Field Monitoring of Curved Girder Bridges with Integral Abutments, TPF-5(169), 2014

Nationally, there are questions regarding the design, fabrication, and erection of horizontally curved steel girder bridges due to unpredicted girder displacements, fit-up, and locked-in stresses. One reason for the concerns is that up to one-quarter of steel girder bridges are being designed with horizontal curvature. There is also an urgent need to reduce bridge maintenance costs by eliminating or reducing deck joints, which can be achieved by expanding the use of integral abutments to include curved girder bridges. However, the behavior of horizontally curved bridges with integral abutments during thermal loading is not well known nor understood. The purpose of this study was to investigate the behavior of horizontal curved bridges with integral abutment (IAB) and semi-integral abutment bridges (SIAB) with a specific interest in the response to changing temperatures. The long-term objective of this effort is to establish guidelines for the use of integral abutments with curved girder bridges. The primary objective of this work was to monitor and evaluate the behavior of six in-service, horizontally curved, steel-girder bridges with integral and semi-integral abutments. In addition, the influence of bridge curvature, skew and pier bearing (expansion and fixed) were also part of the study. Two monitoring systems were designed and applied to a set of four horizontally curved bridges and two straight bridges at the northeast corner of Des Moines, Iowa—one system for measuring strains and movement under long term thermal changes and one system for measuring the behavior under short term, controlled live loading. A finite element model was developed and validated against the measured strains. The model was then used to investigate the sensitivity of design calculations to curvature, skew and pier joint conditions. The general conclusions were as follows: (1) There were no measurable differences in the behavior of the horizontally curved bridges and straight bridges studied in this work under thermal effects. For preliminary member sizing of curved bridges, thermal stresses and movements in a straight bridge of the same length are a reasonable first approximation. (2) Thermal strains in integral abutment and semi-integral abutment bridges were not noticeably different. The choice between IAB and SIAB should be based on life – cycle costs (e.g., construction and maintenance). (3) An expansion bearing pier reduces the thermal stresses in the girders of the straight bridge but does not appear to reduce the stresses in the girders of the curved bridge. (4) An analysis of the bridges predicted a substantial total stress (sum of the vertical bending stress, the lateral bending stress, and the axial stress) up to 3 ksi due to temperature effects. (5) For the one curved integral abutment bridge studied at length, the stresses in the girders significantly vary with changes in skew and curvature. With a 10⁰ skew and 0.06 radians arc span length to radius ratio, the curved and skew integral abutment bridges can be designed as a straight bridge if an error in estimation of the stresses of 10% is acceptable.

Field Observation of Five Lightweight Aggregate Pretensioned Prestressed Bridge Beams, HR-104, 1969

The use of lightweight aggregates in pretensioned prestressed concrete beams is becoming more advantageous as our design criteria dictate longer span concrete bridges. Bridge beams of greater lengths have been restricted from travel on many of our highways because the weight of the combined beams and transporting vehicle was excessive, making hauls of any distance prohibitive. This, along with the fact that new safety requirements necessitate the use of longer spans in grade separation structures over major highways, prompted the State of Iowa to investigate the use of lightweight aggregate bridge beams. The objective of this project is the collection of field deflection measurements for five pretensioned prestressed lightweight aggregate concrete bridge beams fabricated by conventional plant processes; also the comparison of the actual cambers and deflections of the beams with that predicted from the design assumptions.

Investigation of Two Bridge Alternatives for Low Volume Roads Volume 2 of 2 Concept 2: Beam In Slab Bridge, HR-382, 1997

