Abstract
In vitro yeast phagocytosis by haemocytes of the compound ascidian Botryllus schlosseri was studied, with particular attention to interactions among different immunocyte types. It is demonstrated that the supernatant from haemocyte cultures matched with yeast cells contains factor(s) able to enhance yeast ingestion by Botryllus phagocytes. The increase in phagocytosis is not the consequence of yeast opsonisation, as the phagocytic index does not significantly increase when yeast cells, previously incubated in the culture media, are washed and re-suspended in filtered sea water. When haemocytes were fractionated by density gradient centrifugation and each band was incubated with yeast, the ability to stimulate phagocytosis was found in the supernatants from haemocyte cultures of fractions rich in morula cells (MC). Previous studies have demonstrated that MC express molecules recognised by anti-cytokine antibodies, as a consequence of the recognition of foreign molecules or cells. Our results indicate that molecules immunoreactive with anti-cytokine antibodies are required for modulating phagocyte activity, as the above-reported enhancing effect is completely absent in the presence of anti-IL-1-α and anti-TNF-α, but not of anti-rabbit-IgG antibodies, and they also highlight the presence of ‘cross-talk‘ between MC and phagocytes. A new scenario is therefore sketched, in which MC actively recognise non-self molecular patterns and, upon this recognition, release humoural factor(s) recognised by phagocytes, which modulate phagocytosis.
Similar content being viewed by others
References
Abbas AK, Lichtman AH, Pober JS (2000) Cellular and molecular immunology. WB Saunders Company, Philadelphia
Azumi K, Ishimoto R, Fujita T, Nonaka M, Yokosawa H (2000) Opsonin-independent and—dependent phagocytosis in the ascidian Halocynthia roretzi: galactose-specific lectin and complement C3 functions as target-dependent opsonins. Zool Sci 17:625–632
Azumi K, Kuribayashi F, Kanegasaki S, Yokosawa H (2002) Zymosan induces production of superoxide anions by hemocytes of the solitary ascidian Halocynthia roretzi. Comp Biochem Physiol 133C:567–574
Azumi K, De Santis R, De Tomaso A, Rigoutsos I, Yoshizaki F, Pinto MR, Marino R, Shida K, Ikeda M, Ikeda M, Arai M, Inoue Y, Shimizu T, Satoh N, Rokhsar DS, Du Pasquier L, Kasahara M, Satake M, Nonaka M (2003) Genomic analysis of immunity in a urochordate and the emergence of the vertebrate immune system: “waiting for Godot”. Immunogenetics 55:570–581
Ballarin L, Cima F (2005) Cytochemical properties of Botryllus schlosseri haemocytes: indications for morpho-functional characterisation. Eur J Histochem 49 (in press)
Ballarin L, Cima F, Sabbadin A (1993) Histoenzymatic staining and characterization of the colonial ascidian Botryllus schlosseri hemocytes. Boll Zool 60:19–24
Ballarin L, Cima F, Sabbadin A (1994) Phagocytosis in the colonial ascidian Botryllus schlosseri. Dev Comp Immunol 18:467–481
Ballarin L, Cima F, Sabbadin A (1995) Morula cells and histocompatibility in the colonial ascidian Botryllus schlosseri. Zool Sci 12:757–764
Ballarin L, Cima F, Sabbadin A (1998) Phenoloxidase and cytotoxicity in the compound ascidian Botryllus schlosseri. Dev Comp Immunol 22:479–492
Ballarin L, Tonello C, Guidolin L, Sabbadin A (1999) Purification and characterization of a humoral opsonin, with specificity for d-galactose, in the colonial ascidian Botryllus schlosseri. Comp Biochem Physiol 123B:115–123
Ballarin L, Tonello C, Sabbadin A (2000) Humoral opsonin from the colonial ascidian Botryllus schlosseri as a member of the galectin family. Mar Biol 136:813–822
Ballarin L, Franchini A, Ottaviani E, Sabbadin A (2001) Morula cells as the main immunomodulatory haemocytes in ascidians: evidences from the colonial species Botryllus schlosseri. Biol Bull 201:59–64
Ballarin L, Scanferla M, Cima F, Sabbadin A (2002a) Phagocyte spreading and phagocytosis in the compound ascidian Botryllus schlosseri: evidence for an integrin-like, RGD-dependent recognition mechanism. Dev Comp Immunol 26:39–48
Ballarin L, Cima F, Floreani M, Sabbadin A (2002b) Oxidative stress induces cytotoxicity during rejection reaction in the compound ascidian Botryllus schlosseri. Comp Biochem Physiol 133C:411–418
Ballarin L, Menin A, Franchi N, Bertoloni G, Cima F (2005) Morula cells and non-self recognition in the compound ascidian Botryllus schlosseri. Invertebr Surv J 2:1–5
Beck G (1998) Macrokines: invertebrate cytokine-like molecules? Front Biosci 3:D559–D569
Beck G, Vasta GR, Marchalonis JJ, Habicht GS (1989) Characterization of interleukin-1 activity in tunicates. Comp Biochem Physiol 92B:93–98
Beck G, O’Brien RF, Habicht GS, Stillman DL, Cooper EL, Raftos DA (1993) Invertebrate cytokines III: invertebrate interleukin-1-like molecules stimulate phagocytosis by tunicate and echinoderm cells. Cell Immunol 146:284–299
Beck G, Cardinale S, Wang L, Reiner M, Sugumaran M (1996) Characterization of a defense complex consisting of interleukin I and phenol oxidase from the hemolymph of the tobacco hornworm, Manduca sexta. J Biol Chem 271:11035–11038
