Early diagnosis of MAP infection is a pressing need to enable efficient intervention with the spread of MAP infection in herds. Hence, study of lymphocyte subsets and their expressed adhesion molecules could contribute in defining a distinct diagnostic marker (or markers) at the subclinical period of the infection that could in turn facilitate the development of effective diagnostic approach. In accordance with this objective, milk and blood samples were collected from two groups of cattle naturally infected with MAP and their corresponding negative controls. Group (C) comprised 3-4 year-old ELISA negative/PCR positive-cattle that were considered as subclinical seronegative low shedder group (early stage). Group (A) included 6-8 year-old ELISA positive-cattle, which were considered as a clinical seropositive group (late stage). Flow cytometry of B cells, CD8(+), CD4(+) and gammadelta cells and the adhesion molecules CD44(+), CD62L, LFA-1 and LPAM-1 indicated increase in CD4(+) and B cells levels, with higher levels in blood than milk of group A, and significant expression of CD44(+) in blood and milk and LPAM-1 in blood only. The CD8(+) cells count in milk was higher than blood in the late stage. The peculiar feature of the early stage (group C) was the high level of gammadelta cells in the blood and milk, with tendency to express high level of CD62L. Compelling evidence could support the assumption that the dominant gammadelta cells at early stage of MAP infection could be of CD8CD2(-)WC+1(+) phenotype. gammadelta cells appear as promising markers in defining early changes of MAP infection due to their important role in priming innate and cell mediated immunity. Possible utilization of these peculiar changes in the gammadelta cells level in the early diagnosis of MAP infection should be the subject of further research.