ena-STUDY-HIV Pathogenesis Research Unit-16-11-2020-12:42:03:912-455
The founder virus HIV envelope was developed based on the consensus sequence of 1894 subtype C founder virus sequences. The protein has been used as an immunogen has as a SOSIP.664 trimer shows antigenically relevant epitopes and improved immunogenicity.
This study includes the development and characterization of a putative immunogen that resembles the HIV-1 native spike, is antigenically relevant and is able to produce broadly neutralizing antibodies. Previous work done in our lab has shown that HIV-1 envelopes (Env) in complex with a 2dCD4-S60C protein produced a "super-immunogen" able to induce broadly neutralizing antibodies in rabbits, following immunization. This project looks to improve on previous experiments by producing a soluble, homogeneous and cleaved native-like HIV-1 trimer (termed SOSIP.664 gp140 trimers) based on an in-silico designed founder virus consensus C Env. Subtype-C being the clade responsible for more than 50% of infections worldwide and also the most prevalent subtype in Africa and Southeast Asia, the use of a consensus C Env ensures some genetic diversity coverage. The immunogen development aspect of the project involves the design, expression and purification of the consensus C trimerized Env and compares it to the widely characterized BG505 SOSIP.664 trimer. Covalent coupling of the Env to the 2dCD4-S60C protein followed by further characterization including hydrogen-deuterium exchange experiments to study the molecular and structural conformation as well as the dynamics of this complex.The immunogenicity of the complex was interrogated in immunization studies and the resultant breadth and potency of neutralization response from the determined from the animal sera.
This study includes the development and characterization of a putative immunogen that resembles the HIV-1 native spike, is antigenically relevant and is able to produce broadly neutralizing antibodies. Previous work done in our lab has shown that HIV-1 envelopes (Env) in complex with a 2dCD4-S60C protein produced a "super-immunogen" able to induce broadly neutralizing antibodies in rabbits, following immunization. This project looks to improve on previous experiments by producing a soluble, homogeneous and cleaved native-like HIV-1 trimer (termed SOSIP.664 gp140 trimers) based on an in-silico designed founder virus consensus C Env. Subtype-C being the clade responsible for more than 50% of infections worldwide and also the most prevalent subtype in Africa and Southeast Asia, the use of a consensus C Env ensures some genetic diversity coverage. The immunogen development aspect of the project involves the design, expression and purification of the consensus C trimerized Env and compares it to the widely characterized BG505 SOSIP.664 trimer. Covalent coupling of the Env to the 2dCD4-S60C protein followed by further characterization including hydrogen-deuterium exchange experiments to study the molecular and structural conformation as well as the dynamics of this complex.The immunogenicity of the complex was interrogated in immunization studies and the resultant breadth and potency of neutralization response from the determined from the animal sera.