Identification of B cell antigenome in Mycobacterium bovis by immunoproteomic analysis

Bovine tuberculosis (bTB) is a common zoonosis prevalent in many countries with grave economic consequences. Most developed and developing countries have implemented the test-and-slaughter policy to protect public health and reduce economic losses in the cattle industry. The official diagnosis of bTB is based on assays dependent on cell-mediated immunity (CMI). CMI-based diagnosis demonstrates diagnostic incapability at late stages of infection, which could be overcome by diagnosis based on humoral immunity (HI). Therefore, there is an urgent need to identify and define the B cell antigenome of Mycobacterium bovis. In this study, the B cell antigenome of culture filtrate proteins (CFP) was defined by mass spectrometry-based proteomics technology. Four spots were detected on 2-dimensional gel electrophoresis (2-DE) against M. bovis-positive serum in an immunoblotting experiment. Twenty-one proteins were identified in four spots by proteomic tools, such as Mb2900, Mb2898, Mb0448, Mb3834c, Mb1918c, Mb0134c, Mb0358 and Mb1868c, which are known B cell antigens, including 13 new proteins, i.e. Mb3751, Mb2006c, Mb3276c, Mb2244, Mb1164c, Mb2553c, Mb2946c, Mb1849c, Mb1511c, Mb1034c, Mb2616c, Mb0854c and Mb2267. These new proteins identified by 2-DE and immunoblotting were the B cell antigens used in developing serological diagnostic methods based on HI to bTB.