Die Rolle von Plasminogen bei der Immunevasion der Gram-negativen Bakterien Haemophilus influenzae und Borrelia burgdorferi

Der Mensch nutzt zur Erkennung und zur Abwehr von infektiösen Erregern eine Reihe von hocheffizienten Abwehrmechanismen. Das Gerinnungs- und das Komplementsystem übernehmen dabei eine wichtige Rolle und repräsentieren die erste Verteidigungslinie bei der Abwehr von infektiösen Erregern. Das Gerinnungssystem wird nach Verletzung bzw. beim Eindringen eines Erregers umgehend aktiviert und verhindert so die Ausbreitung des Erregers. Ebenfalls hat das Komplementsystem eine wichtige Rolle bei der Erkennung und Eliminierung von infektiösen Erregern. Pathogene Mikroorganismen haben im Laufe der Evolution ein Repertoire an Virulenzfaktoren entwickelt und sie sind so in der Lage in einem immunkompetenten Wirt zu überleben und eine Infektion hervorzurufen. Die Bindung von Komplementregulatoren ist eine effiziente Strategie der Immunevasion pathogener Mikroorganismen. In dieser Arbeit konnte ich eine weitere Strategie zur Evasion beschreiben und einen neuen Zusammenhang zwischen dem Komplement- und dem Gerinnungssystem darstellen. Plasminogen (Plg) ist das Schlüsselenzym der Fibrinolyse und wird von einer großen Anzahl von pathogenen Erregern an die Oberfläche gebunden. Die beiden Gram-negativen Bakterien Haemophilus influenzae und Borrelia burgdorferi rekrutieren Plg durch Protein E (PE) und durch CRASP-3, -4 und -5 („complement regulator-acquiring surface proteins“) an ihrer Oberfläche, das durch uPa zu Plasmin aktiviert wird. Das Protein E (PE) von H. influenzae ist das Hauptbindeprotein für Plg. Gebundenes Plasmin spaltet die natürlichen Substrate, das Opsonin C3b, C5 und das EZM-Protein Fibrinogen, wodurch die Komplementkaskade unterbrochen und die stabile EZM zerstört wird. Somit verhilft Plg den beiden Gram-negativen Bakterien H. influenzae und B. burgdorferi zu einer effizienten Strategie der Immunevasion und Gewebsinvasion

Staphylococcus aureus proteins Sbi and Efb recruit human plasmin to degrade complement C3 and C3b

Upon host infection, the human pathogenic microbe Staphylococcus aureus (S. aureus) immediately faces innate immune reactions such as the activated complement system. Here, a novel innate immune evasion strategy of S. aureus is described. The staphylococcal proteins surface immunoglobulin-binding protein (Sbi) and extracellular fibrinogen-binding protein (Efb) bind C3/C3b simultaneously with plasminogen. Bound plasminogen is converted by bacterial activator staphylokinase or by host-specific urokinase-type plasminogen activator to plasmin, which in turn leads to degradation of complement C3 and C3b. Efb and to a lesser extend Sbi enhance plasmin cleavage of C3/C3b, an effect which is explained by a conformational change in C3/C3b induced by Sbi and Efb. Furthermore, bound plasmin also degrades C3a, which exerts anaphylatoxic and antimicrobial activities. Thus, S. aureus Sbi and Efb comprise platforms to recruit plasmin(ogen) together with C3 and its activation product C3b for efficient degradation of these complement components in the local microbial environment and to protect S. aureus from host innate immune reactions

Disentangling the AGN and star-formation contributions to the radio-X-ray emission of radio-loud quasars at 1<z<21<z<2

To constrain the emission mechanisms responsible for generating the energy powering the active galactic nuclei (AGN) and their host galaxies, it is essential to disentangle the contributions from both as a function of wavelength. Here we introduce a state-of-the-art AGN radio-to-X-ray spectral energy distribution fitting model (ARXSED). ARXSED uses multiple components to replicate the emission from the AGN and their hosts. At radio wavelengths, ARXSED accounts for radiation from the radio structures (e.g., lobes,jets). At near-infrared to far-infrared wavelengths, ARXSED combines a clumpy medium and a homogeneous disk to account for the radiation from the torus. At the optical-UV and X-ray, ARXSED accounts for the emission from the accretion disk. An underlying component from radio to UV wavelengths accounts for the emission from the host galaxy. Here we present the results of ARXSED fits to the panchromatic SEDs of 20 radio-loud quasars from the 3CRR sample at $1<z\lesssim2$. We find that a single power-law is unable to fit the radio emission when compact radio structures (core, hot spots) are present. We find that the non-thermal emission from the quasars' radio structures contributes significantly ($>70\%$) to the submm luminosity in half the sample, impacting the submm-based star formation rate estimates. We present the median intrinsic SED of the radio-loud quasars at $z>1$ and find that the median SED of \cite{Elvis1994} is unable to describe the SED of the radio-selected AGN at $z>1$. The AGN torus and accretion disk parameters inferred from our fitting technique agree with those in the literature for similar samples. We find that the orientation of the torus/accretion disk does not line up with the inclination of the radio jets in our sample

Beyond Simple AGN Unification with Chandra-observed 3CRR Sources at 0.5 &lt; z &lt; 1

