Figure 4.

Kaposi’s sarcoma–associated herpesvirus (KSHV) infection of primary human neural cultures. Primary human neuron cultures were established from triturated mixed cell suspensions from neural tissue specimens. Infection with rKSHV.219, which expresses green fluorescent protein (GFP) from the EF1-a promoter, was conducted at an multiplicity of infection of 0.5. Cultures were micrographed at 488 nM excitation 72 hours after infection. A, Three representative panels show cells with apparent neuron morphology that are expressing GFP, indicative of KSHV infection. B, Similar cultures were fixed in 4% buffered paraformaldehyde and subjected to immunofluorescence-based detection of the indicated antigens or of recombinant KSHV–derived GFP in situ. C, An identically treated mock-infected control culture from the tissue donor.