Identification of Antigenic Proteins Encoded by Human Herpesvirus 8 and Seroprevalence in the General Population and among Patients with and without Kaposi's Sarcoma

ABSTRACT To establish a sensitive and specific antibody assay, potent antigenic proteins encoded by human herpesvirus 8 (HHV8) were studied. Fifteen recombinant HHV8-encoded proteins were produced as glutathioneS-transferase fusion proteins. The sera from AIDS-associated Kaposi's sarcoma (KS) patients reacted with four proteins encoded by open reading frames (ORFs) K8.1, 59, 65, and 73 in a Western blot assay. An enzyme-linked immunosorbent assay (ELISA) using these four proteins as antigens (mixed-antigen ELISA) revealed that all 26 sera derived from KS patients (24 with and 2 without human immunodeficiency virus infection) became positive for anti-HHV8 antibodies. The presence of HHV8 was demonstrated in 14 (1.4%) of 1,004 sera from the Japanese general population and 10 (1.9%) of 527 sera from patients without HHV8-associated diseases. The presence of immunoglobulin G (IgG) and IgM antibodies against HHV8 examined further by the mixed-antigen ELISA and Western blotting revealed IgG antibody in all ELISA-positive sera, while IgM antibody against ORF K8.1 was absent. These data suggest that the ORF 73 and 65 proteins are potent antigens for a sensitive serological assay.

[1]  S. Sakurada,et al.  Expression and antigenicity of human herpesvirus 8 encoded ORF59 protein in AIDS‐associated Kaposi's sarcoma , 1999, Journal of medical virology.

[2]  H. Satoh,et al.  Establishing and characterizing a CD30‐positive cell line harboring HHV‐8 from a primary effusion lymphoma , 1999, Journal of medical virology.

[3]  S. Mori,et al.  High expression of HHV-8-encoded ORF73 protein in spindle-shaped cells of Kaposi's sarcoma. , 1999, The American journal of pathology.

[4]  B. Chandran,et al.  Comparison of human sera reactivities in immunoblots with recombinant human herpesvirus (HHV)-8 proteins associated with the latent (ORF73) and lytic (ORFs 65, K8.1A, and K8.1B) replicative cycles and in immunofluorescence assays with HHV-8-infected BCBL-1 cells. , 1999, Virology.

[5]  J. Dillner,et al.  Human serum antibodies to a major defined epitope of human herpesvirus 8 small viral capsid antigen. , 1999, The Journal of infectious diseases.

[6]  R. Desrosiers,et al.  Identification and Characterization of Kaposi’s Sarcoma-Associated Herpesvirus K8.1 Virion Glycoprotein , 1999, Journal of Virology.

[7]  K. Tadokoro,et al.  Seroprevalence of human herpesvirus 8 in human immunodeficiency virus 1‐positive and human immunodeficiency virus 1‐negative populations in Japan , 1999, Journal of medical virology.

[8]  A. Haase,et al.  A Cluster of Latently Expressed Genes in Kaposi’s Sarcoma-Associated Herpesvirus , 1998, Journal of Virology.

[9]  B. Chandran,et al.  Human herpesvirus-8 ORF K8.1 gene encodes immunogenic glycoproteins generated by spliced transcripts. , 1998, Virology.

[10]  L. Cuevas,et al.  Prevalence and transmission of Kaposi's sarcoma‐associated herpesvirus (human herpesvirus 8) in Ugandan children and adolescents , 1998, International journal of cancer.

[11]  K. Begtrup,et al.  Risk factors for Kaposi's‐sarcoma‐associated herpesvirus (KSHV/HHV‐8) seropositivity in a cohort of homosexual men, 1981–1996 , 1998, International journal of cancer.

[12]  R. Biggar,et al.  Interassay correlation of human herpesvirus 8 serologic tests. HHV-8 Interlaboratory Collaborative Group. , 1998, The Journal of infectious diseases.

[13]  M. Kaplan,et al.  Detection and titration of human herpesvirus-8-specific antibodies in sera from blood donors, acquired immunodeficiency syndrome patients, and Kaposi's sarcoma patients using a whole virus enzyme-linked immunosorbent assay. , 1998, Blood.

[14]  P. Pellett,et al.  Mapping and Serodiagnostic Application of a Dominant Epitope within the Human Herpesvirus 8 ORF 65-Encoded Protein , 1998, Journal of Clinical Microbiology.

[15]  T. Harr,et al.  High Human Herpesvirus 8 Seroprevalence in the Homosexual Population in Switzerland , 1998, Journal of Clinical Microbiology.

[16]  F. Neipel,et al.  The immunogenic glycoprotein gp35-37 of human herpesvirus 8 is encoded by open reading frame K8.1. , 1998, Journal of virology.

[17]  B. Chandran,et al.  Reactivities of human sera with human herpesvirus-8-infected BCBL-1 cells and identification of HHV-8-specific proteins and glycoproteins and the encoding cDNAs. , 1998, Virology.

[18]  L. Rainbow,et al.  The 222- to 234-kilodalton latent nuclear protein (LNA) of Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8) is encoded by orf73 and is a component of the latency-associated nuclear antigen , 1997, Journal of virology.

[19]  J. Goedert,et al.  Detection of serum antibodies to a Kaposi's sarcoma-associated herpesvirus-specific peptide. , 1997, The Journal of infectious diseases.

[20]  R. Sun,et al.  Identification, expression, and immunogenicity of Kaposi's sarcoma-associated herpesvirus-encoded small viral capsid antigen , 1997, Journal of virology.

[21]  R. Greenblatt,et al.  The prevalence of serum antibody to human herpesvirus 8 (Kaposi sarcoma-associated herpesvirus) among HIV-seropositive and high-risk HIV-seronegative women. , 1997, JAMA.

[22]  H. Zeichhardt,et al.  Detection of antibodies against viral capsid proteins of human herpesvirus 8 in AIDS-associated Kaposi’s sarcoma , 1997, Journal of Molecular Medicine.

[23]  J. Russo,et al.  Nucleotide sequence of the Kaposi sarcoma-associated herpesvirus (HHV8). , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[24]  P. Simmonds,et al.  Prevalence of Kaposi's sarcoma associated herpesvirus infection measured by antibodies to recombinant capsid protein and latent immunofluorescence antigen , 1996, The Lancet.

[25]  J. Phair,et al.  Seroconversion to antibodies against Kaposi's sarcoma-associated herpesvirus-related latent nuclear antigens before the development of Kaposi's sarcoma. , 1996, The New England journal of medicine.

[26]  E. Cesarman,et al.  Primary effusion lymphoma: a distinct clinicopathologic entity associated with the Kaposi's sarcoma-associated herpes virus. , 1996, Blood.

[27]  F. Sigaux,et al.  Kaposi's sarcoma-associated herpesvirus-like DNA sequences in multicentric Castleman's disease. , 1995, Blood.

[28]  P. Moore,et al.  Detection of herpesvirus-like DNA sequences in Kaposi's sarcoma in patients with and those without HIV infection. , 1995, The New England journal of medicine.

[29]  E. Cesarman,et al.  Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma. , 1994, Science.

[30]  D. Smith,et al.  Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. , 1988, Gene.