Identification of ectopic anionic trypsin I in rat lungs potentiating pneumotropic virus infectivity and increased enzyme level after virus infection

Towatari, Takae; Ide, Mikiko; Ohba, Kumiko; Chiba, Yu; Murakami, Meiko; Shiota, Mayumi; Kawachi, Miki; Yamada, Hiroshi; Kido, Hiroshi

doi:10.1046/j.1432-1033.2002.02937.x

Cited by 39 publications

(22 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The HA 0 processing proteases reported for human IAV in animals and humans that recognize single basic motif are pancreatic trypsin (Klenk et al, 1975), plasmin from calf and chicken serum (Lazarowitz et al, 1973), blood clotting factor Xa from chick embryo (Gotoh et al, 1990), tryptase Clara from rat lungs (Kido et al, 1992), mini-plasmin from rat lungs (Murakami et al, 2001), ectopic anionic trypsin from rat lungs (Towatari et al, 2002), porcine mast cell tryptase (Chen et al, 2000), tryptase TC30 from porcine lungs (Sato et al, 2003) and transmembrane protease serine (TMPRSS) 2 and type II membrane protein human airway trypsin-like protease (HAT) (Böttcher et al, 2006) (Table 1). Why various trypsin-type HA 0 processing proteases capable of potentiating IAV infections exist in the airways?…”

Section: Ha0 Processing Proteases In the Airway For Human Seasonal Inmentioning

confidence: 99%

Host envelope glycoprotein processing proteases are indispensable for entry into human cells by seasonal and highly pathogenic avian influenza viruses

Kido¹,

Okumura²,

Takahashi³

2009

J Mol Genet Med

Self Cite

View full text Add to dashboard Cite

Influenza A virus (IAV) is one of the most common infectious pathogens in humans and causes considerable morbidity and mortality. The recent spread of highly-pathogenic avian IAV H5N1 viruses has reinforced the importance of pandemic preparedness. In the pathogenesis of IAV infection, cellular proteases play critical roles in the process of viral entry into cells that subsequently leads to tissue damage in the infected organs. Since there are no processing protease for the viral membrane fusion glycoprotein hemagglutinin precursor (HA 0 ) in IAV, entry of the virus into cells is determined primarily by the host cellular HA 0 processing proteases that proteolytically activate membrane fusion activity. HA 0 of seasonal human IAV has the consensus cleavage site motif Q(E)-T/X-R and is selectively processed by at least seven different trypsintype processing proteases identified to-date in animal model experiments using mouse-adapted IAV or gene expression system in MDCK cells. As is the case for the highly pathogenic avian influenza (HPAI) A virus, endoproteolytic processing of the HA 0 occurs through ubiquitous cellular processing proteases, which selectively recognize the multi-basic consensus cleavage site motifs, such as R-X-K/R-R, and K-X-K/R-R. The cleavage enzymes for the R-X-K/R-R motif, but not K-X-K/R-R motif, have been reported to be furin and pro-protein convertase (PC)5/6 in the trans-Golgi network. Here we report new members of type II transmembrane serine proteases of the cell membrane, mosaic serine protease large form (MSPL) and its splice variant TMPRSS13, which recognize and cleave both R-X-K/R-R and K-X-K/R-R motifs without calcium. Furthermore, IAV infection significantly up-regulates a latent ectopic pancreatic trypsin, one of the potent HA processing proteases, and pro-matrix metalloprotease-9, in various organs. These proteases may synergistically damage the blood-brain barrier in the brain and basement membrane of blood vessels in various organs, resulting in severe edema and multiple organ failure. In this review, we discuss these proteases as new drug target molecules for IAV treatment acting by inhibition of IAV multiplication and prevention of multiple organ failure, other than anti-viral agents, viral neuraminidase inhibitors.

show abstract

Section: Ha0 Processing Proteases In the Airway For Human Seasonal Inmentioning

confidence: 99%

Host envelope glycoprotein processing proteases are indispensable for entry into human cells by seasonal and highly pathogenic avian influenza viruses

Kido¹,

Okumura²,

Takahashi³

2009

J Mol Genet Med

Self Cite

View full text Add to dashboard Cite

show abstract

“…Less is known about proteases that cleave influenza viruses under conditions of natural infection. Several trypsin-like proteases isolated from rat and swine lung were shown to support replication of influenza viruses in vitro (3,9,18,25). However, it remains unclear whether these proteases play a role in in vivo infection.…”

