At present, there are no approved vaccines or treatments for the prevention or control of EEEV infection in the general human population, and EEEV is an emergent threat to human health . One of the most virulent viruses in North America is the Eastern equine encephalitis virus (EEEV). Currently, there are no approved vaccines or antiviral remedies for this infection, which has recently experienced a rise in human cases . Currently, there is no specific treatment for EEE, and a formalin-killed vaccine based on the NA-EEEV strain (PE-6) is available for use in horses and emus. However, this vaccine does not induce significant neutralizing anti-SA-EEEV and anti-E2 antibodies . The VEEV TC-83 strain is the only licensed live attenuated vaccine for protecting horses in endemic regions. To develop this vaccine strain, the Trinidad donkey (TrD) IA/B strain underwent 83 passages in guinea pig heart cells . Currently, there is no approved human VEEV vaccine; however, military personnel and laboratory workers administer the TC-83 live attenuated vaccine .