Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies

作者：Porcino Gabriane Nascimento; Carvalho Campos Cristiane; Ribeiro Gomes Maia Ana Carolina; Detoni Michelle Lima; Faria Pinto Priscila; Coimbra Elaine Soares; Marques Marcos Jose; Juliano Maria Aparecida; Juliano Luiz; Diniz Vanessa Alvaro; Corte Real Suzana; Vasconcelos Eveline Gomes^*

来源：Experimental Parasitology, 2012, 132(2): 293-299.

DOI：10.1016/j.exppara.2012.08.009

• Apyrase
• ATP diphosphohydrolase
• NTPase
• Peptide
• Leishmaniasis
• Immunocytochemical

Nucleoside triphosphate diphosphohydrolase (NTPDase) activity was recently characterized in Leishmania (Viannia) braziliensis promastigotes (Lb), and an antigenic conserved domain (r82-121) from the specific NTPDase 1 isoform was identified. In this work, mouse polyclonal antibodies produced against two synthetic peptides derived from this domain (LbB1LJ, r82-103; LbB2LJ, r102-121) were used. The anti-LbB1LJ or anti-LbB2LJ antibodies were immobilized on protein A-sepharose and immunoprecipitated the NTPDase 1 of 48 kDa and depleted approximately 40% of the phosphohydrolytic activity from detergent-homogenized Lb preparation. Ultrastructural immunocytochemical microscopy identified the NTPDase 1 on the parasite surface and in its subcellular cytoplasmic vesicles, mitochondria, kinetoplast and nucleus. The ATPase and ADPase activities of detergent-homogenized Lb preparation were partially inhibited by anti-LbB1LJ antibody (43-79%), which was more effective than that inhibition (18-47%) by anti-LbB2LJ antibody. In addition, the immune serum anti-LbB1LJ (67%) or anti-LbB2LJ (33%) was cytotoxic, significantly reducing the promastigotes growth in vitro. The results appoint the conserved domain from the L braziliensis NTPDase as an important target for inhibitor design and the potential application of these biomolecules in experimental protocols of disease control.

• 出版日期2012-10