Inflammatory epithelial cytokines after<i>in vitro</i>respiratory syncytial viral infection are associated with reduced lung function

Duan, Wenming; Cen, Yuchen; Lin, Cindy; Ouyang, Hong; Du, Kai; Kumar, Anushree; Wang, Borui; Avolio, Julie; Grasemann, Hartmut; Moraes, Theo J.

doi:10.1183/23120541.00365-2021

Cited by 7 publications

(3 citation statements)

References 61 publications

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“…A likely reason why MeV, and other related paramyxoviruses and pneumoviruses, can gain control over the host IFN pathway is by expressing proteins responsible for antagonizing host IFN specifically ( 61 – 67 ). Despite expression of IFN combative proteins, related viruses still induce high levels of IFN in cells compared to our findings in this study ( 43 , 68 – 70 ). Thus, the question remains of how MeV is so much more contagious than related viruses containing almost identical genomes ( 1 ).…”

Section: Discussioncontrasting

confidence: 77%

Interferon-independent processes constrain measles virus cell-to-cell spread in primary human airway epithelial cells

Durnell,

Hippee,

Cattaneo

et al. 2023

Microbiol Spectr

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Amplification of measles virus (MeV) in human airway epithelia may contribute to its extremely high contagious nature. We use well-differentiated primary cultures of human airway epithelial cells (HAE) to model ex vivo how MeV spreads in human airways. In HAE, MeV spreads cell-to-cell for 3–5 days, but then, infectious center growth is arrested. What stops MeV spread in HAE is not understood, but interferon (IFN) is known to slow MeV spread in other in vitro and in vivo models. Here, we assessed the role of type I and type III IFN in arresting MeV spread in HAE. The addition of IFN-β or IFN-λ1 to the medium of infected HAE slowed MeV infectious center growth, but when IFN receptor signaling was blocked, infectious center size was not affected. In contrast, blocking type-I IFN receptor signaling enhanced respiratory syncytial virus spread. HAE were also infected with MeV mutants defective for the V protein. The V protein has been demonstrated to interact with both MDA5 and STAT2 to inhibit activation of innate immunity; however, innate immune reactions were unexpectedly muted against the V-defective MeV in HAE. Minimal innate immunity activation was confirmed by deep sequencing, quantitative RT-PCR, and single-cell RNA-seq analyses of the transcription of IFN and IFN-stimulated genes. We conclude that in HAE, IFN-signaling can contribute to slowing infectious center growth; however, IFN-independent processes are most important for limiting cell-to-cell spread. IMPORTANCE Fundamental biological questions remain about the highly contagious measles virus (MeV). MeV amplifies within airway epithelial cells before spreading to the next host. This final step likely contributes to the ability of MeV to spread host-to-host. Over the course of 3–5 days post-infection of airway epithelial cells, MeV spreads directly cell-to-cell and forms infectious centers. Infectious center formation is unique to MeV. In this study, we show that interferon (IFN) signaling does not explain why MeV cell-to-cell spread is ultimately impeded within the cell layer. The ability of MeV to spread cell-to-cell in airway cells without appreciable IFN induction may contribute to its highly contagious nature. This study contributes to the understanding of a significant global health concern by demonstrating that infectious center formation occurs independent of the simplest explanation for limiting viral transmission within a host.

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Section: Discussioncontrasting

confidence: 77%

Interferon-independent processes constrain measles virus cell-to-cell spread in primary human airway epithelial cells

Durnell,

Hippee,

Cattaneo

et al. 2023

Microbiol Spectr

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“…We then examined qPCR amplifications of RGPD and RANBP2 on RANBP2specific RT samples compared with classical RT using poly dT and random primers, and found that the RANBP2-specific RT successfully eliminated the detection of RGPD transcripts without impacting the detection of RANBP2 (Figures 1C,D). Of note, because of the presence of RANBP2 paralogs in humans (9), the sequencing of RANBP2 would also need to be performed using RNA and a RANBP2-specific primer for first-strand synthesis (1). However, in contrast to previous work which positioned the primer binding site in the 3′ untranslated region of RANBP2 (1), we designed the RANBP2-RT primer to bind to the closest region to the 5′ end of the coding sequence to increase the efficiency and accuracy of the RT.…”

Section: Resultsmentioning

confidence: 99%

Rapid and inexpensive bedside diagnosis of RAN binding protein 2-associated acute necrotizing encephalopathy

Gouy,

Decorsière,

Desgraupes

et al. 2023

Front. Neurol.

