Skip to main content
SpringerLink
Log in

Proteins ofNippostrongylus brasiliensis analyzed by immunoblotting

  • Original Investigations
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

Antigenic proteins were characterized by the immunoblotting technique with sera from rats and mice after infection as well as hyperimmune sera. The immune response of infected animals was mainly directed toward five proteins of adult worms (190, 118, 110, 98, and 52 kDa) and four proteins of the third larval stage (L3; 92, 78, 58, and 24 kDa). The immunoblots indicated that stage-specific proteins of the homogeneates were recognized. Three stage-specific proteins of L3 larvae (150, 135, and 125 kDa) and three proteins typical to the adult worm (100, 82, and 67 kDa) were identified. The majority of the worm proteins elicited an IgG response. IgE synthesis was induced by living and dead parasites and was directed towards four proteins (190, 150, 125, and 98 kDa). Three proteins additionally induced an IgG or IgM antibody response. The immune response as shown by the immunoblotting technique seems to be directed towards (1) antigens that are present for the duration of an infection and (2) stage-specific antigens that are expressed for only a short time during the life cycle of the parasite.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

EDTA:

ethylenediaminetetraacetate

IMS:

sera from infected mice

IRS:

sera from infected rats

kDa:

kilodalton

L3:

larvae, third-stage larvac

MS imm. L3:

sera from mice immunized with dead L3 larvae

MS imm. W:

sera from mice immunized with dead adult worms

PBS:

phosphate-buffered saline

PCMB:

p-chloromercuribenzoate

PMSF:

phenylmethylsulfonylfuoride

RMS:

sera from reinfected mice

RRS:

sera from reinfected rats

TBS:

TRIS-buffered saline

TLCK:

p-tosyl-l-lysine-chloromethylketone

TPCK:

l-1-tosylamido-2-phenylethyl-chloromethylketone

TRIS:

tris(hydroxymethyl)-aminomethane

SDS:

sodium dodecyl sulfate

References

  • Ambler J, Orr TSC (1972) Studies on an allergenic component extracted fromNippostrongylus brasiliensis. Immunochemistry 9:263–272

    Google Scholar 

  • Befus AD, Bienenstock J (1982) Factors involved in symbiosis and host resistance at the mucus parasite interface. Prog Allergy 31:76–177

    Google Scholar 

  • Bohn A, König W (1982) Generation of monoclonal murine anti-DNP IgE, IgM and IgG1 antibodies: biochemical and biological characterization. Immunology 47:297–311

    Google Scholar 

  • Bohn A, König W (1985) Characterization of antigens of the nematodeNippostrongylus brasiliensis by monoclonal antibodies. Z Parasitenkd 71:663–672

    Google Scholar 

  • Bohn A, König W (1986) Generation of monoclonal antibodies to characterize antigens of the nematodeNippostrongylus brasiliensis. Zentralbl Bakteriol Parasitenkd Infektionskr Hyg Abt Orig Reihe A 261:119–132

    Google Scholar 

  • Bust JS, Ogilvie BM (1975) In vitro maintenance of nematode parasites assayed by acteylcholinesterase and allergen secretions. Exp Parasitol 38:75–82

    Google Scholar 

  • Capron A, Dessaint JP (1985) Effector and regulatory mechanisms in immunity to schistosomes: a heuristic view. Ann Rev Immunol 3:455–476

    Google Scholar 

  • Capron A, Dessaint J-P, Capron M, AmeisenJC, Tonnel AB (1986) From parasites of allergy: a second receptor for IgE. Immunol Today 7:15–18

    Google Scholar 

  • Clegg JA, Smith MA (1978) Prospects for the development of dead vaccines against helminths. Adv Parasitol 16:165–218

    Google Scholar 

  • Conrad DH, Keegan A, Rao M, Lee WT (1987a) Synthesis and regulation of the IgE receptor on B lymphocyte cell lines. Int Arch Allergy Appl Immunol 82:402–404

    Google Scholar 

  • Conrad DH, Waldschmidt TJ, Lee WT, Rao M, Keegan AD, Noelle RJ, Lynch RG, Kehry MR (1987b) Effect of B cell stimulatory factor-1 (interleukin 4) on Fcε and Fcγ receptor expression on murine B lymphocytes and B cell lines. J Immunol 139:2290–2296

    Google Scholar 

  • Engvall E (1980) Enzyme immunoassay: ELISA and EMIT. Methods Enzymol 70:419–439

    Google Scholar 

  • Finkelman FD, Ohra J, Goroff DK, Smith J, Villacreses N, Mond JJ, Paul WE (1986) Production of BFS-1 during an in vitro, T-dependent immune response. J Immunol 137:2878–2885

    Google Scholar 

  • Finkelman FD, Urban JF, Ohara J, Snapper CM, Mountz JD, Paul WE, Katona JM (1987a) Requirement for BSF-1 during in vivo IgE production. Fed Proc 46:1200–1206

    Google Scholar 

  • Finkelman FD, Snapper CM, Mountz JD, Katona IM (1987b) Polyclonal activation of the murine immune system by a goat antibody to mouse IgD: IX. Induction of a polyclonal IgE response. J Immunol 138:2826–2830

