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Research Project: Molecular Characterization of Arbovirus-Mosquito-Vertebrate Host Interactions

Location: Foreign Arthropod Borne Animal Disease Research

Project Number: 3022-32000-024-005-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Sep 1, 2021
End Date: Aug 31, 2025

The long-term goal of this project is to characterize the effects of the vertebrate and invertebrate host on virus protein packaging. Our previous collaboration demonstrated that there are differences in the proteins package in the Rift Valley fever (RVF) virion when propagated in C6/36 mosquito vs Vero mammalian cell lines. This generated the hypothesis that propagation of RVF virus (RVFV) in mosquitoes would affect the initial cell tropism during mammalian infection. The objective of this agreement is to determine if this previous observation is consistent with other mosquito and mammalian cell lines.

To determine if the differential expression is consistent among RVFV isolates and hosts, the analysis will be repeated with at least two additional virus strains, at least two additional mosquito and two mammalian cell lines. The number of strains and cell lines will depend on the consistency of the results observed. That is, if 3 of the mosquito and mammalian cell lines examined produce the same viral protein profile then additional cell lines may not need to be examined. However, if there is inconsistency in the protein profiles of RVFV propagated within various insect or mammalian derived cell-lines then additional cell lines will be examined. Mosquito cell lines include: CxTr Culex tarsalis (ABADRU); Aag2 an Aedes aegypti (innate immune competent cell line), Toxorhynchites amboinensi Tra-171 (ATCC CRL-1591), Ae albopictus (ATCC CCL-126), and Ae aegypti (ATCC CCL-125). Cell lines from species known to be susceptible to RVFV infection will be used such as; bovine kidney cells (MDBK) and ovine kidney cells (OEK). Other RVFV virus strains include: rZinga, rSA51, SA01 and Ken06. Virions will be purified from infected cell lines and their viral structural proteins analyzed by western blot with available specific antibodies. Additional protein characterization such as Mass Spectrometry analysis will be performed for confirmation.