Quantitation of complement and leukocyte binding to a parasitic helminth species

Journal of Immunological Methods
Paul R GiacominLindsay A Dent

Abstract

Methods used to quantify complement deposition and cell adherence to parasitic helminths usually involve subjective visual comparisons of immunofluorescence or time-consuming manual counting of bound cells. Such targets are relatively large and, generally, few individual organisms can be analysed. More objective and efficient radiometric assays are available, but these also have considerable disadvantages. We have developed an improved immunofluorescence-based method for quantitation of complement deposition on viable third-stage larvae of the nematode Nippostrongylus brasiliensis (L3). A similar strategy was also applied to measuring leukocyte adherence to the parasite. Fluorescein isothiocyanate (FITC)-conjugated antibodies were used to detect complement on serum-treated larvae. The adherence of carboxyfluorescein diacetate succinimidyl ester (CFSE)-labelled mouse leukocytes to larvae was investigated using the same basic approach. Images of fluorescent larvae or fluorescent cells attached to larvae were generated with a Bio-Rad Molecular Imager FX and fluorescence intensity was quantified. Hundreds of larvae can be analysed simultaneously in multiple samples, and these strategies allow rapid and sensitive quantitation that i...Continue Reading

References

Apr 1, 1977·Experimental Parasitology·R Leventhal, E J Soulsby
Oct 1, 1979·The Australian Journal of Experimental Biology and Medical Science·S J ProwseC R Jenkin
Nov 1, 1990·The Journal of Experimental Medicine·L A DentC J Sanderson
Nov 1, 1985·Parasite Immunology·R ChandrashekarD Subrahmanyam
Jun 27, 2002·The Journal of Infectious Diseases·Taru MeriSeppo Meri

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Citations

Jun 17, 2009·Infection and Immunity·Karita HaapasaloT Sakari Jokiranta
Mar 21, 2007·The Journal of Immunology : Official Journal of the American Association of Immunologists·Ljubov SimsonChristopher R Parish

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