<i>N. meningitidis</i> and <i>N. gonorrhoeae</i> isolates.

<p><i>N. meningitidis</i> and <i>N. gonorrhoeae</i> isolates.</p

Immunogenicity of Ghfp.

<p>(<b>A</b>) Detection of fHbp variants in whole cell lysates of <i>N. meningitidis</i> by Western blot analysis using anti-Ghfp serum. Recognition of recombinant V1 (<b>B</b>), and V2and V3 (<b>C</b>) fHbps by anti-Ghfp serum by ELISA. (<b>D</b>) SBA responses for anti-Ghfp serum.</p

Ghfp is not expressed on the surface of <i>N. gonorrhoeae</i>.

<p>(<b>A</b>) Western blot analysis of Ghfp expression by <i>N. gonorrhoeae</i> strains F62, F62Δ<i>ghfp</i> and FA1090, and fHbp V3.28 expressed by <i>N. meningitidis</i> strains M1239 and M1239Δ<i>fhbp</i> using anti-Ghfp serum. (<b>B</b>) Western blot analysis of Ghfp expressed by a panel of clinical <i>N. gonorrhoeae</i> isolates (GC1-11, inclusive). Surface expression of Ghfp (<b>C</b>) and fHbp V3.28 (<b>D</b>) was assessed by flow cytometry analysis using anti-Ghfp serum. Error bars, SEM of three separate experiments; ** p<0.05 and NS (not significant, Student's <i>t</i>-test). Representative flow cytometry overlays are shown below the graphs. Bacteria incubated without anti-Ghfp serum are shown as the grey filled areas. (<b>E</b>) Bacteria were exposed to proteinase K (3 ng/ml and serial three-fold dilutions) and the effect on proteins (shown) determined by Western blot analysis using anti-Ghfp, anti-RecA and anti- α-Lst serum.</p

Oligonucleotides used in this study.

<p>Oligonucleotides used in this study.</p

Immune responses against isogenic <i>N. meningitidis</i> strains.

<p>(<b>A</b>) Western blot analysis of whole cell lysates of fHbp expressed by isogenic MC58Δ<i>fhbp</i> strains detected by anti-Ghfp serum. (<b>B</b>) Surface expression of different fHbps in the isogenic MC58Δ<i>fhbp</i> strains detected by flow cytometry. Graph shows the mean ± SEM of three separate experiments. (<b>C</b>) Representative corresponding flow cytometry overlay of MC58 (grey hatched area), MC58Δ<i>fhbp</i> and MC58Δ<i>fhbp</i>+<i>fhbp</i> V1.1 detected by anti-Ghfp by flow cytometry. (<b>D</b>) SBA responses against isogenic MC58 strains for anti-Ghfp, fHbp V1.1, and fHbp V3.45 serum using rabbit complement; NK, no killing. (<b>E</b>) SBA responses against isogenic H44/76 strains with anti-Ghfp serum using rabbit complement.</p

fH binding to modified fHbp V3.45.

<p>(<b>A</b>) Analysis of fH binding to modified V3.45 fHbp by far Western using normal human serum as the source of fH. Molecular mass is shown in kDa. (B) SPR values of fH_6–7 binding to wild type and modified V3.45 fHbp; NBD, no binding detected. (<b>C</b>) Detection of full length fH (5 nM) binding to wild-type and modified V3.45 fHbp by ELISA. Data represents the mean ± SEM of three different experiments.</p

fH binding capacity of Ghfp.

<p>(<b>A-C</b>) fH binding to wild type and modified Ghfp and fHbp V3.45 was assessed by far Western analysis using normal human serum as the source of fH. Western blots are representatives of three separate experiments. Molecular mass is shown in kDa. (<b>D</b>) Typical equilibrium fit for binding of fH_6–7 to Ghfp^M4–5. (<b>E</b>) SPR was performed with Ghfp, Ghfp^M4 (R288H), Ghfp^M5 (D318G) and Ghfp^M4–5 (R288H/D318G); NBD, no binding detected. (<b>F</b>) Detection of full length fH (5 nM) binding to wild-type and modified Ghfp by ELISA. Error bars, SEM of three separate experiments; *** p<0.01 and * p<0.1 (Student's <i>t</i>-test). (<b>G</b>) Cartoon presentation of the predicted structure of Ghfp (Yellow) and fH (Blue). Residues M1 (R176), M2 (D199) and M3 (D212) are shown in green while those amino acids that are important for fH binding, M4 (R288) and M5 (D318G), are shown in red.</p

Characteristics of included studies.

<p>RCT: Randomised controlled trial. GS: Gilead Sciences. R: Roche. GSK: GlaxoSmithKline. NS: not stated.</p>a<p>Copies/mL converted to IU/mL by dividing by 5.</p>b<p>The limit of detection of the HBV viral load assays used fell during the course of follow up in two studies.</p>c<p>Individual patient data available.</p>d<p>National Institutes of Health/National Institute on Drug Abuse.</p>e<p>Commonwealth Department of Health and Ageing (Canberra, Australia).</p>f<p>supported in part by National Institute of Allergy and Infectious Diseases.</p>g<p>Medical Student Summer Research Training Program, supported through grants from the National Institutes of Health; Wake Forest University School of Medicine Departments, Centers, and Institutes; and private gifts.</p>h<p>Swiss National Science Foundation through the Swiss HIV Cohort Study, the Wilsdorf, Sidaide, and de Brocard Foundations, Geneva, from the Departments of Social Affairs and Economics, Geneva.</p>i<p>In part by the Adult AIDS Clinical Trials Group (ACTG) funded by the National Institute of Allergy and Infectious Diseases; virology support funding by the NIH/NIAID and the Adult ACTG Central Group; the Birmingham VA Medical Center, UAB CFAR core clinic and laboratory facilities; and NIDDK UCSF Liver Center.</p>j<p>Italian Ministry of University.</p>k<p>In part by the German BMBF Network of Competence for Viral Hepatitis (Hep Net).</p

Log of HBV viral load assay cut-off against proportion undetectable at one year.

<p>Log of HBV viral load assay cut-off against proportion undetectable at one year.</p

Percentage with undetectable HBV viral load over time, by HBeAg status.

<p>Percentage with undetectable HBV viral load over time, by HBeAg status.</p