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AIMS the aims of this work were to evaluate the protective role of 250 kDa polypeptide band of Naegleria fowleri. METHODS AND RESULTS  We designed an immunization strategi in Balb/c mice which were inoculated by i.n. route with an electroluted 250 kDa polypeptide band of N. fowleri. We observed that 250 kDa band induced 80% of protection, whereas the coadministration with Cholera Toxin induced a 100% of protection. Moreover, high levels of IgA and IgG specific antibodies were detected by ELISA assay. We also analyzed migration molecules (α4β1 and LFA-1) on T and B lymphocytes in nose-associated lymphoid tissue (NALT), cervical lymph nodes (CN) and nasal passages (NP) by flow cytometry. We observed that the percentage of B cells (B220/ α4β1) and T cells (CD4/α4β1) in NP was higher in all immunized groups compared with the other compartments analyzed. Finally, we detected by immunohistochemistry ICAM-1 and V-CAM-1 in the nasal cavity. CONCLUSION the immunization with 250 kDa polypeptide band, protect mice against N. fowleri challenge and modifies migration molecules and its ligands.