Tick-borne encephalitis virus NS1 glycoprotein during acute and persistent infection of cells.
Tick-borne encephalitis virus (TBEV) was propagated in porcine embryo kidney (PS) cells until 48 h whereas human kidney (RH) cells maintained the virus persistence during at least 2 months. One of possible reasons of flavivirus chronic infection might be abnormal NS1 gene expression. Immunoblotting with monoclonal antibodies (MAbs) revealed the similarity of the intracellular and secreted NS1 nonstructural glycoprotein size and linear antigenic determinants in both the infected cell lines. However, according to the competitive binding of MAbs with the TBEV NS1 extracellular glycoprotein, its contiguous epitopes differed for acute or persistent infection. To map the TBEV NS1 glycoprotein antigenic determinants its recombinant analogues were used. All the studied MAbs could bind with the full-length NS1 recombinant protein. Deletion of the TBEV NS1 gene internal region resulted in defective NS1d1 protein without the region between 269 and 333 a.a. Lack of NS1d1 binding with 20B4 MAb and diminished binding with 22H8 and 17C3 MAbs permitted to map their antigenic determinants within or nearby deleted region, respectively. Interaction of other MAbs with the NS1 and NS1d1 recombinant proteins did not differ, suggesting that their epitopes were located in the region of N-terminal 268 a.a. or C-terminal 19 a.a. of the TBEV NS1 protein. The second NS1d2 truncated protein contained the first N-terminal 33 a.a. of the TBEV NS1 protein and was able to bind with 29G9 MAb. Taken together the data stand for the differences in the N-terminal structure of the TBEV NS1 multimers secreted from the acute and persistent infected cells whereas the intracellular and secreted monomer processing was the same. The modified NS1 protein oligomers in the RH cellular line might slow virus replication and could result in the TBEV persistence.
Bugrysheva
J V
JV
Novosibirsk Institute of Bioorganic Chemistry of Siberian Branch of Russian Academy of Sciences, Lavrentyev's Prospect 8, 630090 -90, Novosibirsk, Russia.
Matveeva
V A
VA
Dobrikova
E Y
EY
Bykovskaya
N V
NV
Korobova
S A
SA
Bakhvalova
V N
VN
Morozova
O V
OV
D000818:Animals; D002460:Cell Line; D004669:Encephalitis Viruses, Tick-Borne / Q000254:growth & development; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D006023:Glycoproteins / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D013552:Swine; D017361:Viral Nonstructural Proteins / Q000032:analysis* / Q000276:immunology; D017735:Virus Latency
D018985:Epitopes, B-Lymphocyte; D006023:Glycoproteins; C100315:NS1 protein, Flavivirus; D017361:Viral Nonstructural Proteins