Establishment and application of a specific ELISpot detection method for rat tetanus vaccine
XING Xi-xi
MIAO Feng
TAO Ting-lei
LIU Pan-pan
XING Qing-qing
QIU Yun-liang
Abstract：Objective:To establish an ELISpot method for detection of antigen-specific T cell immune response of rats to tetanus vaccine.Methods:Based on the SD rat animal model,a dynamic monitoring strategy was adopted after immunization with a single dose of vaccine.Blood samples were collected before the first day of administration and on Day 14 and 28 to isolate PBMC,and key parameters such as the concentration of peptide stimulus and the ratio of positive controls were systematically optimized to establish an interferon-γ ELISpot detection method(IFN-γ ELISpot).The level of T cell immune response to the vaccine was quantitated.Results:The optimal concentration of peptide stimulus was determined to be 5 μg·mL-1·peptide-1,and the optimal positive control stimulus and concentration was20ng·mL-1 PMA+0.4μg·mL-1 Ionomycin.Nine dominant polypeptide sequences that could effectively stimulate T cell response were identified.Conclusion:The established method shows good performance in detecting the cellular immunity level after immunization with tetanus vaccine.
Keywords：tetanus vaccineELISpotT cell immunity responseIFN-γpeptide screeningefficacy evaluation of vaccine
Publication Date：2026-01-30
Online Publishing Date：2026-08-14(First online date of this platform, not the publication date of the document)
Pages：7( 193-199 )