ABSTRACT Kaposi’s sarcoma-associated herpesvirus (KSHV) is associated with Kaposi’s sarcoma and several B cell malignancies. K8.1, the major antigenic component of the KSHV virion, has been reported to play a critical role in the infection of certain B cells, but otherwise its function remains enigmatic. We created a K8.1 knockout virus (KSHVΔK8.1) in the BAC16 genetic background and analyzed its infectivity on a range of adherent cells. We observed a strong defect on several epithelial cells, e.g. the HaCaT keratinocyte model cell line, HEK 293T and A549 lung epithelial cells, but no such defect on other cells, among them e.g. lymphatic and blood endothelial cells. Mechanistically, we found that reduced infectivity of the K8.1 knockout virus correlated with reduced attachment to HaCaT cells. The defect in infectivity of KSHVΔK8.1 could be rescued by complementation through expression of K8.1 in KSHVΔK8.1 producing cells by means of a lentiviral vector. In a coculture infection model, KSHVΔK8.1 was highly efficient at infecting the BJAB B cell line but was significantly impaired at infecting the MC116 B cell line, in line with a previous report. In fusion assays together with the gH/gL glycoprotein complex and gB, the components of the conserved herpesviral core fusion machinery, we did not observe activation of membrane fusion by K8.1 or its R8.1 homolog of the rhesus monkey rhadinovirus. In summary, we found K8.1 to function in a highly cell-specific manner during KSHV entry at the attachment step, playing an important role in the infection of epithelial cells.

Kaposi’s sarcoma-associated herpesvirus glycoprotein K8.1 is critical for infection in a cell-specific manner and functions at the attachment step on keratinocytes