Recent reports have indicated that 23.5% of the nation's highway bridges are structurally deficient and 17.7% are functionally obsolete. A significant number of these bridges are on the Iowa secondary road system where over 86% of the rural bridge management responsibilities are assigned to the counties. Some of the bridges can be strengthened or otherwise rehabilitated, but many more are in need of immediate replacement. In a recent investigation (HR-365 "Evaluation of Bridge Replacement Alternatives for the County Bridge System") several types of replacement bridges that are currently being used on low volume roads were identified. It was also determined that a large number of counties (69%) have the ability and are interested in utilizing their own forces to design and construct short span bridges. After reviewing the results from HR-365, the research team developed one "new" bridge replacement concept and a modification of a replacement system currently being used. Both of these bridge replacement alternatives were investigated in this study, the results of which are presented in two volumes. This volume (Volume 2) presents the results of Concept 2 - Modification of the Beam-in-Slab Bridge. Concept 1 - Steel Beam Precast Units is presented in Volume 1. Concept 2 involves various laboratory tests of the Beam-in-Slab bridge (BISB) currently being used by Benton County and several other Iowa counties. In this investigation, the behavior and strength of the BISB were determined; a new method of obtaining composite action between the steel beams and concrete was also tested. Since the Concept 2 bridge is primarily intended for use on low-volume roads, the system can be constructed with new or used beams. In the experimental part of the investigation, there were three types of laboratory tests: push-out tests, service and ultimate load tests of models of the BISB, and composite beam tests utilizing the newly developed shear connection. In addition to the laboratory tests, there was a field test in which an existing BISB was service load tested. An equation was developed for predicting the strength of the shear connection investigated; in addition, a finite element model for analyzing the BISB was also developed. Push-out tests were completed to determine the strength of the recently developed shear connector. A total of 36 specimens were tested, with variables such as hole diameter, hole spacing, presence of reinforcement, etc. being investigated. In the model tests of the BISB, two and four beam specimens [L=9,140 mm (30 ft)] were service load tested for behavior and load distribution data. Upon completion of these tests, both specimens were loaded to failure. In the composite beam tests, four beams, one with standard shear studs and three using the shear connection developed, were tested. Upon completion of the service load tests, all four beams were loaded to failure. The strength and behavior of the beams with the new shear connection were found to be essentially the same as that of the specimen with standard shear studs.

Investigation of Two Bridge Alternatives for Low Volume Roads - Phase II Volume 2 of 2, Concept 2: Beam in Slab Bridge, TR-410, 2000

This project continues the research which addresses the numerous bridge problems on the Iowa secondary road system. It is a continuation (Phase 2) of Project HR-382, in which two replacement alternatives (Concept 1: Steel Beam Precast Units and Concept 2: Modification of the Benton County Beam-in-Slab Bridge) were investigated. In previous research for concept 1, a precast unit bridge was developed through laboratory testing. The steel-beam precast unit bridge requires the fabrication of precast double-tee (PCDT) units, each consisting of two steel beams connected by a reinforced concrete deck. The weight of each PCDT unit is minimized by limiting the deck thickness to 4 in., which permits the units to be constructed off-site and then transported to the bridge site. The number of units required is a function of the width of bridge desired. Once the PCDT units are connected, a cast-in-place reinforced concrete deck is cast over the PCDT units and the bridge railing attached. Since the steel beam PCDT unit bridge design is intended primarily for use on low-volume roads, used steel beams can be utilized for a significant cost savings. In previous research for concept 2, an alternate shear connector (ASC) was developed and subjected to static loading. In this investigation, the ASC was subjected to cyclic loading in both pushout specimens and composite beam tests. Based on these tests, the fatigue strength of the ASC was determined to be significantly greater than that required in typical low volume road single span bridges. Based upon the construction and service load testing, the steel-beam precast unit bridge was successfully shown to be a viable low volume road bridge alternative. The construction process utilized standard methods resulting in a simple system that can be completed with a limited staff. Results from the service load tests indicated adequate strength for all legal loads. An inspection of the bridge one year after its construction revealed no change in the bridge's performance. Each of the systems previously described are relatively easy to construct. Use of the ASC rather than the welded studs significantly simplified the work, equipment, and materials required to develop composite action between the steel beams and the concrete deck.

