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Immunological and Functional Studies on a Campylobacter Jejuni Invasion Antigen:
Peb1a
I.
Background/Objective
Campylobacter jejuni is a common cause of bacterial gastroenteritis that often
results from consuming undercooked poultry. Data suggest that the
Campylobacter Peb1a protein is utilized as an adhesin allowing attachment to
human intestinal epithelial cells leading to invasion and pathogenesis. Little is
known about the function of the Peb1a gene or its expression amongst C. jejuni
strains, not to mention it’s definitive role in bacterial invasion of intestinal cells. To
address these issues the production of monoclonal antibodies against the
antigen is our main priority in order to further analyze the antigen naturally found
on C. Jejuni
II.
Methods
The Peb1a gene was cloned, expressed in E. coli, and purified using epitope
affinity capture. Mice were immunized with Peb1a, and the sera collected were
tested for reactivity against different C. jejuni strains. Mouse serum analyses
revealed positive reactivity for all strains, with each individual strain presenting
different quantities of expressed Peb1a. Cell fusion techniques were then
employed to generate hybridoma clones from the fusion of the immunized mouse
B-cells and myeloma cells. Resultant monoclonal antibody producing clones are
being employed for immunological assays of the Peb1a antigen, and its putative
role in bacterial invasion of human cells by C. jejuni via tissue culture
methodologies.
III.
Results
Based off of ELISA, Western Blots/PAGE and other immunoassay data produced from
reacting mouse serum and hybridoma supernatants against Peb1a and C. Jejuni strains,
the data suggests that monoclonal antibodies were produced against the peb1a antigen
found on C. Jejuni.
IV.
Discussion/Conclusion
Because monoclonal antibodies was produced against Peb1A, we can now utilize this
antibody for further downstream analysis of the function of the peb1A antigen found on
C. Jejuni and further study its role in adhesion and invasion.