Effect of specific cellular immune response induced by anti-Aedes albopictus salivary protein rAlb-34k2-1 on DENV-2 replication in BMDM cells
- Author:
QIN Yanchun
;
LI Ying
;
HU Huan
- Publication Type:Journal Article
- Keywords:
rAlb-34k2-1 protein;
DENV-2;
cellular immune response
- From:
China Tropical Medicine
2024;24(8):907-
- CountryChina
- Language:Chinese
-
Abstract:
Abstract: Objective To investigate the effect of Aedes albopictus salivary protein rAlb-34k2-1-induced mouse splenocytes immune response on dengue virus type 2 (DENV-2)-infected bone marrow-derived macrophages (BMDM) in vitro. Methods The specific cell proliferation response of splenic cells from immunized/non-immunized mice to Alb34k2-1 protein was analyzed using the CCK-8 assay. The antigen-presenting cells (APCs) loaded with mosquito saliva proteins rAlb-34k2-1 protein/rAalb_CTL1 protein were prepared and co-cultured with lymphocytes from spleens of immunized/non-immunized mice. On the one hand, the levels of DENV-2 nucleic acids in BMDM treated with cell supernatants from mixed lymphocyte co-cultures were analyzed by qRT-PCR, and the effect of specific T cell response induced by rAlb-34k2-1 on DENV-2 replication in BMDM was analyzed. On the other hand, the expression of the DENV-2 E gene in BMDM pretreated with rAlb-34k2-1 protein was also evaluated to determine the direct effect of immunized mouse spleen lymphocytes co-cultured at 48 h and 72 h and the effect of spleen lymphocytes from immunized or non-immunized mice on DENV-2 replication in pretreated BMDMs was analyzed. Results Compared to the non-immunized group, the rAlb-34k2-1 protein in vitro caused a specific proliferation of immunized-splenocytes, with a proliferation index (SI) >2 (P<0.000 1).Mixed lymphocyte culture supernatants loaded with rAlb-34k2-1 protein inhibited DENV-2 replication in BMDM, while the mixed lymphocyte culture supernatants loaded with rAalb_CTL1 protein had no such effect (2-ΔΔCT<0.5, P<0.05). There was no difference in DENV-2 intracellular replication in rAlb-34k2-1 pretreated BMDM after co-culture with either immunized-lymphocytes or non-immunized-lymphocytes. Conclusions The supernatant induced by the mouse T cell response to the rAlb-34k2-1 protein can inhibit DENV-2 replication in BMDM cells, providing preliminary experimental evidence to explore the role of mosquito salivary protein-induced immune responses in the pathogenesis of mosquito-borne viruses.
- Full text:20251107104326833124.Effect of specific cellular immune response induced by anti-Aedes albopictus salivary protein rAlb-34k2-1 on DENV-2 replication in BMDM cells.pdf
