Production and characterization of monoclonal antibodies reactive with the mycelial and yeast phases of Penicillium marneffei

A definitive tissue diagnosis of Penicillium marneffei is hampered by a microscopic similarity to the yeast form of Histoplasma capsulatum and Cryptococcus neoformans. In order to obtain a better discrimination for accurate diagnosis, monoclonal antibodies (MAbs) were produced from hybridomas raised from Balb/c mice immunized with mycelial culture filtrate. By indirect immunofluorescent or immunoblot analyses, one immunoglobulin (Ig) G₁ (3C2) MAb and three IgM (8B11, 3B9 and 8C3) MAbs were found to react strongly with P. marneffei antigens. In the immunoblots, the MAbs 8B11 and 3B9 reacted most strongly with a high molecular weight component ( > 200 kDa) produced during either the mycelial or yeast phase of fungal growth. The immunoreactive epitopes for these two IgM MAbs were most likely associated with carbohydrate moieties, judging from their susceptibility to periodate oxidation and concanavalin A binding. This is in contrast to the immunoreactive epitopes for the MAbs 8C3 and 3C2, which were resistant to destruction by periodate treatment and did not bind to the lectin. Judging from immunofluorescent intensity, the three IgM MAbs could react strongly with the yeast cells present in the tissue biopsies of patients with P. marneffei infection.