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PubMed2006

Dichotomic phylogenetic tree of the pyruvate kinase family: K+ -dependent and -independent enzymes.

K+ dependence was assumed to be a feature of all pyruvate kinases until it was discovered that some enzymes express K+ -independent activity. Almost all the K+ -independent pyruvate kinases have Lys at position 117, instead of the Glu present in the K+ -dependent muscle enzyme. Mutagenesis studies show that the internal positive charge substitutes for the K+ requirement (Laughlin, L. T. & Reed, G. H. (1997) Arch. Biochem. Biophys. 348, 262-267). In this work a phylogenetic analysis of pyruvate kinase was performed to ascertain the abundance of K+ -independent activities and to explore whether the K+ activating effect is related to the evolutionary history of the enzyme. Of the 230 studied sequences, 46% have Lys at position 117, and the rest have Glu. Pyruvate kinases with Lys117 and Glu117 are separated in two clusters. All of the enzymes of the Glu117 cluster that have been characterized are K+ -dependent, whereas those of the Lys117 cluster are K+ -independent. Thus, there is a strict correlation between the dichotomy of the tree and the dependence of activity on K+. 77% of the pyruvate kinases that possess Lys117 have Lys113/Gln114; they also have Ile, Val, or Leu at position 120. These residues are replaced by Glu117 and Thr113/Lys114/Thr120 in 80% of K+ -dependent pyruvate kinases. Structural analysis indicates that these residues are in a hinge region involved in the acquisition of the catalytic conformation of the enzyme. The route of conversion from K+ -independent to K+ -dependent pyruvate kinases is described. A plausible explanation of how enzymes developed K+ dependence is put forth.

Amino Acid Sequence↗

PubMed1999

Language-independent automatic acquisition of morphological knowledge from synonym pairs.

Medical words exhibit a rich and productive morphology. Beyond simple inflection, derivation and composition are a common way to form new words. Morphological knowledge is therefore very important for any medical language processing application. Whereas rich morphological resources are available for the English medical language with the UMLS Specialist Lexicon, no such resources are publicly available for French or most other languages. We propose a simple and powerful method to help acquire automatically such knowledge. This method takes advantage of the synonym terms present in medical terminologies. In a bootstrapping step, it detects morphologically related words from which it learns "derivation rules". In an expansion step, it then applies these rules to the whole vocabulary available. Our goal is to acquire data for French and other languages for which they are not available. However, to evaluate the efficiency of the method, we tested it on English in a setting which is close to that prevailing for French, and we confronted its results to those obtained with the Specialist lexical variant generation tool.

Electronic Data Processing↗

PubMed2005

Ceftiofur-resistant Salmonella strains isolated from dairy farms represent multiple widely distributed subtypes that evolved by independent horizontal gene transfer.

Salmonella is the leading cause of known food-borne bacterial infections in the United States, with an incidence rate of approximately 15 cases per 100,000 people. The rise of antimicrobial-resistant Salmonella subtypes, including the appearance of subtypes resistant to ceftriaxone, represents a particular concern. Ceftriaxone is used to treat invasive cases of Salmonella in children and is closely related to ceftiofur, an antibiotic commonly used to treat diseases of cattle. In order to develop a better understanding of the evolution and transmission of ceftiofur resistance in Salmonella, we characterized ceftiofur-resistant and -sensitive Salmonella isolates from seven New York dairy farms. A total of 39 isolates from these seven farms were analyzed for evolutionary relatedness (by DNA sequencing of the Salmonella genes fimA, manB, and mdh), antibiotic resistance profiles, and the presence of bla(CMY-2), a beta-lactamase gene associated with resistance to cephalosporins. Our data indicate that (i) resistance to ceftriaxone and ceftiofur was highly correlated with the presence of bla(CMY-2); (ii) ceftiofur-resistant Salmonella strains were geographically widespread, as shown by their isolation from farms located throughout New York State; (iii) ceftiofur-resistant Salmonella strains isolated from farms represent multiple distinct subtypes and evolutionary lineages, as determined by serotyping, DNA sequence typing, and antimicrobial-resistance profiles; and (iv) ceftiofur-resistant Salmonella strains evolved by multiple independent acquisitions of an identical bla(CMY-2) allele and by clonal spread of ceftiofur-resistant subtypes.

Alleles↗

PubMed1998

Molecular phylogeny of arthropods and their relatives: polyphyletic origin of arthropodization.

