Publications by authors named "Timo Wolf"

58 Publications

Analysis of Humoral Immune Responses in Patients With Severe Acute Respiratory Syndrome Coronavirus 2 Infection.

J Infect Dis 2021 01;223(1):56-61

Paul-Ehrlich-Institut, Department of Virology, Langen, Germany.

Background: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has caused a pandemic with tens of millions of cases and hundreds of thousands of deaths. The infection causes coronavirus disease 2019 (COVID-19), a disease of the respiratory system of divergent severity. In the current study, humoral immune responses were characterized in a cohort of 143 patients with COVID-19 from the University Hospital Frankfurt am Main, Germany.

Methods: SARS-CoV-2-specific-antibodies were detected by enzyme-linked immunosorbent assay (ELISA). SARS-CoV-2 and human coronavirus NL63 neutralization activity was analyzed with pseudotyped lentiviral vectors.

Results: The severity of COVID-19 increased with age, and male patients encountered more serious symptoms than female patients. Disease severity was correlated with the amount of SARS-CoV-2-specific immunoglobulin (Ig) G and IgA and the neutralization activity of the antibodies. The amount of SARS-CoV-2-specific IgG antibodies decreased with time after polymerase chain reaction conformation of the infection, and antibodies directed against the nucleoprotein waned faster than spike protein-directed antibodies. In contrast, for the common flu coronavirus NL63, COVID-19 disease severity seemed to be correlated with low NL63-neutralizing activities, suggesting the possibility of cross-reactive protection.

Conclusion: The results describe the humoral immune responses against SARS-CoV-2 and might aid the identification of correlates of protection needed for vaccine development.
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http://dx.doi.org/10.1093/infdis/jiaa680DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7665662PMC
January 2021

Lung Ultrasound Findings in Patients with COVID-19.

SN Compr Clin Med 2020 Oct 1:1-7. Epub 2020 Oct 1.

Department for Infectious Diseases, Centre of Internal Medicine II, University Hospital Frankfurt, Theodor-Stern-Kai 7 (H68-2), 60590 Frankfurt am Main, Germany.

The current SARS-CoV-2 outbreak leads to a growing need of point-of-care thoracic imaging that is compatible with isolation settings and infection prevention precautions. We retrospectively reviewed 17 COVID-19 patients who received point-of-care lung ultrasound imaging in our isolation unit. Lung ultrasound was able to detect interstitial lung disease effectively; severe cases showed bilaterally distributed B-Lines with or without consolidations; one case showed bilateral pleural plaques. Corresponding to CT scans, interstitial involvement is accurately depicted as B-Lines on lung ultrasound. Lung ultrasound might be suitable for detecting interstitial involvement in a bedside setting under high security isolation precautions.
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http://dx.doi.org/10.1007/s42399-020-00553-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7529355PMC
October 2020

A maltose-regulated large genomic region is activated by the transcriptional regulator MalT in Actinoplanes sp. SE50/110.

Appl Microbiol Biotechnol 2020 Nov 28;104(21):9283-9294. Epub 2020 Sep 28.

Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615, Bielefeld, Germany.

Actinoplanes sp. SE50/110 is the industrially relevant producer of acarbose, which is used in the treatment of diabetes mellitus. Recent studies elucidated the expression dynamics in Actinoplanes sp. SE50/110 during growth. From these data, we obtained a large genomic region (ACSP50_3900 to ACSP50_3950) containing 51 genes, of which 39 are transcribed in the same manner. These co-regulated genes were found to be stronger transcribed on maltose compared with glucose as a carbon source. The transcriptional regulator MalT was identified as an activator of this maltose-regulated large genomic region (MRLGR). Since most of the genes are poorly annotated, the function of this region is farther unclear. However, comprehensive BLAST analyses indicate similarities to enzymes involved in amino acid metabolism. We determined a conserved binding motif of MalT overlapping the -35 promoter region of 17 transcription start sites inside the MRLGR. The corresponding sequence motif 5'-TCATCC-5nt-GGATGA-3' displays high similarities to reported MalT binding sites in Escherichia coli and Klebsiella pneumoniae, in which MalT is the activator of mal genes. A malT deletion and an overexpression mutant were constructed. Differential transcriptome analyses revealed an activating effect of MalT on 40 of the 51 genes. Surprisingly, no gene of the maltose metabolism is affected. In contrast to many other bacteria, MalT is not the activator of mal genes in Actinoplanes sp. SE50/110. Finally, the MRLGR was found partly in other closely related bacteria of the family Micromonosporaceae. Even the conserved MalT binding site was found upstream of several genes inside of the corresponding regions. KEY POINTS : • MalT is the maltose-dependent activator of a large genomic region in ACSP50_WT. • The consensus binding motif is similar to MalT binding sites in other bacteria. • MalT is not the regulator of genes involved in maltose metabolism in ACSP50_WT.
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http://dx.doi.org/10.1007/s00253-020-10923-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7567727PMC
November 2020

Antimicrobial Susceptibility and Phylogenetic Relations in a German Cohort Infected with Mycobacterium abscessus.

