Influenza H5N1 virus infection of polarized human alveolar epithelial cells and lung microvascular endothelial cells

Background: Highly pathogenic avian influenza (HPAI) H5N1 virus is entrenched in poultry in Asia and Africa and continues to infect humans zoonotically causing acute respiratory disease syndrome and death. There is evidence that the virus may sometimes spread beyond respiratory tract to cause disseminated infection. The primary target cell for HPAI H5N1 virus in human lung is the alveolar epithelial cell. Alveolar epithelium and its adjacent lung microvascular endothelium form host barriers to the initiation of infection and dissemination of influenza H5N1 infection in humans. These are polarized cells and the polarity of influenza virus entry and egress as well as the secretion of cytokines and chemokines from the virus infected cells are likely to be central to the pathogenesis of human H5N1 disease.Aim: To study influenza A (H5N1) virus replication and host innate immune responses in polarized primary human alveolar epithelial cells and lung microvascular endothelial cells and its relevance to the pathogenesis of human H5N1 disease.Methods: We use an in vitro model of polarized primary human alveolar epithelial cells and lung microvascular endothelial cells grown in transwell culture inserts to compare infection with influenza A subtype H1N1 and H5N1 viruses via the apical or basolateral surfaces.Results: We demonstrate that both influenza H1N1 and H5N1 viruses efficiently infect alveolar epithelial cells from both apical and basolateral surface of the epithelium but release of newly formed virus is mainly from the apical side of the epithelium. In contrast, influenza H5N1 virus, but not H1N1 virus, efficiently infected polarized microvascular endothelial cells from both apical and basolateral aspects. This provides a mechanistic explanation for how H5N1 virus may infect the lung from systemic circulation. Epidemiological evidence has implicated ingestion of virus-contaminated foods as the source of infection in some instances and our data suggests that viremia, secondary to, for example, gastro-intestinal infection, can potentially lead to infection of the lung. HPAI H5N1 virus was a more potent inducer of cytokines (e.g. IP-10, RANTES, IL-6) in comparison to H1N1 virus in alveolar epithelial cells, and these virus-induced chemokines were secreted onto both the apical and basolateral aspects of the polarized alveolar epithelium.Conclusion: The predilection of viruses for different routes of entry and egress from the infected cell is important in understanding the pathogenesis of influenza H5N1 infection and may help unravel the pathogenesis of human H5N1 disease. © 2009 Chan et al; licensee BioMed Central Ltd.published_or_final_versio

Early apoptosis of porcine alveolar macrophages limits avian influenza virus replication and proinflammatory dysregulation

Pigs are evidently more resistant to avian than swine influenza A viruses, mediated in part through frontline epithelial cells and alveolar macrophages (AM). Although porcine AM (PAM) are crucial in influenza virus control, their mode of control is unclear. To gain insight into the possible role of PAM in the mediation of avian influenza virus resistance, we compared the host effects and replication of two avian (H2N3 and H6N1) and three mammalian (swine H1N1, human H1N1 and pandemic H1N1) influenza viruses in PAM. We found that PAM were readily susceptible to initial infection with all five avian and mammalian influenza viruses but only avian viruses caused early and extensive apoptosis (by 6 h of infection) resulting in reduced virus progeny and moderated pro- inflammation. Full length viral PB1-F2 present only in avian influenza viruses is a virulence factor that targets AM for mitochondrial associated apoptotic cell death. With the use of reverse genetics on an avian H5N1 virus, we found that full length PB1-F2 contributed to increased apoptosis and pro-inflammation but not to reduced virus replication. Taken together, we propose that early apoptosis of PAM limits the spread of avian influenza viruses and that PB1-F2 could play a contributory role in the process

Proteomic analysis of the Plasmodium male gamete reveals the key role for glycolysis in flagellar motility.