Because the enigmatic arthropod relatives, such as tardigrades, onychophorans, pentastomids, and pycnogonids, have features characteristic of not only both major arthropod groups but also several other animal phyla, analysis of their phylogenetic positions may provide important clues for metazoan phylogeny in general. From analyses of complete or nearly complete 18S rDNA sequences, arthropod monophyly including all the traditional arthropod relatives was well supported. For detailed analyses within arthropods without outgroup effects, we made two reduced alignments. From the analyses of these reduced data sets, arthropods were divided into two clades: one was chelicerates and myriapods, whereas the other included crustaceans plus insects, onychophorans plus pentastomids, and tardigrades. The pycnogonids were grouped closely with the chelicerates. The Cambrian fossil record supports that myriapod-chelicerate and crustacean-insect (including tardigrades, onychophorans, and pentastomids) stems arose separately from a protoarthropod (perhaps nonarthropodized animal, such as a lobopod which has noncalcified cuticles, and nonjointed legs). Thus, the acquisition of truly arthropod characters occurred independently in the two stems. In this case, nonarthropodized characters of tardigrades, onychophorans, and pentastomids are assumed to be primitive.

Animals↗

PubMed2000

Longitudinal risk of std acquisition in adolescent girls using a generalized estimating equations model

Background: To date, few studies have used a longitudinal approach in examining risk factors for STD acquisition in teenage girls. In the present study, we examined risk of STD acquisition in adolescent girls over a three-year period using longitudinal analyses.Methods: One hundred and seventy-four girls between the ages of 12 and 16 (mean age = 14.5) participated in a longitudinal study of adolescent romantic relationships. The racial composition of the subject pool was 80% African-American and 20% Caucasian. These girls were followed for three years at six-month intervals. By the end of the study, 127 of the girls were sexually experienced, and thus, were included in the analyses regarding risk for STDs. The risk for history of STD was evaluated using a generalized estimating equations (GEE) model, which allows for the inclusion of data from subjects with missing visits. The following independent variables were entered into the model: age, race, screening IQ score, qualitative cognitive functioning, perceptions of STD prevalence among female friends, and whether a condom was used at last intercourse. Additional independent variables included number of lifetime partners (4 or more lifetime partners, 2 to 3 lifetime partners, only one partner) and age of sexual debut (less than 14 years, 14-16 years, and 17 years and older), five factors from the Family Environment Scale, and three types of parental monitoring (direct, direct when with peers, and indirect).Results: The results of the longitudinal (GEE) model indicated that having a history of an STD was significantly related to a younger age of sexual debut (p <.01), having more partners (p <.0001), being African-American (p =. 01), and having a lower screening IQ score (p <.01).Conclusions: These results suggest that risk of STD is associated with both modifiable and non-modifiable variables. Race and intellectual functioning independently contributed to STD risk, stressing the importance of utilizing prevention programs that are culturally and developmentally specific. For example, less intelligent girls may need to have information presented in a more concrete manner, with a focus on the present rather than the future. The targets of these prevention programs should continue to emphasize delaying initiation, as well as partner selection. Finally, the fact that variables for STD acquisition in this longitudinal model are comparable to previous cross-sectional studies of STD acquisition suggests that these are potent predictors of risk.

Journal Article↗

PubMed1986

Influence of V1 and V3 isomyosins on the mechanical behaviour of rat papillary muscle as studied by pseudo-random binary noise modulated length perturbations.

Experiments were done on four-week-old rats, containing biochemically verified V1 only, and thyroidectomized adult rats, treated with propylthiouracil, verified to contain V3 only. Contracture tension was induced in isolated papillary muscles either by high potassium solution or 0.5 mmol l-1 Ba2+. Small amplitude length perturbations with peak-to-peak value not exceeding 0.15% L0 were applied to the activated muscle. Both the applied length perturbations and the corresponding resulting force changes were analysed by computer for dynamic stiffness and phase values. In order to reduce data acquisition time, pseudo-random binary noise length changes, rather than the conventional sinusoidal length changes, were used. The plot of the dynamic stiffness against frequency displays a minimum, akin to a resonance phenomenon. The frequency, fmin, at which this resonance occurs, reflects crossbridge kinetics. It was found that the fmin values for the two types of papillary muscles differed by a factor of two. Experiments were also done on chemically skinned muscles containing V1 or V3 isomyosin activated by different concentrations of either barium or calcium ions. It was found that fmin values of skinned fibres were higher than those obtained from intact fibres. However, for each type of muscle the fmin was independent of the activator used as well as the level of activation. The ratio of fmin for V3 to that for V1 remained the same as for intact preparations. We conclude that the difference in mechanical parameters did not arise a possible difference in excitation-contraction coupling mechanism, but rather is a difference in the dynamic properties of the two types of crossbridges.

