TY  - JOUR
A1  - Carvalho-Pereira, Joana
A1  - Fernandes, Filipa
A1  - Araújo, Ricardo
A1  - Springer, Jan
A1  - Loeffler, Juergen
A1  - Buitrago, María José
A1  - Pais, Célia
A1  - Sampaio, Paula
T1  - Multiplex PCR based strategy for detection of fungal pathogen DNA in patients with suspected invasive fungal infections
JF  - Journal of Fungi
N2  - A new and easy polymerase chain reaction (PCR) multiplex strategy, for the identification of the most common fungal species involved in invasive fungal infections (IFI) was developed in this work. Two panels with species-specific markers were designed, the Candida Panel for the identification of Candida species, and the Filamentous Fungi Panel for the identification of Aspergillus species and Rhizopusarrhizus. The method allowed the correct identification of all targeted pathogens using extracted DNA or by colony PCR, showed no cross-reactivity with nontargeted species and allowed identification of different species in mixed infections. Sensitivity reached 10 to 1 pg of DNA and was suitable for clinical samples from sterile sites, with a sensitivity of 89% and specificity of 100%. Overall, the study showed that the new method is suitable for the identification of the ten most important fungal species involved in IFI, not only from positive blood cultures but also from clinical samples from sterile sites. The method provides a unique characteristic, of seeing the peak in the specific region of the panel with the correct fluorescence dye, that aids the ruling out of unspecific amplifications. Furthermore, the panels can be further customized, selecting markers for different species and/or resistance genes.
KW  - molecular diagnosis
KW  - fungal infections
KW  - multiplex PCR
KW  - Candida sp.
KW  - Aspergillus sp.
KW  - Rhizopus arrhizus
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-219392
SN  - 2309-608X
VL  - 6
IS  - 4
ER  - 
TY  - JOUR
A1  - Sánchez-Maldonado, Jose Manuel
A1  - Moñiz-Díez, Ana
A1  - ter Horst, Rob
A1  - Campa, Daniele
A1  - Cabrera-Serrano, Antonio José
A1  - Martínez-Bueno, Manuel
A1  - Garrido-Collado, María del Pilar
A1  - Hernández-Mohedo, Francisca
A1  - Fernández-Puerta, Laura
A1  - López-Nevot, Miguel Ángel
A1  - Cunha, Cristina
A1  - González-Sierra, Pedro Antonio
A1  - Springer, Jan
A1  - Lackner, Michaela
A1  - Alcazar-Fuoli, Laura
A1  - Fianchi, Luana
A1  - Aguado, José María
A1  - Pagano, Livio
A1  - López-Fernández, Elisa
A1  - Clavero, Esther
A1  - Potenza, Leonardo
A1  - Luppi, Mario
A1  - Moratalla, Lucia
A1  - Solano, Carlos
A1  - Sampedro, Antonio
A1  - Cuenca-Estrella, Manuel
A1  - Lass-Flörl, Cornelia
A1  - Canzian, Federico
A1  - Loeffler, Juergen
A1  - Li, Yang
A1  - Einsele, Hermann
A1  - Netea, Mihai G.
A1  - Vázquez, Lourdes
A1  - Carvalho, Agostinho
A1  - Jurado, Manuel
A1  - Sainz, Juan
T1  - Polymorphisms within the TNFSF4 and MAPKAPK2 loci influence the risk of developing invasive aspergillosis: a two-stage case control study in the context of the aspBIOmics consortium
JF  - Journal of Fungi
N2  - Here, we assessed whether 36 single nucleotide polymorphisms (SNPs) within the TNFSF4 and MAPKAPK2 loci influence the risk of developing invasive aspergillosis (IA). We conducted a two-stage case control study including 911 high-risk patients diagnosed with hematological malignancies that were ascertained through the aspBIOmics consortium. The meta-analysis of the discovery and replication populations revealed that carriers of the TNFSF4\(_{rs7526628T/T}\) genotype had a significantly increased risk of developing IA (p = 0.00022). We also found that carriers of the TNFSF4\(_{rs7526628T}\) allele showed decreased serum levels of TNFSF14 protein (p = 0.0027), and that their macrophages had a decreased fungicidal activity (p = 0.048). In addition, we observed that each copy of the MAPKAPK2\(_{rs12137965G}\) allele increased the risk of IA by 60% (p = 0.0017), whereas each copy of the MAPKAPK2\(_{rs17013271T}\) allele was estimated to decrease the risk of developing the disease (p = 0.0029). Mechanistically, we found that carriers of the risk MAPKAPK2\(_{rs12137965G}\) allele showed increased numbers of CD38+IgM-IgD- plasmablasts in blood (p = 0.00086), whereas those harboring two copies of the allele had decreased serum concentrations of thymic stromal lymphopoietin (p = 0.00097). Finally, we also found that carriers of the protective MAPKAPK2\(_{rs17013271T}\) allele had decreased numbers of CD27-IgM-IgD- B cells (p = 0.00087) and significantly lower numbers of CD14+ and CD14+CD16- cells (p = 0.00018 and 0.00023). Altogether, these results suggest a role of the TNFSF4 and MAPKAPK2 genes in determining IA risk.
