Salivary gland hyaluronidase in various species of phlebotomine sand flies (Diptera: psychodidae)
Introduction
Sand flies (Diptera, Phlebotominae), similarly to other blood-sucking arthropods, possess an array of salivary compounds that modulate hemostasis and immunity of the host in order to facilitate blood meal acquisition without inducing an immediate hypersensitivity response (reviewed by Champagne and Valenzuela, 1996, Wikel et al., 1996. Sand fly salivary compounds also have a key role in transmission of parasitic protozoa of the genus Leishmania, serious human pathogens. The immunosuppressive effect of saliva decreases the infective dose of the parasite and has an enhancing effect on Leishmania infection in the vertebrate host (Titus and Ribeiro, 1988, Theodos et al., 1991). Studies carried out on two important vectors, L. longipalpis and P. papatasi, revealed that saliva inhibits activation of T-cells (Titus, 1998) and some macrophage functions, including the oxidative burst (Gillespie et al., 2000) and NO production (Hall and Titus, 1995, Katz et al., 2000). In natural conditions this allows the establishment of the infection from a low number of highly infective metacyclic forms ejected from the sand fly proboscis (Warburg and Schlein, 1986).
One of molecules which may participate in both, a blood meal acquisition and enhancement of Leishmania infection, is a hyaluronidase, an enzyme recently detected in saliva of L. longipalpis and P. papatasi (Charlab et al., 1999, Ribeiro et al., 2000). Hyluronidases cleave hyaluronic acid (HA) which is a high molecular weight glycosaminoglycan and a major component of the extracellular matrix in vertebrates. In addition, most of them also cleave other components of extracellular matrix, particularly chondroitin sulfates. There are, however, at least three types of hyaluronidases hydrolyzing HA via different mechanisms. Testicular hyaluronidases of mammals and venom hyaluronidases of Hymenoptera are endo-N-acetyl-hexosaminidases degrading high molecular weight substrate to tetrasaccharides. Hyaluronidases from leeches and nematodes belong to endo-glucuronidases that specifically hydrolyse hyaluronic acid (with tetrasaccharides as the final product), while bacterial hyaluronidases act via β-elimination, yielding disaccharides as the main end products (reviewed by Kreil (1995).
In venomous snakes and arthropods like spiders, scorpions and hymenopteran insects, hyaluronidases are frequently found in toxins. They are called “spreading factors” as they facilitate the spreading of toxic compounds by degradation of the extracellular matrix (see review by Kreil (1995)). In bloodsucking arthropods other than sand flies the hyaluronidase activity was detected in the tick Amblyomma hebraeum (Neitz et al., 1978) and the black fly Simulium vittatum (Ribeiro et al., 2000). The activity is thought to play an important role in blood-meal acquisition by increasing the permeability of host tissue for other pharmacological compounds present in saliva; by depolymerizing hyaluronic acid it may help diffusion of antihemostatic agents into the vicinity of the feeding lesion or help to enlarge the size of the feeding hematoma.
In this study, we present some physico-chemical properties of sand fly hyaluronidases and compare their activities among six species of the genera Lutzomyia and Phlebotomus.
Section snippets
Materials and methods
Seven colonies of six sand fly species were used: Phlebotomus papatasi Scopoli, P. duboscqi Neveu-Lemaire, P. sergenti Parrot, P. perniciosus Newstead, P. halepensis Theodor and Lutzomyia longipalpis Lutz and Nieva. In P. papatasi, two colonies of different in geographical origin (Turkey and Cyprus) were compared. The flies used to establish the colonies were collected in Sanliurfa, Turkey (P. papatasi and P. sergenti) or were provided by Dr. M. Maroli (P. perniciosus), Prof. R. Ward (L.
Results
Protein content in SGEs differed between species. The highest protein level was observed in P. duboscqi - 0.78 μg per “optimal gland” (SD 0.071), followed by P. papatasi from Cyprus colony - 0.51 μg/gland (SD 0.053), P. halepensis - 0.41 μg/gland (SD 0.055), P. papatasi from Turkish colony - 0.33 μg/gland (SD 0.049) and P. sergenti - 0.23 μg/gland (SD 0.017). The lowest protein content was found in L. longipalpis - 0.18 μg/gland (SD 0.038).
The dot method on gels with copolymerized
Discussion
The pharmacological complexity of sand fly saliva was well-demonstrated by various authors in two sand flies, Lutzomyia longipalpis and P. papatasi, belonging to the New-World and Old-World species, respectively. Although considerable differences have been found in saliva composition of various species (Volf et al., 2000, Valenzuela et al., 2001, Ribeiro and Modi, 2001, Volf and Rohoušová, 2001), hyaluronidase activity is present in all species studied. This supports the hypothesis about the
Acknowledgements
We thank Vera Volfova for excellent dissection of salivary glands. The work was supported by the grants GUK 137/2001, GACR 310/00/0760 and J13/981131-B4.
References (29)
- et al.
A microtiter-based assay for hyaluronidase activity not requiring specialized reagents
Analytical Biochemistry
(1997) - et al.
Improved methodology for analysis and quantitation of proteins on one-dimensional silver-stained slab gels
Analytical Biochemistry
(1983) - et al.
Isolation of maxadilan, a potent vasodilatory peptide from the salivary glands of the sand fly Lutzomyia longipalpis
Journal of Biological Chemistry
(1991) - et al.
Hyaluronan fragments induce nitric-oxide synthase in murine macrophages through a nuclear factor kappa B-dependent mechanism
Journal of Biological Chemistry
(1997) - et al.
Blood-finding strategy of a capillary-feeding sandfly, Lutzomyia longipalpis
Comparative Biochemistry and Physiology
(1986) Salivary gland lysate from the sand fly Lutzomyia longipalpis suppresses the immune response of mice to sheep red blood cells in vivo and concanavalin A in vitro
Experimental Parasitology
(1998)- et al.
Pharmacology of haematophagous arthropod saliva
- et al.
Toward an understanding of the biochemical and pharmacological complexity of the saliva of a hematophagous sand fly Lutzomyia longipalpis
Proceedings of the National Academy of Sciences of the United States of America
(1999) - et al.
Biochemical maturation of Spam1 (PH-20) during epididymal transit of mouse sperm involves modifications of N-linked oligosaccharides
Molecular Reproduction and Development
(1999) - et al.
The immunomodulatory factors of bloodfeeding arthropod saliva
Parasite Immunology
(2000)
Bee venom hyaluronidase is homologous to a membrane protein of mammalian sperm
Proceedings of the National Academy of Sciences of the United States of America
Sand fly vector saliva selectively modulates macrophage functions that inhibit killing of Leishmania major and nitric oxide production
Journal of Immunology
Preliminary characterization of the multiple forms of ram sperm hyaluronidase
Biochemical Journal
Adenosine, AMP and protein phosphatase activity in sandfly saliva
American Journal of Tropical Medicine and Hygiene
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