Carballal MJ, Lopez C, Azevedo C, Villalba A (1997) Enzymes involved in defense functions of hemocytes of mussel Mytilus galloprovincialis. J Invertebr Pathol 70:96–105
Cima F, Ballarin L, Sabbadin A (1996) New data on phagocytes and phagocytosis in the compound ascidian Botryllus schlosseri (Tunicata: Ascidiacea). Ital J Zool 63:357–364
Cima F, Matozzo V, Marin MG, Ballarin L (2000) Haemocytes of the clam Tapes philippinarum: morphofunctional characterisation. Fish Shellfish Immunol 10:677–693
Cima F, Perin A, Burighel P, Ballarin L (2001) Morpho-functional characterisation of haemocytes of the compound ascidian Botrylloides leachi (Tunicata, Ascidiacea). Acta Zool 82:261–274
Cima F, Basso G, Ballarin L (2003) Apoptosis and phosphatidylserine-mediated recognition during the take-over phase of the colonial life-cycle in the ascidian Botryllus schlosseri. Cell Tissue Res 312:369–376
Cima F, Sabbadin A, Ballarin L (2004) Cellular aspects of allorecognition in the compound ascidian Botryllus schlosseri. Dev Comp Immunol 28:881–889
Clow LA, Raftos DA, Gross PS, Courtney Smith L (2004) The sea urchin complement homologue, SpC3, functions as an opsonin. J Exp Biol 207:2147–2155
Coombe DR, Ey PL, Jenkin CR (1984) Particle recognition by haemocytes from the colonial ascidian Botrylloides leachi: evidence that the B. leachi HA-2 is opsonic. J Comp Physiol Bull 154:509–521
Fryer SE, Hull CJ, Bayne CJ (1989) Phagocytosis of yeast by Biomphalaria glabrata: carbohydrate specificity of hemocyte receptors and a plasma opsonin. Dev Comp Immunol 13:9–16
Janeway CA, Medzhitov R (2002) Innate immune recognition. Annu Rev Immunol 20:197–216
Kelly KL, Cooper EL, Raftos DA (1992) A humoral opsonin from the solitary Urochordate Styela clava. Comp Biochem Physiol 103B:749–753
Kelly KL, Cooper EL, Raftos DA (1993) Cytokine-like activity of a humoral opsonin from the solitary urochordate Styela clava . Zool Sci 10:57–64
Lopez C, Carballal MJ, Azevedo C, Villalba A (1997) Morphological characterization of the hemocytes of the clam, Ruditapes decussatus (Mollusca: Bivalvia). J Invertebr Pathol 69:51–57
Michibata H, Hirata J, Uesaka M, Namakunai T (1987) Separation of vanadocytes: determination and characterization of vanadium ions in the separated blood cells of the ascidian, Ascidia ahodori. J Exp Zool 24:33–38
Nonaka M, Azumi K (1999) Opsonic complement system of the solitary ascidian, Halocynthia roretzi. Dev Comp Immunol 23:421–427
Nonaka M, Azumi K, Ji X, Namikawa-Yamada C, Sasaki M, Saiga H, Dodds AW, Sekine H, Homma MK, Matsushita M, Endo Y, Fujita T (1999) Opsonic complement component C3 in the solitary ascidian, Halocynthia roretzi. J Immunol 162:387–391
Ohtake S, Abe T, Shishikura F, Tanaka K (1994) The phagocytes in hemolymph of Halocynthia roretzi and their phagocytic activity. Zool Sci 11:681–691
Raftos DA, Cooper EL, Habicht GS, Beck G (1991) Invertebrate cytokines: tunicate cell proliferation stimulated by an interleukin 1-like molecule. Proc Natl Acad Sci USA 88:9518–9522
Raftos DA, Cooper EL, Stillman DL, Habicht GS, Beck G (1992) Invertebrate cytokines II: release of interleukin-1-like molecules from tunicate hemocytes stimulated with zymosan. Lymphokine Cytokine Res 11:235–240
Raftos DA, Stillman DL, Cooper EL (1998) Chemotactic responses of tunicate (Urochordata, Ascidiacea) hemocytes in vitro. J Invertebr Pathol 72:44–49
Sekine H, Kenjo A, Azumi K, Ohi G, Takahashi M, Kasukawa R, Ichikawa N, Nakata M, Mizuochi T, Matsushita M, Endo Y, Fujita T (2001) An ancient lectin-dependent complement system in an ascidian: novel lectin isolated from the plasma of the solitary ascidian, Halocynthia roretzi. J Immunol 167:4504–4510
Sminia T, Van der Knaap WPW, Edelenbosch P (1979) The role of serum factors in phagocytosis of foreign particles by blood cells of the freshwater snail Lymnaea stagnalis. Dev Comp Immunol 3:37–44
Smith VJ, Peddie CM (1992) Cell cooperation during host defense in the solitary tunicate Ciona intestinalis (L.). Biol Bull 183:211–219
Takahashi H, Azumi K, Yokosawa H (1995) A novel membrane glycoprotein involved in ascidian hemocyte aggregation and phagocytosis. Eur J Biochem 233:778–783
Tripp MR (1992) Phagocytosis by hemocytes of the hard clam, Mercenaria mercenaria. J Invertebr Pathol 59:222–227
Wiesner A, Wittwer D, Götz P (1996) A small phagocytosis stimulating factor is released by and acts on phagocytosing Galleria mellonella haemocytes in vitro. J Insect Physiol 42:829–835
Acknowledgements
This work was supported by the Italian M.I.U.R. (PRIN 2004).
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by R. Cattaneo-Vietti, Genova
Rights and permissions
About this article
Cite this article
Menin, A., Favero, M.d., Cima, F. et al. Release of phagocytosis-stimulating factor(s) by morula cells in a colonial ascidian. Marine Biology 148, 225–230 (2005). https://doi.org/10.1007/s00227-005-0081-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00227-005-0081-7