ABSTRACT
Objective To study the impact of V2 mutations on neutralizing ability of HIV-1-specific neutralizing antibodies. Methods We tested the influence of L175P mutation to the neutralizing ability of V3-specific antibodies by pseudotype virus and the binding affinity of those V3-spesific antibodies to gpl20 monomer by ELISA. Results We found L175P mutation changed the neutralizing ability of V3-specific antibodies. However, L175P mutation showed no effects on the binding affinity of these antibodies to gpl20 monomer. Conclusion Our results revealed the L175P mutation at V2 loop changed the natural trimmer structure of gp120 and enhanced the neutralizing ability of V3-specific antibodies.
ABSTRACT
Serological and molecular characterization of Leptospiral isolates helps us to identify serovar, which is useful, for epidemiological study. Serological characterization is tedious and requires a panel of monoclonal antibodies and expertise to read the results. This study is a preliminary work to evaluate the usefulness of Denaturing Gradient Gel Electrophoresis (DGGE) to identify serovars of leptospira. The V3 region of most conserved 16S rDNA of five pathogenic leptospiral serovars and one saprophytic serovar was characterized. DGGE method was employed to separate the amplified V3 region based on the nucleotide sequence. On DGGE, amplified V3 region of leptospiral serovars, under study, showed bands at different positions indicating DGGE as the effective method of characterization in the future. DNA sequencing of V3 region of the three serovars showed great difference in nucleotide sequence supporting the results of DGGE.