Neonate Intestinal Immune Response to CpG Oligodeoxynucleotide Stimulation

Background

The development of mucosal vaccines is crucial to efficiently control infectious agents for which mucosae are the primary site of entry. Major drawbacks of these protective strategies are the lack of effective mucosal adjuvant. Synthetic oligodeoxynucleotides that contain several unmethylated cytosine-guanine dinucleotide (CpG-ODN) motifs are now recognized as promising adjuvants displaying mucosal adjuvant activity through direct activation of TLR9-expressing cells. However, little is known about the efficacy of these molecules in stimulating the intestinal immune system in neonates.

Methodology/Principal Findings

First, newborn mice received CpG-ODN orally, and the intestinal cytokine and chemokine response was measured. We observed that oral administration of CpG-ODN induces CXC and CC chemokine responses and a cellular infiltration in the intestine of neonates as detected by immunohistochemistry. We next compared the efficiency of the oral route to intraperitoneal administration in stimulating the intestinal immune responses of both adults and neonates. Neonates were more responsive to TLR9-stimulation than adults whatever the CpG-ODN administration route. Their intestinal epithelial cells (IECs) indirectly responded to TLR9 stimulation and contributed to the CXC chemokine response, whereas other TLR9-bearing cells of the lamina-propria produced CC chemokines and Th1-type cytokines. Moreover, we showed that the intestine of adult exhibited a significantly higher level of IL10 at homeostasis than neonates, which might be responsible for the unresponsiveness to TLR9-stimulation, as confirmed by our findings in IL10-deficient mice.

Conclusions/Significance

This is the first report that deciphers the role played by CpG-ODN in the intestine of neonates. This work clearly demonstrates that an intraperitoneal administration of CpG-ODN is more efficient in neonates than in adults to stimulate an intestinal chemokine response due to their lower IL-10 intestinal level. In addition we report the efficiency of the oral route at inducing intestinal chemokine responses in neonate that might be taken into consideration for further vaccine development against neonatal diseases.     

口服乳酸杆菌对实验动物免疫功能及肠道正常菌群的影响

本文报告C_(57)BL/6小鼠口服乳酸杆菌后,脾细胞和胸腺细胞的增殖反应,腹腔巨噬细胞的C_3b受体活性及其对L_(929)细胞的细胞毒性作用都明显增强。停用乳酸杆菌10天后,以上免疫指标又恢复到正常水平。其次,Wister大鼠口服乳酸杆菌后,检查粪便菌群中几种厌氧菌和需氧菌的活菌数目,结果表明对厌氧菌群的生长有扶持作用,对需氧菌群的生长则起限制作用,这提示有利于宿主调整肠道正常菌群的平衡。     

肝脏固有免疫系统对肠源性感染的免疫应答与免疫耐受机制

肝特殊的解剖结构及生理特征使其成为暴露肠源性抗原的主要器官。由于肝具有独特的固有免疫系统,在正常情况下,肝分布多种致耐受的抗原提呈细胞,对持续性表达或递呈于肝的肠源性抗原物质,诱发针对该抗原的系统性免疫耐受,避免肝受到不必要的免疫损伤。当炎症发生及肝脏固有免疫系统活化时,则通过免疫效应细胞及免疫效应因子对肠源性病原体发挥强烈地免疫应答以控制感染。该过程形成机制的研究对肝功能的理解及肝性疾病的预防与治疗至关重要。本文就肝固有免疫系统对肠源性感染的免疫应答与免疫耐受形成机制作一综述。     

Current Research on the Immune Response to Experimental Sporotrichosis

Sporotrichosis is often manifested as a chronic granulomatous infection and the monocytes/macrophages play a central role in the host defense system. Surface components of Sporothrix schenckii have been characterized and suggestions have been made as to their possible role in pathogenicity. Ergosterol peroxide, cell-wall compounds (alkali-insoluble fraction-F1 and lipid extract-LEY), and exoantigen from the yeast form of the fungus have been characterized as virulence factors, activating both innate, by cytotoxins linked to the activation of reactive oxygen and nitrogen species (H₂O₂ and NO), and adaptive immune response to produce cytokines Th1 and Th2 profile. In this study, preliminary results have demonstrated that, in systemic sporotrichosis, TLR-4 triggers the innate immune response, activating an oxidative burst. These data represent the first report of the participation of TLR-4 in murine sporotrichosis, in the presence of lipids from the cell wall of S. schenckii. These results taken together may open new perspectives of study leading to an antifungal agent that could be used to benefit the entire population.     

