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Characterization of a recombinant T cell and B cell reactive polypeptide of Onchocerca volvulus
F Seeber1, N Brattig, P T Soboslay
1Institut für Tropenhygiene, University of Heidelberg, Germany.
Abstract:
To identify potentially protective Ag of the filarial nematode Onchocerca volvulus on the molecular level we screened a cDNA library of O. volvulus with a human serum raised against radiation-attenuated infective larvae of O. volvulus. A cDNA clone of 218 bp (OvL3-1) was selected for further studies. It was expressed in Escherichia coli and affinity purified recombinant polypeptide was tested for its ability to stimulate in vitro PBMC from African onchocerciasis patients and PBMC from chimpanzees experimentally infected with O. volvulus. An enhanced cell proliferation by PBMC was observed in many patients after stimulation with the recombinant OvL3-1 polypeptide. In addition, some patients' PBMC responded to OvL3-1 stimulation with enhanced IL-2 production. Infected chimpanzees also showed an increase in T cell proliferation. Onchocerciasis patients had variable levels of specific antibodies directed to the recombinant polypeptide when sera were tested by ELISA. A mAb directed against the recombinant protein located the native target Ag in the muscles of the adult worm. The molecular mass of native OvL3-1 was found to be 50 kDa on immunoblots. Polymerase chain reaction analysis of RNA from different life stages of the parasite showed that OvL3-1 is transcribed in all parasite stages within the mammalian host. A homologous gene is also present in other filarial parasites. The protein corresponding to OvL3-1, therefore, represents an immunogen present during the whole life-span of the parasite, and because of its B and T cell stimulatory properties, it may be a candidate for a protective Ag in human filariasis.
Insights
Researchers identified a potential protective antigen (OvL3-1) from the filarial worm Onchocerca volvulus. This antigen stimulates immune cells and may be a candidate for developing vaccines against human filariasis.
Area of Science:
- Parasitology
- Immunology
- Molecular Biology
Background:
- Onchocerca volvulus causes onchocerciasis, a significant human filarial disease.
- Identifying protective antigens is crucial for vaccine development against filariasis.
Purpose of the Study:
- To identify and characterize potentially protective antigens of Onchocerca volvulus at the molecular level.
- To evaluate the immunogenicity of a candidate antigen (OvL3-1) in patients and experimental models.
Main Methods:
- Screening of an O. volvulus cDNA library with human serum.
- Expression and purification of recombinant OvL3-1 polypeptide.
- In vitro stimulation of peripheral blood mononuclear cells (PBMCs) and antibody testing (ELISA).
- Immunoblotting and PCR analysis for antigen localization and expression.
Main Results:
- A cDNA clone, OvL3-1, was identified and expressed as a recombinant polypeptide.
- Recombinant OvL3-1 stimulated significant T cell proliferation and IL-2 production in onchocerciasis patients' PBMCs.
- Experimental infection in chimpanzees showed increased T cell proliferation upon stimulation.
- The native OvL3-1 antigen (50 kDa) is located in adult worm muscles and transcribed across all parasite life stages.
- Homologous genes exist in other filarial parasites.
Conclusions:
- OvL3-1 is an immunogenic antigen present throughout the O. volvulus life cycle.
- Its ability to stimulate both B and T cells makes it a promising candidate for a protective antigen in human filariasis vaccine development.