Summary
Five monoclonal antibodies (mABs) against surface antigens on circulating, glass-adherent hemocytes of the snail, Biomphalaria glabrata, were produced by somatic cell hybridization methods. Two mABs (IID2.6-Bg and IID4.8-Bg) are pan-hemocytic, reacting uniformly with epitopes shared by all adherent hemocytes. Determinants recognized by these mABs also are present in soluble form and appear to be associated with a hemoglobin-depleted ultracentrifuged fraction of snail hemolymph. Hybridoma-derived mABs IIC6.8-Bg and VB10.3-Bg recognize hemocyte surface epitopes expressed by only 50–60% of the adherent cell population. These mABs also are reactive with soluble hemolymph antigens but apparently recognize determinants which are different from the IID2.6-Bg and IID4.8-Bg reactive sites. Another antigenically distinct hemocyte subpopulation is recognized by mAB IID7.1-Bg. Epitopes that are reactive with this mAB differ from the previously described determinants by their asymmetrical distribution on the surface of positive cells and the absence of soluble antigenic components in hemolymph. Furthermore, unlike the other mABs, the prevalence of hemocytes staining with IID7.1-Bg antibodies differed between two strains of B. glabrata. Results of this study clearly demonstrate that circulating B. glabrata hemocytes, consisting of a single, predominant population of adherent cells, is composed of several distinct antigenic subpopulations based on the specific binding of anti-hemocyte mAB probes. Our successful application of hybridoma techniques to the study of molluscan hemocyte surface antigens underscores further the great potential usefulness of this method in analysing the molecular basis of hemocyte reactivity.
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References
Anderson RS, Good RA (1976) Opsonic involvement in phagocytosis by mollusk hemocytes. J Invertebr Pathol 27:57–64
Bayne CJ (1983) Molluscan immunobiology. In: Saleuddin ASM, Wilbur KM (eds) Biology of mollusca. Academic Press, New York, in press
Bayne CJ, Buckley PM, DeWan PC (1980) Macrophage-like hemocytes are cytotoxic for sporocysts of Schistosoma mansoni in vitro. J Parasitol 66:413–419
Cheng TC (1975) Functional morphology and biochemistry of molluscan phagocytes. Ann NY Acad Sci 266:343–379
Chorney MJ, Cheng TC (1980) Discrimination of self and non-self in invertebrates. Contemp Top Immunobiol 9:37–54
Damle NK, Hansen JA, Good RA, Gupta S (1981) Monoclonal antibody analysis of human T lymphocyte subpopulations exhibiting autologous mixed lymphocyte reactions. Proc Natl Acad Sci (USA) 78:5096–5098
Kearney JF, Radbruch A, Liesegang B, Rajwesky K (1979) A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol 123:1548–1550
Knaap WPW van der, Boerrigter-Barendsen LH, Hoeven DSP van den, Sminia T (1981) Immunocytochemical demonstration of a humoral defense factor in blood cells (amoebocytes) of the pond snail Lymnaea stagnalis. Cell Tissue Res 219:292–296
Köhler G, Milstein C (1975) Continuous culture of fused cells secreting antibodies of predefined specificity. Nature (London) 256:495–497
Kung PC, Goldstein G, Reinherz EL, Schlossman SF (1979) Monoclonal antibodies defining distinctive human T cell surface antigens. Science 206:347–349
Lackie AM (1980) Invertebrate immunity. Parasitology 80:393–412
Nossal GJV, Szenberg A, Ada GL, Austin CM (1964) Single cell studies on 19s antibody production. J Exp Med 119:485–502
Oi VT, Herzenberg LA (1980) Immunoglobulin-producing hybrid cell lines. In: Mishell BB, Shiigi SM (eds) Selected methods in cellular immunology. WH Freeman and Company, San Francisco, pp 351–372
Ortaldo JR, Sharrow SO, Timonen T, Herberman RB (1981) Determination of surface antigens on highly purified human NK cells by flow cytometry with monoclonal antibodies. J Immunol 127:2401–2409
Prowse RH, Tait NN (1969) In vitro phagocytosis by amoebocytes from the hemolymph of Helix aspersa (Muller) I. Evidence for opsonic factor(s) in serum. Immunology 17:437–443
Ratcliffe NA, Rowley AF (eds) (1981) Invertebrate blood cells. Vols. 1 and 2, Academic Press, London and New York, p 641
Reinherz EL, Kung PC, Pesando JM, Ritz J, Goldstein G, Schlossman SF (1979) Ia determinants on human T-cell subsets defined by monoclonal antibody. J Exp Med 150:1472–1482
Renwrantz LR, Cheng TC (1977) Identification of agglutinin receptors on hemocytes of Helix pomatia. J Invertebr Pathol 29:88–96
Richards CS (1975) Genetic factors in susceptibility of Biomphalaria glabrata for different strains of Schistosoma mansoni. Parasitology 70:231–241
Schoenberg DA, Cheng TC (1980) Lectin-binding specificities of hemocytes from two strains of Biomphalaria glabrata as determined by microhemadsorption assays. Dev Comp Immunol 4:617–628
Schoenberg DA, Cheng TC (1981) Lectin-binding specificities of Bulinus truncatus hemocytes as demonstrated by microhemadsorption. Dev Comp Immunol 4:145–149
Sminia T (1981) Gastropods. In: Ratcliffe NA, Rowley AF (eds) Invertebrate blood cells. Academic Press, London and New York, pp 191–232
Sminia T, Barendsen L (1980) A comparative morphological and enzyme histochemical study on blood cells of the freshwater snails Lymnaea stagnalis, Biomphalaria glabrata, and Bulinus truncatus. J Morphol 165:31–39
Sminia T, Knaap WPW van der, 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
Sminia T, Winsemius AA, Knaap WPW van der (1981) Recognition of foreignness by blood cells of the freshwater snail Lymnaea stagnalis with special reference to the role and structure of the cell coat. J Invertebr Pathol 38:175–183
Ugolini V, Nunez G, Smith RG, Stastny P, Capra JD (1980) Initial characterization of monoclonal antibodies against human monocytes. Proc Natl Acad Sci (USA) 77:6764–6768
Yoshino TP (1981) Comparison of concanavalin A-reactive determinants on hemocytes of two Biomphalaria glabrata snail stocks: Receptor binding and redistribution. Dev Comp Immunol 5:229–240
Yoshino TP (1983a) Surface antigens of Biomphalaria glabrata (Gastropoda) hemocytes: Occurrence of membrane-associated hemolymph-like factors antigenically related to snail hemoglobin. J Invertebr Pathol 41:310–320
Yoshino TP (1983b) Lectins and antibodies as molecular probes of molluscan hemocyte surface membranes. Dev Comp Immunol, in press
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Yoshino, T.P., Granath, W.O. Identification of antigenically distinct hemocyte subpopulations in Biomphalaria glabrata (Gastropoda) using monoclonal antibodies to surface membrane markers. Cell Tissue Res. 232, 553–564 (1983). https://doi.org/10.1007/BF00216428
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DOI: https://doi.org/10.1007/BF00216428