[The relationship between cellular infection by Hantaan virus and expression of beta3 integrin].
OBJECTIVE
To investigate the relationship between cellular entry of Hantaan virus (HTNV) and expression of beta3 integrin in beta3-integrin-deficient and HTNV-insusceptible China hamster ovary (CHO) cells.


METHODS
Eukaryotic expression vector encoding human integrin beta3 and eukaryotic expression vector harboring human integrin alphav or alphaIIb subunit cDNA were transfected into HTNV non-permissive CHO cells individually or collectively. Screening for stable transfectant clones was performed using G418 selective (culture medium. The exogenous gene expression was analyzed qualitatively and quantitatively by immunofluorescence assay (IFA) and flow cytometry (FCM). Various modified CHO cells and untransfected CHO cells were infected using HTNV A9. At various time points after infection, HTNV antigens in infected cells were detected qualitatively and quantitatively by IFA, FCM.


RESULTS
Highly-effective surface expression of beta3 integrin was measured in CHO/alphavbeta3 and CHO/alphaIIbbeta3, while weaker surface expression was detected in CHO/beta3 (P < 0.05). Expression of alphav or alphaIIb integrin in the individually transfected group was significantly lower than in the cotransfected group (P < 0.01) and the sites of localization changed. In contrast, effective surface expression was not seen when pcDNA3 was transfected alone. The infection rate of CHO/alphavbeta3 (60.1%) and CHO/alphaIIbbeta3 (55. 9%) cells were significantly higher than that of CHO/beta3 (38.7%) cells, while the infection rate of CHO/beta3 was significantly higher than that of CHO/alphav, CHO/pcDNA3 and CHO cells respectively. There was a close relationship between the positive percentage of HTNV A9-infected cells and expression of beta3 integrin.


CONCLUSIONS
These results indicated that cellular entry of HTNV was related to the expression of beta3 integrin.
Mou
Dan-Lei
DL
Center of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China.
Bai
Xue-Fan
XF
Huang
Chang-Xing
CX
Wang
Ying-Peng
YP
Pan
Lei
L
Li
Guang-Yu
GY
Sun
Yong-Tao
YT
Nie
Qing-He
QH
Yang
Wei-Song
WS
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D016466:CHO Cells; D006224:Cricetinae; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D018063:Hantaan virus / Q000276:immunology / Q000502:physiology*; D006801:Humans; D039661:Integrin beta3 / Q000235:genetics / Q000378:metabolism*; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D000956:Antigens, Viral; D039661:Integrin beta3; D011991:Receptors, Virus