The effects of mating on egg production in the cotton stainer, Dysdercus fasciatus

Egg production in mated and virgin females of the cotton stainer, Dysdercus fasciatus Signoret, was studied in the laboratory. No significant differences were found in adult longevity, pre-oviposition period, inter-oviposition period, the total number of eggs oviposited during the whole reproductive period, and the mean number of mature eggs retained in the ovaries at each oviposition. Thus, mating has no influence on egg production or oviposition in this insect.

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Résumé L'influence de la copulation sur la fécondité de la Punaise rouge du Coton, Dysdercus fasciatus fut étudiée au laboratoire à la température de 28±1° et à l'humidité relative de 55–65%. Six facteurs furent spécialement étudiés: la longévité des adultes, la durée de la période précédant la ponte, la durée moyenne des périodes comprises entre les pontes, le nombre de pontes déposées par chaque femelle durant sa phase reproductrice, le nombre moyen d'oeufs par ponte et le nombre moyen d'oeufs mûrs retenus dans les ovaires à chaque ponte.L'accouplement n'avait pas d'effet sur ces différents facteurs, excepté les quatrième et cinquième facteurs. Les femelles vierges déposaient davantage de pontes que les femelles ayant copulé, mais ces pontes avaient une plus petite quantité d'oeufs que les autres. Il en résultait que le nombre total des oeufs émis par les deux catégories de femelles durant leur vie était à peu près identique. Ainsi, dans cette espèce, l'accouplement n'avait pas d'influence apparente sur la fécondité relative et globale.

     

The Immune Response of Hemocytes of the Insect Oncopeltus fasciatus against the Flagellate Phytomonas serpens

The genus Phytomonas includes parasites that are etiological agents of important plant diseases, especially in Central and South America. These parasites are transmitted to plants via the bite of an infected phytophagous hemipteran. Despite the economic impact of these parasites, many basic questions regarding the genus Phytomonas remain unanswered, such as the mechanism by which the parasites cope with the immune response of the insect vector. In this report, using a model of systemic infection, we describe the function of Oncopeltus fasciatus hemocytes in the immune response towards the tomato parasite Phytomonas serpens. Hemocytes respond to infection by trapping parasites in nodular structures and phagocytizing the parasites. In electron microscopy of hemocytes, parasites were located inside vacuoles, which appear fused with lysosomes. The parasites reached the O. fasciatus salivary glands at least six hours post-infection. After 72 hours post-infection, many parasites were attached to the salivary gland outer surface. Thus, the cellular responses did not kill all the parasites.     

The Transcriptomic Response of Rat Hepatic Stellate Cells to Endotoxin: Implications for Hepatic Inflammation and Immune Regulation

With their location in the perisinusoidal space of Disse, hepatic stellate cells (HSCs) communicate with all of the liver cell types both by physical association (cell body as well as cytosolic processes penetrating into sinusoids through the endothelial fenestrations) and by producing several cytokines and chemokines. Bacterial lipopolysaccharide (LPS), circulating levels of which are elevated in liver diseases and transplantation, stimulates HSCs to produce increased amounts of cytokines and chemokines. Although recent research provides strong evidence for the role of HSCs in hepatic inflammation and immune regulation, the number of HSC-elaborated inflammatory and immune regulatory molecules may be much greater then known at the present time. Here we report time-dependent changes in the gene expression profile of inflammatory and immune-regulatory molecules in LPS-stimulated rat HSCs, and their validation by biochemical analyses. LPS strongly up-regulated LPS-response elements (TLR2 and TLR7) but did not affect TLR4 and down-regulated TLR9. LPS also up-regulated genes in the MAPK, NFκB, STAT, SOCS, IRAK and interferon signaling pathways, numerous CC and CXC chemokines and IL17F. Interestingly, LPS modulated genes related to TGFβ and HSC activation in a manner that would limit their activation and fibrogenic activity. The data indicate that LPS-stimulated HSCs become a major cell type in regulating hepatic inflammatory and immunological responses by altering expression of numerous relevant genes, and thus play a prominent role in hepatic pathophysiology including liver diseases and transplantation.     

Peculiarities of the epidermal gland system of the cotton stainer Dysdercus fasciatus Signoret (Heteroptera: Pyrrhocoridae)

The epidermal gland system of adult Dysdercus fasciatus, which shows conspicuous sexual dimorphism, is described. Observations on the development of the glands and a discussion of their possible functions are included.     

