Alternative complement pathway inhibition does not abrogate meningococcal killing by serum of vaccinated individuals

Dysregulation of complement activation causes a number of diseases, including paroxysmal nocturnal hemoglobinuria and atypical hemolytic uremic syndrome. These conditions can be treated with monoclonal antibodies (mAbs) that bind to the complement component C5 and prevent formation of the membrane attack complex (MAC). While MAC is involved in uncontrolled lysis of erythrocytes in these patients, it is also required for serum bactericidal activity (SBA), i.e. clearance of encapsulated bacteria. Therefore, terminal complement blockage in these patients increases the risk of invasive disease by Neisseria meningitidis more than 1000-fold compared to the general population, despite obligatory vaccination. It is assumed that alternative instead of terminal pathway inhibition reduces the risk of meningococcal disease in vaccinated individuals. To address this, we investigated the SBA with alternative pathway inhibitors. Serum was collected from adults before and after vaccination with a meningococcal serogroup A, C, W, Y capsule conjugate vaccine and tested for meningococcal killing in the presence of factor B and D, C3, C5 and MASP-2 inhibitors. B meningococci were not included in this study since the immune response against protein-based vaccines is more complex. Unsurprisingly, inhibition of C5 abrogated killing of meningococci by all sera. In contrast, both factor B and D inhibitors affected meningococcal killing in sera from individuals with low, but not with high bactericidal anti-capsular titers. While the anti-MASP-2 mAb did not impair SBA, inhibition of C3 impeded meningococcal killing in most, but not in all sera. These data provide evidence that vaccination can provide protection against invasive meningococcal disease in patients treated with alternative pathway inhibitors

Inhibition of the different complement pathways has varying impacts on the serum bactericidal activity and opsonophagocytosis against Haemophilus influenzae type b

Defense against Haemophilus influenzae type b (Hib) is dependent on antibodies and complement, which mediate both serum bactericidal activity (SBA) and opsonophagocytosis. Here we evaluated the influence of capsule-specific antibodies and complement inhibitors targeting the central component C3, the alternative pathway (AP; fB, fD), the lectin pathway (LP; MASP-2) and the terminal pathway (C5) on both effector functions. Findings may be relevant for the treatment of certain diseases caused by dysregulation of the complement system, where inhibitors of complement factors C3 or C5 are used. Inhibitors against other complement components are being evaluated as potential alternative treatment options that may carry a reduced risk of infection by encapsulated bacteria. Serum and reconstituted blood of healthy adults were tested for bactericidal activity before and after vaccination with the Hib capsule-conjugate vaccine ActHIB. Most sera had bactericidal activity prior to vaccination, but vaccination significantly enhanced SBA titers. Independently of the vaccination status, both C3 and C5 inhibition abrogated SBA, whereas inhibition of the LP had no effect. AP inhibition had a major inhibitory effect on SBA of pre- vaccination serum, but vaccination mitigated this inhibition for all disease isolates tested. Despite this, SBA-mediated killing of some Hib isolates remained retarded. Even for the most serum-resistant isolate, SBA was the dominating defense mechanism in reconstituted whole blood, as addition of blood cells to the serum did not enhance bacterial killing. Limited Fc receptor-mediated opsonophagocytosis was unmasked when bacterial killing by the membrane attack complex was blocked. In the presence of C3 or C5 inhibitors, addition of post-vaccination, but not of pre-vaccination serum to the blood cells triggered opsonophagocytosis, leading to suppression of bacterial multiplication. Taken together, our data indicate that for host defense against Hib, killing by SBA is more efficient than by blood cell opsonophagocytosis. However, additional defense mechanisms, such as bacterial clearance by spleen and liver, may play an important role in preventing Hib-mediated sepsis, in particular for Hib isolates with increased serum-resistance. Results indicate potentially improved safety profile of AP inhibitors over C3 and C5 inhibitors as alternative therapeutic agents in patients with increased susceptibility to Hib infection

Biochemical and molecular diagnosis of mitochondrial respiratory chain disorders

AbstractBiochemical diagnosis of mitochondrial respiratory chain disorders requires caution to avoid misdiagnosis of secondary enzyme defects, and can be improved by the use of conservative diagnostic criteria. Pathogenic mutations causing mitochondrial disorders have now been identified in more than 30 mitochondrial DNA (mtDNA) genes encoding respiratory chain subunits, ribosomal- and t-RNAs. mtDNA mutations appear to be responsible for most adult patients with mitochondrial disease and approximately a quarter of paediatric patients. A family history suggesting maternal inheritance is the exception rather than the norm for children with mtDNA mutations, many of whom have de novo mutations. Prenatal diagnosis and pre-implantation genetic diagnosis can be offered to some women at risk of transmitting a mtDNA mutation, particularly those at lower recurrence risk. Mutations in more than 30 nuclear genes, including those encoding for respiratory chain subunits and assembly factors, have now been shown to cause mitochondrial disorders, creating difficulties in prioritising which genes should be studied by mutation analysis in individual patients. A number of approaches offer promise to guide the choice of candidate genes, including Blue Native-PAGE immunoblotting and microarray expression analysis

Mitochondrial haplotypes reveal low diversity and restricted connectivity in the critically endangered batoid population of a Marine Protected Area

