Author:
Fayaz Shima,Raz Abbasali,Bahrami Fariborz,Fard-Esfahani Pezhman,Parvizi Parviz,Ajdary Soheila
Abstract
AbstractSand fly salivary proteins have immunomodulatory and anti-inflammatory features; hence, they are proven to perform important roles in the early establishment of Leishmania parasite in the vertebrate host. Among them, salivary apyrase with anti-hemostatic properties has a crucial role during the blood meal process. In the present study, a Genome-Walking method was used to characterize a full-length nucleotide sequence of Phlebotomus (P.) kandelakii apyrase (Pkapy). Bioinformatics analyses revealed that Pkapy is a ~ 36 kDa stable and hydrophilic protein that belongs to the Cimex family of apyrases. Moreover, recombinant proteins of Pkapy and P. papatasi apyrase (Ppapy) were over-expressed in Escherichia coli BL2 (DE3) and their antigenicity in BALB/c mice was evaluated. Dot-blot and ELISA results indicated that both recombinant apyrases could induce antibodies in BALB/c. Moreover, a partial cross-reactivity between Pkapy and Ppapy was found. In vitro stimulation of splenocytes from immunized mice with the recombinant proteins indicated cross-reactive T cell proliferative responses. Cytokine analysis revealed significant production of IFN-γ (p < 0.001) and IL-10 (p < 0.01) in response to Pkapy. In conclusion, the full-length nucleotide sequence and molecular characteristics of Pkapy were identified for the first time. Immunologic analyses indicated that Pkapy and Ppapy are immunogenic in BALB/c mice and show partial cross-reactive responses. The immunity to Pkapy was found to be a Th1-dominant response that highlights its potential as a component for an anti-Leishmania vaccine.
Funder
Pasteur Institute of Iran
Publisher
Springer Science and Business Media LLC
Reference37 articles.
1. Steverding, D. The history of leishmaniasis. Parasit. Vectors 10, 82 (2017).
2. Abdeladhim, M., Kamhawi, S. & Valenzuela, J. G. What’s behind a sand fly bite? The profound effect of sand fly saliva on host hemostasis, inflammation and immunity. Infect. Genet. Evol. J. Mol. Epidemiol. Evol. Genet. Infect. Dis. 28, 691–703 (2014).
3. Kamhawi, S., Belkaid, Y., Modi, G., Rowton, E. & Sacks, D. Protection against cutaneous leishmaniasis resulting from bites of uninfected sand flies. Science 290, 1351–1354 (2000).
4. Lestinova, T., Rohousova, I., Sima, M., de Oliveira, C. I. & Volf, P. Insights into the sand fly saliva: Blood-feeding and immune interactions between sand flies, hosts, and Leishmania. PLoS Negl. Trop. Dis. 11, e0005600 (2017).
5. Mozaffari, E. et al. Epidemiology of visceral leishmaniasis with emphasis on the dynamic activity of sand flies in an important endemic focus of disease in Northwestern Iran. J. Arthropod.-Borne Dis. 14, 97–105 (2020).