J Clin Microbiol 2020 11 18;58(12). Epub 2020 Nov 18.

University Center of Infectious Diseases, UCI, University Hospital Frankfurt, Goethe University, Frankfurt am Main, Germany

is a highly antibiotic-resistant opportunistic pathogen causing clinically challenging infections in patients with preexisting lung diseases or under immunosuppression. Hence, reliable antibiotic susceptibility data are required for effective treatment. Aims of this study were to investigate (i) the congruence of genotypic and phenotypic antimicrobial susceptibility testing, (ii) the relationship between resistance profile and clinical course, and (iii) the phylogenetic relations of in a German patient cohort. A total of 39 isolates from 29 patients infected or colonized with underwent genotypic and phenotypic drug susceptibility testing. Clinical data were correlated with susceptibility data. Phylogenetic analysis was performed by means of whole-genome sequencing (WGS) and single-nucleotide polymorphism (SNP) analysis. Macrolide resistance was mainly mediated by functional Erm(41) methyltransferases (T28 sequevars) in subsp. ( = 25) and subsp. ( = 2). It was significantly associated with impaired culture conversion ( = 0.02). According to the core SNP phylogeny, we identified three clusters of closely related isolates with SNP distances below 25. Representatives of all circulating global clones (Absc. 1, Absc. 2, and Mass. 1) were identified in our cohort. However, we could not determine evidence for in-hospital interhuman transmission from clinical data. In our patient cohort, we identified three clusters with closely related isolates and representatives of the previously described international clusters but no human-to-human in-hospital transmission. Macrolide and aminoglycoside susceptibility data are critical for therapeutic decision-making in infections.
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http://dx.doi.org/10.1128/JCM.01813-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7685876PMC
November 2020

Longitudinal Isolation of Potent Near-Germline SARS-CoV-2-Neutralizing Antibodies from COVID-19 Patients.

Cell 2020 08 13;182(4):843-854.e12. Epub 2020 Jul 13.

Laboratory of Experimental Immunology, Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany; German Center for Infection Research, Partner Site Bonn-Cologne, 50931 Cologne, Germany; Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany. Electronic address:

The SARS-CoV-2 pandemic has unprecedented implications for public health, social life, and the world economy. Because approved drugs and vaccines are limited or not available, new options for COVID-19 treatment and prevention are in high demand. To identify SARS-CoV-2-neutralizing antibodies, we analyzed the antibody response of 12 COVID-19 patients from 8 to 69 days after diagnosis. By screening 4,313 SARS-CoV-2-reactive B cells, we isolated 255 antibodies from different time points as early as 8 days after diagnosis. Of these, 28 potently neutralized authentic SARS-CoV-2 with IC as low as 0.04 μg/mL, showing a broad spectrum of variable (V) genes and low levels of somatic mutations. Interestingly, potential precursor sequences were identified in naive B cell repertoires from 48 healthy individuals who were sampled before the COVID-19 pandemic. Our results demonstrate that SARS-CoV-2-neutralizing antibodies are readily generated from a diverse pool of precursors, fostering hope for rapid induction of a protective immune response upon vaccination.
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http://dx.doi.org/10.1016/j.cell.2020.06.044DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7355337PMC
August 2020

Epitopes of Naturally Acquired and Vaccine-Induced Anti-Ebola Virus Glycoprotein Antibodies in Single Amino Acid Resolution.

Biotechnol J 2020 Sep 8;15(9):e2000069. Epub 2020 Jun 8.

Max Planck Institute of Colloids and Interfaces, Department of Biomolecular Systems, Potsdam, 14476, Germany.

The Ebola virus (EBOV) can cause severe infections in humans, leading to a fatal outcome in a high percentage of cases. Neutralizing antibodies against the EBOV surface glycoprotein (GP) can prevent infections, demonstrating a straightforward way for an efficient vaccination strategy. Meanwhile, many different anti-EBOV antibodies have been identified, whereas the exact binding epitopes are often unknown. Here, the analysis of serum samples from an EBOV vaccine trial with the recombinant vesicular stomatitis virus-Zaire ebolavirus (rVSV-ZEBOV) and an Ebola virus disease survivor, using high-density peptide arrays, is presented. In this proof-of-principle study, distinct IgG and IgM antibodies binding to different epitopes of EBOV GP is detected: By mapping the whole GP as overlapping peptide fragments, new epitopes and confirmed epitopes from the literature are found. Furthermore, the highly selective binding epitope of a neutralizing monoclonal anti-EBOV GP antibody could be validated. This shows that peptide arrays can be a valuable tool to study the humoral immune response to vaccines in patients and to support Ebola vaccine development.
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http://dx.doi.org/10.1002/biot.202000069DOI Listing
September 2020

Severe COVID-19 infection in a patient with multiple sclerosis treated with fingolimod.

Mult Scler Relat Disord 2020 Jul 6;42:102180. Epub 2020 May 6.

Department of Anesthesiology, Intensive Care and Pain Therapy, University Hospital, Goethe University, Frankfurt am Main, Germany.