mentioning

confidence: 99%

Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium

Böttcher

Matrosovich

Beyerle

et al. 2006

J Virol

454

447

View full text Add to dashboard Cite

Host cell proteases that cleave the hemagglutinin (HA) of influenza viruses in the human respiratory tract are still not identified. Here we cloned two human type II transmembrane serine proteases with known airway localization, TMPRSS2 and HAT, into mammalian expression vector. Cotransfection of mammalian cells with plasmids encoding HA and either protease resulted in HA cleavage in situ. Transient expression of either protease in MDCK cells enabled multicycle replication of influenza viruses in these cells in the absence of exogenous trypsin. These data suggest that TMPRSS2 and HAT are candidates for proteolytic activation of influenza viruses in vivo.The ability of the hemagglutinin protein (HA) of influenza viruses to mediate fusion between viral and endosomal membranes during virus entry into the cell depends on cleavage of fusion-incompetent precursor HA0 into disulfide-linked subunits HA1 and HA2 by a host endoprotease. Cleavage of HA is essential for infection and determines viral pathogenicity and tissue tropism (reviewed in references 8, 10, 11, and 22). Thus, the highly pathogenic avian influenza viruses of subtypes H5 and H7 are cleaved at the multibasic motif R-X-R/K-R by ubiquitous subtilisin-like cellular proteases (11, 23) and cause lethal systemic infection in birds. All other influenza A viruses, including human epidemic and pandemic strains, have a single arginine at the HA cleavage site; these viruses can only be cleaved in a limited number of tissues, such as the intestinal tract in birds and the respiratory tract in birds and mammals (11,22).Early studies demonstrated that influenza viruses with monobasic cleavage site can be proteolytically activated in cell culture by the addition of trypsin (12, 13). Less is known about proteases that cleave influenza viruses under conditions of natural infection. Several trypsin-like proteases isolated from rat and swine lung were shown to support replication of influenza viruses in vitro (3,9,18,25). However, it remains unclear whether these proteases play a role in in vivo infection. In the case of human influenza, specific proteases responsible for HA cleavage in the human respiratory tract have not been identified thus far.In search of such proteases, we use a new approach. Instead of isolating and characterizing influenza virus-activating enzymes from respiratory tissues, we clone and express genes of trypsin-like proteases known to be present in the human airway epithelium and test them for cleavage of HA. We have analyzed here two such proteases, TMPRSS2 (5, 14, 21) and HAT (human airway trypsin-like protease) (4, 24, 27, 28), given their previous detection in the human airways and the availability of their full-length coding sequences. As the source of human genetic material for cloning, we used differentiated cultures of human airway epithelial cells grown at the air-liquid interface in serum-free, hormone-and growth factor-supplemented medium as previously described (6,15). These cultures support multicycle replication of human influenza virus...

show abstract

“…It has been suggested that soluble proteases might activate human influenza viruses in the infected host (18,19,30,43). However, analysis of cultured human respiratory epithelial cells revealed efficient HA cleavage during HA biogenesis and upon uptake of virions into target cells, suggesting that HA cleavage in the human host is a cell-associated process (47).…”

mentioning

confidence: 99%

TMPRSS2 and TMPRSS4 Facilitate Trypsin-Independent Spread of Influenza Virus in Caco-2 Cells

Bertram

Glowacka

Błażejewska

et al. 2010

J Virol

191

217

View full text Add to dashboard Cite

Proteolysis of influenza virus hemagglutinin by host cell proteases is essential for viral infectivity, but the proteases responsible are not well defined. Recently, we showed that engineered expression of the type II transmembrane serine proteases (TTSPs) TMPRSS2 and TMPRSS4 allows hemagglutinin (HA) cleavage. Here we analyzed whether TMPRSS2 and TMPRSS4 are expressed in influenza virus target cells and support viral spread in the absence of exogenously added protease (trypsin). We found that transient expression of TMPRSS2 and TMPRSS4 resulted in HA cleavage and trypsin-independent viral spread. Endogenous expression of TMPRSS2 and TMPRSS4 in cell lines correlated with the ability to support the spread of influenza virus in the absence of trypsin, indicating that these proteases might activate influenza virus in naturally permissive cells. Indeed, RNA interference (RNAi)-mediated knockdown of both TMPRSS2 and TMPRSS4 in Caco-2 cells, which released fully infectious virus without trypsin treatment, markedly reduced the spread of influenza virus, demonstrating that these proteases were responsible for efficient proteolytic activation of HA in this cell line. Finally, TMPRSS2 was found to be coexpressed with the major receptor determinant of human influenza viruses, 2,6-linked sialic acids, in human alveolar epithelium, indicating that viral target cells in the human respiratory tract express TMPRSS2. Collectively, our results point toward an important role for TMPRSS2 and possibly TMPRSS4 in influenza virus replication and highlight the former protease as a potential therapeutic target.

show abstract

Identification of ectopic anionic trypsin I in rat lungs potentiating pneumotropic virus infectivity and increased enzyme level after virus infection

Cited by 39 publications

References 39 publications

Host envelope glycoprotein processing proteases are indispensable for entry into human cells by seasonal and highly pathogenic avian influenza viruses

Host envelope glycoprotein processing proteases are indispensable for entry into human cells by seasonal and highly pathogenic avian influenza viruses

Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium

TMPRSS2 and TMPRSS4 Facilitate Trypsin-Independent Spread of Influenza Virus in Caco-2 Cells

Contact Info

Product

Resources

About