Biology subjects

Laksono, B. M.

Publications and source records attributed to Laksono, B. M..

3 recordsLinked to original sources

bioRxiv2025

Enterovirus D68 infection of human inducible pluripotent stem cell-derived skeletal muscles resulted in structural destruction, loss of muscle function and hampered muscle regeneration

Enterovirus D68 (EV-D68) is an emerging respiratory virus that commonly causes mild to severe respiratory diseases. EV-D68 infection is also associated with extra-respiratory complications, especially acute flaccid myelitis. However, how the virus invades the central nervous system (CNS) and infects motor neurons is not fully understood. One possible neuroinvasive route is through infection of skeletal muscles, which allows the virus to infect motor neurons via the neuromuscular junction. However, we hypothesise that direct EV-D68 infection of human skeletal muscles can impair muscle function and thus contribute to the development of EV-D68-associated muscle weakness. Here, we inoculated human induced pluripotent stem cell-derived skeletal muscle myotubes grown in 2D with different EV-D68 isolates, which resulted in a productive infection and cell death. We showed, through neuraminidase treatment, that sialic acids facilitate infection of these cells. EV-D68 infection of 3D tissue engineered skeletal muscles led to tissue damage, reduction of contractile force and depletion of muscle and satellite cells. Altogether, we showed that human skeletal muscle can act as an extra-respiratory replication site and infection of skeletal muscles may contribute to EV-D68-associated muscle weakness. Author summaryAfter causing a global outbreak in 2014, enterovirus D68 (EV-D68) - a respiratory virus - has been associated with polio-like paralysis. How the virus spreads from the respiratory system to the central nervous system is poorly understood. One of the possible routes for this spread is by infecting skeletal muscles before spreading to motor neurons. However, it is not known whether the virus can infect human skeletal muscles and whether EV-D68 infection of skeletal muscles can also lead to paralysis. Here, we used 2D and 3D human skeletal muscle models to investigate EV-D68 infection of human skeletal muscles. We found that EV-D68 can replicate in these 2D and 3D models and the infection results in destruction of muscle fibres and loss of muscle function. We also found that the infection results in loss of satellite cells, which are important for muscle regeneration, suggesting that EV-D68 infection hampers skeletal muscle regeneration. Our study provides new insights into the potential role of human skeletal muscles in EV-D68-associated paralysis.

microbiology↗

bioRxiv2022

The potential role of human immune cells in the systemic dissemination of enterovirus-D68

Enterovirus-D68 (EV-D68) often causes mild respiratory infections, but can also cause severe respiratory infections and extra-respiratory complications, including acute flaccid myelitis (AFM). Systemic dissemination of EV-D68 is crucial for the development of extra-respiratory diseases, but it is currently unclear how EV-D68 viremia occurs. We hypothesize that immune cells contribute to the systemic dissemination of EV-D68, as this is a mechanism commonly used by other enteroviruses. Therefore, we investigated the susceptibility and permissiveness of human primary immune cells for different EV-D68 isolates. In human peripheral blood mononuclear cells (PBMC) inoculated with EV-D68, only B cells were susceptible but virus replication was limited. However, B cell-rich cultures, such as Epstein-Barr virus-transformed B-lymphoblastoid cell line (BLCL) and primary lentivirus-transduced B cells, were productively infected. In BLCL, neuraminidase treatment to remove 2,6- and 2,3-linked sialic acids resulted in a significant decrease of EV-D68 infected cells, suggesting that sialic acids are the functional receptor on B cells. Subsequently, we showed that dendritic cells (DCs), particularly immature DCs, are susceptible and permissive for EV-D68 infection and that they can spread EV-D68 to autologous BLCL. Altogether, our findings suggest that immune cells, especially B cells and DCs, play an important role in the development the systemic dissemination of EV-D68 during an infection, which is an essential step towards the development of extra-respiratory complications. Author summaryEnterovirus D68 (EV-D68) is an emerging respiratory virus that has caused outbreaks worldwide since 2014. EV-D68 infects primarily respiratory epithelial cells and the infection commonly results in mild respiratory diseases. However, EV-D68 infection is also associated with complications outside the respiratory tract, including a polio-like paralysis. Despite the severity of these extra-respiratory complications, it is unclear how EV-D68 is able to spread outside the respiratory tract and infect other organs, like the central nervous system (CNS). To understand this, we investigated if immune cells play a role in the extra-respiratory spread of EV-D68. We showed that EV-D68 can infect and replicate in specific immune cells, i.e. B cells and dendritic cells (DCs), and that the virus can be transferred from DCs to B cells. Our findings suggest that lymphoid tissues, which harbor many immune cells, can be a secondary replication site for EV-D68, from where virus is released in the circulation. Our data reveal the importance of immune cells in the systemic spread of EV-D68, which is essential for infection of extra-respiratory tissues. Intervention strategies that prevent EV-D68 infection of immune cells will therefore potentially prevent virus spread from the respiratory tract to other organs.

microbiology↗

bioRxiv2019

Measles skin rash: infection of lymphoid and myeloid cells in the dermis precedes viral dissemination to keratinocytes in the epidermis

Measles is characterised by fever and a maculopapular skin rash, which is associated with immune clearance of measles virus (MV)-infected cells. Histopathological analyses of skin biopsies from humans and non-human primates (NHPs) with measles rash have identified MV-infected keratinocytes and mononuclear cells in the epidermis, around hair follicles and near sebaceous glands. Here, we address the pathogenesis of measles skin rash by combining data from experimentally infected NHPs, ex vivo infection of human skin sheets and in vitro infection of primary human keratinocytes. Longitudinal analysis of the skin of experimentally MV-infected NHPs demonstrated that infection in the skin precedes onset of rash by several days. MV infection was initiated in lymphoid and myeloid cells in the dermis before dissemination to the epidermal keratinocytes. These data were in good concordance with ex vivo MV infections of human skin sheets, in which dermal cells were more targeted than the epidermal ones. To address viral dissemination to the epidermis and to determine whether the dissemination is receptor-dependent, we performed experimental infections of primary keratinocytes collected from healthy or nectin-4-deficient donors. These experiments demonstrated that MV infection of keratinocytes is nectin-4-dependent, and nectin-4 expression was higher in differentiated than in proliferating keratinocytes. Based on these data, we hypothesise that measles skin rash is initiated by migrating MV-infected lymphocytes that infect dermal skin-resident CD150+ immune cells. The infection is subsequently disseminated from the dermal papillae to nectin-4+ keratinocytes in the basal epidermis. Lateral spread of MV infection is observed in the superficial epidermis, most likely due to the higher level of nectin-4 expression on differentiated keratinocytes. Finally, MV-infected cells are cleared by infiltrating immune cells, causing hyperaemia and oedema, which give the appearance of morbilliform skin rash. Author SummarySeveral viral infections are associated with skin rash, including parvovirus B19, human herpesvirus type 6, dengue virus and rubella virus. However, the archetype virus infection that leads to skin rash is measles. Although all of these viral exanthemata often appear similar, their pathogenesis is different. In the case of measles, the appearance of skin rash is a sign that the immune system is clearing MV-infected cells from the skin. How the virus reaches the skin and is locally disseminated remains unknown. Here we combine observations and expertise from pathologists, dermatologists, virologists and immunologists to delineate the pathogenesis of measles skin rash. We show that MV infection of dermal myeloid and lymphoid cells precedes viral dissemination to the epidermal keratinocytes. We speculate that immune-mediated clearance of these infected cells results in hyperaemia and oedema, explaining the redness of the skin and the slightly elevated spots of the morbilliform rash.

microbiology↗