Background: Fingolimod is used for immune therapy in patients with multiple sclerosis. Long-term treatment is associated with a small increase in the risk of herpes virus reactivation and respiratory tract infections. Patients with coronavirus disease 2019 (COVID-19) under Fingolimod treatment have not been described.

Methods And Results: We report a 57-year old female patient with a relapsing remitting multiple sclerosis under fingolimod treatment who experienced a severe COVID-19 infection in March 2020 (Extended Disability Status Scale: 2.0). Having peripheral lymphopenia typical for fingolimod treatment (total lymphocytes 0.39/nL [reference range 1.22-3.56]), the patient developed bilateral interstitial pneumonia with multiple ground-glass opacities on chest CT. Fingolimod medication was stopped. On the intensive care unit, non-invasive ventilation was used to provide oxygen and ventilation support regularly. Over the following two days, oxygenation improved, and the patient was transferred to a normal ward five days after admission.

Conclusion: The implications fingolimod has on COVID-19 are complex. As an S1P analogue, fingolimod might enhance lung endothelial cell integrity. In addition, in case of a so-called cytokine storm, immunomodulation might be beneficial to reduce mortality. Future studies are needed to explore the risks and therapeutic effects of fingolimod in COVID-19 patients.
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http://dx.doi.org/10.1016/j.msard.2020.102180DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7202802PMC
July 2020

Fifty years of imported Lassa fever: a systematic review of primary and secondary cases.

J Travel Med 2020 Jul;27(4)

Health Protection Authority, Municipality of Frankfurt am Main, Frankfurt, Germany.

Rationale For Systematic Review: Lassa fever is the most common cause of imported haemorrhagic fevers cases in non-endemic countries. As a disease with a high case fatality rate that has regularly caused clusters of nosocomial transmission in endemic areas, prompt diagnosis is vital. We conducted a systematic review of imported cases of the last 50 years with the aim of defining the clinical and epidemiological characteristics that will enhance early diagnosis, prompt initiation of treatment and an appropriate public health response to Lassa fever cases.

Methods: We performed a retrospective, systematic review of 36 primary and two secondary cases of Lassa fever in non-endemic countries outside West Africa by searching the PubMed database. This yielded 56 relevant publications that were included in our analysis.

Results: The case fatality rate of 35.1% for imported cases was higher than that reported for endemic countries. The majority of patients showed clinical features consistent with Lassa fever and had a typical exposure. There was a considerable delay in diagnosis in imported cases with high associated numbers of contacts. Ribavirin was rarely used for post-exposure prophylaxis. Only two secondary transmissions occurred. Thirty-one percent of patients received Lassa fever-specific treatment and five required intensive care.

Conclusions: Although importation of Lassa fever to non-endemic countries is a rare event, it has repeatedly happened over five decades. Suspicion of Lassa fever should be based on careful consideration of clinical features and exposure history in order to assist early diagnosis in returning travellers from West Africa.
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http://dx.doi.org/10.1093/jtm/taaa035DOI Listing
July 2020

Comparative analysis of phenotypic and genotypic antibiotic susceptibility patterns in Mycobacterium avium complex.

Int J Infect Dis 2020 Apr 5;93:320-328. Epub 2020 Mar 5.

Institute of Medical Microbiology and Infection Control, University Hospital Frankfurt, Goethe University, Frankfurt am Main, Germany.

Objective: Phenotypic (Sensititre Myco, pDST) and genotypic drug susceptibility testing (GenoType NTM DR, gDST) in M. avium complex (MAC) have become available as standardized assays, but comparable data is needed. This study aimed to investigate the phenotypic and genotypic drug susceptibility patterns in MAC clinical isolates.

Methods: Overall, 98 isolates from 85 patients were included. pDST and gDST were performed on all isolates and results compared regarding specificity and sensitivity using pDST as a reference method. The impact of drug instability on pDST results was studied using a biological assay over 14 days. In addition, the evolution of antimicrobial resistance was investigated in sequential isolates of 13 patients.

Results: Macrolide resistance was rare, 1.2% (95% CI 0.7-7.3) of isolates in the base cohort. No aminoglycoside resistances were found, but 14.1% of the studied isolates (95% CI 7.8-23.8) showed intermediate susceptibility. The GenoType NTM DR identified two out of four macrolide-resistant isolates. Antibiotic stability was demonstrated to be poor in rifampicin, rifabutin, and doxycycylin.

Conclusions: pDST results in NTM for unstable antibiotics must be interpreted with care. A combination of pDST and gDST will be useful for the guidance of antimicrobial therapy in MAC-disease.
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http://dx.doi.org/10.1016/j.ijid.2020.02.059DOI Listing
April 2020

Two cases of airport-associated falciparum malaria in Frankfurt am Main, Germany, October 2019.

Euro Surveill 2019 Dec;24(49)

These authors contributed equally to this work and share last authorship.

Two cases of presumably airport-acquired falciparum malaria were diagnosed in Frankfurt in October 2019. They were associated with occupation at the airport, and parasites from their blood showed genetically identical microsatellite and allele patterns. Both had severe malaria. It took more than a week before the diagnosis was made. If symptoms are indicative and there is a plausible exposure, malaria should be considered even if patients have not travelled to an endemic area.
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http://dx.doi.org/10.2807/1560-7917.ES.2019.24.49.1900691DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6905295PMC
December 2019

Polyclonal and convergent antibody response to Ebola virus vaccine rVSV-ZEBOV.

