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C-reactive protein (CRP) is an acute phase serum protein secreted by liver hepatocytes. Besides its presence in the serum, CRP was also found on the surface of human leukocytes. However, the binding ability of CRP to rabbit leukocytes has never been previously investigated. The objective of our study was to optimize the detection of rabbit CRP binding leukocytes in order to observe the acute phase of immune response by flow cytometry after Complete Freund´s Adjuvant (CFA) administration. Blood samples were analysed using ELISA assay and flow cytometry immediately before and 2 days after the CFA administration. Significant (P<0.01) increase in the proportion of CRP+ leukocytes (up to 10%) and in their subsets (lymphocytes up to 14% and granulocytes up to 8%) were observed in samples after CFA immunization. ELISA also revealed significantly (P<0.01) higher CRP concentration in rabbit blood plasma after CFA immunization (about 3 mg/L) compared to control samples (about 1.5 mg/L) before immunization. In conclusion, the increase in level of CRP protein during the immune response in rabbits could be measured beside the ELISA also using flow cytometry via CRP binding leukocytes. This novel assay could be therefore successfully applied in further biomedical and veterinary research.

Novel assay for the detection of CRP protein in rabbit leukocytes using flow cytometry