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Brain and Nerve No to Shinkei Volume 42, Issue 4 （April 1990）

Brain and Nerve 脳と神経 42巻4号（1990年4月発行）

Japanese English

原著

単純ヘルペス脳炎剖検脳におけるウイルス抗原，浸潤細胞およびサイトカイン産生細胞の検索 THE DETECTION AND QUANTITATIVE ANALYSIS OF VIRAL ANTIGENS, INFILTRATING MONONUCLEAR CELLS, IFN-γ, LT AND IL-6 BEARING CELLS IN THE FROZEN BRAIN TISSUE SECTIONS FROM A PATIENT WITH HERPES SIMPLEX ENCEPHALITIS BY IMMUNOCYTOCHEMISTRY 中村正三 ¹ , 島崎茂 ¹ , 永野功 ¹ , 吉岡勝 ¹ , 関沢剛 ¹ , 小暮久也 ¹ Shozo Nakamura ¹ , Shigeru Shimazaki ¹ , Isao Nagano ¹ , Masaru Yoshioka ¹ , Tsuyoshi Sekizawa ¹ , Kyuya Kogure ¹ ¹東北大学医学部脳研神経内科 ¹Department of Neurology, Institute of Brain Diseases, Tohoku University School of Medicine キーワード： herpes simplex encephalitis , cytokines , viral antigen , immunocytochemistry Keyword: herpes simplex encephalitis , cytokines , viral antigen , immunocytochemistry pp.377-382

発行日 1990年4月1日 Published Date 1990/4/1

DOI https://doi.org/10.11477/mf.1406900046

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Abstract 文献概要
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　単純ヘルペス脳炎剖検凍結脳を用いて，ウイルス抗原（viral antigen：VA），浸潤細胞の同定，さらにサイトカイン産生細胞の検出を免疫組織化学法により行ない，宿主の急性期免疫反応を検討した。また，免疫組織化学法とin situハィブリダイゼーション法により，Ia抗原とHSV−1mRNAの同時検出も行った。

　症例は58歳男子で第12病日に死亡した。実験には頭頂葉の一部を用いた。本組織ではVA陽性細胞およびmRNAが多数検出された。浸潤細胞数はLeu−11.6％，Leu−2a0.4％，Leu−3a0.9％，Leu−70.7％，HLA-DR（Ia）10％でLeu−12は検出されなかった。さらにVA陽性細胞の多い皮質部分ではIFN—γ，LT IL−6陽性細胞いずれも多数検出され，これらはアストロサイト，マクロファージおよび感染した神経細胞であった。

　この結果からヘルペス脳炎急性期患者の一部脳のVAの多い時期にはリンパ球浸潤が少ない時期でもIa陽性細胞が出現し，さらに非特異的免疫反応が惹起され，そこにはIFN—γ，LT, IL−6産生細胞が多数出現し，急性期反応に関与していると考えられた。

To identify viral antigens, the types of infilt- rating mononuclear cells and cytokine bearing cells, the frozen brain tissue sections form a pa-tient with herpes simplex encephalitis who died on 12 th hospital days, were examined by immu-nocytochemical methods and combined immunocy-tochemistry and in situ hybridization.

The avidin-biotin peroxidase complx (ABC) tec-hniques were applied for the detection of antigens. All monoclonal antibodied to Leu series and polyc-lonal antisera to lymphotoxin (LT), interlukin-6 (IL-6) and interferon-γ (IFN-γ) were purchased form Becton Dickinson Co., and Genzyme Co., (USA) respectively.

A large number of neurons and glial cells stai-ning positively HSV-1 antigens were found in the gray matter (Fig. 1). Moreover, although a mo-derate number of HLA-DR (Ia) positive cells were found in the parenchyma, there were few cells dis-playing positively for Leu-3 a, Leu-2 a and Leu-7 respectively (Fig. 2). To evaluate the number of positive cells appeared in the brain tissues, Leu stain for 4, 2 a, 3 a, 7, 12 and HLA-DR demonst-rated 1. 6%, 0.4%, 0.9%, 0.7% and 10% respec-tively.

In addition, numerous number of IFN-T positive cells were detected arround the lesion and ran-domly distributed throughly the lesion (Fig. 3 a). IL-6 positive cells (Fig. 3 b) and LT positive cells (Fig. 3 c) were also similar in distribution to IFN-γ positive cells.

Moreover, in simultaneous detection of HLA-DR and HSV-1 mRNA by the combined immu-nocytochemistry and in situ hybridization, there were seen glial cells staining positively for HLA-DR (Ia) and several cells with mRNA (Fig. 4 a and b).

From the results, it may be considered that in a part of brain tissue at the early stage of herpes simplex encephalitis, these cytokine bearing cells play an important role at the early inflammatory process of host immune response prior to appea-rance of mononuclear cells.