共查询到20条相似文献,搜索用时 15 毫秒
1.
Salcedo SP Marchesini MI Lelouard H Fugier E Jolly G Balor S Muller A Lapaque N Demaria O Alexopoulou L Comerci DJ Ugalde RA Pierre P Gorvel JP 《PLoS pathogens》2008,4(2):e21
Brucella is an intracellular pathogen able to persist for long periods of time within the host and establish a chronic disease. We show that soon after Brucella inoculation in intestinal loops, dendritic cells from ileal Peyer's patches become infected and constitute a cell target for this pathogen. In vitro, we found that Brucella replicates within dendritic cells and hinders their functional activation. In addition, we identified a new Brucella protein Btp1, which down-modulates maturation of infected dendritic cells by interfering with the TLR2 signaling pathway. These results show that intracellular Brucella is able to control dendritic cell function, which may have important consequences in the development of chronic brucellosis. 相似文献
2.
Activation of Rho and Rab GTPases dissociates Brucella abortus internalization from intracellular trafficking 总被引:1,自引:0,他引:1
Chaves-Olarte E Guzmán-Verri C Méresse S Desjardins M Pizarro-Cerdá J Badilla J Gorvel JP Moreno E 《Cellular microbiology》2002,4(10):663-676
Brucella abortus is an intracellular pathogen that relies on unconventional virulence factors to infect hosts. In non-professional phagocytes, Rho GTPases-activation by the Escherichia coli cytotoxic necrotizing factor (CNF) promoted massive Brucella entrance by membrane ruffling, a mechanism that differs from the common mode of entrance used by this bacterium in non-treated cells. Cytotoxic necrotizing factor treatment, however, did not alter the intracellular route followed by the wild type or non-virulent defined mutants. In contrast, expression of a constitutively active Rab5Q79L GTPase did not alter cell-invasion by Brucella but hampered its ability to reach the endoplasmic reticulum. The CNF-induced Brucella super-infection did not reduce the ability of host cells to synthesize DNA and progress through the cell cycle. Furthermore, CNF-treatment increased the isolation of Brucella-containing compartments by a factor of 15. These results demonstrate that in non-professional phagocytic cells, Brucella manipulates two different sets of GTPases during its biogenesis, being internalization and intracellular trafficking two consecutive but independent processes. Besides, CNF-induced super-infection demonstrates that Brucella does not interfere with crucial cellular processes and has shown its potential as tool to characterize the intracellular compartments occupied by this bacterium. 相似文献
3.
Uterine epithelial and eosinophil estrogen receptors in rats during the estrous cycle 总被引:3,自引:0,他引:3
S. H. Lee 《Histochemistry and cell biology》1982,74(4):443-452
Summary In the uterus of the adult female rats, the luminal epithelial cells and the eosinophil leukocytes are rich in cytoplasmic estrogen receptors. During the estrous cycle, the epithelial estrogen receptor concentration reaches its peak level, in proestrus, drops precipitously in estrus, and hits the trough, at metestrus. Repopulation of the cytoplasm with estrogen binding sites occurs during diestrus. This pattern of cyclic change is indicative of a rapid turnover of estrogen receptors in the epithelial cells and its regulation by endogenous estrogens. The concentration of estrogen receptors in the cytoplasm of the eosinophils does not appear to fluctuate during the cycle. But the intrauterine, distribution of these leukocytes is clearly cyclic in pattern, ostensibly influenced by estrogens. While progesterone binding activity is consistently demonstrated in tandem with estrogen receptors in the cytoplasm of the epithelial cells, it has not been observed in the eosinophil leukocytes. These findings support the claim that there are two estrogen receptor systems in the rat uterus, one mediating the intracellular events of the genomic response to estrogens, and the other being concerned with non-genomic responses.Abbreviations BSA
Bovine serum albumin
- FITC
fluorescein isothiocyanate
- TMRITC
tetramethylrhodamine isothiocyanate
- CMO
O-carboxymethyl oxime 相似文献
4.
