Identification of differential protein recognition pattern between Naegleria fowleri and Naegleria lovaniensis

Abstract

Many pathogenicity factors are involved in the development of primary amoebic meningoencephalitis (PAM) caused by N fowleri. However, most of them are not exclusive for N fowleri and they have not even been described in other nonpathogenic Naegleria species. Therefore, the objective of this work was to identify differential proteins and protein pattern recognition between Naegleria fowleri and Naegleria lovaniensis using antibodies anti‐N fowleri as strategy to find vaccine candidates against meningoencephalitis. Electrophoresis and Western blots conventional and 2‐DE were performed for the identification of antigenic proteins, and these were analysed by the mass spectrometry technique. The results obtained in 2‐DE gels and Western blot showed very notable differences in spot intensity between these two species, specifically those with relative molecular weight of 100, 75, 50 and 19 kDa. Some spots corresponding to these molecular weights were identified as actin fragment, myosin II, heat shock protein, membrane protein Mp2CL5 among others, with differences in theoretical post‐translational modifications. In this work, we found differences in antigenic proteins between both species, proteins that could be used for a further development of vaccines against N fowleri infection.

Identification of Immunogenic Antigens of Naegleria fowleri Adjuvanted by Cholera Toxin

“…As we mentioned above, in a recently published work [11], we found differential proteins between N. fowleri and N. lovaniensis, specifically using IgG antibodies by 2-DE Western blot assay. However, the analysis did not include an immunization scheme, with or without CT as an adjuvant, in order to assess survival rates with different numbers of immunizations.…”

“…Interestingly, many immunogenic bands, including these six bands, were adjuvanted by CT, where some proteins were identified, such as myosin II heavy chain (Q25561), actin (B5M6J9), ATP synthase F1 subunit alpha (M4H5H9), amino acid decarboxylase (C6L6E3), membrane protein (Q95UJ2), Hsp70 (Q6B3P1), and glyceraldehyde 3 phosphate dehydrogenase (D2W142). These are proteins that had been previously reported by Gutiérrez-Sanchez et al (2020) [11], with differential expression between N. fowleri and N. lovaniensis.…”

“…Moreover, mass spectrometry also identified differences in proteins, such as actin fragment, myosin II, heat shock protein, membrane protein Mp2CL5 regarding sequence coverage, and the number of post-translational modifications, that had not been previously reported. [11]. However, the identification of immunogenic antigens recognized by both local and systemic specific antibodies, including a scheme with a different number of immunizations and the use or not of CT as an adjuvant, has not yet been analyzed.…”

Isolation and Identification of Naegleria Species in Irrigation Channels for Recreational Use in Mexicali Valley, Mexico

“…The results obtained showed very notable differences in spot intensity between these two species by 2-DE gels and 2-DE Western blot, specifically those with relative molecular weights of 100, 75, 50 and 19 kDa. Some spots corresponding to these molecular weights were identified as actin fragment, myosin II, heat shock protein, and membrane protein Mp2CL5, among others, with differences in theoretical post-translational modifications [ 34 ]. Interestingly, it has been shown that some pathogenicity factors are not exclusive to N fowleri , since they have also been reported in nonpathogenic Naegleria species with different levels of expression [ 35 , 36 , 37 , 38 ].…”

Primary Amoebic Meningoencephalitis by Naegleria fowleri: Pathogenesis and Treatments

“…Furthermore, another study has revealed a membrane protein called Mp2CL5 (17 kDa) in pathogenic Naegleria species. It has been proposed that it plays a role in the amoeba’s cellular recognition and adhesion [ 37 ].…”