Jingjing Zhang

Dr. Jingjing Zhang


Research Focus & Metrics

Academic Career Length

2Years (2025-Present)

(Calculated once Scilit publication data is synced: current year − year of first publication + 1)

Representative Publications

View All Publications
26 November 2025 in Vaccines
Verified
Journal Article
Open access
PDF file
ZZZhixiao Zhang
Zhixiao Zhang
JZJingjing Zhang
Jingjing Zhang
CMChangyong Mu
Changyong Mu
KMKaili Ma
Kaili Ma
DGDongxiu Gao
Dongxiu Gao
QLQihan Li
Qihan Li
Background: Rabies virus (RABV) causes approximately 59,000 human deaths annually. Current pre- and post-exposure vaccination relies on inactivated vaccines (INVs) with limited yield and immunogenicity. We engineered a dual-cationic LNP-based nucleocapsid-like nanostructure (NLS) that co-encapsulates RABV G-mRNA and recombinant RABV-N to engage MHC-I/II pathways and enhance protection. Methods: A pVAX-RABV-G plasmid containing 5′/3′UTRs, Kozak, and poly(A) was transcribed in vitro. RABV-N with an N-terminal 6× His tag was expressed in E. coli BL21(DE3) and purified by Ni-Sepharose affinity chromatography. Dual-cationic LNPs (DHA, DOTAP Cl, mPEG-DTA2K, DOPC) were formulated by microfluidics at a 4:1 (G-mRNA:RABV-N) mass ratio. Vaccine quality was assessed by encapsulation efficiency, DLS, PDI, zeta potential, and TEM. Mice received empty LNPs, INV, G-mRNA, or NLS under varied schedules and doses. ELISA measured RABV-G/N-IgG; RFFIT determined neutralizing antibody (nAb) titers; ELISPOT quantified CTL response; qPCR assessed T-cell activation genes. On day 35 after the first immunization of vaccines, mice were challenged intramuscularly with 25 LD50 of CVS-24. Results: G-mRNA purity was >95% and drove strong RABV-G expression in 293T cells. Purified RABV-N was approximately 52 kDa, >90% pure, and reactive to anti-His and anti-N antibodies. NLS achieved >95% encapsulation, a diameter of 136.9 nm, PDI 0.09, and a +18.7 mV zeta potential. A single dose yielded approximately 10 IU mL−1 nAb by day 7; two doses peaked at approximately 1000 IU mL−1. Mice showed 100% survival and no viral rebound in brain, spinal cord, and sciatic nerve. NLS induced stronger MHC-I/II-linked cellular immunity and higher RABV G/N-specific IFN-γ spot frequencies than G-mRNA or INV. Conclusions: The dual-antigen NLS vaccine co-delivering G-mRNA and RABV-N via dual-cationic LNPs robustly activates MHC-I/II, rapidly generates high-titer nAb (≥10 IU mL−1 within 1 week), and sustains potent CD8+ CTL and CD4+ Th responses. A two-dose regimen (days 0 and 21) conferred complete protection, supporting the NLS platform as a next-generation rabies vaccine candidate.

22 October 2025 in Preprints.org
Verified
Preprint
Open access
PDF file
ZZZhixiao Zhang
Zhixiao Zhang
JZJingjing Zhang
Jingjing Zhang
CMChangyong Mu
Changyong Mu
KMKaili Ma
Kaili Ma
DGDongxiu Gao
Dongxiu Gao
QLQihan Li
Qihan Li

Based on a dual-cationic lipid nanoparticle (LNP) platform, we developed a novel rabies virus (RABV) nucleocapsid-like nanostructure (NLS) vaccine. The antigen comprises two nucleocapsid components: a mRNA encoding the RABV glycoprotein (G-mRNA) and a recombinant RABV nucleoprotein (RABV-N) expressed in Escherichia coli (E. coli). These components are co-encapsulated within dual-cationic LNPs to form the RABV NLS vaccine. Compared with the G-mRNA vaccine and inactivated vaccine (INV), the NLS vaccine elicits significantly stronger humoral and cellular immune responses. Seven days after a single dose, the NLS vaccine elicited neutralizing antibody titers of ~10 IU mL-1. Following two doses on days 0 and 21, the peak neutralizing antibody titer reached 1,000 IU mL-1, and 100% of mice survived a challenge with 25 LD₅₀ of the fixed RABV strain on day 14 post-boost.