Clinical proteomics : a focus on transformed human T and B lymphocytes

Résumé large public La protéomique clinique est une discipline qui vise l'étude des protéines dans un but diagnostique ou thérapeutique. Nous avons utilisé cette approche pour étudier les lymphocytes T «tueurs » ou cytotoxiques qui font partie des globules blancs du système sanguin et agissent dans la lutte contre les infections et les tumeurs. Ces cellules sont impliquées dans l'immunothérapie cellulaire qui se fonde sur la capacité naturelle des ces lymphocytes à repérer les cellules tumorales et à les détruire. L'introduction du gène de la télomérase dans les lymphocytes T résulte en une prolongation de leur longévité, ce qui en ferait des candidats intéressants pour l'immunothérapie cellulaire. Il subsiste cependant des doutes quant aux conséquences de l'utilisation de ces lymphocytes «immortalisés ». Pour répondre à cette question, nous avons comparé le profile protéique de lymphocytes T cytotoxiques «jeunes » et vieux » avec celui des lymphocytes «immortalisés ». Nous avons trouvé que ces derniers présentent une double face et partagent à la fois les caractéristiques de la jeunesse et de la vieillesse. Dans une seconde étude de protéomique clinique, nous nous sommes penchés sur les lymphocytes B «immortalisés » cette fois-ci non pas avec la télomérase, mais avec le virus d'Epstein-Barr. Ces derniers sont utilisés comme modèle dans l'étude de la leucodystrophie, une maladie génétique rare qui affecte le cerveau. Notre but est d'identifier des marqueurs biologiques potentiels qui pourraient aider le diagnostic et le traitement de cette maladie neurodégénérative. Nous avons pour ce faire comparé les profiles protéiques des lymphocytes B «immortalisés » provenant d'individus sains et malades. Malheureusement, notre analyse n'a pas révélé de différences notoires entre ces deux classes de lymphocytes. Ceci nous permet toutefois de conclure que la maladie n'affecte pas la synthèse des protéines de manière prépondérante dans ces cellules sanguines. En résumé, le travail présenté dans cette thèse montre à la fois le potentiel et les limites de l'analyse des protéines lymphocytaires, dans différentes situations biologiques. Résumé La protéomique clinique ouvre la porte vers de multiples horizons relatifs au traitement de diverses maladies. Ce domaine particulier alliant la protéomique à la médecine, implique l'intervention de la biologie moléculaire et cellulaire. Dans notre étude, nous nous sommes d'abord intéressés aux lymphocytes T CD8+ cytotoxiques dans le contexte de l'immunothérapie adoptive. Le fondement de cette thérapie repose sur la capacité naturelle de ces lymphocytes à reconnaître les cellules tumorales et à les détruire chez les patients atteints de cancer. L'introduction du gène de la transcriptase réverse de la télomérase (hTERT) dans les lymphocytes T humains permet de rallonger leur durée de vie, sans toutefois induire d'altérations liées à la transformation. Cependant, des incertitudes subsistent quant à la ressemblance physiologique et biochimique entre ces cellules surexprirnant la télomérase et les cellules normales. Afin de répondre à cette question, nous avons comparé l'expression des protéines de lymphocytes humains T CD8+ «jeunes » et «vieux »avec celle de lymphocytes transduits avec hTERT. Nous avons trouvé que les lymphocytes T surexprimant la télomérase ont un profile protéique intermédiaire, avec certaines expressions protéiques similaires aux jeunes cellules T et d'autres se rapprochant des cellules vieilles. Dans la seconde partie de notre étude, nous nous sommes intéressés aux lymphocytes B transformés avec le virus d'Epstein-Barr provenant de patients atteints d'une maladie génétique rare du cerveau, la leucodystrophie. Dans cette maladie, des mutations dans le facteur de transcription eIF2B, impliqué dans la synthèse protéique, ont été trouvées. Afin d'analyser les conséquences de ces mutations et de trouver des biomarqueurs spécifiques à cette maladie, nous avons effectué une analyse protéomique des lymphoblastes provenant de malades et d'individus sains. Nous avons trouvé que les mutations dans le complexe ubiquitaire eIF2B n'affectent pas de manière significative l'expression des protéines des lymphoblastes mutés. En conclusion, notre travail illustre le potentiel et les limitations des technologies protéomiques utilisées pour disséquer l'implication des protéines dans différentes situations biologiques. Summary Clinical proteomics opens the door to multiple applications related to the treatment of diseases. This particular field is at the crossroad of proteomics and medicine and involves tools from cellular and molecular biology. We focused first our investigations on cytotoxic T cells in the context of adoptive immunotherapy, which is an interesting and evolving field. The basis of this therapy relies on the natural capacity of cytotoxic CD8+ T lymphocytes in recognizing tumor cells and destroying them in cancer patients. As their number is reduced, the idea would be to use transformed T lymphocytes with extended life span. Overexpression of telomerase into human T lymphocytes results in the extension of their replicative life span, but it still remains unclear whether these cells are physiologically indistinguishable from normal ones. To address this question, we compared the proteome of young and aged CD8+ T lymphocytes with that of T cells transduced with hTERT and found that the latter cells displayed an intermediate protein pattern, sharing similar protein expression with young, but also with aged T cells. We were then interested in studying Epstein-Barr virus transformed B lymphocytes in the context of a rare human brain genetic disorder called leukodystrophy. In this disease, mutations in the ubiquitous factor eIF2B involved in protein synthesis and its regulation have been reported. In order to analyze the functional consequences of the mutations and to find out specific biomarkers of eIF2B-related disorders, proteomic and peptidomic studies were carried out on lymphoblasts from eIF2Bmutated patients versus healthy patients. Following two-dimensional gel electrophoresis and mass fingerprints, mutations in the eIF2B complex did not appear to significantly affect the proteome of the mutated lymphoblasts extracts. To conclude, our work emphasizes the potentials and the limitations of the proteomic technologies used to analyze the role of lymphocyte proteins in different biological situations.