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Acute necrotizing encephalopathy 1 (ANE1) is a very rare disorder associated with a dominant heterozygous mutation in the RANBP2 (RAN binding protein 2) gene. ANE1 is frequently triggered by a febrile infection and characterized by serious and irreversible neurological damage. Although only a few hundred cases have been reported, mutations in RANBP2 are only partially penetrant and can occur de novo, suggesting that their frequency may be higher in some populations. Genetic diagnosis is a lengthy process, potentially delaying definitive diagnosis. We therefore developed a rapid bedside qPCR-based tool for early diagnosis and screening of ANE1 mutations. Primers were designed to specifically assess RANBP2 and not RGPD (RANBP2 and GCC2 protein domains) and discriminate between wild-type or mutant RANBP2. Nasal epithelial cells were obtained from two individuals with known RANBP2 mutations and two healthy control individuals. RANBP2-specific reverse transcription followed by allele-specific primer qPCR amplification confirmed the specific detection of heterozygously expressed mutant RANBP2 in the ANE1 samples. This study demonstrates that allele-specific qPCR can be used as a rapid and inexpensive diagnostic tool for ANE1 using preexisting equipment at local hospitals. It can also be used to screen non-hospitalized family members and at risk-population to better establish the frequency of non-ANE-associated RANBP2 mutations, as well as possible tissue-dependent expression patterns.Systematic review registrationThe protocol was registered in the international prospective register of systematic reviews (PROSPERO– CRD42023443257).

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“…For qPCR, technical triplicates of 6 μL reactions (for use in Applied Biosystems ViiA 7 Real-Time PCR 384-well system or QuantStudio™7 384-well System [4368814, ThermoFisher]) were prepared with 1 μL of 2X or 300X diluted cDNA. All primers were PrimeTime™ Standard probes from Integrated DNA Technologies (Coralville, USA) ( Table 3 ) and RSV primers were custom-designed using a previously validated sequence for NS1 [ 36 ] ( Table 4 ). TaqMan™ Fast Advanced Master Mix was used (4444556, ThermoFisher) as per manufacturer’s instructions.…”

Section: Methodsmentioning

confidence: 99%

M1-like, but not M0- or M2-like, macrophages, reduce RSV infection of primary bronchial epithelial cells in a media-dependent fashion

Ronaghan

Soo²,

Pena³

et al. 2022

PLoS ONE

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Respiratory syncytial virus (RSV) is a common childhood infection that in young infants can progress into severe bronchiolitis and pneumonia. Disease pathogenesis results from both viral mediated and host immune processes of which alveolar macrophages play an important part. Here, we investigated the role of different types of alveolar macrophages on RSV infection using an in vitro co-culture model involving primary tissue-derived human bronchial epithelial cells (HBECs) and human blood monocyte-derived M0-like, M1-like, or M2-like macrophages. It was hypothesized that the in vitro model would recapitulate previous in vivo findings of a protective effect of macrophages against RSV infection. It was found that macrophages maintained their phenotype for the 72-hour co-culture time period and the bronchial epithelial cells were unaffected by the macrophage media. HBEC infection with RSV was decreased by M1-like macrophages but enhanced by M0- or M2-like macrophages. The medium used during the co-culture also impacted the outcome of the infection. This work demonstrates that alveolar macrophage phenotypes may have differential roles during epithelial RSV infection, and demonstrates that an in vitro co-culture model could be used to further investigate the roles of macrophages during bronchial viral infection.

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Inflammatory epithelial cytokines afterin vitrorespiratory syncytial viral infection are associated with reduced lung function

Cited by 7 publications

References 61 publications

Interferon-independent processes constrain measles virus cell-to-cell spread in primary human airway epithelial cells

Interferon-independent processes constrain measles virus cell-to-cell spread in primary human airway epithelial cells

Rapid and inexpensive bedside diagnosis of RAN binding protein 2-associated acute necrotizing encephalopathy

M1-like, but not M0- or M2-like, macrophages, reduce RSV infection of primary bronchial epithelial cells in a media-dependent fashion

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