KW  - invasive aspergillosis
KW  - TNFSF4
KW  - MAPKAPK2
KW  - genetic susceptibility
KW  - B cells
KW  - monocytes
KW  - serum biomarkers
KW  - TSLP
KW  - TNFSF14
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-220107
SN  - 2309-608X
VL  - 7
IS  - 1
ER  - 
TY  - JOUR
A1  - Weiss, Esther
A1  - Schlegel, Jan
A1  - Terpitz, Ulrich
A1  - Weber, Michael
A1  - Linde, Jörg
A1  - Schmitt, Anna-Lena
A1  - Hünniger, Kerstin
A1  - Marischen, Lothar
A1  - Gamon, Florian
A1  - Bauer, Joachim
A1  - Löffler, Claudia
A1  - Kurzai, Oliver
A1  - Morton, Charles Oliver
A1  - Sauer, Markus
A1  - Einsele, Hermann
A1  - Loeffler, Juergen
T1  - Reconstituting NK Cells After Allogeneic Stem Cell Transplantation Show Impaired Response to the Fungal Pathogen Aspergillus fumigatus
JF  - Frontiers in Immunology
N2  - Delayed natural killer (NK) cell reconstitution after allogeneic stem cell transplantation (alloSCT) is associated with a higher risk of developing invasive aspergillosis. The interaction of NK cells with the human pathogen Aspergillus (A.) fumigatus is mediated by the fungal recognition receptor CD56, which is relocated to the fungal interface after contact. Blocking of CD56 signaling inhibits the fungal mediated chemokine secretion of MIP-1α, MIP-1β, and RANTES and reduces cell activation, indicating a functional role of CD56 in fungal recognition. We collected peripheral blood from recipients of an allograft at defined time points after alloSCT (day 60, 90, 120, 180). NK cells were isolated, directly challenged with live A. fumigatus germ tubes, and cell function was analyzed and compared to healthy age and gender-matched individuals. After alloSCT, NK cells displayed a higher percentage of CD56\(^{bright}\)CD16\(^{dim}\) cells throughout the time of blood collection. However, CD56 binding and relocalization to the fungal contact side were decreased. We were able to correlate this deficiency to the administration of corticosteroid therapy that further negatively influenced the secretion of MIP-1α, MIP-1β, and RANTES. As a consequence, the treatment of healthy NK cells ex vivo with corticosteroids abrogated chemokine secretion measured by multiplex immunoassay. Furthermore, we analyzed NK cells regarding their actin cytoskeleton by Structured Illumination Microscopy (SIM) and flow cytometry and demonstrate an actin dysfunction of NK cells shown by reduced F-actin content after fungal co-cultivation early after alloSCT. This dysfunction remains until 180 days post-alloSCT, concluding that further actin-dependent cellular processes may be negatively influenced after alloSCT. To investigate the molecular pathomechansism, we compared CD56 receptor mobility on the plasma membrane of healthy and alloSCT primary NK cells by single-molecule tracking. The results were very robust and reproducible between tested conditions which point to a different molecular mechanism and emphasize the importance of proper CD56 mobility.
KW  - natural killer cell
KW  - stem cell transplantation
KW  - corticosteroids
KW  - CCL3
KW  - CCL4
KW  - CCL5
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-212581
SN  - 1664-3224
VL  - 11
ER  -