A mouse hybridoma secreting a monoclonal antibody (MAb) that bound
a noncapsular epitope expressed on C. neoformans was developed
by immunizing BALB/c mice with formalin-killed serotype A yeasts. The hybridoma,
designated CSFi, secreted an immunoglobulin G2b MAb that reacted with all
C. neoformans serotypes tested, including the acapsular mutant ATCC
52817 (Cap67). Postsectioned immune electron microscopy revealed extensive
binding of the MAb to the cell walls of both encapsulated and acapsular
yeasts. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western
blot analysis of secreted antigens recovered from concentrated culture
supernatants from both encapsulated and acapsular strains was conducted.
The results showed that this MAb bound predominantly to antigens with molecular
masses of approximately 75 and 100 kDa. A competitive enzyme-linked immunosorbent
assay was used to demonstrate that the MAb was not cross-reactive with
purified glucuronoxylomannan derived from either serotypes A or D. Experiments
conducted with mouse peritoneal phagocytes and the mouse phagocyte-like
cell line, J774A.1, demonstrated that the CSFi MAb opsonized the yeasts
and increased their adherence to both types of phagocytic cells. We conclude,
therefore, that antibodies directed at noncapsular epitopes can serve as
opsonins and may have a role in modulating cryptococcal infection.