Biology subjects

Netherton, C. L.

Publications and source records attributed to Netherton, C. L..

4 recordsLinked to original sources

bioRxiv2026

Suidae iPSC-derived macrophages as models for investigating susceptibility and resilience to African swine fever virus

African swine fever virus (ASFV) causes a lethal haemorrhagic fever in pigs and spread of this disease threatens many pig species (Suidae) globally. By contrast, ASFV infections in the natural evolved hosts, the warthog and bushpig, are subclinical. The macrophage (M{varphi}) is the primary target of ASFV and species-dependent responses in M{varphi}s are presumed to influence disease susceptibility. In an attempt to model these differences in vitro, we generated transgene-regulated induced pluripotent stem cells (iPSCs) from domestic pig, wild boar, red river hog and warthog, and confirmed that their corresponding iPSC-derived M{varphi}s (iPSCdMs) supported infection and replication of ASFV. In contrast to the other species, however, warthog iPSCdMs did not induce interferon upon infection by either virulent or attenuated ASFV. iPSCdMs may therefore represent an experimental system to understand how ASFV infection of M{varphi}s contributes to disease and aid development of strategies to combat this economically and environmentally devastating pathogen.

developmental biology↗

bioRxiv2025

Early disruption of the innate-adaptive immune axis in vivo after infection with virulent Georgia2007/1 ASFV

Effective immune defence and pathogen clearance requires coordination between innate and adaptive immune responses. However, virulent African swine fever virus (ASFV), which has a high case fatality rate in pigs, causes severe disease by exploiting multiple immune evasion strategies to suppress host responses. The global spread of Georgia2007/1 and its derivatives poses a significant threat to the pig industry and global food security. Although modified live virus vaccines for ASF exist, multiple safety concerns have restricted their use internationally. Conversely, subunit vaccine candidates have not matched the protective efficacy of modified live virus vaccines. This highlights the need to further investigate ASFV-induced immunopathology to support the development of next-generation ASF vaccines. Immune dynamics in whole blood and lymphoid tissues were examined over time after oronasal infection with Georgia 2007/1. CD4+ T cells, {gamma}{delta}-TCR+ T cells and CD21+ B cells were impacted by lymphopenia, and initial immune activation was detected. However, as the disease progressed, impaired maintenance and depletion of adaptive immune cells, such as CD4+ T cells and professional antigen-presenting dendritic cells and macrophages was observed. This depletion of cells may have compromised the innate-adaptive immune axis, weakening host ability to mount a robust adaptive immune response and potentially contributing to disease progression. Differential ASFV infection profiles within the spleen were also detected, highlighting the diversity of ASFV cellular tropism. Further investigation into the innate-adaptive immune axis is needed to better understand its role in ASFV infection. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=168 SRC="FIGDIR/small/660190v1_ufig1.gif" ALT="Figure 1"> View larger version (53K): org.highwire.dtl.DTLVardef@12b96a4org.highwire.dtl.DTLVardef@af2722org.highwire.dtl.DTLVardef@13237e4org.highwire.dtl.DTLVardef@1f371f1_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

bioRxiv2024

Exploring virus-host interactions through combined proteomic approaches identifies BANF1 as a new essential factor for African Swine Fever Virus.

