High Throughput Sequencing and Proteomics to Identify Immunogenic Proteins of a New Pathogen: The Dirty Genome Approach

BACKGROUND:With the availability of new generation sequencing technologies, bacterial genome projects have undergone a major boost. Still, chromosome completion needs a costly and time-consuming gap closure, especially when containing highly repetitive elements. However, incomplete genome data may be sufficiently informative to derive the pursued information. For emerging pathogens, i.e. newly identified pathogens, lack of release of genome data during gap closure stage is clearly medically counterproductive. METHODS/PRINCIPAL FINDINGS:We thus investigated the feasibility of a dirty genome approach, i.e. the release of unfinished genome sequences to develop serological diagnostic tools. We showed that almost the whole genome sequence of the emerging pathogen Parachlamydia acanthamoebae was retrieved even with relatively short reads from Genome Sequencer 20 and Solexa. The bacterial proteome was analyzed to select immunogenic proteins, which were then expressed and used to elaborate the first steps of an ELISA. CONCLUSIONS/SIGNIFICANCE:This work constitutes the proof of principle for a dirty genome approach, i.e. the use of unfinished genome sequences of pathogenic bacteria, coupled with proteomics to rapidly identify new immunogenic proteins useful to develop in the future specific diagnostic tests such as ELISA, immunohistochemistry and direct antigen detection. Although applied here to an emerging pathogen, this combined dirty genome sequencing/proteomic approach may be used for any pathogen for which better diagnostics are needed. These genome sequences may also be very useful to develop DNA based diagnostic tests. All these diagnostic tools will allow further evaluations of the pathogenic potential of this obligate intracellular bacterium

Imaging of Bubonic Plague Dynamics by In Vivo Tracking of Bioluminescent Yersinia pestis

Yersinia pestis dissemination in a host is usually studied by enumerating bacteria in the tissues of animals sacrificed at different times. This laborious methodology gives only snapshots of the infection, as the infectious process is not synchronized. In this work we used in vivo bioluminescence imaging (BLI) to follow Y. pestis dissemination during bubonic plague. We first demonstrated that Y. pestis CO92 transformed with pGEN-luxCDABE stably emitted bioluminescence in vitro and in vivo, while retaining full virulence. The light produced from live animals allowed to delineate the infected organs and correlated with bacterial loads, thus validating the BLI tool. We then showed that the first step of the infectious process is a bacterial multiplication at the injection site (linea alba), followed by a colonization of the draining inguinal lymph node(s), and subsequently of the ipsilateral axillary lymph node through a direct connection between the two nodes. A mild bacteremia and an effective filtering of the blood stream by the liver and spleen probably accounted for the early bacterial blood clearance and the simultaneous development of bacterial foci within these organs. The saturation of the filtering capacity of the spleen and liver subsequently led to terminal septicemia. Our results also indicate that secondary lymphoid tissues are the main targets of Y. pestis multiplication and that colonization of other organs occurs essentially at the terminal phase of the disease. Finally, our analysis reveals that the high variability in the kinetics of infection is attributable to the time the bacteria remain confined at the injection site. However, once Y. pestis has reached the draining lymph nodes, the disease progresses extremely rapidly, leading to the invasion of the entire body within two days and to death of the animals. This highlights the extraordinary capacity of Y. pestis to annihilate the host innate immune response

La Distribution

Au début des années 1980, une enquête menée auprès de metteurs en scène pose la question des modalités, concrètes et symboliques, de la distribution. Mis à part cette enquête, exclusivement ancrée dans le champ théâtral, aucune étude systématique n’a été consacrée à la distribution dans les arts de la scène, alors même que celle-ci engage et articule des questionnements décisifs, au croisement de l’artistique, du politique et de l’économique. (…

Louis Pasteur. Papiers. V — CORRESPONDANCE. CLXXV-CLXXXVI Correspondance de Louis Pasteur. CLXXX-CLXXXVI Lettres reçues. CLXXXVI Suzor - Zévort.