Animals↗

PubMed2005

Bayesian phylogenetic analysis reveals two-domain topology of S-adenosylhomocysteine hydrolase protein sequences.

S-Adenosylhomocysteine hydrolase (SahH) is involved in the degradation of the compound which inhibits methylation reactions. Using a Bayesian approach and other methods, we reconstructed a phylogenetic tree of amino acid sequences of this protein originating from all three major domains of living organisms. The SahH sequences formed two major branches: one composed mainly of Archaea and the other of eukaryotes and majority of bacteria, clearly contradicting the three-domain topology shown by small subunit rRNA gene. This topology suggests the occurrence of lateral transfer of this gene between the domains. Poor resolution of eukaryotes and bacteria excluded an ultimate conclusion in which out of the two domains this gene appeared first, however, the congruence of the secondary branches with SS rRNA and/or concatenated ribosomal protein datasets phylogenies suggested an "early" acquisition by some bacterial and eukaryotic phyla. Similarly, the branching pattern of Archaea reflected the phylogenies shown by SS rRNA and ribosomal proteins. SahH is widespread in Eucarya, albeit, due to reductive evolution, it is missing in the intracellular parasite Encephalitozoon cuniculi. On the other hand, the lack of affinity to the sequences from the alpha-Proteobacteria and cyanobacteria excludes a possibility of its acquisition in the course of mitochondrial or chloroplast endosymbioses. Unlike Archaea, most bacteria carry MTA/SAH nucleosidase, an enzyme involved also in metabolism of methylthioadenosine. However, the double function of MTA/SAH nucleosidase may be a barrier to ensure the efficient degradation of S-adenosylhomocysteine, specially when the intensity of methylation processes is high. This would explain the presence of S-adenosylhomocysteine hydrolase in the bacteria that have more complex metabolism. On the other hand, majority of obligate pathogenic bacteria due to simpler metabolism rely entirely on MTA/SAH nucleosidase. This could explain the observed phenetic pattern in which bacteria with larger (>6 Mb-million base pairs) genomes carry SAH hydrolase, whereas bacteria that have undergone reductive evolution usually carry MTA/SAH nucleosidase. This suggests that the presence or acquisition of S-adenosylhomocysteine hydrolase in bacteria may predispose towards higher metabolic, and in consequence, higher genomic complexity. The good examples are the phototrophic bacteria all of which carry this gene, however, the SahH phylogeny shows lack of congruence with SSU rRNA and photosyntethic genes, implying that the acquisition was independent and presumably preceded the acquisition of photosyntethic genes. The majority of cyanobacteria acquired this gene from Archaea, however, in some species the sahH gene was replaced by a copy from the beta- or gamma-Proteobacteria.

Adenosylhomocysteinase↗

PubMed1994

Phylogeny of the malarial genus Plasmodium, derived from rRNA gene sequences.

Malaria is among mankind's worst scourges, affecting many millions of people, particularly in the tropics. Human malaria is caused by several species of Plasmodium, a parasitic protozoan. We analyze the small subunit rRNA gene sequences of 11 Plasmodium species, including three parasitic to humans, to infer their evolutionary relationships. Plasmodium falciparum, the most virulent of the human species, is closely related to Plasmodium reichenowi, which is parasitic to chimpanzee. The estimated time of divergence of these two Plasmodium species is consistent with the time of divergence (6-10 million years ago) between the human and chimpanzee lineages. The falciparum-reichenowi clade is only remotely related to two other human parasites, Plasmodium malariae and Plasmodium vivax, which are also only remotely related to each other. Thus, the parasitic associations of the Plasmodium species with their human hosts are phylogenetically independent. The remote phylogenetic relationship between the two bird parasites, Plasmodium gallinaceum and Plasmodium lophurae, and any of the human parasites provides no support for the hypothesis that infection by Plasmodium falciparum is a recent acquisition of humans, possibly coincident with the onset of agriculture.

Animals↗

PubMed1979

Human rotavirus in an adult population with travelers' diarrhea and its relationship to the location of food consumption.

