Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2008 Feb 13;3(2):e1582.
doi: 10.1371/journal.pone.0001582.

RNA-containing cytoplasmic inclusion bodies in ciliated bronchial epithelium months to years after acute Kawasaki disease

Anne H Rowley  1 , Susan C Baker, Stanford T Shulman, Francesca L Garcia, Linda M Fox, Ian M Kos, Susan E Crawford, Pierre A Russo, Rashid Hammadeh, Kei Takahashi, Jan M Orenstein
Affiliations

RNA-containing cytoplasmic inclusion bodies in ciliated bronchial epithelium months to years after acute Kawasaki disease

Anne H Rowley et al. PLoS One. .

Abstract

Background: Kawasaki Disease (KD) is the most common cause of acquired heart disease in children in developed nations. The KD etiologic agent is unknown but likely to be a ubiquitous microbe that usually causes asymptomatic childhood infection, resulting in KD only in genetically susceptible individuals. KD synthetic antibodies made from prevalent IgA gene sequences in KD arterial tissue detect intracytoplasmic inclusion bodies (ICI) resembling viral ICI in acute KD but not control infant ciliated bronchial epithelium. The prevalence of ICI in late-stage KD fatalities and in older individuals with non-KD illness should be low, unless persistent infection is common.

Methods and principal findings: Lung tissue from late-stage KD fatalities and non-infant controls was examined by light microscopy for the presence of ICI. Nucleic acid stains and transmission electron microscopy (TEM) were performed on tissues that were strongly positive for ICI. ICI were present in ciliated bronchial epithelium in 6/7 (86%) late-stage KD fatalities and 7/27 (26%) controls ages 9-84 years (p = 0.01). Nucleic acid stains revealed RNA but not DNA within the ICI. ICI were also identified in lung macrophages in some KD cases. TEM of bronchial epithelium and macrophages from KD cases revealed finely granular homogeneous ICI.

Significance: These findings are consistent with a previously unidentified, ubiquitous RNA virus that forms ICI and can result in persistent infection in bronchial epithelium and macrophages as the etiologic agent of KD.

PubMed Disclaimer

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Light microscopic studies of tissues from KD patient 1.
A, H&E of ciliated bronchial epithelium, demonstrating amphophilic supranuclear ICI (arrows); B, IHC using synthetic antibody J of ciliated bronchial epithelium, showing ICI (arrows); C, methyl green pyronin stain of ciliated bronchial epithelium, demonstrating red ICI (arrows) indicating the presence of RNA; D, IHC using synthetic antibody J on myocardium lesion, demonstrating antigen-positive macrophages (thin arrows) infiltrating an area that has undergone myocardial dropout near a vessel (thick arrow shows single viable cardiac myocyte in this field). A–D = 40X.
Figure 2
Figure 2. Light microscopic studies of ciliated bronchial epithelium from KD patient 3.
A, immunohistochemistry (IHC) using synthetic antibody J, examples of specific staining of ICI indicated with arrows; B, IHC using control antibody I, no specific staining; C, H&E stain demonstrating that ICI are visible as amphophilic bodies (arrows); D, methyl green pyronin stain of paraffin section of a tangentially sectioned bronchus showing purple nuclei (DNA) and red ICI (RNA, arrows); E, higher-power image of Figure A, showing intense brown staining of multiple clusters of intracytoplasmic inclusion bodies (ICI, arrows); F, IHC from another bronchus, demonstrating fewer ICI (block arrows) and an IHC-positive macrophage in the lumen of the bronchus (thin arrow); G, frozen section of lung stained with methyl green pyronin; this unfixed tissue section shows typical blue-green nuclei (open arrows) and shows clusters of (thin arrow) and individual (black block arrows) ICI staining red, characteristic of RNA. A,B taken with 10X objective, E = 20X, C, D, F,G = 40X.
Figure 3
Figure 3. Peribronchial lymph node from KD patient 2.
A, IHC using synthetic antibody J, showing antigen-positive macrophages containing spheroid bodies (arrows); B, transmission electron microscopy (TEM) of macrophage, showing two large, finely granular ICI (arrows). A = 40X, B = 20,000X.
Figure 4
Figure 4. TEM of formalin-fixed, paraffin-embedded lung from KD patient 1.
A, ciliated bronchial epithelium demonstrating electron-dense apical ICI (block arrows). B, alveolar macrophage, demonstrating perinuclear, finely granular spheroid bodies similar to those seen within the bronchial epithelium (block arrow). N = nucleus, C = cilia. A = 9,500X, B = 26,000X.
Figure 5
Figure 5. Transmission electron microscopy (TEM) of ciliated bronchial epithelium from glutaraldehyde-fixed, previously frozen lung from KD patient 3.
A) Lower power image demonstrating finely granular, variably electron-dense ICI (block arrows). N = nucleus, L = lysosomes, M = mitochondria, C = cilia. B) Higher power image of ICI (block arrows). Note rough endoplasmic reticulum (thin arrow). ICI are spherical and do not appear membrane-bound. A = 14,000X, B = 52,000X.
See this image and copyright information in PMC

References

    1. Rowley AH, Shulman ST. Kawasaki Disease. In: Kliegman R, Behrman R, Jenson H, Stanton B, editors. Nelson Textbook of Pediatrics 18th edition. Philadelphia: Elsevier; 2007. pp. 1036–42.
    1. Rowley AH, Shulman ST. New developments in the search for the etiologic agent of Kawasaki disease. Current Opinion in Pediatrics. 2007;19:71–74. - PubMed
    1. Rowley AH, Eckerley CA, Jack HM, Shulman ST, Baker SC. IgA plasma cells in vascular tissue of patients with Kawasaki syndrome. J Immunol. 1997;159:5946–55. - PubMed
    1. Rowley AH, Shulman ST, Mask CA, Finn LS, Terai M, et al. IgA plasma cell infiltration of proximal respiratory tract, pancreas, kidney, and coronary artery in acute Kawasaki disease. J Infect Dis. 2000;182:1183–91. - PubMed
    1. Rowley AH, Shulman ST, Spike BT, Mask CA, Baker SC. Oligoclonal IgA response in the vascular wall in acute Kawasaki disease. J Immunol. 2001;166:1334–43. - PubMed

Publication types

MeSH terms