Experimental studies of migration in bugs of the genus Dysdercus

Summary Using a tethered flight technique, migration was studied in the African cotton stainer bugs Dysdercus fasciatus Sign., D. nigrofasciatus Stål, and D. superstitiosus (F.) (Heteroptera: Pyrrhocoridae). The flight system of males of all three species matures in 4–6 days post-eclosion, and flight can continue throughout life. Fed females of D. fasciatus do not fly, but rather histolyse the flight muscles and develop oocytes. Fed females of D. nigrofasciatus and D. superstitiosus exhibit some flight at 4 days post-eclosion, but flight is essentially absent at 6 days when flight muscle histolysis and oögenesis are in progress. Starved females of all three species undertake considerable flight and do not histolyse the flight muscles or develop oöcytes. In starved pairs coupling frequency is lowered permitting increased migration. The three species are thus facultative migrants using starvation as a releasing cue. D. fasciatus feeds on baobab (Adansonia) fruits which are seasonally abundant and is the most opportunistic species with the highest reproductive potential and no flight in the presence of food. D. nigrofasciatus feeds on a variety of herbaceous annuals and perennials; it follows an intermediate migration and reproductive strategy. Finally D. superstitiosus is the most catholic in its food habits and has the lowest reproductive potential. Both of the latter species retain some flight capacity even when fed, presumably so that individuals can locate suitable host plants.     

Makisterone C: A 29-carbon ecdysteroid from developing embryos of the cotton stainer bug,Dysdercus fasciatus

An ecdysteroid RIA was used to determine the ecdysteroid titer in developing embryos of Dysdercus fasciatus and revealed that peak titer occurred approximately 120 h post-oviposition. Analysis of neutral sterols at this time indicated sitosterol to be the predominant neutral sterol with lesser amounts of campesterol. Embryonic sterols were highly reflective of the sterols found in the cotton seed diet upon which previous generations of the bugs had fed. Analysis of the embryonic extract for ecdysteroids indicated the presence of both makisterone A and the 29-carbon ecdysteroid makisterone C. Isolation of these compounds was accomplished by reversed-phase and silica HPLC in conjunction with RIA, and the identification of both compounds was confirmed by mass spectrometry. No ecdysone or 20-hydroxyecdysone was detected in the embryonic sample.     

短期盐胁迫下盐穗木的转录组分析

盐穗木（Halostachys caspica）是荒漠盐碱地广泛分布的盐生植物,具有极强的耐盐性。为揭示盐胁迫下盐穗木基因组层面的基因表达变化特性,通过对300和500mmol·L^-1 NaCl胁迫3h的盐穗木同化枝进行了转录组测序。有效序列组装共得到153298条平均长度为643bp的unigenes,进行GO和KEGG功能聚类,分别获得47个GO功能小类和118个KEGG通路。差异表达基因分析显示,短期低盐（300mmol·L^-1）响应基因有4432个,高盐（500mmol·L^-1）响应基因有2580个,两个胁迫的共差异基因有1245个,主要富集在细胞过程、代谢过程和响应刺激等类别中。从短期盐胁迫下盐穗木转录组筛选出渗透调节和活性氧清除的相关基因,大多为上调基因。说明盐穗木能够通过促进渗透调节和增强活性氧清除提高短期的盐胁迫适应能力。     

Transcriptomic Response to Yersinia pestis:RIG-I Like Receptor Signaling Response Is Detrimental to the Host against Plague

Bacterial pathogens have evolved various mechanisms to modulate host immune responses for successful infection. In this study, RNA- sequencing technology was used to analyze the responses of human monocytes THP1 to Yersinia pestis infection. Over 6000 genes were differentially expressed over the 12 h infection. Kinetic responses of pathogen recognition receptor signaling pathways, apoptosis, antigen processing, and presentation pathway and coagulation system were analyzed in detail. Among them, RIG-I-like receptor （RLR） signaling pathway, which was established for antiviral defense, was significantly affected. Mice lacking MAVS, the adaptor of the RLR signaling pathway, were less sensitive to infection and exhibited lower IFN-13 production, higher Thl-type cytokines IFN-γ and IL-12 production, and lower Th2-type cytokines IL-4 and IL-13 production in the serum compared with wild-type mice. Moreover, infection of pathogenic bacteria other than E pestis also altered the expression of the RLR pathway, suggesting that the response of RLR pathway to bacterial infection is a universal mechanism.