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Heparan sulfate proteoglycan: an arbovirus attachment factor integral to mosquito salivary gland ducts
Kristen A Ciano1, Jason J Saredy2, Doria F Bowers3
1Department of Biology, University of North Florida, Jacksonville, FL 32224, USA. kristenaciano@gmail.com.
Abstract:
Variants of the prototype Alphavirus, Sindbis (SINV), were used in per os infections of adult female mosquitoes to investigate arbovirus interaction with the salivary gland (SG). Infection of Aedine mosquitoes with AR339, a heparan sulfate proteoglycan (HSPG)-dependent variant, resulted in gross pathology in the SG lateral lobes while infection with TR339, a HSPG-independent variant, resulted in minimal SG pathology. HSPG was detected in the internal ducts of the SG lateral lobes by immunolabeling but not in the median lobe, or beyond the triad structure and external ducts. Reports that human lactoferrin interacts with HSPG, suggested an interference with virus attachment to receptors on vertebrate cells. Pre-incubation of Aedes albopictus cultured C7-10 cells with bovine lactoferrin (bLF) followed by adsorption of SINV resulted in earlier and greater intensity of cytopathic response to TR339 compared with AR339. Following pre-treatment of C7-10 cells with bLF, plaques from tissue culture-adapted high-titer SINVTaV-GFP-TC were observed at 48 h post-infection (p.i.), while plaques from low-titer SINVTaV-GFP-TC were not observed until 120 h p.i. Confocal optics detected this reporter virus at 30 days p.i. in the SG proximal lateral lobe, a region of HSPG-immunolocalization. Altogether these data suggest an association between SINV and HSPG in the host mosquito.
Insights
Sindbis virus (SINV) interaction with mosquito salivary glands was studied. Heparan sulfate proteoglycan (HSPG) presence correlated with pathology, suggesting a role in arbovirus infection.
Area of Science:
- Arthropod-borne virus research
- Mosquito-borne disease vectors
- Viral pathogenesis
Background:
- Alphaviruses, like Sindbis virus (SINV), are arboviruses transmitted by mosquitoes.
- The interaction between arboviruses and mosquito salivary glands is crucial for transmission.
- Heparan sulfate proteoglycans (HSPG) are implicated in cellular processes and viral infections.
Purpose of the Study:
- To investigate the interaction of Sindbis virus (SINV) variants with mosquito salivary glands (SG).
- To determine the role of heparan sulfate proteoglycan (HSPG) in SINV infection and pathology within the mosquito SG.
- To explore the potential interference of lactoferrin with SINV attachment to host cells.
Main Methods:
- Per os infection of Aedine mosquitoes with SINV variants (AR339 and TR339).
- Immunolabeling to detect HSPG distribution in mosquito SG.
- In vitro cell culture experiments using Aedes albopictus cells pre-incubated with bovine lactoferrin (bLF) followed by SINV infection.
- Confocal microscopy to visualize virus presence in SG.
Main Results:
- HSPG-dependent SINV variant (AR339) caused significant SG pathology, while the HSPG-independent variant (TR339) showed minimal pathology.
- HSPG was localized in specific regions of the SG lateral lobes.
- Pre-incubation with bovine lactoferrin enhanced cytopathic effects of SINV in cell cultures, particularly for the HSPG-independent variant.
- Reporter SINV was detected in HSPG-immunolocalized regions of the SG.
Conclusions:
- These findings suggest a significant association between Sindbis virus (SINV) and heparan sulfate proteoglycan (HSPG) in the mosquito host.
- HSPG may play a role in SINV pathogenesis within the salivary glands.
- Lactoferrin might interfere with SINV attachment, offering potential insights into transmission blocking strategies.
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