The role of human rotavirus in adult diarrhea was evaluated in 164 newly arrived US students attending summer school at an urban Mexican university. Rotavirus was identified in stool samples by electron microscopy. Rotavirus was found in 26 of 109 students with diarrhea (24%) and in 8 of 55 asymptomatic control students (15%). Although bacterial pathogens were recovered from virus positive students with diarrhea, viral shedding also occurred independently of other agents. Clinical disease in students excreting only rotavirus tended to be mild and was accompanied by a low density of viral shedding. Food consumption in the home and at public eating establishments was examined the week before illness. While the location of food consumption was found to be important in the acquisition of diarrhea, there was no apparent relationship of the site where meals were eaten and the acquisition of rotavirus by students newly arrived in Mexico. These data support our previous study in a US student population residing in a rural setting in Mexico and implicate rotavirus as a cause of diarrhea among students traveling to Mexico from the United States. The present study offers additional evidence that rotavirus infection in this population might be spread by a nonfood vehicle of transmission which differs from spread of enterotoxigenic E coli, Shigella, or Salmonella strains in the same population.

Adolescent↗

PubMed1998

High-resolution venography of the brain using magnetic resonance imaging.

The purpose of this study was to evaluate a non-flow related magnetic resonance imaging method to visualize small veins independent of arteries in the human brain. A long TE, high-resolution 3D gradient echo MR acquisition was used to highlight venous information. The method is based on the paramagnetic property of deoxyhemoglobin and the resulting phase difference between veins and brain parenchyma at long echo times. The MR magnitude images were masked with a phase mask filter to enhance small structure visibility. Venous information down to sub-pixel vessel diameters of several hundred microns is visible. Venous data are displayed in an angiographic manner using a minimum intensity projection algorithm. Both superficial veins and deep white matter veins are visible. The method has been successfully applied in volunteers. Preliminary results in patients with cerebral arteriovenous malformations indicate its potential in clinical applications. The proposed method is easy to implement and does not require administration of a contrast agent or application of specially designed rf pulses to highlight the veins. Rather it exploits the intrinsic magnetic properties (BOLD-effect) and the prolonged T2* of venous blood. The method may be of diagnostic potential in the assessment of arteriovenous malformations or other vascular venous lesions.

Adolescent↗

PubMed1994

Clinical experience of Hokkaido University-PACS and FCR-angiography.

Three years' experience with Hokkaido University-PACS (HU-PACS) is reported. In particular, this paper describes the suitability of FCR-angiography for HU-PACS, which has been in clinical use since March 1991. Image quality of FCR-arteriograms was evaluated in the head-and-face region and the abdominal region independently. The image quality in both regions was excellent. Quality of transferred images to image workstation for HU-PACS with 10:1 data compression was also evaluated, and no appreciable image degradation or loss of information was found in the transferred images. There was no significant difference in the examination time required for one patient in abdominal angiography between conventional angiography and FCR-angiography. In summary, FCR-angiography is suitable for HU-PACS as its image acquisition modality.

Abdomen↗

PubMed1991

Personal relevance and the human right hemisphere.

Brain damage can selectively disrupt or distort information and ability across the range of human behaviors. One domain that has not been considered as an independent attribute consists of acquisition and maintenance of personal relevant entities such as "familiar" faces, persons, voices, names, linguistic expressions, handwriting, topography, and so on. In experimental studies of normal mentation, personal relevance is revealed in studies of emotion, arousal, affect, preference and familiarity judgments, and memory. Following focal brain damage, deficits and distortions in the experience of personal relevance, as well as in recognizing formerly personally relevant phenomena, are well known to occur. A review and interpretation of these data lead to a proposal that the right hemisphere has a special role in establishing, maintaining, and processing personally relevant aspects of the individual's world.

Agnosia↗

PubMed1991

Computer software for the on-line measurement of the left ventricular end-systolic pressure-dimension relationship.

A microcomputer system was designed to measure the end-systolic pressure-dimension relationship (ESPDR), an index of cardiac contractility that is independent of preload, afterload, and heart rate. To test the system, pressure-dimension data were obtained from swine left ventricles and from a mathematical model of the heart. Algorithms for filtering, location of end-systole, selection of the measurement interval, and calculation of the ESPDR were evaluated on the basis of speed, precision, accuracy, and robustness. The resulting program runs on an IBM-AT and measures ESPDR on-line within 60 seconds of the start of data acquisition. By reducing the time spent in data analysis and providing rapid feedback of information, the on-line software has increased productivity and facilitated improvements in experimental technique.

Algorithms↗

PubMed2006

Novel fast semi-automated software to segment cartilage for knee MR acquisitions.

