scholarly journals Development of an Immunofluorescence Assay Using Recombinant Proteins Expressed in Insect Cells To Screen and Confirm Presence of Human Herpesvirus 8-Specific Antibodies

2008 ◽  
Vol 15 (8) ◽  
pp. 1259-1264 ◽  
Author(s):  
Veenu Minhas ◽  
Lynsey N. Crosby ◽  
Kay L. Crabtree ◽  
Saul Phiri ◽  
Tendai J. M'soka ◽  
...  
Keyword(s):  
Insect Cells ◽  
Human Herpesvirus ◽  
Immunofluorescence Assay ◽  
Human Herpesvirus 8 ◽  
Herpesvirus 8 ◽  
Specific Antibodies ◽  
Combined Use ◽  
Immunodeficiency Virus ◽  
Low Levels

ABSTRACT Human herpesvirus 8 (HHV-8), or Kaposi's sarcoma (KS)-associated herpesvirus, has been linked to all forms of KS. The results of most current serological assays for the detection of HHV-8-specific antibodies have low levels of concordance among themselves. To establish a sensitive and specific testing strategy that can be used to screen for HHV-8-specific antibodies, three HHV-8 proteins, ORF65, ORF73, and K8.1A, were expressed by using baculoviral vectors in insect cells and incorporated into a monoclonal antibody-enhanced immunofluorescence assay (mIFA) termed the Sf9 three-antigen mIFA. The results obtained by this mIFA were compared to those obtained by a standard mIFA with an HHV-8-infected B-cell line (BC3 mIFA). Test sera were obtained from patients diagnosed with KS, human immunodeficiency virus type 1-infected patients at high risk for HHV-8 infection, and healthy controls from a local blood bank. The combined use of both assays had a sensitivity of 94% and a specificity of 96%. The performance of these two assays when they were used together indicates that they may be useful for the reliable detection of HHV-8-specific immunoglobulin G antibodies in a population.

scholarly journals Searching for Human Herpesvirus 8 Antibodies in Serum Samples from Patients Infected with Human Immunodeficiency Virus Type 1 and Blood Donors from São Paulo, Brazil

1999 ◽  
Vol 179 (6) ◽  
pp. 1591-1592 ◽  
Author(s):  
Adele Caterino‐de‐Araujo ◽  
Maria Luisa Calabrò ◽  
Elizabeth de los Santos‐Fortuna ◽  
Jamal Suleiman ◽  
Luigi Chieco‐Bianchi
Keyword(s):  
Human Immunodeficiency Virus ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Blood Donors ◽  
Human Herpesvirus ◽  
Human Herpesvirus 8 ◽  
Serum Samples ◽  
Herpesvirus 8 ◽  
Immunodeficiency Virus

scholarly journals Reciprocal Regulatory Interaction between Human Herpesvirus 8 and Human Immunodeficiency Virus Type 1

2001 ◽  
Vol 276 (16) ◽  
pp. 13427-13432 ◽  
Author(s):  
Li-Min Huang ◽  
Miao-Fen Chao ◽  
Mao-Yuan Chen ◽  
Hsiu-ming Shih ◽  
Yu-Ping Chiang ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Human Herpesvirus ◽  
Human Herpesvirus 8 ◽  
Regulatory Interaction ◽  
Herpesvirus 8 ◽  
Immunodeficiency Virus

scholarly journals Human herpesvirus-8 (Kaposi’s sarcoma-associated herpesvirus) ORF50 interacts synergistically with the tat gene product in transactivating the human immunodeficiency virus type 1 LTR

2001 ◽  
Vol 82 (8) ◽  
pp. 1965-1970 ◽  
Author(s):  
Elisabetta Caselli ◽  
Paola Menegazzi ◽  
Arianna Bracci ◽  
Monica Galvan ◽  
Enzo Cassai ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Human Herpesvirus ◽  
Biologically Active ◽  
Human Herpesvirus 8 ◽  
Herpesvirus 8 ◽  
Immunodeficiency Virus ◽  
Hiv 1

Human herpesvirus-8 (HHV-8) is a lymphotropic virus associated with several AIDS-related neoplasms. Two ORFs play a critical role in the regulation of virus replication: ORF50, encoding an immediate-early transcriptional activator, and ORF57, encoding a post-transcriptional regulator. We analysed their effects on the activation of the human immunodeficiency virus type 1 (HIV-1) LTR. ORF50 interacted synergically with tat, inducing a 10-fold enhancement of HIV-1 LTR transactivation. This effect occurred both in BCBL-1 cells, latently infected with HHV-8, and in HL3T1 cells, an epithelial cell line non-permissive to HHV-8 infection. Also, ORF57 enhanced tat-induced transactivation of HIV-1 LTR, but only in BCBL-1 cells, suggesting that its action was likely mediated by the induction of other viral functions. Finally, when both ORFs were expressed, the enhancement of transactivation induced by ORF50 was partially inhibited. The findings suggest that ORF57 can modulate ORF50 activity and that ORF50 may render biologically active small amounts of tat.