Dynamics of Highway Bridges, HR-67, 1961

The AASHO specifications for highway bridges require that in designing a bridge, the live load must be multiplied by an impact factor for which a formula is given, dependent only upon the length of the bridge. This formula is a result of August Wohler's tests on fatigue in metals, in which he determined that metals which are subjected to large alternating loads will ultimately fail at lower stresses than those which are subjected only to continuous static loads. It is felt by some investigators that this present impact factor is not realistic, and it is suggested that a consideration of the increased stress due to vibrations caused by vehicles traversing the span would result in a more realistic impact factor than now exists. Since the current highway program requires a large number of bridges to be built, the need for data on dynamic behavior of bridges is apparent. Much excellent material has already been gathered on the subject, but many questions remain unanswered. This work is designed to investigate further a specific corner of that subject, and it is hoped that some useful light may be shed on the subject. Specifically this study hopes to correlate, by experiment on a small scale test bridge, the upper limits of impact utilizing a stationary, oscillating load to represent axle loads moving past a given point. The experiments were performed on a small scale bridge which is located in the basement of the Iowa Engineering Experiment Station. The bridge is a 25 foot simply supported span, 10 feet wide, supported by four beams with a composite concrete slab. It is assumed that the magnitude of the predominant forcing function is the same as the magnitude of the dynamic force produced by a smoothly rolling load, which has a frequency determined by the passage of axles. The frequency of passage of axles is defined as the speed of the vehicle divided by the axle spacing. Factors affecting the response of the bridge to this forcing function are the bridge stiffness and mass, which determine the natural frequency, and the effects of solid damping due to internal structural energy dissipation.

Feasibility Study of Dynamic Overload and Ultimate Load Tests of Full-Scale Highway Bridges, HR-160, 1973

As a result of the construction of the Saylorville Dam and Reservoir on the Des Moines River, six highway bridges are scheduled for removal. Five of these are old high-truss single-lane bridges, each bridge having several simple spans. The other bridge is a fairly modern (1955) double 4-span continuous beam-and-slab composite highway bridge. The availability of these bridges affords an unusual opportunity for study of the behavior of full-scale bridges. Because of the magnitude of the potential testing program, a feasibility study was initiated and the results are presented in this two-part final report. Part I summarizes the findings and Part II presents the supporting detailed information.