Nat Med 2019 10 7;25(10):1589-1600. Epub 2019 Oct 7.

Laboratory of Experimental Immunology, Institute of Virology, Faculty of Medicine and University Hospital of Cologne, University of Cologne, Cologne, Germany.

Recombinant vesicular stomatitis virus-Zaire Ebola virus (rVSV-ZEBOV) is the most advanced Ebola virus vaccine candidate and is currently being used to combat the outbreak of Ebola virus disease (EVD) in the Democratic Republic of the Congo (DRC). Here we examine the humoral immune response in a subset of human volunteers enrolled in a phase 1 rVSV-ZEBOV vaccination trial by performing comprehensive single B cell and electron microscopy structure analyses. Four studied vaccinees show polyclonal, yet reproducible and convergent B cell responses with shared sequence characteristics. EBOV-targeting antibodies cross-react with other Ebolavirus species, and detailed epitope mapping revealed overlapping target epitopes with antibodies isolated from EVD survivors. Moreover, in all vaccinees, we detected highly potent EBOV-neutralizing antibodies with activities comparable or superior to the monoclonal antibodies currently used in clinical trials. These include antibodies combining the IGHV3-15/IGLV1-40 immunoglobulin gene segments that were identified in all investigated individuals. Our findings will help to evaluate and direct current and future vaccination strategies and offer opportunities for novel EVD therapies.
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http://dx.doi.org/10.1038/s41591-019-0602-4DOI Listing
October 2019

Species distribution and clinical features of infection and colonisation with non-tuberculous mycobacteria in a tertiary care centre, central Germany, 2006-2016.

Infection 2019 Oct 15;47(5):817-825. Epub 2019 May 15.

Department of Infectious Diseases, University Hospital Frankfurt, Frankfurt, Germany.

Purpose: NTM are ubiquitous bacteria that can cause colonisation and infection in immunocompetent and compromised hosts. The aim of this study was to elucidate the epidemiology of infection or colonisation with NTM for the metropolitan region of Frankfurt, Germany.

Methods: All patients from whom NTM were isolated within the period from 2006 to 2016 were included in this retrospective analysis. Patient data were retrieved using the local patient data management system. Different groups were formed according to clinical manifestations, underlying diseases and mycobacterial species. They were compared in regard to mortality, duration of infection/colonisation and their geographical origins.

Results: A total of 297 patients with a median of 28 new patients each year were included. Most patients suffered from lung infection or colonisation (72.7%, n = 216), followed by disseminated mycobacteriosis (12.5%, n = 37). The majority were HIV-positive, suffering from malignoma or cystic fibrosis (29.3%, n = 87, 16.2%, n = 48, and 13.8%, n = 41, respectively). 17.2% of patients showed no predisposing condition (n = 51). Mycobacterium avium complex (MAC) species were most frequently isolated (40.7%, n = 121). Infection/colonisation was longest in CF patients (median of 1094 days). The mortality was highest in malignoma patients (52.4%), while CF patients had the lowest overall mortality rate (5.3%). But mortality analysis showed non-significant results within different mycobacterial species and clinical manifestations.

Conclusion: NTM remain rare but underestimated pathogens in lung and disseminated disease. MAC were the species most frequently isolated. Depending on species and underlying predispositions, the duration of infection/colonisation can be unexpectedly long.
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http://dx.doi.org/10.1007/s15010-019-01317-2DOI Listing
October 2019

Clinical characteristics in a sentinel case as well as in a cluster of tularemia patients associated with grape harvest.

Int J Infect Dis 2019 Jul 6;84:116-120. Epub 2019 May 6.

University Hospital Frankfurt, Frankfurt, Germany.

Background: Tularemia is caused by Francisella tularensis and can occasionally establish foodborne transmission.

Methods: Patients were identified by active case detection through contact with the treating physicians and consent for publication was obtained. Clinical data were accumulated through a review of the patient charts. Serology, culture, and PCR methods were performed for confirmation of the diagnosis.

Case Cluster: A 46-year-old patient was hospitalised in the University Hospital Frankfurt (a tertiary care hospital) for pharyngitis and cervical lymphadenitis with abscess. A diagnosis of tularemia was made serologically, but treatment with ciprofloxacin initially failed. F. tularensis was detected in pus from the lymph node using a specific real-time PCR. The use of RD1 PCR led to the identification of the subspecies holarctica. Antibiotic therapy with high-dose ciprofloxacin and gentamicin was administered and was subsequently changed to ciprofloxacin and rifampicin. During a must-tasting, five other individuals became infected with tularemia by ingestion of contaminated must. All patients required treatment durations of more than 14 days.

Conclusions: Mechanically harvested agricultural products, such as wine must, can be a source of infection, probably due to contamination with animal carcasses. The clinical course of tularemia can be complicated and prolonged and requires differentiated antibiotic treatment.
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http://dx.doi.org/10.1016/j.ijid.2019.04.031DOI Listing
July 2019

New filovirus disease classification and nomenclature.