Heterologous expression of functional, nicotinic acetylcholine receptors (nAChR) in mammalian cells has been difficult to achieve or optimize, even for nAChR containing only one kind of subunit. In this study, we determined effects of lowered temperature or of exposure to the protein synthesis inhibitor cycloheximide (CHX) on cell surface expression of homomeric alpha7-nAChR in transfected SH-EP1 human epithelial cells. We found that incubation of cells for 2 days at 25 degrees C or in the presence of 0.5-2 microg/mL of CHX caused approximately four- or approximately eight-fold increases, respectively, in surface binding sites for 125I-labeled alpha-bungarotoxin (I-Bgt). These increases were accompanied by increases in peak whole-cell current responses to nicotinic agonists. Either treatment lowered protein synthesis and cell proliferation, but experiments using puromycin indicated that a reduction in protein synthesis or cell proliferation per se was not sufficient to increase surface binding. I-Bgt binding to whole-cell membrane pools increased in response to either treatment, suggesting that the increase in surface binding was due, at least in part, to an increase in intracellular receptor levels. The cyclophilin inhibitor cyclosporin A reduced surface expression in untreated as well as CHX- or 25 degrees C-treated cells. The results suggest practical means for increasing cell surface and functional expression of alpha7-nAChR. Although these effects are not simply due to protein synthesis inhibition or reduced cell proliferation, they do involve an increase in intracellular receptor pool size. 相似文献
5.
Pieter P. E. van Lierop Sigrid M. Swagemakers Charlotte I. de Bie Sabine Middendorp Peter van Baarlen Janneke N. Samsom Wilfred F. J. van IJcken Johanna C. Escher Peter J. van der Spek Edward E. S. Nieuwenhuis 《PloS one》2013,8(11)
Objective
In current clinical practice, optimal treatment of inflammatory bowel disease (IBD) aims at the induction and maintenance of clinical remission. Clinical remission is apparent when laboratory markers of inflammation are normal and clinical symptoms are absent. However, sub-clinical inflammation can still be present. A detailed analysis of the immune status during this inactive state of disease may provide a useful tool to categorize patients with clinical remission into subsets with variable states of immune activation.Design
By using Affymetrix GeneChips, we analysed RNA gene expression profiles of peripheral blood leukocytes from pediatric IBD patients in clinical remission and controls. We performed (un)supervised clustering analysis of IBD-associated genes and applied Ingenuity® pathway software to identify specific molecular profiles between patients.Results
Pediatric IBD patients with disease in clinical remission display heterogeneously distributed gene expression profiles that are significantly distinct from controls. We identified three clusters of IBD patients, each displaying specific expression profiles of IBD-associated genes.Conclusion
The expression of immune- and IBD-associated genes in peripheral blood leukocytes from pediatric IBD patients in clinical remission was different from healthy controls, indicating that sub-clinical immune mechanisms are still active during remission. As such, RNA profiling of peripheral blood may allow for non-invasive patient subclassification and new perspectives in treatment regimes of IBD patients in the future. 相似文献6.
《Free radical research》2013,47(3):183-188
The relationship between glutathione peroxidase (GSH-Px) activity and opsonized zymosan-induced chemiluminescence (CL) has been studied with exudate leukocytes obtained at different times after induction of inflammatory responses in the mouse peritoneal cavity with heat-killed Corynebacterium parvum and in the rat pleural cavity with I-carrageenin. GSH-Px activity in mouse peritoneal exudate cells fell markedly after 2–4h, returning to normal within 1–2 days. The lowered enzyme activity was associated with an increased ability of the cells to generate CL. Rat pleural exudate cells exhibited a slight fall in GSH-Px activity after 6h which increased to supranormal levels within 1–2 days. During this period the ability of the cells to generate CL continually increased. The data indicate that during the early phase of increased generation of reactive oxygen species (ROS) by inflammatory leukocytes, the intracellular protective mechanism, represented by GSH-Px, is compromised. Subsequently, GSH-Px activity increases to or above initial levels possibly due to the presence of mononuclear cells and/or as a response to the increased generation of ROS. 相似文献
7.