Problems of Bridge Supporting and Expansion Devices and an Experimental Comparison of the Dynamic Behavior of Rigid and Elastomeric Bearings, HR-105, 1965

The design of satisfactory supporting and expansion devices for highway bridges is a problem which has concerned bridge design engineers for many years. The problems associated with these devices have been emphasized by the large number of short span bridges required by the current expanded highway program of expressways and interstate highways. The initial objectives of this investigation were: (1) To review and make a field study of devices used for the support of bridge superstructures and for provision of floor expansion; (2) To analyze the forces or factors which influence the design and behavior of supporting devices and floor expansion systems; and (3) To ascertain the need for future research particularly on the problems of obtaining more economical and efficient supporting and expansion devices, and determining maximum allowable distance between such devices. The experimental portion was conducted to evaluate one of the possible simple and economical solutions to the problems observed in the initial portion. The investigation reported herein is divided into four major parts or phases as follows: (1) A review of literature; (2) A survey by questionnaire of design practice of a number of state highway departments and consulting firms; (3) Field observation of existing bridges; and, (4) An experimental comparison of the dynamic behavior of rigid and elastomeric bearings.

Structural Characterization of UHPC Waffle Bridge Deck and Connections, TR-614, 2014

The AASHTO strategic plan in 2005 for bridge engineering identified extending the service life of bridges and accelerating bridge construction as two of the grand challenges in bridge engineering. These challenges have the objective of producing safer and more economical bridges at a faster rate with a minimum service life of 75 years and reduced maintenance cost to serve the country’s infrastructure needs. Previous studies have shown that a prefabricated full-depth precast concrete deck system is an innovative technique that accelerates the rehabilitation process of a bridge deck, extending its service life with reduced user delays and community disruptions and lowering its life-cycle costs. Previous use of ultra-high performance concrete (UHPC) for bridge applications in the United States has been considered to be efficient and economical because of its superior structural characteristics and durability properties. Full-depth UHPC waffle deck panel systems have been developed over the past three years in Europe and the United States. Subsequently, a single span, 60-ft long and 33-ft wide prototype bridge with full-depth prefabricated UHPC waffle deck panels has been designed and built for a replacement bridge in Wapello County, Iowa. The structural performance characteristics and the constructability of the UHPC waffle deck system and its critical connections were studied through an experimental program at the structural laboratory of Iowa State University (ISU). Two prefabricated full-depth UHPC waffle deck (8 feet by 9 feet 9 inches by 8 inches) panels were connected to 24-ft long precast girders, and the system was tested under service, fatigue, overload, and ultimate loads. Three months after the completion of the bridge with waffle deck system, it was load tested under live loads in February 2012. The measured strain and deflection values were within the acceptable limits, validating the structural performance of the bridge deck. Based on the laboratory test results, observations, field testing of the prototype bridge, and experience gained from the sequence of construction events such as panel fabrication and casting of transverse and longitudinal joints, a prefabricated UHPC waffle deck system is found to be a viable option to achieve the goals of the AASHTO strategic plan.

The temporoparietal fascia flap: a versatile tool in head and neck reconstruction.

PURPOSE OF REVIEW: The article reviews recent significant advances and current applications of the temporoparietal fascia flap (TPFF) in head and neck surgery. RECENT FINDINGS: The recent literature describes a wide span of new applications of the TPFF in many areas. Significant developments and refinements in the reconstruction of orbitomaxillary composite defects and orbital exenteration cavities are reported. The TPFF combined with alloplastic framework is gaining in importance in external ear reconstruction. Innovative prefabricated skin or soft-tissue grafts based on the TPFF are used to restore facial contour or in the reconstruction of complex facial defects. The free TPFF finds a role in laryngotracheal reconstruction as a vascular carrier to support cartilage grafts. SUMMARY: Owing to its reliability and unequalled structural properties, the TPFF still plays a central role in facial reconstruction. Future investigation will likely incorporate the free TPFF as a vascular carrier of bioengineered tissues, such as cartilage and mucosa, for various head and neck indications.

