Mucopolysaccharide Material Resulting from the Interaction of Treponema pallidum (Nichols Strain) with Cultured Mammalian Cells

During incubation of Treponema pallidum (Nichols strain) with cultured mammlian cells derived from normal rabbit testes (NRT), an amorphous material accumulated at the surface of the cultured cells. This material was randomly distributed on all tissue cells within the culture chambers. The amount of amorphous material was dependent on the treponemal inocula. With 3 × 10 8 organisms per ml, this material was readily apparent within 2 days; with 4 × 10 7 organisms per ml, this material was detectable within 4 to 5 days; with lower inocula, the accumulation of amorphous material was far less apparent. Deposition of this surface-associated material required attachment of treponemes to the cultured cells, and the amount deposited was related to the number of treponemes attached per cell. This amorphous material was not detected when NRT cells were incubated with preparations of T. pallidum that were heat or air inactivated. In addition, the accumulaton of amorphous material was not due to a soluble component from host testicular tissue or to a soluble component developing during treponemal infection. This was demonstrated by the inability of membrane filtered preparations of T. pallidum to induce the deposition of amorphous material at the surface of the cultured cells. The nature of this material appeared to be acidic mucopolysaccharide as indicated by its metachromatic staining properties, its stainability with ruthenium red, and its partial degradation by bovine and streptomyces hyaluronidase. This amorphous material that accumulated in vitro at the surface of cultured cells may be similar to the mucoid material that accumulates in vivo during syphilitic infection.