Nat Rev Microbiol 2019 05;17(5):261-263

World Health Organization Regional Office for Africa, Brazzaville, Democratic Republic of the Congo.

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http://dx.doi.org/10.1038/s41579-019-0187-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6637750PMC
May 2019

HLA-B57.01 shields people living with HIV for significantly better periodontal health.

J Periodontol 2018 08 20;89(8):966-972. Epub 2018 Jul 20.

University Hospital Frankfurt, J.W. Goethe-University, Department for Infectious Diseases, Internal Medicine II, Frankfurt, Germany.

Background: To assess the association between HLA-B57.01 (Human leukocyte antigen) and clinical parameters of chronic periodontitis in people living with HIV (PLWHIV).

Methods: All patients were recruited from the HIVCENTER at the University Hospital Frankfurt during April 2014 and July 2015. Periodontal examination included Periodontal Screening Index (PSI), Gingivalindex (GI), Bleeding on Probing Index (BOP), Periodontal Probing Depth (PD), Clinical Attachment Level (CAL) and DMF-T Score (decayed, missing, filled teeth). Associations among periodontitis, HLA-system and additional risk factors in PLWHIV were evaluated in multivariate analyses.

Results: One hundred PLWHIV were enrolled. Forty-five patients were naive, meaning that these patients never took antiretroviral (ARV) drugs before, 55 patients treated with combined antiretroviral therapy (cART). Nineteen patients presented a positive HLA-B 57.01 status. PLWHIV who were carriers of HLA-B 57.01 had significantly lower PSI-scores (Grade 3 or higher; 0/19 [0%] versus 16/41 [39%] versus 17/40 [42%]; p = < 0.001), GI-scores (Grade 2 or higher; 0/19 [0%] versus 19/41 [46%] versus 28/40 [70%]; p = < 0.001) and BOP-Scores (2/19 [1%] versus 38/41 [92%] versus 40/40 [100%]; p = < 0.001) in comparison to naive PLWHIV and PLWHIV receiving cART, who were both not carriers of HLA-B 57.01. A lower value of PSI-, GI- and BOP-Score is associated with improved periodontal health. The adjusted odds ratio (OR) of periodontitis was decreased in patients who were carriers of HLA-B 57.01 by measurement of PSI-Score (OR = 0.006, 95% confidence interval (CI) = 0.001 to 0.026), GI-Score (OR = 0.018, 95% confidence interval (CI) = 0.003 to 0.104) and BOP-Score (OR = 0.003, 95% confidence interval (CI) = < 0.001 to 0.011).

Conclusions: HLA-B 57.01 is an independent resistance indicator for generalized periodontitis in PLWHIV with respect to established cofactors.
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http://dx.doi.org/10.1002/JPER.17-0532DOI Listing
August 2018

Incidence and risk factors for relapses in HIV-associated non-Hodgkin lymphoma as observed in the German HIV-related lymphoma cohort study.

Haematologica 2018 05 8;103(5):857-864. Epub 2018 Feb 8.

University of Schleswig Holstein, Campus Kiel, Kiel, Germany.

Outcome of HIV-infected patients with AIDS-related lymphomas has improved during recent years. However, data on incidence, risk factors, and outcome of relapses in AIDS-related lymphomas after achieving complete remission are still limited. This prospective observational multicenter study includes HIV-infected patients with biopsy- or cytology-proven malignant lymphomas since 2005. Data on HIV infection and lymphoma characteristics, treatment and outcome were recorded. For this analysis, AIDS-related lymphomas patients in complete remission were analyzed in terms of their relapse- free survival and potential risk factors for relapses. In total, 254 of 399 (63.7%) patients with AIDS-related lymphomas reached a complete remission with their first-line chemotherapy. After a median follow up of 4.6 years, 5-year overall survival of the 254 patients was 87.8% (Standard Error 3.1%). Twenty-nine patients relapsed (11.4%). Several factors were independently associated with a higher relapse rate, including an unclassifiable histology, a stage III or IV according to the Ann Arbor Staging System, no concomitant combined antiretroviral therapy during chemotherapy and R-CHOP-based compared to more intensive chemotherapy regimens in Burkitt lymphomas. In conclusion, complete remission and relapse rates observed in our study are similar to those reported in HIV-negative non-Hodgkin lymphomas. These data provide further evidence for the use of concomitant combined antiretroviral therapy during chemotherapy and a benefit from more intensive chemotherapy regimens in Burkitt lymphomas. Modifications to the chemotherapy regimen appear to have only a limited impact on relapse rate.
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http://dx.doi.org/10.3324/haematol.2017.180893DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5927994PMC
May 2018

Optimization of the EC26-2A4 Epitope in the gp41 Membrane Proximal External Region Targeted by Neutralizing Antibodies from an Elite Controller.

AIDS Res Hum Retroviruses 2018 04 25;34(4):365-374. Epub 2018 Jan 25.

1 Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy , Frankfurt, Germany .