OBJECTIVE: Validation of a new fast software technique to segment the cartilage on knee magnetic resonance (MR) acquisitions. Large studies of knee osteoarthritis (OA) will require fast and reproducible methods to quantify cartilage changes for knee MR data. In this report we document and measure the reproducibility and reader time of a software-based technique to quantify the volume and thickness of articular cartilage on knee MR images. METHODS: The software was tested on a set of duplicate sagittal three-dimensional (3D) dual echo steady state (DESS) acquisitions from 15 (8 OA, 7 normal) subjects. The repositioning, inter-reader, and intra-reader reproducibility of the cartilage volume (VC) and thickness (ThC) were measured independently as well as the reader time for each cartilage plate. The root-mean square coefficient of variation (RMSCoV) was used as metric to quantify the reproducibility of VC and mean ThC. RESULTS: The repositioning RMSCoV was as follows: VC=2.0% and ThC=1.2% (femur), VC=2.9% and ThC=1.6% (medial tibial plateau), VC=5.5% and ThC=2.4% (lateral tibial plateau), and VC=4.6% and ThC=2.3% (patella). RMSCoV values were higher for the inter-reader reproducibility (VC: 2.5-8.6%) (ThC: 1.9-5.2%) and lower for the intra-reader reproducibility (VC: 1.6-2.5%) (ThC: 1.2-1.9%). The method required an average of 75.4min per knee. CONCLUSIONS: We have documented a fast reproducible semi-automated software method to segment articular cartilage on knee MR acquisitions.

Cartilage, Articular↗

PubMedDate not supplied

[Further comment on the coccidian nature of cryptosporidia (Sporozoa: Apicomplexa)].

The coccidian nature of the genus Cryptosporidium was undoubtedly accepted by Tyzzer who was the first to describe this sporozoan parasite in 1907. Electron microscopic studies made in 70-90s demonstrated the intracellular, although extracytoplasmic localization of Cryptosporidium spp. The pattern of Cryptosporidium life cycle fits well that of other intestinal homogeneous coccidian genera of the suborder Eimeriina: macro- and microgamonts develop independently, a microgamont gives rise to numerous male gametes, oocysts serving for parasite's spreading in the environment. Along with these characters, Cryptosporidium spp. demonstrate some secondary peculiarities (an endogenous phase of development in microvilli of epithelial surfaces, two morphofunctional types of oocysts, the smallest number of sporozoites per oocyst, a multi-membraneous "feeder" organelle etc.), which may be due presumably to their early acquisition of specialization in the course of evolution. The recent studies based on molecular sequence data (18S rRNA) applied to 8 eimeriid and isosporid coccidian genera (Morrison, Ellis, 1997), suggested that the subclass Coccidia (class, according to Morrison and Ellis) be considered monophylic if Cryptosporidium were excluded, and this genus was regarded as the sister group to the rest of the Apicomplexa, or as the sister to the suborder (class) Hematozoa within the Apicomplexa. Either of these placements of Cryptosporidium definitely conflicts with both the generally accepted taxonomic scheme by Levine (1982) and the phenotypically based phylogeny of the phylum Apicomplexa (Barta e. a., 1990). The author's opinion is that the differences between the examined eimeriid and isosporid coccidia, on the one hand, and Cryptosporidium, on the other hand, provided by molecular sequence data, may testify primarily to the well known morphofunctional dissimilarities between the compared organisms, rather than cast doubt on the coccidian nature of Cryptosporidium. Again, these data can hardly prove that Cryptosporidium does not belong to the coccidia. Thus, the modern molecular sequence data, despite their obvious scientific value, would make sense for phylogeny estimation only, if they are critically analysed and considered in combination with results of the relevant basic research.

Animals↗

PubMed1990

Tracking the spread of the HIV infection epidemic among young adults in the United States: results of the first four years of screening among civilian applicants for U.S. military service.

Because the period from infection to clinically apparent disease is long, variable, and changing as new therapies are developed and applied, AIDS data are inadequate for tracking current values of critical parameters of HIV infection epidemics: prevalence of infection, rate of acquisition of new infections (incidence rate), and direction and rate of change of infection incidence over time (acceleration). These "vital signs" of infection epidemics can be tracked using serial cross-sectional seroprevalence data, however. From October 1985 through September 1989, more than 2.3 million applicants for U.S. military service were screened for antibody to HIV. The overall seroprevalence was 1.31 per 1,000 (3,014/2,300,675). Seroprevalences were highest near urban centers of the AIDS epidemic and were independently associated with age, race/ethnicity, and gender. Based on age seroprevalence trends, it was crudely estimated that at least one of 2,000 young men and one of 7,000 young women are infected with HIV annually in the U.S. Infection incidence rates, estimated from age and temporal trends, were estimated to be highest among black males (1.40/1,000/year) and lowest among white females (0.03/1,000/year). Poisson regression analysis of seroprevalence trends suggested that infection incidence rates accelerated among black females during the first 3 years of screening. Since selection factors undoubtedly changed over the period, estimates based on these data probably underestimate actual values in the general population, particularly near urban AIDS epicenters. Nonetheless, even crude estimates of these critical values, particularly among adolescents and young adults, are useful to guide policy development, to allocate resources, and to monitor program effects.