Iowa Historic Property Study: Iowa-Illinois Memorial Bridge Carrying I-74 and U.S. 6 over Mississippi River between Bettendorf, Scott County, Iowa and Moline, Rock Island County, Illinois, 2012

The 1935 Iowa-Illinois Memorial Bridge is being documented at this time to fulfill the requirements of the Memorandum of Agreement regarding the removal of the Iowa-Illinois Memorial Bridge and the Iowana Farms Milk Company Building for the proposed improvements to Interstate 7 4 in Bettendorf, Iowa, and Moline, Illinois.1 The 1959 twin suspension bridge will be removed as well, but it was determined to be ineligible for the National Register of Historic Places. Discussion of the history of the 1959 twin span is included, however, in the current report as part of the overall history of the Iowa-Illinois Memorial Bridge. Fieldwork for the documentation occurred in November 2009 and October 2010 (Fig. 1). Limitations on photography included limited shoreline access on the Illinois side, making good views of the bridge from the south somewhat challenging. Also, photographs on the bridge deck were not possible because of interstate traffic and prohibitions on pedestrian traffic. Within the last few years, online primary sources have proliferated, along with historical materials regarding the Iowa-Illinois Memorial Bridge. Sources available online for this report included numerous historical photographs, as well as historical Davenport, Iowa, and U.S. newspapers that document the bridge planning and construction. Additional primary source material was found at the University of Iowa Libraries, the State Historical Society of Iowa in Iowa City, the Bettendorf Public Library, the Richardson-Sloane Special Collections Center at the Davenport Public Library, and the Iowa State University Special Collections in Ames.

Symptomatic intracranial hemorrhage after stroke thrombolysis: comparison of prediction scores.

BACKGROUND AND PURPOSE: Several prognostic scores have been developed to predict the risk of symptomatic intracranial hemorrhage (sICH) after ischemic stroke thrombolysis. We compared the performance of these scores in a multicenter cohort. METHODS: We merged prospectively collected data of patients with consecutive ischemic stroke who received intravenous thrombolysis in 7 stroke centers. We identified and evaluated 6 scores that can provide an estimate of the risk of sICH in hyperacute settings: MSS (Multicenter Stroke Survey); HAT (Hemorrhage After Thrombolysis); SEDAN (blood sugar, early infarct signs, [hyper]dense cerebral artery sign, age, NIH Stroke Scale); GRASPS (glucose at presentation, race [Asian], age, sex [male], systolic blood pressure at presentation, and severity of stroke at presentation [NIH Stroke Scale]); SITS (Safe Implementation of Thrombolysis in Stroke); and SPAN (stroke prognostication using age and NIH Stroke Scale)-100 positive index. We included only patients with available variables for all scores. We calculated the area under the receiver operating characteristic curve (AUC-ROC) and also performed logistic regression and the Hosmer-Lemeshow test. RESULTS: The final cohort comprised 3012 eligible patients, of whom 221 (7.3%) had sICH per National Institute of Neurological Disorders and Stroke, 141 (4.7%) per European Cooperative Acute Stroke Study II, and 86 (2.9%) per Safe Implementation of Thrombolysis in Stroke criteria. The performance of the scores assessed with AUC-ROC for predicting European Cooperative Acute Stroke Study II sICH was: MSS, 0.63 (95% confidence interval, 0.58-0.68); HAT, 0.65 (0.60-0.70); SEDAN, 0.70 (0.66-0.73); GRASPS, 0.67 (0.62-0.72); SITS, 0.64 (0.59-0.69); and SPAN-100 positive index, 0.56 (0.50-0.61). SEDAN had significantly higher AUC-ROC values compared with all other scores, except for GRASPS where the difference was nonsignificant. SPAN-100 performed significantly worse compared with other scores. The discriminative ranking of the scores was the same for the National Institute of Neurological Disorders and Stroke, and Safe Implementation of Thrombolysis in Stroke definitions, with SEDAN performing best, GRASPS second, and SPAN-100 worst. CONCLUSIONS: SPAN-100 had the worst predictive power, and SEDAN constantly the highest predictive power. However, none of the scores had better than moderate performance.