Studies have been carried out on the levels of serum and urine colony stimulating activity (CSA) and peripheral blood and bone marrow colony forming cell numbers in children with acute lymphocytic leukemia (ALL) during various phases of their disease. These studies have suggested that serum and urine levels of colony stimulating factor are reduced during the inital or relapse phase of the disease compared to levels found during remission. It has also been found that the number of bone marrow colony forming cells is reduced in relapse or before treatment and elevated during remission while the number of peripheral blood colony forming cells is increased during relapse or before treatment and normal during remission. It has also been shown that mixing of serum or leukemic cells with normal human bone marrow cells inhibits colony formation. 相似文献
8.
Bacteria of the genus Brucella are Gram-negative pathogens of several animal species that cause a zoonotic disease in humans known as brucellosis or Malta fever. Within their hosts, brucellae reside within different cell types where they establish a replicative niche and remain protected from the immune response. The aim of this article is to discuss recent advances in the field in the specific context of the Brucella intracellular 'lifestyle'. We initially discuss the different host cell targets and their relevance during infection. As it represents the key to intracellular replication, the focus is then set on the maturation of the Brucella phagosome, with particular emphasis on the Brucella factors that are directly implicated in intracellular trafficking and modulation of host cell signalling pathways. Recent data on the role of the type IV secretion system are discussed, novel effector molecules identified and how some of them impact on trafficking events. Current knowledge on Brucella gene regulation and control of host cell death are summarized, as they directly affect intracellular persistence. Understanding how Brucella molecules interplay with their host cell targets to modulate cellular functions and establish the intracellular niche will help unravel how this pathogen causes disease. 相似文献
9.
Posadas DM Ruiz-Ranwez V Bonomi HR Martín FA Zorreguieta A 《Cellular microbiology》2012,14(6):965-982
Brucella is an intracellular pathogen responsible of a zoonotic disease called brucellosis. Brucella survives and proliferates within several types of phagocytic and non-phagocytic cells. Like in other pathogens, adhesion of brucellae to host surfaces was proposed to be an important step in the infection process. Indeed, Brucella has the capacity to bind to culture human cells and key components of the extracellular matrix, such as fibronectin. However, little is known about the molecular bases of Brucella adherence. In an attempt to identify bacterial genes encoding adhesins, a phage display library of Brucella suis was panned against fibronectin. Three fibronectin-binding proteins of B. suis were identified using this approach. One of the candidates, designated BmaC was a very large protein of 340 kDa that is predicted to belong to the type I (monomeric) autotransporter family. Microscopy studies showed that BmaC is located at one pole on the bacterial surface. The phage displaying the fibronectin-binding peptide of BmaC inhibited the attachment of brucellae to both, HeLa cells and immobilized fibronectin in vitro. In addition, a bmaC deletion mutant was impaired in the ability of B. suis to attach to immobilized fibronectin and to the surface of HeLa and A549 cells and was out-competed by the wild-type strain in co-infection experiments. Finally, anti-fibronectin or anti-BmaC antibodies significantly inhibited the binding of wild-type bacteria to HeLa cells. Our results highlight the role of a novel monomeric autotransporter protein in the adhesion of B. suis to the extracellular matrix and non-phagocytic cells via fibronectin binding. 相似文献
10.
Brucella strains encounter oxygen deprivation during their intracellular replication in host cells, and the capacity of these bacteria to utilize NO(3) as an alternative electron acceptor for respiration plays an important role in their successful adaption to their intracellular niche. In this issue of Molecular Microbiology, Carrica et al (2012). report that NtrY and NtrX comprise a redox-responsive two-component regulator in Brucella abortus 2308 that responds to decreasing levels of O(2) and induces the expression of this strain's denitrification genes. Thus, NtrYX joins the increasing number of genetic regulators that contribute to the metabolic versatility required for the virulence of Brucella strains in their mammalian hosts. 相似文献
11.