    Google Scholar 

  • Grzych J-M, Capron M, Dissous C, Capron A (1984) Blocking activity of rat monoclonal antibodies in experimental schistosomiasis. J Immunol 133:998–1004

    Google Scholar 

  • Haque A, Lefebvre MN, Ogilvie BM, Capron A (1978a)Dipetalonema viteae in hamsters: effect of antisera to immunization with parasite extracts on production of microfilariae. Parasitology 76:61–76

    Google Scholar 

  • Haque A, Chassoux D, Ogilvie BM, Capron A (1978b)Dipetalonema viteae infection in hamsters: enhancement and suppression of microfilaraemia. Parasitology 76:77–84

    Google Scholar 

  • Hofstetter M, Pointdexter RW, Ruiz-Tizben E, Ottesen E (1982) Modulation of the host response in human schistosomiasis: III. Blocking antibodies specifically inhibit immediate hypersensitivity responses to parasite antigens. Immunology 46:777–785

    Google Scholar 

  • Hogarth-Scott RS (1968) Naturally occuring antibodies to the cuticle of nematodes. Parasitology 58:221–226

    Google Scholar 

  • Hussain R, Ottesen EA (1985) IgE response in human filariasis: III. Specificities of IgE and IgG antibodies compared by immunoblotting analysis. J Immunol 135:1415–1520

    Google Scholar 

  • Hussian R, Bradbury SM, Strejan G (1973) Hypersensitivity ofAscaris antigens: VIII. Characterization of highly purified allergen. J Immunol 111:260–268

    Google Scholar 

  • Khalife J, Capron M, Capron M, Grzych J-M, Butterworth AE, Dunne DW, Ouma JH (1986) Immunity in human schistosomiasis mansoni. Regulation of protective immune mechanisms by IgM blocking antibodies. J Exp Med 164:1626–1640

    Google Scholar 

  • Maizels RM, Philipp M, Ogilvie BM (1982) Molecules on the surface of parasitic nematodes as probes of the immune response in infection. Immunol Rev 61:109–136

    Google Scholar 

  • Mogbel R, Miller HRP, Wäkelin D, Mac Donald AJ, Kay AB (1987) Leukotrienes and intestinal worms: Allergy and Inflammation Kay AB (ed) Acad Press, pp 367–387.

  • Ogilvie BM (1971)Nippostrongylus brasiliensis in mice: an explanation for the failure to induce worm expulsion from passively immunized animals. Int J Parasitol 1:161–167

    Google Scholar 

  • Ogilivie BM, Rothwell TL, Bremner KC, Schnitzerling HJ, Nolan J, Keith RK (1973) Acelylcholinesterase secretion by parasitic nematodes: I. Evidence for secretion of the enzyme by a number of species. Int J Parasitol 3:589–597

    Google Scholar 

  • Pery P, Petit A, Poulain J, Luffau G (1974) Phosphorylcholine bearing components in homogenates of nematodes. Eur I Immunol 4:637–639

    Google Scholar 

  • Pery P, Luffau G, Charley J, Petit A, Rouze P, Bernard S (1979) Phosphorylcholine antigens fromNippostrongylus brasiliensis: II. Isolation and partial characterization of phosphorylcholine antigens from adult worms. Ann Immunol (Paris) 130C:889–899

    Google Scholar 

  • Petit A, Pery P, Luffau G (1980) Purification of an allergen from culture fluids ofNippostrongylus brasiliensis. Mol Immunol 17:1341–1349

    Google Scholar 

  • Pfeiffer P, König W, Bohn A (1983) Genetic dependence of IgE antibody production in mice infected with the nematodeNippostrongylus brasiliensis. Int Arch Allergy Immunol 72:347–355

    Google Scholar 

  • Philipp M, Rumjaneck FD (1984) Antigenic and dynamic properties of helminth surface structures. Mol Biochem Parasitol 10:245–268

    Google Scholar 

  • Sanderson BE, Ogilvie BM (1971) A study of acetylcholinesterase throughout the life cycle ofNippostrongylus brasiliensis. Parasitology 62:367–373

    Google Scholar 

  • Tarleton RL, Kemp WM (1981) Demonstration of IgG-Fe and C3 receptors on adultSchistosoma mansoni. J Immunol 126:376–384

    Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

  • Wakelin D (1985) Genetic control of immunity to helminth infections. Parasitol Today 1:17–23

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Mediinische Mikrobiologie und Immunologie, Arbeitsgruppe für Infektabwehrmechanismen, Ruhr-Universität Bochum, D-4630, Bochum, Germany

    U. Dorzok, A. Bohn & W. König

Authors
  1. U. Dorzok
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Bohn
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. W. König
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supported by Deutsche Forschungsgemeinschaft Bo 740/1-2; the work by U. Dorzok comprises the partial fulfillment of a PhD thesis

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dorzok, U., Bohn, A. & König, W. Proteins ofNippostrongylus brasiliensis analyzed by immunoblotting. Parasitol Res 75, 482–487 (1989). https://doi.org/10.1007/BF00930977

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00930977

Keywords

Search

Navigation