scholarly journals Human Herpesvirus 8-Encoded vGPCR Activates Nuclear Factor of Activated T Cells and Collaborates with Human Immunodeficiency Virus Type 1 Tat

2003 ◽  
Vol 77 (10) ◽  
pp. 5759-5773 ◽  
Author(s):  
Shibani Pati ◽  
James S. Foulke ◽  
Oxana Barabitskaya ◽  
Jynho Kim ◽  
B. C. Nair ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
T Cells ◽  
Human Herpesvirus ◽  
Human Herpesvirus 8 ◽  
Nuclear Factor ◽  
Activated T Cells ◽  
Herpesvirus 8 ◽  
Immunodeficiency Virus ◽  
Hiv 1

ABSTRACT Human herpesvirus 8 (HHV-8), the etiologic agent of Kaposi's sarcoma (KS), encodes a chemokine receptor homologue, the viral G protein-coupled receptor (vGPCR), that has been implicated in KS pathogenesis. Expression of vGPCR constitutively activates several signaling pathways, including NF-κB, and induces the expression of proinflammatory and angiogenic factors, consistent with the inflammatory hyperproliferative nature of KS lesions. Here we show that vGPCR also constitutively activates the nuclear factor of activated T cells (NF-AT), another transcription factor important in regulation of the expression of inflammatory cytokines and related factors. NF-AT activation by vGPCR depended upon signaling through the phosphatidylinositol 3-kinase-Akt-glycogen synthetase kinase 3 (PI3-K/Akt/GSK-3) pathway and resulted in increased expression of NF-AT-dependent cell surface molecules (CD25, CD29, Fas ligand), proinflammatory cytokines (interleukin-2 [IL-2], IL-4), and proangiogenic factors (granulocyte-macrophage colony-stimulating factor GMCSF and TNFα). vGPCR expression also increased endothelial cell-T-cell adhesion. Although infection with HHV-8 is necessary to cause KS, coinfection with human immunodeficiency virus type 1 (HIV-1), in the absence of antiretroviral suppressive therapy, increases the risk of KS by many orders of magnitude. NF-AT and NF-κB activation by vGPCR was greatly increased by the HIV-1 Tat protein, although Tat alone had little effect on NF-AT. The enhancement of NF-AT by Tat appears to be mediated through collaborative stimulation of the PI3-K/Akt/GSK-3 pathway by vGPCR and Tat. Our data further support the idea that vGPCR contributes to the pathogenesis of KS by a paracrine mechanism and, in addition, provide the first evidence of collaboration between an HIV-1 protein and an HHV-8 protein.

scholarly journals Phylogenetic analysis of human herpesvirus-8 in South Africa and identification of a novel subgroup

2000 ◽  
Vol 81 (8) ◽  
pp. 2029-2038 ◽  
Author(s):  
Lee Alagiozoglou ◽  
Freddy Sitas ◽  
Lynn Morris
Keyword(s):  
South Africa ◽  
Genetic Diversity ◽  
Human Immunodeficiency Virus ◽  
Human Herpesvirus ◽  
Human Herpesvirus 8 ◽  
Single Strand Conformational Polymorphism ◽  
Pathogenic Potential ◽  
Herpesvirus 8 ◽  
Immunodeficiency Virus

The incidence of Kaposi’s sarcoma in South Africa is increasing in parallel with the human immunodeficiency virus type 1 epidemic. An 804 bp region in the ORF75 gene of 40 human herpesvirus-8 (HHV-8) isolates from South Africa was sequenced and the phylogenetic relationships were compared to published sequences. Nineteen strains clustered with subgroup B and 11 with subgroup A; however, the bootstrap values supporting these subgroups were not significant. Three strains grouped significantly with the C subgroup, while eight sequences did not cluster with any of the previously classified subgroups and were termed novel (N). The N subgroup differed from the A, B and C subgroups by DNA distances of 4·8, 4·2 and 4·5%, respectively, although within the N subgroup there was only 0·4% variation. The inclusion of this subgroup increased the number of previously described subgroup-specific polymorphisms from 17 to 47 over an 804 bp region. There was sufficient inter-subgroup genetic diversity for a single-strand conformational polymorphism assay to be used to identify them rapidly. Thus, based on analysis of the ORF75 gene, a unique HHV-8 subgroup, termed N, is present in South Africa, which accounts for 20% of circulating strains. Further studies are required to determine the degree of genetic divergence, distribution and pathogenic potential of this novel subgroup.

Sign in / Sign up

Export Citation Format

Share Document