ACKNOWLEDGEMENTS This study was supported by NatureScot, Scottish Government project SP02B, a Heredity Fieldwork Grant of the Genetics Society, and Save Our Seas Foundation project SOSF 470. We would like to thank Leigh Taylor, Ronnie Campbell and Roger Eaton for skippering the sampling charters in the Marine Protected Area and all anglers who provided skate recapture data. Thanks to Fenella Wood and Danielle Sloan for assisting on charter trips. Further, thanks go to Marine Scotland Science (Francis Neat), the Centre for Environment Fisheries and Aquaculture Science (Vicky Bendall and Stewart Hetherington), and the University of St Andrews for providing tissue samples and Lauren Smith and Dan Wise for contributing samples of egg cases.Peer reviewedPublisher PD

In vaccinated individuals serum bactericidal activity against B meningococci is abrogated by C5 inhibition but not by inhibition of the alternative complement pathway

INTRODUCTION: Several diseases caused by the dysregulation of complement activation can be treated with inhibitors of the complement components C5 and/or C3. However, complement is required for serum bactericidal activity (SBA) against encapsulated Gram-negative bacteria. Therefore, C3 and C5 inhibition increases the risk of invasive disease, in particular by Neisseria meningitidis. As inhibitors against complement components other than C3 and C5 may carry a reduced risk of infection, we compared the effect of inhibitors targeting the terminal pathway (C5), the central complement component C3, the alternative pathway (FB and FD), and the lectin pathway (MASP-2) on SBA against serogroup B meningococci. METHODS: Serum from adults was collected before and after vaccination with the meningococcal serogroup B vaccine 4CMenB and tested for meningococcal killing. Since the B capsular polysaccharide is structurally similar to certain human polysaccharides, 4CMenB was designed to elicit antibodies against meningococcal outer membrane proteins. RESULTS: While only a few pre-vaccination sera showed SBA against the tested B meningococcal isolates, 4CMenB vaccination induced potent complement-activating IgG titers against isolates expressing a matching allele of the bacterial cell surface-exposed factor H-binding protein (fHbp). SBA triggered by these cell surface protein-specific antibodies was blocked by C5 and reduced by C3 inhibition, whereas alternative (factor B and D) and lectin (MASP-2) pathway inhibitors had no effect on the SBA of post-4CMenB vaccination sera. DISCUSSION: Compared to the SBA triggered by A,C,W,Y capsule polysaccharide conjugate vaccination, SBA against B meningococci expressing a matching fHbp allele was remarkably resilient against the alternative pathway inhibition

Outbursts on normal stars. FH Leo misclassified as a novalike variable

We present high resolution spectroscopy of the common proper motion system FH Leo (components HD 96273 and BD+07 2411B), which has been classified as a novalike variable due to an outburst observed by Hipparcos, and we present and review the available photometry. We show from our spectra that neither star can possibly be a cataclysmic variable, instead they are perfectly normal late-F and early-G stars. We measured their radial velocities and derived the atmospheric fundamental parameters, abundances of several elements including Fe, Ni, Cr, Co, V, Sc, Ti, Ca and Mg, and we derive the age of the system. From our analysis we conclude that the stars do indeed constitute a physical binary. However, the observed outburst cannot be readily explained. We examine several explanations, including pollution with scattered light from Jupiter, binarity, microlensing, background supernovae, interaction with unseen companions and planetary engulfment. While no explanation is fully satisfactory, the scattered light and star-planet interaction scenarios emerge as the least unlikely ones, and we give suggestions for further study.Comment: 8 pages, 7 figures. Accepted for publication in A&

Unusually Weak Diffuse Interstellar Bands toward HD 62542

As part of an extensive survey of diffuse interstellar bands (DIBs), we have obtained optical spectra of the moderately reddened B5V star HD 62542, which is known to have an unusual UV extinction curve of the type usually identified with dark clouds. The typically strongest of the commonly catalogued DIBs covered by the spectra -- those at 5780, 5797, 6270, 6284, and 6614 A -- are essentially absent in this line of sight, in marked contrast with other lines of sight of similar reddening. We compare the HD 62542 line of sight with others exhibiting a range of extinction properties and molecular abundances and interpret the weakness of the DIBs as an extreme case of deficient DIB formation in a dense cloud whose more diffuse outer layers have been stripped away. We comment on the challenges these observations pose for identifying the carriers of the diffuse bands.Comment: 20 pages, 4 figures; aastex; accepted by Ap

Effect of Neutrino Heating on Primordial Nucleosynthesis

We have modified the standard code for primordial nucleosynthesis to include the effect of the slight heating of neutrinos by $e^\pm$ annihilations. There is a small, systematic change in the $^4$He yield, $\Delta Y \simeq +1.5\times 10^{-4}$, which is insensitive to the value of the baryon-to-photon ratio $\eta$ for 10^{-10}\la \eta \la 10^{-9}. We also find that the baryon-to-photon ratio decreases by about 0.5\% less than the canonical factor of 4/11 because some of the entropy in $e^\pm$ pairs is transferred to neutrinos. These results are in accord with recent analytical estimates.Comment: 14 pages/4 Figs (upon request