Biology subjects

Ciomperlik-Patton, J.

Publications and source records attributed to Ciomperlik-Patton, J..

4 recordsLinked to original sources

bioRxiv2025

Enhancing Tissue-Specific Antiviral Immunity to Disrupt Arbovirus Transmission by Mosquitoes

Arboviruses, including dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV), pose a significant global health and economic burden, with Aedes aegypti serving as their primary vector. Arbovirus infection in Ae. aegypti progresses sequentially through the midgut (MG), carcass (CA), and salivary glands (SG), with each tissue exhibiting distinct antiviral responses. Here, we investigate tissue-specific antiviral mechanisms, focusing on the small interfering RNA (siRNA) pathway in SGs. Our results reveal that SGs possess weaker antiviral defense and are more susceptible to arboviral infection compared to MGs and CAs. Notably, overexpression of Dicer2 (Dcr2), a key component of the siRNA pathway, in SGs leads to a significant decrease in arboviral replication. Conversely, Dcr2 overexpression in fat bodies, the primary tissue in CAs, only moderately suppresses DENV2 infection and has no notable effect on Mayaro virus (MAYV) infection. Remarkably, the simultaneous overexpression of Dcr2 in both MGs and SGs enhances antiviral activity, effectively blocking the transmission of multiple arboviruses. These findings reveal the tissue-specific dynamics of mosquito antiviral immunity and underscore the potential for targeting SG-specific immunity to disrupt arbovirus transmission, providing a promising approach for controlling mosquito-borne diseases.

microbiology↗

bioRxiv2024

Recombinant B3 clade enterovirus D68 strains are efficiently rescued in 293T cells and infect human spinal cord organoids.

Enterovirus D68 (EV-D68) is associated with respiratory disease in children. Between 2014 and 2018, biennial EV-D68 outbreaks coincided with peaks of a polio-like neurologic condition called acute flaccid myelitis (AFM). We hypothesized that specific mutations within the currently circulating B3 clade of EV-D68 impacted neurovirulence. However, recovery of these strains from infectious clones by published methods proved unsuccessful. Therefore, we tested different cell lines, reagents, and conditions to enhance efficiency of recombinant (r)EV-D68 rescue. In this study, we present a tractable rescue system to define virulence determinants in B3 clade strains. Using this approach, we successfully rescued historic and contemporary rEV-D68 strains to high titer after limited cell culture passage. All strains in our study replicated efficiently in the parental RD cell line and in human respiratory epithelial cell lines, with BEAS-2B cells exhibiting greater permissivity than A549 cells. While B2 and B3 clade strains could infect the SH-SY5Y neuroblastoma cell line, the neurovirulent B2 clade strain rUSA/IL/2014-18952 was more dependent on neuron differentiation than B3 clade strains and replication was not sustained under multi-cycle growth conditions. Conversely, replication of rUSA/IL/2014-18952 was more efficient in human spinal cord organoids, which model the cellular heterogeneity of the spinal cord, while replication of B3 clade strains was more modest. Immunofluorescence staining confirmed infection, with viral antigen colocalized with neurons. These findings suggest shifting dynamics of EV-D68 neurovirulence and provide a critical platform for further assessment of viral determinants of neurovirulence in B3 clade strains. IMPORTANCEEnterovirus D68 (EV-D68) can cause acute flaccid myelitis (AFM), a debilitating neurological condition of the spinal cord in children. Identifying viral determinants of EV- D68 neuropathogenesis is critical to understanding recent shifts in AFM prevalence; however, these investigations are limited to a small subset of infectious clones distantly related to currently circulating B3 clade strains. In this study, we leverage improved rescue strategies to characterize recombinant (r)EV-D68 strains from the dominant B3 clade. While all rEV-D68 replicate efficiently in the parental cell line and in human respiratory epithelial cells, B3 clade strains achieved greater titers in cultured neurons than a neurovirulent B2 clade strain, with less dependence on neuronal differentiation state. All B3 clade strains established infection in human spinal cord organoids, but replication varied between strains. Therefore, our study presents a tractable rescue system to begin to dissect viral determinants of shifting neurotropism within contemporary EV-D68 clades.

microbiology↗

bioRxiv2021

SARS-CoV-2 susceptibility of cell lines and substrates commonly used in diagnosis and isolation of influenza and other viruses

Coinfection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and other viruses is inevitable as the COVID-19 pandemic continues. This study aimed to evaluate cell lines commonly used in virus diagnosis and isolation for their susceptibility to SARS-CoV-2. While multiple kidney cell lines from monkeys were susceptible and permissive to SARS-CoV-2, many cell types derived from human, dog, mink, cat, mouse, or chicken were not. Analysis of MDCK cells, which are most commonly used for surveillance and study of influenza viruses, demonstrated that they were insusceptible to SARS-CoV-2 and that the cellular barrier to productive infection was due to low expression level of the angiotensin converting enzyme 2 (ACE2) receptor and lower receptor affinity to SARS-CoV-2 spike, which could be overcome by over-expression of canine ACE2 in trans. Moreover, SARS-CoV-2 cell tropism did not appear to be affected by a D614G mutation in the spike protein.

microbiology↗

bioRxiv2021

Respiratory and intestinal epithelial cells exhibit differential susceptibility and innate immune responses to contemporary EV-D68 isolates

Enterovirus D68 (EV-D68) has been implicated in outbreaks of severe respiratory illness and acute flaccid myelitis (AFM) and is detected in patient respiratory samples and from stool and wastewater, suggesting both respiratory and enteric routes of transmission. Here, we used a panel of EV-D68 isolates, including a historical isolate and multiple contemporary isolates from AFM outbreak years, to define the dynamics of viral replication and the host response to infection in primary human airway cells and stem cell-derived enteroids. We show that some recent EV-D68 isolates have decreased sensitivity to acid and temperature compared with an earlier isolate and that the respiratory, but not intestinal, epithelium induces a robust type III interferon (IFN) response that restricts infection. Our findings define the differential responses of the respiratory and intestinal epithelium to contemporary EV-D68 isolates and suggest that some isolates have the potential to target both the human airway and gastrointestinal tracts.

microbiology↗