抗病反应;在植物与病原物互作中,植物合成新的抗菌物质植保素,原有的抗菌物质也会增加。植物次生代谢产物的积累受到病原物、发育,环境等多种因素的调节。本文重点介绍次生代谢产物在植物抗病防御中的相关作用以及影响其合成的各种因素。     

Susceptibility and behavioral responses of the dampwood termite Zootermopsis angusticollis to the entomopathogenic nematode Steinernema carpocapsae

Termites exploit microbially rich resources such as decayed wood and soil that are colonized by potentially pathogenic and parasitic fungi, bacteria, viruses, and nematodes. In colonies composed of thousands of individuals, the risk of infection among nestmates is significant, and individual and social behavior could involve various adaptations to resist disease and parasitism. Here we show that the dampwood termite Zootermopsis angusticollis (Hagen) exhibits a dosage dependent susceptibility to the soil nematode Steinernema carpocapsae (Weiser) (Mexican strain) and that this social insect significantly alters its behavior in response to this entomopathogenic roundworm. Relative to their baseline behavior, termites exposed to infective juveniles increased the frequency and duration of allogrooming and vibratory displays as well as two other novel behaviors, abdominal tip-raising and self-scratching. Whereas the first two behaviors likely reflect general adaptations to reduce susceptibility to a variety of pathogens and parasites, the latter behaviors might be specific to nematodes because they have never been observed in Z. angusticollis in any other pathogenic context. Our results support the hypotheses that behavioral responses in termites are important in the control of pathogenic and parasitic microorganisms and that termite susceptibility is socially mediated.     

The effect of ergosterol on the allogrooming behavior of termites in response to the entomopathogenic fungus Metarhizium anisopliae

Termites have physiological and behavioral immunities that make them highly resistant to pathogen infections, which complicates biocontrol efforts. However, the stimuli that trigger the pathogen-avoidance behaviors of termites are still unclear. Our study shows that workers of Coptotermes formosanus exposed to the conidia of Metarhizium anisopliae exhibited a significantly higher frequency and longer duration of allogrooming behaviors compared with untreated termites. Volatile compounds in the cuticle of control termites and termites previously exposed to a suspension of M. anisopliae conidia were analyzed and compared using a gas chromatography-mass spectrometer (GC-MS). Our results showed that the amount of ergosterol differed between the fungus-exposed and control termites. Choice tests showed that termites significantly preferred to stay on filter paper treated with ergosterol (0.05, 0.1, or 1.0 mg/mL) compared with control filter paper. In addition, termites exposed to ergosterol followed by M. anisopliae conidia were allogroomed at a significantly higher frequency and for a longer duration than termites exposed to alcohol (the solvent used with the ergosterol in the ergosterol trials) alone followed by M. anisopliae conidia. These results showed that ergosterol may enhance the allogrooming behavior of termites in the presence of entomopathogenic fungi.     

Novel properties of antimicrobial peptides

Endogenous peptide antibiotics are known as evolutionarily old components of innate immunity. Due to interaction with cell membrane these peptides cause permeabilization of the membrane and lysis of invading microbes. However, some studies proved that antimicrobial peptides are universal multifunctional molecules and their functions extend far beyond simple antibiotics. In this review we present an overview of the general mechanism of action of antimicrobial peptides and discuss some of their additional properties, like antitumour activity, mitogenic activity, role in signal transduction pathways and adaptive immune response.     

Why and how are peptide-lipid interactions utilized for self-defense? Magainins and tachyplesins as archetypes

Animals as well as plants defend themselves against invading pathogenic microorganisms utilizing cationic antimicrobial peptides, which rapidly kill various microbes without exerting toxicity against the host. Physicochemical peptide-lipid interactions provide attractive mechanisms for innate immunity. Many of these peptides form cationic amphipathic secondary structures, typically alpha-helices and beta-sheets, which can selectively interact with anionic bacterial membranes by the aid of electrostatic interactions. Rapid, peptide-induced membrane permeabilization is an effective mechanism of antimicrobial action. This review article summarizes interactions with lipid bilayers of magainins (alpha-helix) and tachyplesins (beta-sheet) discovered in frog skin and horseshoe crab hemolymph, respectively, as archetypes, emphasizing that the mode of interaction is strongly dependent on the physicochemical properties not only of the peptide, but also of the target membrane.     