Oliveira SC de Oliveira FS Macedo GC de Almeida LA Carvalho NB 《Microbes and infection / Institut Pasteur》2008,10(9):1005-1009
Research into intracellular sensing of microbial products is an up and coming field in innate immunity. Toll-like receptors (TLRs) recognize Brucella spp. and bacterial components and initiate mononuclear phagocyte responses that influence both innate and adaptive immunity. Recent studies have revealed the intracellular signaling cascades involved in the TLR-initiated immune response to Brucella infection. TLR2, TLR4 and TLR9 have been implicated in host interactions with Brucella; however, TLR9 has the most prominent role. Further, the relationship between specific Brucella molecules and various signal transduction pathways needs to be better understood. MyD88-dependent and TRIF-independent signaling pathways are involved in Brucella activation of innate immune cells through TLRs. We have recently reported the critical role of MyD88 molecule in dendritic cell maturation and interleukin-12 production during B. abortus infection. This article discusses recent studies on TLR signaling and also highlights the contribution of NOD and type I IFN receptors during Brucella infection. The better understanding of the role by such innate immune receptors in bacterial infection is critical in host-pathogen interactions. 相似文献
12.
《Theriogenology》1996,45(8):1553-1559
Radioimmunoassay (RIA) was used to measure plasma eCG binding in dairy goats (n = 524) at the beginning of a progestagen/eCG treatment and 25 d after eCG administration. The eCG binding was not dependent on the age of the females but increased with the number of treatments they had previously received (3.4 % ± 4.8, n = 47 vs 9.6 % ± 13.2, n = 249; mean ± SD; P < 0.01 for goats treated 0 and 1 time vs those treated 2 to 5 times, respectively). The synchronization treatment led to an increase in the binding of eCG (7.1 % ± 10.9 before vs 28.3 ± 24.5 after treatment; P < 0.01). When eCG binding before treatment was higher than 5 % the onset of estrus was delayed: 37.9 % of goats came into estrus more than 30 h after sponge removal vs 7.4 % when eCG binding was lower than 5 % (P < 0.01). Fertility was significantly decreased when eCG binding was higher than 10 %. These results show that the repetition of treatment with eCG to induce estrus in goats increases eCG binding. This could explain the lowered efficiency of the hormonal treatment to synchronize estrus and the associated decrease in fertility when goats are inseminated at a predetermined time. 相似文献
13.
Mohamed N. Seleem Neeta Jain Nikorn Pothayee Ashish Ranjan J. S. Riffle & Nammalwar Sriranganathan 《FEMS microbiology letters》2009,294(1):24-31
Treatment and eradication of intracellular pathogens such as Brucella is difficult because infections are localized within phagocytic cells and most antibiotics, although highly active in vitro , do not actively pass through cellular membranes. Thus, an optimum strategy to treat these infections should address targeting of active drugs to the intracellular compartment where the bacteria replicate, and should prolong the release of the antibiotics so that the number of doses and associated toxicity can be reduced. We incorporated streptomycin and doxycycline into macromolecular nanoplexes with anionic homo- and block copolymers via cooperative electrostatic interactions among the cationic drugs and anionic polymers. The approach enabled simultaneous binding of both antibiotics into the nanoplexes, and their use resulted in an improvement in performance as compared with the free drugs. Administration of two doses of the nanoplexes significantly reduced the Brucella melitensis load in the spleens and livers of infected BALB/c mice. The nanoplexes were more effective than free drugs in the spleens (0.72-log and 0.51-log reductions, respectively) and in the livers (0.79-log and 0.42-log reductions, respectively) of the infected mice. Further research regarding the design of optimum nanoplex structures will be directed towards alterations in both the core and the shell properties to investigate the effects of the rates and pathways of entry into immune cells where the brucellae replicate. 相似文献
14.