BACKGROUND: Gametogenesis and fertilization play crucial roles in malaria transmission. While male gametes are thought to be amongst the simplest eukaryotic cells and are proven targets of transmission blocking immunity, little is known about their molecular organization. For example, the pathway of energy metabolism that power motility, a feature that facilitates gamete encounter and fertilization, is unknown. METHODS: Plasmodium berghei microgametes were purified and analysed by whole-cell proteomic analysis for the first time. Data are available via ProteomeXchange with identifier PXD001163. RESULTS: 615 proteins were recovered, they included all male gamete proteins described thus far. Amongst them were the 11 enzymes of the glycolytic pathway. The hexose transporter was localized to the gamete plasma membrane and it was shown that microgamete motility can be suppressed effectively by inhibitors of this transporter and of the glycolytic pathway. CONCLUSIONS: This study describes the first whole-cell proteomic analysis of the malaria male gamete. It identifies glycolysis as the likely exclusive source of energy for flagellar beat, and provides new insights in original features of Plasmodium flagellar organization

The regional distribution of different types of influenza receptors in cultured human alveolar epithelial cells and correlation with in vitro infection

BACKGROUND: Sialic acid (Sia) linked glycoproteins are the classical influenza receptors for influenza virus haemagglutinin to bind. The distribution of Sia on cell surfaces is one of the determinants of host tropism, and understanding its expression on human cells and tissues is important for understanding influenza pathogenesis. Previous research has shown the differences in apical versus basolateral infection and release of different influenza virus from polarized epithelial cells1 and correlated this with sialic acid distribution in the human respiratory tract. Moreover, mass spectrometric analysis was recently employed to elucidate the glycans present in the tissue in a higher resolution in human lung.2 The objective of this study was to examine in detail the distribution of these Sia-linked glycans at the cellular level by the use of confocal microscopy …link_to_OA_fulltex

Replication and innate host response of influenza A virus in lung microvascular endothelial cells: new insights into systemic infection and pathogenesis

INTRODUCTION: Though influenza A virus replication kinetics and host responses have been previously studied in umbilical vein endothelial cell or transformed endothelial cell lines, the tropism of influenza A virus including H5N1 and pandemic H1N1pdm for primary human lung microvascular endothelial cell has not been well defined.1 In this study we employed primary human lung microvascular endothelial cells, which are more physiologically relevant for understanding pathogenesis of influenza in the lung as to obtain a better understanding of the links of endothelial cell infection to systematic virus dissemination and multiple organ involvement in severe human influenza …link_to_OA_fulltex

Replication of avian and seasonal influenza viruses in human bronchus and lung

BACKGROUND: Pandemics of 1957 and 1968 were believed to arise from avian influenza viruses.1 The tropism of avian and human seasonal influenza viruses for the human lower respiratory tract deserves investigation. The target cell types that support replication of avian influenza A viruses in the human respiratory tract in the early stages of clinical infection have not well defined. In a previous autopsy studies of human H5N1 disease, influenza A virus were found to infect alveolar epithelial cells2 and macrophages.3 In this study, viral infectivity and replication competence of human and high and low pathogenic avian influenza viruses were systematically investigated in the human conducting and lower respiratory tract using ex vivo organ cultures. We compared the replication kinetics of human seasonal influenza viruses (H1N1 and H3N2), low pathogenic avian influenza viruses (H9N2, H5N8) with that of the highly pathogenic H5N1 viruses isolated from human H5N1 disease …link_to_OA_fulltex

Tropism and innate host response of the 2009 pandemic H1N1 influenza virus compared with related swine influenza viruses and reassortants in ex vivo and in vitro cultures of the human respiratory tract and conjunctiva

BACKGROUND: Pandemic influenza H1N1 (H1N1pdm) virus of swine-origin causes mild disease, but occasionally is associated with acute respiratory distress syndrome and death.1,2 It is important to understand the pathogenesis of this new disease. Previously we showed a comparable virus tropism and host innate immune responses between H1N1pdm and seasonal H1N1 influenza virus in the human respiratory tract,3 however H1N1pdm virus differed from seasonal H1N1 influenza virus in its ability to replicate in human conjunctiva, suggesting subtle differences in receptor-binding profile and highlighting the potential role of the conjunctiva as an additional route of infection. We now compare the tropism and host responses elicited by pandemic H1N1 with that of related swine influenza viruses and a pandemic-swine reassortant virus in ex vivo and in vitro cultures of the human respiratory tract and conjunctiva. We have used recombinant virus to investigate the role of the hemagglutinin (HA) and neuraminidase (NA) of H1N1pdm virus in its conjunctival tropism. These findings are relevant for understanding transmission and therapy …link_to_OA_fulltex