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Case Reports
. 2005 Jul;11(7):1036-41.
doi: 10.3201/eid1107.041313.

Influenza A H5N1 replication sites in humans

Mongkol Uiprasertkul  1 , Pilaipan Puthavathana, Kantima Sangsiriwut, Phisanu Pooruk, Kanittar Srisook, Malik Peiris, John M Nicholls, Kulkanya Chokephaibulkit, Nirun Vanprapar, Prasert Auewarakul
Affiliations
Case Reports

Influenza A H5N1 replication sites in humans

Mongkol Uiprasertkul et al. Emerg Infect Dis. 2005 Jul.

Abstract

Tissue tropism and pathogenesis of influenza A virus subtype H5N1 disease in humans is not well defined. In mammalian experimental models, H5N1 influenza is a disseminated disease. However, limited previous data from human autopsies have not shown evidence of virus dissemination beyond the lung. We investigated a patient with fatal H5N1 influenza. Viral RNA was detected by reverse transcription-polymerase chain reaction in lung, intestine, and spleen tissues, but positive-stranded viral RNA indicating virus replication was confined to the lung and intestine. Viral antigen was detected in pneumocytes by immunohistochemical tests. Tumor necrosis factor-? mRNA was seen in lung tissue. In contrast to disseminated infection documented in other mammals and birds, H5N1 viral replication in humans may be restricted to the lung and intestine, and the major site of H5N1 viral replication in the lung is the pneumocyte.

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Figures

Figure 1
Figure 1
Microscopic shape of the lung showing proliferative phase of diffuse alveolar damage and interstitial pneumonia with reactive hyperplasia of pneumocytes (magnification x100).
Figure 2
Figure 2
A) Detection of H5 influenza viral RNA in lungs, intestines, and spleen by reverse transcription-polymerase chain reaction (RT-PCR). B) Strand-specific RT-PCR detected positive-stranded viral RNA only in lungs and intestines but not in spleen. +/–, total RNA; –, negative-stranded RNA; +, positive-stranded RNA. RT-PCR products of an infected cell culture pellet and supernatant are shown as a control for proper amplification of the specific strands (lower panel). C) Tumor necrosis factor-α (TNF-α) mRNA was detected by RT-PCR only in lung tissue of the patient but not in lung tissue from a healthy control.
Figure 3
Figure 3
Immunohistochemical analysis showing influenza A antigen-specific staining in nuclei of cells lining the alveoli (A). To identify the cell type, slides from consecutive sections were stained with anti-influenza A antibody (B) and double-stained with antiinfluenza A and antisurfactant antibodies (C). The sections were mapped, and the same area in each section was examined. Viral antigen-positive cells were stained both intranuclearly with antiinfluenza antibody and intracytoplasmically with antisurfactant antibody, indicating that the viral antigen-positive cells were type II pneumocytes. Viral antigen-positive cell are marked by circles (magnification x400).
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