C Forestier E Moreno J Pizarro-Cerda J P Gorvel 《Journal of immunology (Baltimore, Md. : 1950)》1999,162(11):6784-6791
In this study, we detailed in a time-dependent manner the trafficking, the recycling, and the structural fate of Brucella abortus LPS in murine peritoneal macrophages by immunofluorescence, ELISA, and biochemical analyses. The intracellular pathway of B. abortus LPS, a nonclassical endotoxin, was investigated both in vivo after LPS injection in the peritoneal cavity of mice and in vitro after LPS incubation with macrophages. We also followed LPS trafficking after infection of macrophages with B. abortus strain 19. After binding to the cell surface and internalization, Brucella LPS is routed from early endosomes to lysosomes with unusual slow kinetics. It accumulates there for at least 24 h. Later, LPS leaves lysosomes and reaches the macrophage cell surface. This recycling pathway is also observed for LPS released by Brucella S19 following in vitro infection. Indeed, by 72 h postinfection, bacteria are degraded by macrophages and LPS is located inside lysosomes dispersed at the cell periphery. From 72 h onward, LPS is gradually detected at the plasma membrane. In each case, the LPS present at the cell surface is found in large clusters with the O-chain facing the extracellular medium. Both the antigenicity and heterogenicity of the O-chain moiety are preserved during the intracellular trafficking. We demonstrate that LPS is not cleared by macrophages either in vitro or in vivo after 3 mo, exposing its immunogenic moiety toward the extracellular medium. 相似文献
15.
H V Thommasen B A Martin B R Wiggs M Quiroga E M Baile J C Hogg 《Journal of applied physiology (Bethesda, Md. : 1985)》1984,56(4):966-974
The effect of pulmonary blood flow on leukocyte uptake and release by the lung was examined in 10 anesthetized spontaneously breathing dogs. Pulmonary arterial and pulmonary venous blood was sampled with catheters placed into the right ventricle and aorta, respectively. Pulmonary blood flow was lowered by inflating a balloon catheter located in the inferior vena cava. In five experiments simultaneous blood samples were drawn from the right ventricle and aorta at 10-s intervals during a control period, a 2- to 3-min period of low flow, and a recovery period. In five additional experiments, less frequent samples were taken over periods of 15-60 min. Total leukocyte concentrations and differential counts were determined for each blood sample. The study shows that large numbers of leukocytes become sequestered within the lung when pulmonary blood flow is low and that an equivalent number of cells are released from the lung after deflation of the balloon catheter. Both the polymorphonuclear leukocytes and the lymphocytes were taken up by the lung when pulmonary blood flow was reduced. We conclude that pulmonary blood flow has a marked effect on the uptake and release of leukocytes by the dog lung. 相似文献
16.
The intracellular phosphoribosyl diphosphate (prpp) levels of Ehrlich ascites tumor cells increased during glucose supplementation and decreased during glucose deprivation, while the numbers of glucose carriers as determined by glucose-reversible cytochalasin-B binding changed in an opposite manner relating to the extracellular glucose concentrations and the intracellular prpp levels of Ehrlich ascites tumor cells in vitro. Incubation of cells with hypoxanthine or 2,4-dinitrophenol lowered the intracellular prpp levels and resulted in an increase in numbers of glucose carriers. 相似文献
17.
DATUS M. HAMMOND RONALD FAYER MERTHYR L. MINER 《The Journal of eukaryotic microbiology》1969,16(2):298-302
SYNOPSIS. Eimeria bovis merozoites occurred in tissue culture medium removed from Leighton tube cultures of embryonic bovine tracheal cells beginning 12-14 days after inoculation with 270,000-369,000 sporozoites per tube. The number of merozoites produced in these cultures increased daily until a peak was reached 18-21 days after inoculation. In 3 experiments an average of 2.0–15.6 million merozoites per tube was produced during the 20-day observation period. When such merozoites were frozen in liquid nitrogen and stored 26–42 days, some were motile upon thawing. These merozoites as well as others freshly obtained from cell cultures and from calves were inoculated into 11 different types of cultured mammalian cells including primary, cell line and established cell line cultures. Some merozoites were exposed to substances normally found in the lumen of the gut, before or at the time of inoculation. Altho small numbers of intracellular merozoites were found, no further development was observed. Gametocytes were observed in the cecum of a calf 4 days after merozoites from cell cultures were introduced into a ligated cecum of the calf. 相似文献
18.