The Flexural Fatigue Strength of Prestressed Steel I-Beams, HR-73, 1962

The purpose of this investigation was to study the flexural fatigue strength of two prestressed steel I-beams which had previously been fabricated in connection with a jointly sponsored project under the auspices of the Iowa State Highway Commission. The beams were prestressed by deflecting them under the action of a concentrated load at the center of a simple span, then welding unstressed high strength steel plates to the top and bottom flanges to retain a predetermined amount of prestress. The beams were rolled sections of A36 steel and the plates were USS "T-1" steel. Each of the two test specimens were subjected to an identical repeated loading until a fatigue failure occurred. The loading was designed to produce stresses equivalent to those which would have occurred in a simulated bridge and amounted to 84 percent of a standard H-15 live load including impact. One of the beams sustained 2,469,100 repetitions of load to failure and the other sustained 2,756,100 cycles. Following the fatigue tests, an experimental study was made to determine the state of stress that had been retained in the prestressed steel beams. This information, upon which the calculated stresses of the test could be superimposed, provided a method of correlating the fatigue strength of the beams with the fatigue information available on the two steels involved.

Live Load Deflections in a Prestressed Steel Beam Bridge, HR-74, 1963

This report describes the measurement of dynamic (live load) deflections in a 240' x 30' three span continuous prestressed steel bridge, skewed 30 degrees. The design assumptions and prestressing procedure are described briefly, and the instrumentation and loading are discussed. The actual deflections are presented in tabular form, and the deflections due to the design live load are calculated. The maximum deflections are presented as a ratio of the span length, and the further use of prestressed steel beams is recommended.

