Affiliations: [a] Department of Immunology, University of Kiel, Germany | [b] Department of Microbiology, University of Kiel, Germany
Correspondence:
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Correspondence and reprint requests to: Hermann Alexander, MD, Department of Immunology, University of Kiel, Brunswikerstr. 2-4, 2300 Kiel, Germany.
Abstract: Human monoclonal antibodies directed against human cytomegalovirus were generated by fusion of in vitro stimulated human spleen lymphocytes from an HCMV-seropositive 53-year-old organ donor with the mouse myeloma cell line Ag8.653. Fourteen human/mouse hybridomas producing anti-cytomegalovirus IgG were screened by an ELISA technique and four selected clones have been estabished since March 1988, generating about 5–40 μg/24 h IgG per ml culture supernatant. Reference and local cytomegalovirus strains were stained by the antibodies without showing cross-reactivity to other herpes viruses. Three monoclonal antibodies, A4B4 (IgG11), A6B3 (IgG1k) and A6A2 (IgG1k), immunoprecipitated a 68 kDa early viral protein which appears during the infectious cycle, first in the nucleus (18–24 h) and then also in the cytoplasm (24–96 h) of infected cells. Inhibition of DNA replication restricted the detection of the 68 kDa viral protein to the nucleus of infected cells. Staining of unfixed infected cells showed that two of the antibodies bound at the surface of a few cells. The fourth monoclonal antibody A3C5 (IgG11) immunoprecipitated a 34/38 kDa late viral protein which appears in the nucleus (48–72 h) of infected cells. These antibodies enable us to study the human host response to human cytomegalovirus and to elucidate the functions of human antibodies especially in their interaction with the T-cell response.
Keywords: Human monoclonal antibodies, human cytomegalovirus, IgG1 antibodies, early viral protein, late viral protein