African swine fever virus (ASFV) causes a highly lethal disease in pigs and represents a significant threat to the global pork industry due to the lack of effective vaccines or treatments. Despite intensive research, many ASFV proteins remain uncharacterized. This study aimed to elucidate the functions of two ASFV proteins, MGF360-21R and A151R, through comprehensive analysis of their interactions with host proteins. Using affinity purification-mass spectrometry and yeast two-hybrid screening approaches, we identified the host protein barrier- to-autointegration factor 1 (BANF1) as a key interactor of both viral proteins. Biochemical and colocalization assays confirmed these interactions and demonstrated that MGF360-21R and A151R expression leads to cytoplasmic relocalization of BANF1. Functionally, BANF1 silencing significantly reduced ASFV replication, indicating its proviral role. Given BANF1s established function in regulating the cGAS/STING-dependent type I interferon (IFN-I) response, we postulated that A151R and MGF360-21R could inhibit this pathway. Using different strategies, we showed that both A151R and MGF360-21R did indeed inhibit IFN-I induction. Generation of ASFV deficient of A151R or MGF360-21R showed that both mutant viruses enhanced the host IFN response in primary porcine macrophages compared to wild-type virus. However, their capacity to inhibit this pathway could occur through mechanisms independent of BANF1. Proteomic analysis of BANF1 interactors during ASFV infection highlighted potentially roles in chromatin remodeling, nuclear transport, and innate immune response pathways. Altogether, our data provide new insights into ASFV-host interactions, identifying BANF1 as an important new host factor required for replication and uncovering novel functions for A151R and MGF360-21R. Author SummaryAfrican swine fever virus (ASFV) is a highly contagious and deadly disease affecting pigs worldwide, for which there are currently no effective vaccines or treatments. Despite extensive research, many ASFV proteins remain poorly understood. Our study investigated two ASFV proteins, MGF360-21R and A151R, to better understand their functions and interactions with host proteins. Using proteomic approaches, we found both these viral proteins interact with a host protein called barrier-to-autointegration factor 1 (BANF1). Importantly, BANF1 silencing significantly reduced ASFV replication, indicating its important role in the viral life cycle. We also showed that MGF360-21R and A151R help the virus evade the immune system by blocking the production of interferons, which are key defensive molecules against viral infections. However, this immune evasion does not seem to depend on their interaction with BANF1. Additionally, our analysis of BANF1s interactions during ASFV infection revealed potential roles in chromatin remodeling, nuclear transport, and the innate immune response. These findings provide new insights into how ASFV interacts with its host and highlight BANF1 as a critical factor in viral replication and immune evasion. Our work contributes to a better understanding of ASFV and could pave the way for developing more effective strategies to fight this virus.

microbiology↗

bioRxiv2019

Identification of a functional small non-coding RNA encoded by African swine fever virus

African swine fever virus (ASFV) causes a lethal haemorrhagic disease of domestic pigs, to which there is no vaccine available. ASFV has a large, double-stranded DNA genome that encodes over 150 proteins. Replication takes place in the cytoplasm of the cell and involves complex interactions with host cellular components including small non-coding RNAs (sncRNAs). A number of DNA viruses are known to manipulate sncRNA either by encoding their own or disrupting host sncRNA. In order to investigate the interplay between ASFV and sncRNAs, study of host and viral small RNAs extracted from ASFV-infected primary porcine macrophages (PAMs) was undertaken. We discovered that ASFV infection had only a modest effect on host miRNAs, with only 6 miRNAs differentially expressed during infection. The data also revealed 3 potential novel small RNAs encoded by ASFV, ASFVsRNA1-3. Further investigation of ASFVsRNA2 detected it in lymphoid tissue from pigs with ASF. Overexpression of ASFVsRNA2 led to up to a 1 log reduction in ASFV growth indicating that ASFV utilises a virally-encoded small RNA to disrupt its own replication. This study describes the modest impact of ASFV on host sncRNAs and the identification of a functional ASFV-encoded sncRNA. ImportanceAfrican swine fever (ASF) poses a major threat to pig populations and food security worldwide. The disease is endemic in Africa and Eastern Europe and rapidly emerging into Asia where it has led to the deaths of millions of pigs in the past 12 months. The development of safe and effective vaccines to protect pigs against ASF has been hindered by lack of understanding of the complex interactions between ASFV and the host cell. We focused our work on characterising the interactions between ASFV and sncRNAs. We found only modest changes to host sncRNA abundance after ASFV infection, and discovered a functional ASFV-encoded sncRNA. The knowledge from this study can be exploited to develop more effective ASFV vaccines that take advantage of the sncRNA system.

molecular biology↗