Author:
Julien Alex, Katherine González, Gauri Gangapurwala, Antje Vollrath, Zoltán Cseresnyés, Christine Weber, Justyna A. Czaplewska, Stephanie Hoeppener, Carl-Magnus Svensson, Thomas Orasch, Thorsten Heinekamp, Carlos Guerrero-Sánchez, Marc Thilo Figge, Ulrich S. Schubert, Axel A. Brakhage
Abstract:
Objectives: In immunocompromised patients, conidia of Aspergillus fumigatus can avoid their elimination inside of alveolar macrophages by interfering with the maturation of phagolysosomes. Currently used antifungals in clinics often prove to be inefficient to treat intracellular persisting pathogens because the drugs have to cross two membranes, i.e., the cytoplasmic and the phagosomal membrane. The utilization of drug delivery systems bears the potential to alter the internalization route of antifungals and thus, reach intracellular persistent microorganisms inside phagosomes or phagolysosomes more efficiently compared to the pristine drug. The aim of this study was to elucidated whether polymeric particles (PP) can target intracellular persistent pathogens.
Methods: We formulated dye-labeled PPs via single emulsion technique to generate PP sizes large enough for their phagocytosis by macrophages. The covalent conjugation of dye molecules to the polymer was confirmed by size exclusion chromatography. PPs were characterized by means of dynamic light scattering and scanning electron microscopy. The uptake of PPs into RAW 264.7 macrophages was analyzed by imaging flow cytometry. Studies with regards to the intracellular localization of PPs was executed via immunofluorescent staining of Rab7 and Lamp1 (confocal laser scanning microscopy) and transmission electron microscopy.
Results: RAW 264.7 macrophages internalized PPs efficiently. Co-localization of PPs and conidia in the same phagolysosome was observed after PPs were added to prior infected macrophages. The fusion of phagolysosomes containing PPs with phagolysosomes containing conidia was confirmed. The number of phagolysosomes containing both conidia and PPs was increased at elevated PP concentrations or after utilization of the fusion enhancer Vacuolin-1.
Conclusion: Our study suggests the fusion of conidia- and PP- containing phagolysosomes as the underlaying mechanism for PPs reaching intracellular A. fumigatus. Methods for increasing fusion events were established in order to exploit the investigated delivery route of PPs. These results represent the requisite for the development of advanced delivery systems reaching intracellular persistent pathogens.
Abstract Number: 84
Conference Year: 2024
Conference abstracts, posters & presentations
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Title
Author
Year
Number
Poster
-
v
Tianyu Lianga, Nir Osherovb, Ruoyu Lia, Qiqi Wanga, Wei Chena, Zhe Wana, Wei Liua*
2024
n/a
-
v
Jillian Simard, MD,1 Christopher Melani, MD,2
Rahul Lakhotia, MD,1
Michail S. Lionakis, MD
ScD,*,3
Stefania Pittaluga, MD PhD,4
James D. Phelan, PhD,*,1 Jagan R. Muppidi, MD
PhD,*,1
Lydia L. Chou, CRNP,*,1
Matthias Holdhoff, MD PhD,*,5 Michael Glantz, MD PhD,*,6
John A. Butman, MD PhD,*,7 Andrea Nicole Lucas, RN,*,8
Seth M. Steinberg, PhD,*,9
Elaine
S. Jaffe, MD,4
Louis M. Staudt, MD PhD,1
Wyndham H. Wilson, MD PhD,1 Mark Roschewski, MD102020
n/a
-
v
Mark Roschewski, MD,1
Christopher Melani, MD,2
Rahul Lakhotia, MD,3
Stefania Pittaluga, MD
PhD,4
James D. Phelan, PhD,*,3
Cody Peer, PhD,*,5 Michail S. Lionakis, MD ScD,*,6 Lydia
L. Chou, CRNP,*,3 Matthias Holdhoff, MD PhD,*,7 Michael Glantz, MD PhD,*,8 Jan Drappatz, MD,*,9
Catherine Lai, MD MPH,10 John A. Butman, MD PhD,*,11 Andrea Nicole Lucas, RN,*,12 Seth
M. Steinberg, PhD,*,13 William D. Figg, PhD,*,14 Elaine S. Jaffe, MD,4
S. Percy Ivy, MD,*,15
Richard F. Little, MDMPH,16 Louis M. Staudt, MD PhD,3
Wyndham H. Wilson, MD PhD32020
n/a
-
v
Binav Baral, MD,*,1 Prasanth Lingamaneni,*,2 Trilok Shrivastava, MD,*,3 Maryam Zia,4 Ishaan Vohra, MD,*,2
Krishna Rekha Moturi*,32020
903
n/a
-
v
Maxwell Green1, Scout Treadwell1, Geetha Gowda1, John Carlson, MD, PhD2
2023
610
n/a
-
v
Catherine Blackwood, PhD1 , Angela Lemons, MS2 , Rachael Rush3 , Walter McKinney2 , Dori Germolec4 , Donald Beezhold, PhD2 , Brett Green, PhD5 , Tara Croston, PhD6
2023
583
n/a
-
v
Catherine Blackwood, PhD1 , Angela Lemons, MS2 , Rachael Rush3 , Walter McKinney2 , Dori Germolec4 , Donald Beezhold, PhD2 , Brett Green, PhD5 , Tara Croston, PhD6
2023
583
n/a
-
v
Jennifer Roper, MD1, Sebastian Ochoa, MD2, Anahita
Agharahimi, MSN, CRNP2, Justina Pfister, RN2, Magdalena
Walkiewicz, PhD2, Amanda Urban, DNP, CRNP3, Rajarshi
Ghosh, PhD2, Michail Lionakis, MD, ScD2, Lisa Kohn, MD, PhD4, Kenneth Olivier, MD, MPH2, Alexandra Freeman, MD22023
516
n/a
-
v
Utkucan Acar, Gloria Sheng, Terrie Ahn, Lisa Kohn, Maria Garcia Lloret, Manish Butte
2022
82
n/a
-
v
C Gutierrez Perez1*, S Dhingra1,2, SM Kwansy3, TJ Opperman3, RA Cramer1
2022
93