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Rhipicephalus microplus and Ixodes ovatus cystatins in tick blood digestion and evasion of host immune response.

Parizi LF, Sabadin GA, Alzugaray MF, Seixas A, Logullo C, Konnai S, Ohashi K, Masuda A, da Silva Vaz I - Parasit Vectors (2015)

Bottom Line: Finally, cross-antigenicity assays revealed that antibodies against the JpIocys2a peptide recognize native and recombinant R. microplus cystatins.The presence of these proteins in different tissues and their ability to differentially inhibit cathepsins suggest distinct roles for JpIocys2a, BrBmcys2b, and BrBmcys2c in blood digestion, egg and larvae development, and modulation of host immune response in tick physiology.The cross-antigenicity between native and recombinant cystatins supports further experiments using JpIocys2a, BrBmcys2b, and BrBmcys2c as vaccine antigens.

View Article: PubMed Central - PubMed

Affiliation: Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Avenida Bento Gonçalves, 9500, Prédio 43421, Porto Alegre, 91501-970, , RS, Brazil. luisfparizi@cbiot.ufrgs.br.

ABSTRACT

Background: Cystatins are a group of cysteine protease inhibitors responsible for physiological proteolysis regulation and present in a wide range of organisms. Studies about this class of inhibitors in parasites have contributed to clarify their roles in important physiological processes, like blood digestion and modulation of host immune response during blood feeding. Thus, cystatins are a subject of research on the development of new parasite control methods. Additionally, the characterization of proteins shared by different parasite species represents a valuable strategy to find potential targets in multi-species control methods. However, cystatin functions in ticks remain undetermined, especially in Rhipicephalus microplus and Ixodes ovatus, two species that affect livestock and human health, respectively.

Methods: Here we report the inhibitory profile of two R. microplus (BrBmcys2b and BrBmcys2c) and one I. ovatus (JpIocys2a) cystatins to commercial cathepsins B, C, and L. The presence of native cystatins in R. microplus tissues was analyzed using sera against recombinant BrBmcys2b and BrBmcys2c. Also, a peptide from JpIocys2a was synthesized for rabbit immunization, and this serum was used to analyze the cross antigenicity between R. microplus and I. ovatus cystatins.

Results: Enzymatic inhibition profile of tick cystatins shows a distinct modulation for cathepsins related to tick blood digestion and evasion of host immune response. Furthermore, BrBmcys2b was detected in saliva and different tissues along tick stages, while BrBmcys2c was detected mainly in gut from partially engorged R. microplus females, demonstrating a distinct pattern of cystatin expression, secretion and traffic between tick tissues. Moreover, phylogenetic analysis suggests that JpIocys2a belongs to the group of tick gut secreted cystatins. Finally, cross-antigenicity assays revealed that antibodies against the JpIocys2a peptide recognize native and recombinant R. microplus cystatins.

Conclusion: The presence of these proteins in different tissues and their ability to differentially inhibit cathepsins suggest distinct roles for JpIocys2a, BrBmcys2b, and BrBmcys2c in blood digestion, egg and larvae development, and modulation of host immune response in tick physiology. The cross-antigenicity between native and recombinant cystatins supports further experiments using JpIocys2a, BrBmcys2b, and BrBmcys2c as vaccine antigens.

No MeSH data available.


Conserved and antigenic tick cystatin regions for JpIocys2a peptide selection. Antigenic index plots for tick cystatins were predicted using the Jameson–Wolf algorithm. Graphic increased positivity shows predictive antigenic sites. Alignment shows conserved regions between JpIocys2a and R. microplus cystatins. Black boxes indicate conserved and antigenic amino acid region for each sequence. Asterisk indicate the selected region for peptide synthesis.
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Fig2: Conserved and antigenic tick cystatin regions for JpIocys2a peptide selection. Antigenic index plots for tick cystatins were predicted using the Jameson–Wolf algorithm. Graphic increased positivity shows predictive antigenic sites. Alignment shows conserved regions between JpIocys2a and R. microplus cystatins. Black boxes indicate conserved and antigenic amino acid region for each sequence. Asterisk indicate the selected region for peptide synthesis.

Mentions: The alignment of JpIocys2a with R. microplus cystatins showed conserved regions among the predicted amino acid sequences (Figure 2). The amino acid sequence STQVEGREYYDTVL from JpIocys2a (STQpep) was selected for peptide synthesis in accordance with the highest identity and antigenic region among all cystatins analyzed.Figure 2


Rhipicephalus microplus and Ixodes ovatus cystatins in tick blood digestion and evasion of host immune response.

