Production and Characterization of Monoclonal Antibodies Against Seoul Virus

ABSTRACT

Hantaviruses are members of the family Bunyaviridae, the etiologic agents of Hemorrhagic fever with renal syndrome (HFRS) and Hantavirus Pulmonary Syndrome (HPS). They are negative-sense, single-stranded RNA viruses possessing a large (L), medium (M) and small (S) genomic segment which encodes a viral polymerase, envelope glycoproteins (G1 and G2) and a nucleocapsid (N) protein, respectively. Seoul (SEO) virus, the causative agent of clinically mild HFRS in worldwide, was isolated from lung tissues of urban rat (Rattus norvegicus) captured in Seoul, Korea, 1982. To clarify the antigenic characteristics and the differentiation of serotypes of hantavirus, 8 hybridoma cell lines secreting monoclonal antibodies (MAbs) against SEOV 80-39 strain were produced by fusion of SP2/0-Ag14 mouse myeloma cells with spleen cells of BALB/c mice, immunized with SEOV. Reactivities of these MAbs were examined by the indirect immunofluorescence assay (IFA), enzyme linked immunosorbent assay (ELISA), immunoblot, high density particle agglutination (HDPA) and plaque reduction neutralization test (PRNT). Eight out of 235 hybridomas secreted MAbs of Seoul virus 80-39 continuously, and these eight MAbs were all IgG. The isotypes of these 8 MAbs are; one clone (F3-3C) was IgG1, six (F1-1B9B, F1-3B, F1-3D, F4-1E, F4-3F, F4- 6C) were IgG2a and one (F1-1B9F) was IgG2b. Seven MAbs (F1-1B9B, F1-1B9F, F1-3B, F1- 3D, F3-3C, F4-1E, F4-3F) reacted with nucleocapsid protein (M.W. 50K) of SEOV by immunoblot. All eight MAbs were cross-reacted with Hantaan (HTN) virus, one (F4-3F) was cross-reacted with Puumala (PUU) virus and two (F1-1B9B, F1-3B) were cross-reacted with Prospect Hill (PH) virus by IFAT. None of these 8 MAbs had neutralizing activity.