Croquis. Louis PasteurLaboratoire. Expériences de Louis PasteurPasteur, Louis, de l'Académie française. PapiersPasteur, Louis, de l'Académie française. Cahier(s) de laboratoirePasteur, Louis, de l'Académie française. CroquisPasteur, Marie Laurent, Mme Louis. Portrait(s)Pasteur Vallery-Radot (Professeur et Mme Louis). Manuscrit(s) provenant d'euxSciences. Louis PasteurContient : Pasteur, Louis, de l'Académie française. Document(s) le concernant ; Terrel des Chênes, E., viticulteur. Lettre(s) ; Thiers, Adolphe, de l'Académie française, homme politique. Lettre(s) ; Thomas, Ambroise, compositeur. Lettre(s) ; Thuillier, Alexandre. Lettre(s) ; Thuillier, Françoise. Lettre(s) ; Thuillier, Louis. Lettre(s) ; Thuillier, Louis. Portrait(s) ; Thuillier, Louis. Lettre(s) reçue(s) ; Verrier, vétérinaire à Provins. Lettre(s) ; Tisserand, Louis-Eugène. Lettre(s) ; Guillemot, Gabriel, pseud. Guy Tomel. Lettre(s) ; Toussaint, Docteur Henry. Lettre(s) ; Tyndall, John, physicien. Lettre(s) ; Strachey, Général, de l'India Office. Lettre(s) ; Tyndall, John, physicien. Lettre(s) reçue(s) ; Brown, Horatio F., historien. Lettre(s) ; Simon, Docteur John. Lettre(s) ; Vaillant, Jean-Baptiste-Philibert, maréchal de France. Lettre(s) ; Vaillard, Docteur Louis, professeur au Val-de-Grâce. Lettre(s) ; Vallery-Radot, Camille. Lettre(s) ; Vallery-Radot, René, écrivain. Lettre(s) ; Roux, Docteur Émile, membre de l'Institut. Lettre(s) ; Van Tieghem, Philippe-Édouard-Léon, membre de l'Institut. Lettre(s) ; Velten, E., brasseur à Marseille. Lettre(s) ; Vercel, Jules. Lettre(s) ; Vergnette de Lamotte, Vicomte Alfred, agronome. Lettre(s) ; Viant, Joseph-Justin. Lettre(s) ; Viala, Eugène, préparateur de Pasteur. Lettre(s) ; Marès, Henri-Pierre-Louis, chimiste. Lettre(s) ; Vialla, Louis, président de la Société d'agriculture de l'Hérault. Lettre(s) ; Ministère de l'Instruction publique. Lettre(s) reçue(s) ; Vignon, Léo, chimiste. Lettre(s) ; Villemin, Docteur Jean-Antoine, de l'Académie de médecine. Lettre(s) ; Violette, Henri, commissaire à la Raffinerie impériale de salpêtre de Lille. Lettre(s) ; Viollette, Charles, physicien. Lettre(s) ; Virchow, Rudolf. Lettre(s) ; Vivier, L., chargé de cours de mathématiques au lycée de Coutances. Lettre(s) ; Vogt, Carl Christoph, naturaliste. Lettre(s) ; Vogüé, Vicomte Eugène-Melchior de, de l'Académie française. Lettre(s) ; Wallon, Henri, membre de l'Institut. Lettre(s) ; Weber, Docteur. Lettre(s) ; Woehrlin, pharmacien-chimiste à Strasbourg. Lettre(s) ; Wurtz, Docteur Adolphe, membre de l'Institut, chimiste. Lettre(s) ; Yersin, Docteur Alexandre-Émile-John. Lettre(s) ; Zévort, Désiré. Lettre(s) ; Zévort, Charles. Lettre(s) ; Zévort, Edgar, recteur d'académie. Lettre(s) ; Forthomme, Camille-Pierre-Guillaume, physicien. Lettre(s) ; Lafon, E.. Lettre(s) ; Vincent, Alexandre-Joseph-Hidulphe, membre de l'Institut. Lettre(s) ; Terquem, Alfred. Lettre(s) ; Hermann, A., professeur. Lettre(s) ; Fauré, professeur de mathématiques au collège de Melun. Lettre(s) ; Robin, P.. Lettre(s) ; Cailly, Félix. Lettre(s) ; Valatour, Martial. Lettre(s) ; Houël, Jules, mathématicien. Lettre(s) ; Leyritz, Albert-Louis, professeur. Lettre(s) ; Berger, Charles-Hippolyte, professeur. Lettre(s) ; Chevilliet. Lettre(s) ; Lamy, Claude-Auguste. Lettre(s) ; Nimier, professeur de physique. Lettre(s)Aux f. 355-386, lettres réunies par Pasteur : « Paquet de lettres d'anciens élèves de l'École relatives à ma nomination de membre de l'Académie des Sciences.