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Isolation and analysis of nephritic-producing immune complexes in Plasmodium berghei-infected mice
Abstract:
A nephritic condition was developed by infecting Swiss Webster albino mice with the malarial parasite Plasmodium berghei NK 65. These animals were tested for urinary protein and the presence of circulating immune complexes using reagent strips and a polyethylene glycol (PEG) precipitation assay. The circulating immune complexes were isolated from the sera using both affinity chromatography and PEG precipitation and from the kidney by acid elution. The isolated complexes were dissociated into their individual components and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The components of the complexes were transferred to nitrocellulose sheets and probed for the presence of malarial antigens using a rabbit anti-P berghei antisera. The overall humoral response to the malarial parasite was evaluated using a radial immunodiffusion assay. The present study confirmed that the malarial-infected animals not only developed the nephritic condition (as evident by the high levels of proteinuria) but also, as indicated by the PEG assay, have the presence of high levels of circulating immune complexes in their serum. The apparent absence in the SDS gels of any abnormal protein bands followed by the inability of the Western blot to reveal any malarial antigens provides some of the strongest evidence to date that these malarial proteins are not directly involved in the circulating immune complexes believed to be responsible for producing this nephritic condition.
Insights
Malaria infection in mice caused kidney disease and circulating immune complexes. However, malarial proteins were not found in these complexes, suggesting they are not the direct cause of the nephritic condition.
Area of Science:
- Immunology
- Pathology
- Parasitology
Background:
- Malaria infection can lead to kidney disease.
- Circulating immune complexes are implicated in the pathogenesis of various kidney disorders.
Purpose of the Study:
- To investigate the role of malarial antigens in the formation of circulating immune complexes and subsequent nephritic condition in infected mice.
Main Methods:
- Induction of nephritic condition in mice via Plasmodium berghei NK 65 infection.
- Quantification of proteinuria and circulating immune complexes using reagent strips and polyethylene glycol (PEG) precipitation.
- Isolation and dissociation of immune complexes from serum and kidney.
- Analysis of complex components by SDS-PAGE and Western blot for malarial antigens.
- Evaluation of humoral response using radial immunodiffusion.
Main Results:
- Malarial infection led to significant proteinuria and high levels of circulating immune complexes.
- SDS-PAGE revealed no abnormal protein bands in the complexes.
- Western blot analysis failed to detect malarial antigens within the isolated complexes.
Conclusions:
- Malarial infection induces a nephritic condition associated with circulating immune complexes in mice.
- Malarial proteins are unlikely to be directly involved in the formation of these immune complexes.
- The exact nature and origin of the antigens forming these immune complexes require further investigation.