Role of lymph node fibroblasts in T cell priming

SummarySecondary lymphoid organs, such as lymph nodes or spleen, are the only places in our body where primary adaptive immune responses are efficiently elicited. These organs have distinct Β and Τ cell rich zones and Τ lymphocytes constantly migrate from the bloodstream into Τ zones to scan dendritic cells (DCs) for antigens they present. Specialized fibroblasts, the Τ zone reticular cells (HR.Cs), span the Τ zone in the form a three-dimensional network. lK.Cs guide incoming Τ cells in their migration, both chemically, by the secretion of the chemokines CCL19 and CCL21, and physically, by construction of a road system to which also DCs adhere. In this way TRCs are thought to facilitate encounters of Τ cells with antigen-bearing DCs and thereby accelerate the selection of rare antigen-specific Τ cells. The resulting Τ cell activation, proliferation and differentiation all take place within the TRC network. However, the influence of TRCs on Τ cell activation has so fer not been elucidated with the possible reasons being that TRCs represent a relative rare cell population and that mice devoid of TRCs have not been described.To circumvent these technical limitations, we established TRC clones and lines to have an abundant source to functionally characterize TRCs. Both the clones and lines show a fibroblastic phenotype, express a surface marker profile comparable to ex vivo TRCs and produce extracellular matrix molecules. However, expression of Ccl19, Ccl21 and ZL-7 is lost and could not be restored by cytokine stimulation. When these TRC clones or lines were cultured in a three-dimensional cell culture system, their morphology changed and resembled that of in vivo TRCs as they formed networks. By adding Τ cells and antigen-loaded DCs to these cultures we successfully reconstructed lymphoid Τ zones that allowed antigen-specific Τ cell activation.To characterize the role of TRCs in Τ cell priming, TRCs were co-cultured with antigen-specific Τ cells in the presence antigen-loaded DCs. Surprisingly, the presence of TRC lines and ex vivo TRCs inhibited rather than enhanced CD8+ Τ cell activation, proliferation and effector cell differentiation. TRCs shared this feature with fibroblasts from non-lymphoid tissues as well as mesenchymal stromal cells. TRCs were identified as a strong source of nitric oxide (NO) thereby directly dampening Τ cell expansion as well as reducing the Τ cell priming capacity of DCs. The expression of inducible NO synthase (iNOS) was up- regulated in a subset of TRCs by both DC-signals as well as interferon-γ produced by primed CD8+ Τ cells. Importantly, iNOS expression was induced during viral infection in vivo in both lymph node TRCs and DCs. Consistent with a role for NO as a negative regulator, the primary Τ cell response was exaggerated in iNOS-/- mice. Our findings highlight that in addition to their established positive roles in Τ cell responses TRCs and DCs cooperate in a negative feedback loop to attenuate Τ cell expansion during acute inflammation.RésuméLes organes lymphoïdes secondaires, comme les ganglions lymphoïdes ou la rate, sont les seuls sites dans notre corps où la réponse primaire des lymphocytes Β et Τ est initiée efficacement. Ces organes ont des zones différentes, riches en cellules Β ou T. Des lymphocytes Τ circulent constamment du sang vers les zones T, où ils échantillonent la surface des cellules dendritiques (DCs) pour identifier les antigènes qu'ils présentent. Des fibroblastes spécialisés - nommés Τ zone reticular cells (TRCs)' forment un réseau tridimensionnel dans la zone T. Les TRCs guident la migration des cellules Τ par deux moyens: chimiquement, par la sécrétion des chimiokines CCL19 et CCL21 et physiquement, par la construction d'un réseau routier en trois dimensions, auquel adhèrent aussi des DCs. Dans ce? cas, on pense que la présence des TRCs facilite les rencontres entre les cellules Τ et les DCs chargées de l'antigène et accélère la sélection des rares cellules Τ spécifiques. Ensuite, l'activation de cellules T, ainsi que la prolifération et la différenciation se produisent toutes à l'intérieur du réseau des TRCs. L'influence des TRCs sur l'activation des cellules T n'est que très peu caractérisée, en partie parce que les TRCs représentent une population rare et que les souris déficientes dans les TRCs n'ont pas encore été découvertes.Pour contourner ces limitations techniques, nous avons établi des clones et des lignées cellulaires de TRC pour obtenir une source indéfinie de ces cellules permettant leur caractérisation fonctionnelle. Les clones et lignées établis ont un phénotype de fibroblaste, ils expriment des molécules de surface similaires aux TRCs ex vivo et produisent de la matrice extracellulaire. Mais l'expression de Ccl19, Ccl21 et 11-7 est perdue et ne peut pas être rétablie par stimulation avec différentes cytokines. Les clones TRC ou les lignées cultivées en un système tridimensionnel de culture cellulaire, montrent une morphologie changée, qui ressemble à celle de TRC ex vivo inclus la construction de réseaux tridimensionnels.Pour caractériser le rôle des TRC dans l'activation des cellules T, nous avons cultivé des TRCs avec des cellules T spécifiques et des DCs chargées avec l'antigène. Etonnamment, la présence des TRC (lignées et ex vivo) inhibait plutôt qu'elle améliorait l'activation, la prolifération et la différenciation des lymphocytes T CDS+. Les TRCs partageaient cette fonction avec des fibr-oblastes des organes non lymphoïdes et des cellules souches du type mésenchymateux. Dans ces conditions, les TRCs sont une source importante d'oxyde nitrique (NO) et par ce fait limitent directement l'expansion des cellules T et réduisent aussi la capacité des DCs à activer les cellules T. L'expression de l'enzyme NO synthase inductible (ïNOS) est régulée à la hausse par des signaux dérivés des DCs et par l'interféron-γ produit par des cellules T de type CD8+ activées. Plus important, l'expression d'iNOS est induite pendant une infection virale in vivo, dans les TRCs et dans les DCs. Par conséquent, la réponse primaire de cellules T est exagérée dans des souris iNOS-/-. Nos résultats mettent en évidence qu'en plus de leur rôle positif bien établi dans la réponse immunitaire, les TRCs et les DCs coopèrent dans une boucle de rétroaction négative pour atténuer l'expansion des cellules T pendant l'inflammation aigiie pour protéger l'intégrité et la fonctionnalité des organes lymphoïdes secondaires.