K B Kurmanova R Zh Ishchanova S Sh Sakhisheva V K Studentsova L E Tsirel'son G U Alshinbaeva 《Antibiotiki i khimioterapii͡a》1990,35(7):35-38
Experimental studies and clinical trials were performed on possible increase of antibiotic therapy efficacy in brucellosis patients by correction of the immunity disorders with vitamin A. It was experimentally shown that vitamin A increased cellular immunity and accelerated sanation of guinea pigs sensitized with Brucella abortus 19 BA. The clinical trials demonstrated that the use of vitamin A in a dose of 33,000 IU thrice a day for 10 to 12 days during the complex treatment of patients with acute (36 persons) and subacute (57 persons) brucellosis lowered the average period of manifestation of the disease clinical signs and formation of the antibodies, increased the skin allergic sensitivity, the lymphocyte blast cell transformation, the total number and subpopulations of the active T-cells, theophylline-resistant lymphocytes, phagocytic and metabolic activity of neutrophils, showed 1.5- and 2-fold increased in the frequency of the infection transformation into a chronic process in patients with acute or subacute brucellosis, respectively. 相似文献
19.
Samuelsen JT Schwarze PE Huitfeldt HS Thrane EV Låg M Refsnes M Skarpen E Becher R 《American journal of physiology. Lung cellular and molecular physiology》2007,292(1):L232-L239
To elucidate the role of cAMP and different cAMP-dependent protein kinases (PKA; A-kinase) in lung cell proliferation, we investigated rat alveolar type 2 cell proliferation in relation to activation or inhibition of PKA and PKA regulatory subunits (RIIalpha and RIalpha). Both the number of proliferating type 2 cells and the level of different regulatory subunits varied during 7 days of culture. The cells exhibited a distinct peak of proliferation after 5 days of culture. This proliferation peak was preceded by a rise in RIIalpha protein level. In contrast, an inverse relationship between RIalpha and type 2 cell proliferation was noted. Activation of PKA increased type 2 cell proliferation if given at peak RIIalpha expression. Furthermore, PKA inhibitors lowered the rate of proliferation only when a high RII level was observed. An antibody against the anchoring region of RIIalpha showed cell cycle-dependent binding in contrast to antibodies against other regions, possibly related to altered binding to A-kinase anchoring protein. Following activation of PKA, relocalization of RIIalpha was confirmed by immunocytochemistry. In conclusion, it appears that activation of PKA II is important in regulation of alveolar type 2 cell proliferation. 相似文献
20.
Comparison of proteoglycans synthesized by porcine normal and polycystic renal tubular epithelial cells in vitro 总被引:2,自引:0,他引:2
L A Beavan F A Carone S Nakamura J K Jones J F Reindel R G Price 《Archives of biochemistry and biophysics》1991,284(2):392-399
Newly synthesized porcine tubular epithelial cell proteoglycans were labeled in vitro with Na2[35S]SO4. At the beginning of the labeling period (24 h) [35S] sulfate incorporated into macromolecules was measured following PD-10 chromatography. There was a significant reduction in the amount of 35S-labeled macromolecules isolated from polycystic cells compared to that from normal cells. The distribution of recovered radiolabeled material among the medium, cell surface, and intracellular fractions was similar for both normal and polycystic cells. Analysis of the proteoglycans in polycystic cells demonstrated that 86 and 73% of 35S-labeled macromolecules were of the heparan sulfate type in polycystic and normal cells, respectively. The remainder was chondroitin sulfate. Proteoglycans were characterized using DEAE-Sephacel ion-exchange chromatography, chondroitinase ABC, heparitinase, and nitrous acid digestion followed by Sepharose CL-4B gel permeation chromatography. The majority of radiolabeled material in the medium, cell surface, and intracellular fractions eluted between 0.35 and 0.39 M NaCl. However, a second peak (peak II) that eluted at 0.25 M NaCl was found in the medium from polycystic cells. This peak accounted for 27% of the total macromolecules secreted into the medium. Proteoglycans in the major peak were susceptible to nitrous acid and chondroitinase ABC digestion. A similar proportion of peak II was degraded by chondroitinase ABC. However, the remainder was only slightly susceptible to treatment with nitrous acid or heparitase. In normal cells a small amount of material eluted at a similar low charge; the proteoglycans were the same as those found in the major peak and appeared as a shoulder on this peak. 相似文献