Generation Of Humanized Mice Model And Analysis Of The Feasibility Of Using It For Studying RV144 AIDS Vaccine

Since human found AIDS, which caused by HIV in 1983, AIDS has become a world-widely infectious diseases. Every year,2.4 million people are newly infected with HIV-1. Hence, a safe and effective HIV-1 vaccine is urgently needed to stop HIV-1 transmission and to curtail the HIV pandemic. However, due to the genetic diversity and mutability of HIV-1, HIV-1 vaccine development over the past three decades had been disappointed until RV144 Thai HIV-1 vaccine trial. For the first time, a large scale trial revealed a HIV-1 vaccine has 31.2% efficacy against HIV-1 acquisition. Fully understanding the immune response induced by this vaccine can point out a direction for more effective HIV-1 vaccine design.However, all the studies regarding RV144 Thai HIV-1 vaccine are based on Human PBMCs and the immune responses at portal of virus entry cannot be studied because of lymphatic and mucosal tissues cannot be procured for the operational and ethical reasons. Thus, the mechanistic correlation of protection of this vaccine remains elusive. Therefore, an animal model which can be infected with HIV-1 is urgently needed to study this vaccine instead of Human.Humanized-BLT (bone marrow, liver and thymus) mice offer an opportunity to evaluate the adaptive immune response in mucosa and secondary lymphatic tissues to RV144 HIV-1 vaccine. We are going to generate humanized-BLT mice through transplanting Human fetal thymus/liver and CD34+cell into NOD/SCID immunodeficient mice and detect the reconstitution result of this animal model. Thereafter, we will vaccinate the humanized mice with RV144 Thai HIV-1 vaccine and analyze the immune response induced by this vaccine in humanized mice which have functional Human immune system.By detecting with flow cytometry, we found that some humanized mice containing Human CD45+lymphocytes higher than 88% in the whole CD45+lymphocytes in mice PBMCs, which demonstrated the success of generating humanized NOD/SCID BLT mice. We also adjusted the vaccination schedule and dosage for humanized mice due to its smaller body weight compare to Human. ELISA and Western Blot results showed that RV144 vaccine can induce humoral immune response in humanized mice. Furthermore, in order to confirm the intracellular staining system (ICS) can be used to detect cellular immune response in vaccinated humanized mice, we used HIV-1 infected mice’s PBMCs to test the feasibility of the ICS system and workability of those intracellular staining fluorescent antibodies. Results showed that this system can be used to detect cellular immune response in humanized mice.All in all, we have successfully generated humanized NOD/SCID BLT mice and preliminarily demonstrated it can be used to study RV144 Thai HIV-1 vaccine.