Skip to main content
Log in

Litomosoides carinii: extraction of the microfilarial sheath components and antigenicity of the sheath fractions

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

Abstract

Microfilarial sheaths ofLitomosoides carinii were isolated and extracted with 2% sodium dodecyl sulfate (SDS) and 5% 2-mercaptoethanol (2ME). Extraction with SDS alone did not alter the ultrastructure of the sheaths and yielded five polypeptides (27–67 kDa) that were not recognized by antibodies of infected hosts but reacted with antibodies to host-serum proteins. 2ME treatment caused partial solubilization of the sheaths (45% as determined by amino acid analysis), which could be further improved by combining 2ME with SDS. The remainder showed filamentous/threadlike structures on electron microscopic examination. As compared with whole sheaths, the insoluble proportion was markedly enriched in alanine and cysteine but contained less galactosamine, serine, and threonine. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) of 2ME/SDS-extractable components showed 12–16 bands of 14>120 kDa. A predominant component had an apparent molecular mass of 22 kDa. Two bands (42 and 120 kDa) could be stained with Coomassie blue but showed “negative” staining when gels were stained with silver. Several components (but not the 22-kDa polypeptide) bore phosphocholine epitopes. Apart from the negatively staining bands, most of the 2ME-soluble sheath components were recognized by antibodies ofL. carinii-infectedMastomys coucha. Except for several polypeptides that had been unspecifically recognized by IgM, the antibody response to sheath components started at the end of the prepatent period.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bardehle G, Klonisch T, Schott H-H, Stirm S, Zahner H (1987) Isolation of pure sheaths ofLitomosoides carinii microfilariae. Parasitol Res 74:188–190

    Google Scholar 

  • Bardehle G, Jepp-Libutzki A, Linder D, Moehnle K, Schott H-H, Zahner H, Zähringer U, Stirm S (1992) Chemical composition ofLitomosoides earinii microfilarial sheaths. Acta Trop (Basel) 50:237–247

    Google Scholar 

  • Fuhrman JA (1990) Biochemistry of microfilarial exsheathment. Exp Parasitol 70:363–366

    Google Scholar 

  • Fujimoto D, Kanaya S (1973) A non-collagen structural protein fromAscaris cuticle. Arch Biochem Biophys 157:1–6

    Google Scholar 

  • Geyer E, Zahner H (1987) Occurrence of albumin and immunoglobulins on the sheath of blood microfilariae ofLitomosoides carinii. Trop Med Parasitol 38:66

    Google Scholar 

  • Hancock K, Tsang VCV (1983) India ink staining of proteins on nitrocellulose paper. Anal Biochem 133:157–162

    Google Scholar 

  • Heinrikson RL, Meredith SC (1986) Amino acid analysis by reverse-phase high-performance liquid chromatography: precolumn derivatization with phenylisothiocyanate. Anal Biochem 136:65–74

    Google Scholar 

  • Klonisch T, Bardehle G, Linder G, Boschek B, Schott H-H, Zahner H, Stirm S (1991) The sheath ofBrugia microfilariae: isolation and composition. Parasitol Res 77:448–451

    Google Scholar 

  • Lämmler G, Saupe E, Herzog H (1968) Intektionsversuche mit der BaumwollrattenfilarieLitomosoides carinii beiMastomys natalensis. Z Parasitenkd 30:281–290

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Lal RB, Ottesen EA (1989) Antibodies to phosphocholine-bearing antigens in lymphatic filariasis and changes following treatment with diethylcarbamazine. Clin Exp Immunol 175:52–57

    Google Scholar 

  • Lal RB, Paranjape RS, Briles DE, Nutman TB, Ottesen EA (1987) Circulating parasite antigen(s) in lymphatic filariasis: use of monoclonal antibodies to phosphocholine for immunodiagnosis. J Immunol 138:3454–3460

    Google Scholar 

  • Lal RB, Kumaraswami V, Steel C, Nutman TB (1990) Phosphocholine-containing antigens ofbrugia malayi nonspecifically suppress lymphocyte function. Am J Trop Med Hyg 42:56–64

    Google Scholar 

  • Lawson DA, Wenk P, Storey DM (1989) Identification of a potential protective microfilariae antigen fromLitomosoides carinii (Nematoda, Filarioidea). Trop Med Parasitol 40:42–46

    Google Scholar 

  • Maizels RM, Burke J, Denham DA (1987) Phosphorylcholinebearing antigens in filarial nematode parasites: analysis of somatic extracts, in vitro secretions and infection sera fromBrugia malayi andB. pahangi. Parasite Immunol 9:49–66

    Google Scholar 

  • McGreevy PB, Ratiwayanto S, Tuti S, McGreevy MM, Dennis DT (1980) Relationship between anti-sheath antibodies and microfilariae in natives living in an endemic area of South Kalimantan, Borneo. Am J Trop Med Hyg 29:553–562

    Google Scholar 

  • Philipp M, Worms MJ, McLaren DJ, Ogilvie BM, Park House RM, Taylor PM (1984) Surface proteins of a filarial nematode: a major soluble antigen and a host component on the cuticle ofLitomosoides carinii. Parasite Immunol 6:63–82

    Google Scholar 

  • Pinder M, Dupont A, Egwang TG (1988) Identification of a surface antigen ofLoa loa microfilariae the recognition of which correlates with the amicrofilaremic state in man. J Immunol 141:2480–2486

    Google Scholar 

  • Schägger H, Jagow G von (1987) Tricine-sodium dodecyl sulfatepolyacrylamide gel electrophoresis for the separation of proteins in the range of 1 to 100 kDa. Anal Biochem 166:368–379

    Google Scholar 

  • Schares G (1992) Studien zur Charakterisierung von Immunglobulinen derMastomys coucha und zur Beeinflussung der humoralen Immunreaktion des Versuchstiers auf eine Infektion mitLitomosoides carinii durch filarizide Behandlungen. Dissertation, University of Gießen

  • Schraermeyer U, Peters W, Zahner H (1987) Formation by the uterus of a peripheral layer of the sheath in microfilariae ofLitomosoides carinii andBrugia malayi. Parasitol Res 73:557–564

    Google Scholar 

  • Towbin H, Staekelin 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 

Download references

Author information

Authors and Affiliations

Authors

Additional information

This work was supported by the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 272)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bardehle, G., Hintz, M., Linder, D. et al. Litomosoides carinii: extraction of the microfilarial sheath components and antigenicity of the sheath fractions. Parasitol Res 78, 501–508 (1992). https://doi.org/10.1007/BF00931571

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00931571

Keywords

Navigation