Acquired Immunodeficiency Syndrome↗

PubMed2006

High spatial-resolution CE-MRA of the carotid circulation with parallel imaging: comparison of image quality between 2 different acceleration factors at 3.0 Tesla.

PURPOSE: We sought to evaluate and compare the image quality and vessel delineation of the carotid arteries with high spatial-resolution contrast-enhanced MRA (CE-MRA) at 3.0 T using integrated parallel acquisition (iPAT) with acceleration factors of 2 and 4. MATERIALS AND METHODS: Using an 8-channel neurovascular array coil, we performed prospective high-spatial resolution CE-MRA at 3.0 T of the head and neck on 24 patients (11 men, 13 women, ages 37-89) with suspected arterio-occlusive disease who were assigned randomly to 2 groups. Twelve patients (group A) were examined with a 3D-GRE sequence using iPAT with acceleration factor of 2. For the next 12 patients (group B) a near-identical sequence with an acceleration factor of 4 was applied. Higher iPAT factors were used to increase the spatial-resolution while keeping scan time unchanged. Two volunteers were scanned by both protocols. Phantom measurements were performed to assess the signal-to-noise ratio (SNR). The presence of artifact, noise, image quality of the arterial segments, and the presence and degree of arterial stenosis were evaluated independently by 2 radiologists. Statistical analysis of data was performed by using Wilcoxon rank sum test and 2-sample Student t test (P < 0.05 was indicative a statistically significant difference). The interobserver variability was tested by kappa coefficient. RESULTS: SNR values were significantly lower when iPAT with acceleration factor of 4 was used (P < 0.001). There was no significant difference between 2 groups in regards to image noise (P = 0.67) and artifact (P = 0.8). Both readers visualized the majority of carotid circulation with good image quality in both groups. For smaller intracranial arteries, such as the second-division of anterior and middle cerebral artery, anterior communicating artery, and superior cerebellar artery, the image quality and vessel delineation was significantly better at an iPAT factor of 4 (P < 0.01). The overall interobserver agreement for both the vessel depiction, and detection of arterial stenoses was higher in group B compared with group A. CONCLUSION: Use of parallel acquisition techniques with a high acceleration factor (iPAT-4) results in superior depiction of small intracranial arterial segments. Imaging at higher magnetic field strength, in addition to the use of an optimized 8-channel array coil, provides sufficient SNR to support faster parallel acquisition protocols, leading to improved spatial-resolution. More extensive clinical studies are warranted to establish the range of applications and confirm the accuracy of the technique.

Adult↗

PubMed2006

CD4+ T regulatory cell induction and function in transplant recipients after CD154 blockade is TLR4 independent.

Although the role of CD4(+) T regulatory cells (Treg) in transplantation tolerance has been established, putative mechanisms of Treg induction and function in vivo remain unclear. TLR4 signaling has been implicated in the regulation of CD4(+)CD25(+) Treg functions recently. In this study, we first examined the role of recipient TLR4 in the acquisition of operational CD4(+) Treg following CD154 blockade in a murine cardiac transplant model. Then, we determined whether TLR4 activation in allograft tolerant recipients would reverse alloimmune suppression mediated by CD4(+) Treg. We document that donor-specific immune tolerance was readily induced in TLR4-deficient recipients by a single dose of anti-CD154 mAb, similar to wild-type counterparts. The function and phenotype of CD4(+) Treg in both wild-type and TLR4 knockout long-term hosts was demonstrated by a series of depletion experiments examining their ability to suppress the rejection of secondary donor-type test skin grafts and to inhibit alloreactive CD8(+) T cell activation in vivo. Furthermore, TLR4 activation in tolerant recipients following exogenous LPS infusion in conjunction with donor-type skin graft challenge, failed to break Treg-mediated immune suppression. In conclusion, our data reveals a distinctive property of CD4(+) Treg in tolerant allograft recipients, whose induction and function are independent of TLR4 signaling.

Animals↗