Antimicrobial peptides important in innate immunity

Antimicrobial peptides are present in all walks of life, from plants to animals, and they are considered to be endogenous antibiotics. In general, antimicrobial peptides are determinants of the composition of the microbiota and they function to fend off microbes and prevent infections. Antimicrobial peptides eliminate micro-organisms through disruption of their cell membranes. Their importance in human immunity, and in health as well as disease, has only recently been appreciated. The present review provides an introduction to the field of antimicrobial peptides in general and discusses two of the major classes of mammalian antimicrobial peptides: the defensins and the cathelicidins. The review focuses on their structures, their main modes of action and their regulation.     

Grooming patterns in Verreaux's sifaka

Lemur grooming has received very little attention in the literature. Nevertheless, allogrooming in lemurs has been suggested to be fundamentally different from the grooming of anthropoids. One reason is that lemurs generally rely on oral rather than manual grooming. Lemur allogrooming has also been suggested to serve less of a social function than has been attributed to anthropoid grooming. I analyzed the allogrooming behaviors of 29 Verreaux's sifaka (Propithecus verreauxi) living in five social groups in the Kirindy Forest of Madagascar. Based upon 1,586 observation hours, I found that sifaka, like anthropoids, spend very little time mutual grooming (2±3%). Half of all allogrooming involved parts of the body that could have been easily groomed by the recipient, such as the limbs. Even though ectoparasite loads are expected to be greater during the rainy season, allogrooming did not increase during the rainy season. Allogrooming rates were influenced by both rank and sex, and increased by 50–100% during the mating season. The results of this study suggest that allogrooming in Verreaux's sifaka plays an important social function, even though it is performed with a toothcomb. Am. J. Primatol. 72:254–261, 2010. © 2009 Wiley‐Liss, Inc.     

Mycoplasma lipoproteins are major determinants of neutrophil extracellular trap formation

Neutrophil granulocytes are paramount to innate responses as major effectors of acute inflammation. Among the various strategies enacted by neutrophils to eliminate microbes NETosis is a novel distinct antimicrobial activity in which an interlacement of chromatin fibres rich in granule‐derived antimicrobial peptides and enzymes is extruded (NETs, neutrophils extracellular traps ). NETs contribute to the pathogenesis of acute and chronic inflammatory disorders. The interactions of mycoplasmas and innate immune cells, in particular neutrophil granulocytes, are poorly defined. Here, we describe NET formation in vivo in the mammary gland and milk of sheep naturally infected by Mycoplasma agalactiae. Also, we assess the contribution of liposoluble proteins, the most abundant component of the Mycoplasma membrane, in inducing NETosis. We demonstrate that Mycoplasma liposoluble proteins induce NET release at levels comparable to what observed with other stimuli, such as lipopolysaccharides and phorbol 12‐myristate 13‐acetate. Stimulation of neutrophils with synthetic diacylated lipopeptides based on the M. agalactiae P48, P80, and MAG_1000 proteins, combined in a mix or used individually, suggests that NETosis might not be dependent on a specific lipopeptide sequence. Also, NETosis is partially abolished when TLR2 is blocked with specific antibodies. The results presented in this work provide evidences for the mechanisms underlying NET activation in mycoplasma infections, and on their contribution to pathogenesis of mycoplasmosis.     

When subterranean termites challenge the rules of fungal epizootics

Over the past 50 years, repeated attempts have been made to develop biological control technologies for use against economically important species of subterranean termites, focusing primarily on the use of the entomopathogenic fungus Metarhizium anisopliae. However, no successful field implementation of biological control has been reported. Most previous work has been conducted under the assumption that environmental conditions within termite nests would favor the growth and dispersion of entomopathogenic agents, resulting in an epizootic. Epizootics rely on the ability of the pathogenic microorganism to self-replicate and disperse among the host population. However, our study shows that due to multilevel disease resistance mechanisms, the incidence of an epizootic within a group of termites is unlikely. By exposing groups of 50 termites in planar arenas containing sand particles treated with a range of densities of an entomopathogenic fungus, we were able to quantify behavioral patterns as a function of the death ratios resulting from the fungal exposure. The inability of the fungal pathogen M. anisopliae to complete its life cycle within a Coptotermes formosanus (Isoptera: Rhinotermitidae) group was mainly the result of cannibalism and the burial behavior of the nest mates, even when termite mortality reached up to 75%. Because a subterranean termite colony, as a superorganism, can prevent epizootics of M. anisopliae, the traditional concepts of epizootiology may not apply to this social insect when exposed to fungal pathogens, or other pathogen for which termites have evolved behavioral and physiological means of disrupting their life cycle.