Classification of Leptospira interrogans serovar lai strain 017 by using monoclonal antibodies

Leptospira interrogans serovar lai was identified in China in 1966 as a new serovar of the icterohaemorrhagiae serogroup by cross-absorption tests. In this study, we established three hybridoma cell lines producing monoclonal antibodies (MAbs) of the immunoglobulin G3 subclass (LW1, LW2, and LW3) and of the immunoglobulin M class (LW4a) against serovar lai strain 017 by the cell fusion technique. Immunological reactivities of the MAbs were determined by the microscopic agglutination test, enzyme-linked immunosorbent assay (ELISA), and indirect immunofluorescence assay. MAbs LW1 and LW3 agglutinated cells of serovars lai, birkini, and gem of serogroup icterohaemorrhagiae. LW2 agglutinated various serovars of serogroup icterohaemorrhagiae, except for serovar tonkini. LW4a reacted positively with various Leptospira species, including a new species, Leptospira parva, in the ELISA and indirect immunofluorescence assay. However, LW4a did not react with Leptonema illini 3055. The results of an inhibition ELISA with heated outer envelope (OE) or periodate-oxidized OE suggested that these MAbs recognize a carbohydrate moiety of the OE as the antigenic determinant.