Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 9 3 2014 09 15 382 393 EN Gholamreza Habibi Department of Parasite Vaccine Research and Production, Razi Vaccine and Serum Research Institute, Karaj, Iran. Kasra Esmaeil-Nia Department of Parasite Vaccine Research and Production, Razi Vaccine and Serum Research Institute, Karaj, Iran. Hasan Izadi Department of Foot and Mouth Disease Vaccine production, Razi Vaccine and Serum Research Institute, Karaj, Iran. Orang Ataie Amarloie Department of Clinical Sciences, Faculty of Veterinary Sciences, Islamic Azad University, Karaj, Iran. Asghar Afshari Department of Parasite Vaccine Research and Production, Razi Vaccine and Serum Research Institute, Karaj, Iran. Nadia Bordbar Department of Quality Control of Biological Products, Razi Vaccine and Serum Research Institute, Karaj, Iran. 2015 10 14 Background: In this study, the pilot production of aerobic bioreactor tropical theileriosis vaccine was optimized with the aim of immunological assays for further mass production. Methods: We have shown earlier the delayed type hypersensitivity (DTH) assay could be used for evaluating the immunity and memory cells against specific Theileria antigen in vaccinated animals. In addition, TNF-α is the principle cytokine in modulating the cytotoxic activity of cytotoxic T-lymphocytes (CTL). Immunological analysis of the vaccine was performed by using two cell mediated immunity(CMI) in vitro and in vivo DTH test (Theilerin) and TNF-α assay. Results: The results of immune responses of susceptible immunized cattle by bioreactor vaccine in comparison with conventional flask vaccine revealed a significant stimulation of immune cells by transcription of high level of TNF-α and positive reaction against Theileria antigen in Theilerin skin test (DTH). Conclusion: The equal immunological results achieved in both above mentioned vaccines verified the satisfactory immunity for aerobic bioreactor theileriosis vaccine for advance mass vaccination in the field on a large scale https://ijpa.tums.ac.ir/index.php/ijpa/article/view/388 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/388/356