The analysis of patient derived HIV neutralizing antibodies (nAbs) and their target epitopes in the viral envelope (Env) protein provides important basic information for vaccine design. In this study we optimized an epitope, EC26-2A4, that is targeted by neutralizing antibodies from an elite controller (EC26) and localizes in the membrane-proximal external region from the gp41 transmembrane protein. Due to its overlap with the epitope of the first generation broadly neutralizing monoclonal Ab (mAb) 2F5 associated with autoreactivity, we first defined the minimal core epitope reacting with antibodies from EC26 plasma, but not with mAb 2F5. The optimized minimal epitope, EC26-2A4ΔM, was able to induce neutralizing antibodies in vaccinated mice. We further analyzed the frequency of antibodies against the EC26-2A4ΔM peptide in HIV-positive patient sera from a treated cohort and an untreated long-term nonprogressor (LTNP) cohort. Interestingly, 27% of the LTNP sera reacted with the peptide, whereas only 9% showed reactivity in the treated cohort. Although there was no association between the presence of antibodies against the EC26-2A4ΔM epitope and viral load or CD4 count in these patients, the CD4 nadir in the treated cohort was higher in patients positive for EC26-2A4ΔM antibodies, in particular in patients having such antibodies at an early and a late timepoint after infection.
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http://dx.doi.org/10.1089/AID.2017.0250DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5899297PMC
April 2018

Control measures following a case of imported Lassa fever from Togo, North Rhine Westphalia, Germany, 2016.

Euro Surveill 2017 Sep;22(39)

Department I of Internal Medicine, University of Cologne, Germany.

In a patient transferred from Togo to Cologne, Germany, Lassa fever was diagnosed 12 days post mortem. Sixty-two contacts in Cologne were categorised according to the level of exposure, and gradual infection control measures were applied. No clinical signs of Lassa virus infection or Lassa specific antibodies were observed in the 62 contacts. Thirty-three individuals had direct contact to blood, other body fluids or tissue of the patients. Notably, with standard precautions, no transmission occurred between the index patient and healthcare workers. However, one secondary infection occurred in an undertaker exposed to the corpse in Rhineland-Palatinate, who was treated on the isolation unit at the University Hospital of Frankfurt. After German authorities raised an alert regarding the imported Lassa fever case, an American healthcare worker who had cared for the index patient in Togo, and who presented with diarrhoea, vomiting and fever, was placed in isolation and medevacked to the United States. The event and the transmission of Lassa virus infection outside of Africa underlines the need for early diagnosis and use of adequate personal protection equipment (PPE), when highly contagious infections cannot be excluded. It also demonstrates that larger outbreaks can be prevented by infection control measures, including standard PPE.
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http://dx.doi.org/10.2807/1560-7917.ES.2017.22.39.17-00088DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5709954PMC
September 2017

Favipiravir and Ribavirin Treatment of Epidemiologically Linked Cases of Lassa Fever.

Clin Infect Dis 2017 Sep;65(5):855-859

Department of Medicine, Infectious Diseases Unit, Goethe University Hospital, Frankfurt/Main, Germany.

Two patients with Lassa fever are described who are the first human cases treated with a combination of ribavirin and favipiravir. Both patients survived but developed transaminitis and had prolonged detectable virus RNA in blood and semen, suggesting that the possibility of sexual transmission of Lassa virus should be considered.
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http://dx.doi.org/10.1093/cid/cix406DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5682919PMC
September 2017

The MalR type regulator AcrC is a transcriptional repressor of acarbose biosynthetic genes in Actinoplanes sp. SE50/110.

BMC Genomics 2017 07 25;18(1):562. Epub 2017 Jul 25.

Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615, Bielefeld, Germany.

Background: Acarbose is used in the treatment of diabetes mellitus type II and is produced by Actinoplanes sp. SE50/110. Although the biosynthesis of acarbose has been intensively studied, profound knowledge about transcription factors involved in acarbose biosynthesis and their binding sites has been missing until now. In contrast to acarbose biosynthetic gene clusters in Streptomyces spp., the corresponding gene cluster of Actinoplanes sp. SE50/110 lacks genes for transcriptional regulators.

Results: The acarbose regulator C (AcrC) was identified through an in silico approach by aligning the LacI family regulators of acarbose biosynthetic gene clusters in Streptomyces spp. with the Actinoplanes sp. SE50/110 genome. The gene for acrC, located in a head-to-head arrangement with the maltose/maltodextrin ABC transporter malEFG operon, was deleted by introducing PCR targeting for Actinoplanes sp. SE50/110. Characterization was carried out through cultivation experiments, genome-wide microarray hybridizations, and RT-qPCR as well as electrophoretic mobility shift assays for the elucidation of binding motifs. The results show that AcrC binds to the intergenic region between acbE and acbD in Actinoplanes sp. SE50/110 and acts as a transcriptional repressor on these genes. The transcriptomic profile of the wild type was reconstituted through a complementation of the deleted acrC gene. Additionally, regulatory sequence motifs for the binding of AcrC were identified in the intergenic region of acbE and acbD. It was shown that AcrC expression influences acarbose formation in the early growth phase. Interestingly, AcrC does not regulate the malEFG operon.

