The purpose of this experiment was to study the effect of Newcastle Disease Virus（NDV）on the migration of peripheral blood T lymphocytes in different geneotypes of Yellow ? feathered chickens，and to provide theoretical knowledge for exploring the immune response mechanism of chickens to NDV. Using Transwell co?culture system，the upper chamber and the lower chamber were cultured with lymphocytes and dendritic cells，respectively. After NDV challenging，the lymphocyte suspension was collected at 0，2，4，6 h of each time point. The total T lymphocyte migration rate was obtained by the flow cytometry method and the optimal co ? cultivation time wasdetermined. Then the migration rate of total T lymphocytes in wild?type（TT），hybrid?type（CT）and homozygotic?type （CC）were compared after NDV challenge. The results showed that the migration rate of total T lymphocytes was 45.04% after 2 hours of NDV challenge. In the groups challenged by virus，the migration rate of total T lymphocytes in CT genotype was 60.03% ，and the total T lymphocytes migration rate in TT genotype was 45.40% . There was a significant difference between the two genotypes（P<0.05）. However，there was no significant difference between CT and CC genotype（P>0.05）. In conclusion，the time of optimal lymphocytes culture was 2 hours，which was suitable for cell migration. After NDV challenge，the migration rate of total T lymphocytes in CT genotype was faster than that of TT genotype.