[Simultaneous in situ detection of viral antigens and RNA in brain tissues from a patient with herpes simplex encephalitis by immunocytochemistry and in situ hybridization].
In order to elucidate the correlation between herpes simplex virus (HSV-1) and the central nervous system tissue, we performed the simultaneous detection of viral antigens and RNA in the brain tissue sections from a patient with herpes simplex virus (HSV) encephalitis using immunocytochemistry and in situ hybridization. In the present study the hybridization protocol reported by Brahic M et al. in 1984 were applied for the simultaneous detection of viral RNA and antigens with a few modification. The sections were first immunocytochemically stained to detect HSV-1 antigens by ABC method, and then hybridized with 3H-labelled HSV-1 cDNA probe for the detection of RNA after the acetylation of slides for the prevention of nonspecific bindings of isotope to slides. In the present study, viral antigens were immunocytochemically stained to brown-colored deposits located in the cytoplasm and nucleus whereas viral RNA were detected as the accumulation of many silver grains over the nuclei or cytoplasm. In this case the light microscopic findings in a part of temporal lobe showed multiple areas of necrosis mainly involving the gray matter and a few inflammatory changes such as perivascular cell cuffings. HSV-1 infected Vero cells as positive control demonstrated both antigens and RNA as shown in Fig.1 a. However, no hybridization signals and color deposits were observed in uninfected Vero cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Nakamura
S
S
Department of Neurology, Tohoku University School of Medicine, Sendai, Japan.
Shimazaki
S
S
Yoshioka
M
M
Nagano
I
I
Sekizawa
T
T
Umene
K
K
D000956:Antigens, Viral / Q000032:analysis*; D001921:Brain / Q000276:immunology* / Q000473:pathology; D001923:Brain Chemistry; D002452:Cell Count; D004660:Encephalitis / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D006561:Herpes Simplex / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008722:Methods; D008875:Middle Aged; D009693:Nucleic Acid Hybridization; D012367:RNA, Viral / Q000032:analysis*; D018139:Simplexvirus / Q000235:genetics* / Q000276:immunology
D000956:Antigens, Viral; D012367:RNA, Viral