Conclusions: This study characterizes the first known transcription factor of the acarbose biosynthetic gene cluster in Actinoplanes sp. SE50/110. It therefore represents an important step for understanding the regulatory network of this organism. Based on this work, rational strain design for improving the biotechnological production of acarbose can now be implemented.
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http://dx.doi.org/10.1186/s12864-017-3941-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5526262PMC
July 2017

Genome improvement of the acarbose producer Actinoplanes sp. SE50/110 and annotation refinement based on RNA-seq analysis.

J Biotechnol 2017 Jun 17;251:112-123. Epub 2017 Apr 17.

Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany. Electronic address:

Actinoplanes sp. SE50/110 is the natural producer of acarbose, which is used in the treatment of diabetes mellitus type II. However, until now the transcriptional organization and regulation of the acarbose biosynthesis are only understood rudimentarily. The genome sequence of Actinoplanes sp. SE50/110 was known before, but was resequenced in this study to remove assembly artifacts and incorrect base callings. The annotation of the genome was refined in a multi-step approach, including modern bioinformatic pipelines, transcriptome and proteome data. A whole transcriptome RNA-seq library as well as an RNA-seq library enriched for primary 5'-ends were used for the detection of transcription start sites, to correct tRNA predictions, to identify novel transcripts like small RNAs and to improve the annotation through the correction of falsely annotated translation start sites. The transcriptome data sets were also applied to identify 31 cis-regulatory RNA structures, such as riboswitches or RNA thermometers as well as three leaderless transcribed short peptides found in putative attenuators upstream of genes for amino acid biosynthesis. The transcriptional organization of the acarbose biosynthetic gene cluster was elucidated in detail and fourteen novel biosynthetic gene clusters were suggested. The accurate genome sequence and precise annotation of the Actinoplanes sp. SE50/110 genome will be the foundation for future genetic engineering and systems biology studies.
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http://dx.doi.org/10.1016/j.jbiotec.2017.04.013DOI Listing
June 2017

subsp. Is Capable of Degrading Quinolone Signals.

Front Microbiol 2017 2;8:339. Epub 2017 Mar 2.

Institute for Molecular Microbiology and Biotechnology, University of Münster Münster, Germany.

employs 2-heptyl-3-hydroxy-4(1)-quinolone (the quinolone signal, PQS) and 2-heptyl-4(1)-quinolone (HHQ) as quorum sensing signal molecules, which contribute to a sophisticated regulatory network controlling the production of virulence factors and antimicrobials. We demonstrate that and clinical isolates are capable of degrading these alkylquinolone signals. Genome sequences of 50 clinical isolates indicated the presence of genes, contributing to fast degradation of HHQ and PQS, in subsp. strains, but not in subsp. and subsp. isolates. A subset of 18 . subsp. isolates contained the same five single nucleotide polymorphisms (SNPs) compared to the region of the type strain. Interestingly, representatives of these isolates showed faster PQS degradation kinetics than the type strain. One of the SNPs is located in the predicted promoter region of the gene encoding a putative transcriptional regulator, and two others lead to a variant of the AqdC protein termed AqdC, which differs in two amino acids from AqdC of the type strain. AqdC, the key enzyme of the degradation pathway, is a PQS dioxygenase catalyzing quinolone ring cleavage. While transcription of and is induced by PQS, transcript levels in a representative of the subset of 18 isolates were not significantly altered despite the detected SNP in the promoter region. However, purified recombinant AqdC and AqdC exhibit different kinetic properties, with approximate apparent values for PQS of 14 μM and 37 μM, and values of 61 s and 98 s, respectively, which may (at least in part) account for the observed differences in PQS degradation rates of the strains. In co-culture experiments of PAO1 and , strains harboring the genes reduced the PQS levels, whereas mycobacteria lacking the gene cluster even boosted PQS production. The results suggest that the presence and expression of the genes in lead to a competitive advantage against .
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http://dx.doi.org/10.3389/fmicb.2017.00339DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5332414PMC
March 2017

HIV-Associated Malignant Lymphoma.

Oncol Res Treat 2017 24;40(3):82-87. Epub 2017 Feb 24.

Acquired immune deficiency syndrome (AIDS)-related lymphomas (ARL) still represent a relevant field of clinical research. For most histological subtypes of ARL, no optimal initial therapy has been clearly defined so far. Rituximab plus chemotherapy is feasible and effective and should be offered to all patients with CD20-positive ARL regardless of their CD4 cell count. Combination antiretroviral therapy (cART) should be given concomitantly with chemotherapy, bearing in mind potential drug-drug interactions. Appropriate treatment of ARL is determined by a number of factors such as lymphoma stage, performance status, comorbidities, histological subtype, and immunosuppression. Treatment should principally be the same as in human immunodeficiency virus (HIV)-negative lymphoma patients. In HIV-related Hodgkin's lymphoma, high cure rates have been achieved with stage-adapted treatment approaches, albeit with worse outcomes compared to immunocompetent patients.
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http://dx.doi.org/10.1159/000456036DOI Listing
October 2017

HIV-1 replication in central nervous system increases over time on only protease inhibitor therapy.

