In vitro and in vivo analysis of human herpesvirus-6 U90 protein expression

N. Nishimura, Tetsushi Yoshikawa, T. Ozaki, H. Sun, F. Goshima, Y. Nishiyama, Y. Asano, T. Kurata, T. Iwasaki

Research output: Contribution to journalArticle

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Abstract

In order to establish a reliable method for the detection of human herpesvirus-6 (HHV-6) B antigens in peripheral blood mononuclear cells (PBMCs) collected from HHV-6 infected patients, we created a polyclonal antibody against the HHV-6 B U90 protein (IEA/ex3) and used it to examine the expression of this protein in virus-infected cells and patients' PBMCs. This antibody reacted with 170 and 195 kDa proteins in HHV-6 B-infected cord blood mononuclear cells. The IEA/ex3 antigen was detected in cord blood mononuclear cells at 6 hr post-infection, and the number of infected cells reached its maximum at 48 hr post-infection. The antigen stained in a punctate pattern and partially localized to the promyelocytic leukemia (PML) protein body. We also examined 60 PBMC samples from 60 febrile children (3-19 months old) and detected IEA/ex3 antigen in the PBMCs by laser-scanning microscopy. HHV-6 was isolated from 31 of the 60 samples. The sensitivity and specificity of the antigen detection were 84% (26/31) and 97% (28/29), respectively, in the samples with virus detected. The mean number of antigen-positive PBMCs was 409/106 cells in 20 samples with viral isolation. A significant correlation (r = 0.566; P = 0.008) was observed between the viral load and number of antigen-positive cells. Although IEA/ex3 antigen was detected by laser-scanning microscopy in PBMCs (without cultivation) collected from six patients with isolated virus, it was detected in only one sample by conventional fluorescence microscopy. Increasing the intensity by cultivation (24 hr) resulted in a higher detection rate (5/6) even by conventional fluorescence microscopy, which is available in most hospital laboratories.

Original languageEnglish
Pages (from-to)86-92
Number of pages7
JournalJournal of Medical Virology
Volume75
Issue number1
DOIs
Publication statusPublished - 01-01-2005

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Human Herpesvirus 6
Blood Cells
Antigens
Cercopithecine Herpesvirus 1
Proteins
Viruses
Fetal Blood
Fluorescence Microscopy
Confocal Microscopy
Hospital Laboratories
In Vitro Techniques
Antibodies
Infection
Viral Load
Fever
Cell Count
Sensitivity and Specificity

All Science Journal Classification (ASJC) codes

  • Virology
  • Infectious Diseases

Cite this

Nishimura, N., Yoshikawa, T., Ozaki, T., Sun, H., Goshima, F., Nishiyama, Y., ... Iwasaki, T. (2005). In vitro and in vivo analysis of human herpesvirus-6 U90 protein expression. Journal of Medical Virology, 75(1), 86-92. https://doi.org/10.1002/jmv.20241
Nishimura, N. ; Yoshikawa, Tetsushi ; Ozaki, T. ; Sun, H. ; Goshima, F. ; Nishiyama, Y. ; Asano, Y. ; Kurata, T. ; Iwasaki, T. / In vitro and in vivo analysis of human herpesvirus-6 U90 protein expression. In: Journal of Medical Virology. 2005 ; Vol. 75, No. 1. pp. 86-92.
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Nishimura, N, Yoshikawa, T, Ozaki, T, Sun, H, Goshima, F, Nishiyama, Y, Asano, Y, Kurata, T & Iwasaki, T 2005, 'In vitro and in vivo analysis of human herpesvirus-6 U90 protein expression', Journal of Medical Virology, vol. 75, no. 1, pp. 86-92. https://doi.org/10.1002/jmv.20241

In vitro and in vivo analysis of human herpesvirus-6 U90 protein expression. / Nishimura, N.; Yoshikawa, Tetsushi; Ozaki, T.; Sun, H.; Goshima, F.; Nishiyama, Y.; Asano, Y.; Kurata, T.; Iwasaki, T.

In: Journal of Medical Virology, Vol. 75, No. 1, 01.01.2005, p. 86-92.

Research output: Contribution to journalArticle

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T1 - In vitro and in vivo analysis of human herpesvirus-6 U90 protein expression

AU - Nishimura, N.

AU - Yoshikawa, Tetsushi

AU - Ozaki, T.

AU - Sun, H.

AU - Goshima, F.

AU - Nishiyama, Y.

AU - Asano, Y.

AU - Kurata, T.

AU - Iwasaki, T.

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Y1 - 2005/1/1

N2 - In order to establish a reliable method for the detection of human herpesvirus-6 (HHV-6) B antigens in peripheral blood mononuclear cells (PBMCs) collected from HHV-6 infected patients, we created a polyclonal antibody against the HHV-6 B U90 protein (IEA/ex3) and used it to examine the expression of this protein in virus-infected cells and patients' PBMCs. This antibody reacted with 170 and 195 kDa proteins in HHV-6 B-infected cord blood mononuclear cells. The IEA/ex3 antigen was detected in cord blood mononuclear cells at 6 hr post-infection, and the number of infected cells reached its maximum at 48 hr post-infection. The antigen stained in a punctate pattern and partially localized to the promyelocytic leukemia (PML) protein body. We also examined 60 PBMC samples from 60 febrile children (3-19 months old) and detected IEA/ex3 antigen in the PBMCs by laser-scanning microscopy. HHV-6 was isolated from 31 of the 60 samples. The sensitivity and specificity of the antigen detection were 84% (26/31) and 97% (28/29), respectively, in the samples with virus detected. The mean number of antigen-positive PBMCs was 409/106 cells in 20 samples with viral isolation. A significant correlation (r = 0.566; P = 0.008) was observed between the viral load and number of antigen-positive cells. Although IEA/ex3 antigen was detected by laser-scanning microscopy in PBMCs (without cultivation) collected from six patients with isolated virus, it was detected in only one sample by conventional fluorescence microscopy. Increasing the intensity by cultivation (24 hr) resulted in a higher detection rate (5/6) even by conventional fluorescence microscopy, which is available in most hospital laboratories.

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