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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 J+Virol
2019 ; 93
(19
): ä Nephropedia Template TP
gab.com Text
Twit Text FOAVip
Twit Text #
English Wikipedia
Serial Section Array Scanning Electron Microscopy Analysis of Cells from Lung
Autopsy Specimens following Fatal A/H1N1 2009 Pandemic Influenza Virus Infection
#MMPMID31292247
Kataoka M
; Ishida K
; Ogasawara K
; Nozaki T
; Satoh YI
; Sata T
; Sato Y
; Hasegawa H
; Nakajima N
J Virol
2019[Oct]; 93
(19
): ä PMID31292247
show ga
A/H1N1 2009 pandemic influenza virus (A/H1N1/pdm09) was first identified as a
novel pandemic influenza A virus (IAV) in 2009. Previously, we reported that many
viral antigens were detected in type II alveolar epithelial cells (AEC-IIs)
within autopsied lung tissue from a patient with A/H1N1/pdm09 pneumonia. It is
important to identify the association between the virus and host cells to
elucidate the pathogenesis of IAV pneumonia. To investigate the distribution of
virus particles and morphological changes in host cells, the autopsied lung
specimens from this patient were examined using transmission electron microscopy
(TEM) and a novel scanning electron microscopy (SEM) method. We focused on
AEC-IIs as viral antigen-positive cells and on monocytes/macrophages (Ms/M?s) and
neutrophils (Neus) as innate immune cells. We identified virus particles and
intranuclear dense tubules, which are associated with matrix 1 (M1) proteins from
IAV. Large-scale two-dimensional observation was enabled by digitally "stitching"
together contiguous SEM images. A single whole-cell analysis using a serial
section array (SSA)-SEM identified virus particles in vesicles within the
cytoplasm and/or around the surfaces of AEC-IIs, Ms/M?s, and Neus; however,
intranuclear dense tubules were found only in AEC-IIs. Computer-assisted
processing of SSA-SEM images from each cell type enabled three-dimensional (3D)
modeling of the distribution of virus particles within an ACE-II, a M/M?, and a
Neu.IMPORTANCE Generally, it is difficult to observe IAV particles in postmortem
samples from patients with seasonal influenza. In fact, only a few viral antigens
are detected in bronchial epithelial cells from autopsied lung sections.
Previously, we detected many viral antigens in AEC-IIs from the lung. This was
because the majority of A/H1N1/pdm09 in the lung tissue harbored an aspartic
acid-to-glycine substitution at position 222 (D222G) of the hemagglutinin
protein. A/H1N1/pdm09 harboring the D222G substitution has a receptor-binding
preference for ?-2,3-linked sialic acids expressed on human AECs and infects them
in the same way as H5N1 and H7N9 avian IAVs. Here, we report the first successful
observation of virus particles, not only in AEC-IIs, but also in Ms/M?s and Neus,
using electron microscopy. The finding of a M/M? harboring numerous virus
particles within vesicles and at the cell surface suggests that Ms/M?s are
involved in the pathogenesis of IAV primary pneumonia.