Med Microbiol Immunol 2016 Dec 28;205(6):575-583. Epub 2016 Jul 28.

Infectious diseases Unit, Medical Department, University Hospital Frankfurt, Goethe-University, Theodor Stern Kai 7, 60590, Frankfurt, Germany.

There are concerns about central nervous system (CNS)-replication of HIV-1 in patients on boosted protease inhibitors. Purpose of this study was to compare HIV-1 viral loads (VLs) from patients treated with only boosted dual protease inhibitor (bdPI), versus combination antiretroviral therapy (cART group), containing two nucleoside analogue reverse transcriptase inhibitors (NRTI) and a third partner. All patients from a large German HIV-treatment cohort with available medication, clinical and demographic data, including results from simultaneous HIV-1 viral load (VL) assessments in cerebrospinal fluid (CSF) and blood plasma, were retrospectively evaluated as controlled cross-sectional study. CSF had been obtained from patients with variable neurological symptoms during 2005-2014. Statistical analysis comprised nonparametric tests, regression and correlation techniques accounting for undetectable quantifications. Statistical analysis comprised nonparametric tests, regression and correlation techniques accounting for undetectable quantifications. Overall, 155 patients were evaluable (bdPI: 24; cART: 131). At time of CSF-collection, both groups were comparable in age, gender, CD4-cell counts, or primary HIV-transmission risks, though bdPI patients were clinically more advanced. The proportion of patients with undetectable HIV-1 (<50 copies/ml) in CSF was lower for bdPI group (25 vs 49.6 %; p = 0.026), but similar in plasma (46 vs 41 %). Median CSF-VL was higher in bdPI group (600 vs 50 copies/ml; p = 0.027) and similar in plasma. Mean VL CSF/plasma ratio was 342.91 for bdPI- and 54.48 for cART patients (p < 0.001). Pearson's regression analysis revealed a trend for an elevated VL-ratio over time within bdPI group. HIV-1 replication was higher and more frequently detectable in CSF from bdPI patients, indicating a worse CNS penetration effectiveness of used boosted PI. Within bdPI group, measured CNS-viral replication was increasing over time, suggesting an over time impaired HIV-1 suppression in CSF.
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http://dx.doi.org/10.1007/s00430-016-0469-7DOI Listing
December 2016

Complete Genome Sequence of an Ebola Virus Isolate Imported from Sierra Leone to Germany Determined by Circle Sequencing.

Genome Announc 2016 Oct 6;4(5). Epub 2016 Oct 6.

Institute for Medical Microbiology, Justus-Liebig University of Giessen, Giessen, Germany

We report here a complete genome sequence of Ebola virus Makona from a nonfatal patient sample that originated in Sierra Leone during the last Ebola virus outbreak in West Africa (species Zaire ebolavirus) using a highly accurate circle sequencing (Cir-seq) method.
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http://dx.doi.org/10.1128/genomeA.01011-16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5054307PMC
October 2016

Genome Sequence of Lassa Virus Isolated from the First Domestically Acquired Case in Germany.

Genome Announc 2016 Sep 22;4(5). Epub 2016 Sep 22.

Institute of Virology, Philipps University Marburg, Marburg, Germany German Center for Infection Research (DZIF), Partner Site Giessen-Marburg-Langen, Germany

Lassa virus (LASV) is a zoonotic, hemorrhagic fever-causing virus endemic in West Africa, for which no approved vaccines or specific treatment options exist. Here, we report the genome sequence of LASV isolated from the first case of acquired Lassa fever disease outside of Africa.
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http://dx.doi.org/10.1128/genomeA.00938-16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5034122PMC
September 2016

Targeted genome editing in the rare actinomycete Actinoplanes sp. SE50/110 by using the CRISPR/Cas9 System.

J Biotechnol 2016 Aug 1;231:122-128. Epub 2016 Jun 1.

Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany. Electronic address:

The application of genome editing technologies, like CRISPR/Cas9 for industrially relevant microorganisms, is becoming increasingly important. Compared to other methods of genetic engineering the decisive factor is that CRISPR/Cas9 is relatively easy to apply and thus time and effort can be significantly reduced in organisms, which are otherwise genetically difficult to access. Because of its many advantages and opportunities, we adopted the CRISPR/Cas9 technology for Actinoplanes sp. SE50/110, the producer of the diabetes type II drug acarbose. The functionality of genome editing was successfully shown by the scarless and antibiotic marker-free deletion of the gene encoding the tyrosinase MelC, which catalyzes the formation of the dark pigment eumelanin in the wild type strain. The generated ΔmelC2 mutant of Actinoplanes sp. SE50/110 no longer produces this pigment and therefore the supernatant does not darken. Furthermore, it was shown that the plasmid containing the gene for the Cas9 protein was removed by increasing the temperature due to its temperature-sensitive replication. The precision of the intended mutation was proven and possible off-target effects caused by the genome editing system were ruled out by genome sequencing of several mutants.
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http://dx.doi.org/10.1016/j.jbiotec.2016.05.039DOI Listing
August 2016