Parizi LF, Sabadin GA, Alzugaray MF, Seixas A, Logullo C, Konnai S, Ohashi K, Masuda A, da Silva Vaz I - Parasit Vectors (2015)

Conserved and antigenic tick cystatin regions for JpIocys2a peptide selection. Antigenic index plots for tick cystatins were predicted using the Jameson–Wolf algorithm. Graphic increased positivity shows predictive antigenic sites. Alignment shows conserved regions between JpIocys2a and R. microplus cystatins. Black boxes indicate conserved and antigenic amino acid region for each sequence. Asterisk indicate the selected region for peptide synthesis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4340882&req=5

Fig2: Conserved and antigenic tick cystatin regions for JpIocys2a peptide selection. Antigenic index plots for tick cystatins were predicted using the Jameson–Wolf algorithm. Graphic increased positivity shows predictive antigenic sites. Alignment shows conserved regions between JpIocys2a and R. microplus cystatins. Black boxes indicate conserved and antigenic amino acid region for each sequence. Asterisk indicate the selected region for peptide synthesis.
Mentions: The alignment of JpIocys2a with R. microplus cystatins showed conserved regions among the predicted amino acid sequences (Figure 2). The amino acid sequence STQVEGREYYDTVL from JpIocys2a (STQpep) was selected for peptide synthesis in accordance with the highest identity and antigenic region among all cystatins analyzed.Figure 2

Bottom Line: Finally, cross-antigenicity assays revealed that antibodies against the JpIocys2a peptide recognize native and recombinant R. microplus cystatins.The presence of these proteins in different tissues and their ability to differentially inhibit cathepsins suggest distinct roles for JpIocys2a, BrBmcys2b, and BrBmcys2c in blood digestion, egg and larvae development, and modulation of host immune response in tick physiology.The cross-antigenicity between native and recombinant cystatins supports further experiments using JpIocys2a, BrBmcys2b, and BrBmcys2c as vaccine antigens.

View Article: PubMed Central - PubMed

Affiliation: Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Avenida Bento Gonçalves, 9500, Prédio 43421, Porto Alegre, 91501-970, , RS, Brazil. luisfparizi@cbiot.ufrgs.br.

ABSTRACT

Background: Cystatins are a group of cysteine protease inhibitors responsible for physiological proteolysis regulation and present in a wide range of organisms. Studies about this class of inhibitors in parasites have contributed to clarify their roles in important physiological processes, like blood digestion and modulation of host immune response during blood feeding. Thus, cystatins are a subject of research on the development of new parasite control methods. Additionally, the characterization of proteins shared by different parasite species represents a valuable strategy to find potential targets in multi-species control methods. However, cystatin functions in ticks remain undetermined, especially in Rhipicephalus microplus and Ixodes ovatus, two species that affect livestock and human health, respectively.

Methods: Here we report the inhibitory profile of two R. microplus (BrBmcys2b and BrBmcys2c) and one I. ovatus (JpIocys2a) cystatins to commercial cathepsins B, C, and L. The presence of native cystatins in R. microplus tissues was analyzed using sera against recombinant BrBmcys2b and BrBmcys2c. Also, a peptide from JpIocys2a was synthesized for rabbit immunization, and this serum was used to analyze the cross antigenicity between R. microplus and I. ovatus cystatins.

Results: Enzymatic inhibition profile of tick cystatins shows a distinct modulation for cathepsins related to tick blood digestion and evasion of host immune response. Furthermore, BrBmcys2b was detected in saliva and different tissues along tick stages, while BrBmcys2c was detected mainly in gut from partially engorged R. microplus females, demonstrating a distinct pattern of cystatin expression, secretion and traffic between tick tissues. Moreover, phylogenetic analysis suggests that JpIocys2a belongs to the group of tick gut secreted cystatins. Finally, cross-antigenicity assays revealed that antibodies against the JpIocys2a peptide recognize native and recombinant R. microplus cystatins.

Conclusion: The presence of these proteins in different tissues and their ability to differentially inhibit cathepsins suggest distinct roles for JpIocys2a, BrBmcys2b, and BrBmcys2c in blood digestion, egg and larvae development, and modulation of host immune response in tick physiology. The cross-antigenicity between native and recombinant cystatins supports further experiments using JpIocys2a, BrBmcys2b, and BrBmcys2c as vaccine antigens.

No MeSH data available.