Low-frequency radio selection finds radio-bright galaxies regardless of the amount of obscuration by gas and dust. We report \chandra\ observations of a complete 178~MHz-selected, and so orientation unbiased, sample of 44 $0.5<z<1$ 3CRR sources. The sample is comprised of quasars and narrow-line radio galaxies (NLRGs) with similar radio luminosities, and the radio structure serves as both an age and an orientation indicator. Consistent with Unification, intrinsic obscuration (measured by \nh, X-ray hardness ratio, and X-ray luminosity) generally increases with inclination. However, the sample includes a population not seen in high-$z$ 3CRR sources: NLRGs viewed at intermediate inclination angles with \nh~$<10^{22}$~cm$^{-2}$. Multiwavelength analysis suggests these objects have lower $L/L_{\rm Edd}$ than typical NLRGs at similar orientation. Thus both orientation and $L/L_{\rm Edd}$ are important, and a "radiation-regulated Unification" provides a better explanation of the sample's observed properties. In comparison with the 3CRR sample at $1<z<2$, our lower-redshift sample shows a higher fraction of Compton-thin NLRGs (45\% vs.\ 29\%) but similar Compton-thick fraction (20\%), implying a larger covering factor of Compton-thin material at intermediate viewing angles and so a more "puffed-up" torus atmosphere. We posit that this is due to a range of $L/L_{\rm Edd}$ extending to lower values in this sample. In contrast, at high redshifts the narrower range and high $L/L_{\rm Edd}$ values allowed orientation (and so simple Unification) to dominate the sample's observed properties

Insightful Problem Solving in an Asian Elephant

The “aha” moment or the sudden arrival of the solution to a problem is a common human experience. Spontaneous problem solving without evident trial and error behavior in humans and other animals has been referred to as insight. Surprisingly, elephants, thought to be highly intelligent, have failed to exhibit insightful problem solving in previous cognitive studies. We tested whether three Asian elephants (Elephas maximus) would use sticks or other objects to obtain food items placed out-of-reach and overhead. Without prior trial and error behavior, a 7-year-old male Asian elephant showed spontaneous problem solving by moving a large plastic cube, on which he then stood, to acquire the food. In further testing he showed behavioral flexibility, using this technique to reach other items and retrieving the cube from various locations to use as a tool to acquire food. In the cube's absence, he generalized this tool utilization technique to other objects and, when given smaller objects, stacked them in an attempt to reach the food. The elephant's overall behavior was consistent with the definition of insightful problem solving. Previous failures to demonstrate this ability in elephants may have resulted not from a lack of cognitive ability but from the presentation of tasks requiring trunk-held sticks as potential tools, thereby interfering with the trunk's use as a sensory organ to locate the targeted food

Haemophilus influenzae Uses the Surface Protein E To Acquire Human Plasminogen and To Evade Innate Immunity

Pathogenic microbes acquire the human plasma protein plasminogen to their surface. In this article, we characterize binding of this important coagulation regulator to the respiratory pathogen nontypeable Haemophilus influenzae and identify the Haemophilus surface protein E (PE) as a new plasminogen-binding protein. Plasminogen binds dose dependently to intact bacteria and to purified PE. The plasminogen-PE interaction is mediated by lysine residues and is also affected by ionic strength. The H. influenzae PE knockout strain (nontypeable H. influenzae 3655 Delta pe) bound plasminogen with similar to 65% lower intensity as compared with the wild-type, PE-expressing strain. In addition, PE expressed ectopically on the surface of Escherichia coli also bound plasminogen. Plasminogen, either attached to intact H. influenzae or bound to PE, was accessible for urokinase plasminogen activator. The converted active plasmin cleaved the synthetic substrate S-2251, and the natural substrates fibrinogen and C3b. Using synthetic peptides that cover the complete sequence of the PE protein, the major plasminogen-binding region was localized to a linear 28-aa-long N-terminal peptide, which represents aa 41-68. PE binds plasminogen and also vitronectin, and the two human plasma proteins compete for PE binding. Thus, PE is a major plasminogen-binding protein of the Gram-negative bacterium H. influenzae, and when converted to plasmin, PE-bound plasmin aids in immune evasion and contributes to bacterial virulence. The Journal of Immunology, 2012, 188: 379-385

Dihydrolipoamide Dehydrogenase of Pseudomonas aeruginosa Is a Surface-Exposed Immune Evasion Protein That Binds Three Members of the Factor H Family and Plasminogen

The opportunistic human pathogen Pseudomonas aeruginosa causes a wide range of diseases. To cross host innate immune barriers, P. aeruginosa has developed efficient strategies to escape host complement attack. In this study, we identify the 57-kDa dihydrolipoamide dehydrogenase (Lpd) as a surface-exposed protein of P. aeruginosa that binds the four human plasma proteins, Factor H, Factor H-like protein-1 (FHL-1), complement Factor H-related protein 1 (CFHR1), and plasminogen. Factor H contacts Lpd via short consensus repeats 7 and 18-20. Factor H, FHL-1, and plasminogen when bound to Lpd were functionally active. Factor H and FHL-1 displayed complement-regulatory activity, and bound plasminogen, when converted to the active protease plasmin, cleaved the chromogenic substrate S-2251 and the natural substrate fibrinogen. The lpd of P. aeruginosa is a rather conserved gene; a total of 22 synonymous and 3 nonsynonymous mutations was identified in the lpd gene of the 5 laboratory strains and 13 clinical isolates. Lpd is surface exposed and contributes to survival of P. aeruginosa in human serum. Bacterial survival was reduced when Lpd was blocked on the surface prior to challenge with human serum. Similarly, bacterial survival was reduced up to 84% when the bacteria was challenged with complement active serum depleted of Factor H, FHL-1, and CFHR1, demonstrating a protective role of the attached human regulators from complement attack. In summary, Lpd is a novel surface-exposed virulence factor of P. aeruginosa that binds Factor H, FHL-1, CFHR1, and plasminogen, and the Lpd-attached regulators are relevant for innate immune escape and most likely contribute to tissue invasion. The Journal of Immunology, 2012, 189: 4939-4950