A case of immunohistochemical false positive staining caused by incompatibility between a CD4 antibody and an autostainer

Precise immunophenotyping of tumor cells by immunohistochemistry is complementary to morphological examination. It is critical for the correct histopathological diagnosis of lymphomas. In this paper, we report a case of T-cell lymphoma whose histopathological diagnosis was confounded by an immunohistochemical pitfall: a false positive caused by incompatibility between an antibody and an autostainer. In this case, based on CD4 immunohistochemistry of the affected lymph nodes, the T-cell lymphoma was diagnosed as CD4-positive at the onset, while it appeared discordantly to be CD4-negative at the second relapse. We noticed that CD4 antibodies and autostainers of different suppliers (designated as suppliers X and Y) were used in an unqualified combination in immunohistochemistry at the onset: that is, the combination of an antibody supplied by X and an autostainer supplied by Y (designated as X-Y combination) was used at the onset. On the other hand, the Y-Y combination was at the second relapse. At the second relapse, flow cytometry of the affected lymph node showed infiltration of CD4-negative T-cell lymphoma. We reasoned that CD4 immunonegativity obtained by the Y-Y combination at the second relapse was specific, while CD4 immunopositivity by the X-Y combination at the onset was false positive. Immunohistochemical reexamination of the lymph node at the onset proved to be CD4-negative by not only the Y-Y but also X-X combinations, confirming our final diagnosis of nodal relapse of CD4-negative T-cell lymphoma. This case illustrates the importance of using compatible combinations of antibodies and autostainers in diagnostic immunohistochemistry.