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经乳汁传播的小鼠乳腺肿瘤病毒感染的瑞士白化小鼠中病毒抗原的检测:年龄、性别和生殖状态的影响。I. 乳腺组织和其他器官中四种抗原的免疫荧光定位

Detection of virus antigens in Swiss albino mice infected by milk-borne mouse mammary tumour virus: the effect of age, sex and reproductive status. I. Localization by immunofluorescence of four antigens in mammary tissues and other organs.

作者信息

Kozma S, Calberg-Bacq C M, François C, Osterrieth P M

出版信息

J Gen Virol. 1979 Oct;45(1):27-40. doi: 10.1099/0022-1317-45-1-27.

Abstract

The indirect immunofluorescence technique was used to investigate the expression of virus antigens in a Swiss albino mouse strain infected by mouse mammary tumour virus (MMTV). The antisera used were monospecific for: the glycopolypeptides gp160 and gp47, and the internal polypeptides p28 and p8. In mice infected with milk-borne MMTV, all four antigens were detected in the mammary gland during the first pregnancy and then throughout life, and in mammary tumours. Antigen gp160, located at the secretory border of all glandular cells, is shared by the plasma membrane of the mammary cell, whether virus-producing or not, and by the virus envelope, as shown by the use of absorbed antisera. The other three antigens are virus-specific. Anti-p47 serum stained the secretory border of cells whereas the fluorescence related to p28 and p8 consisted of a few large intracytoplasmic spots. The specific antibodies for p28 and p8 were only absorbed by disrupted virions, confirming that these polypeptides are internal antigens. With serial sections of glands from pregnant and old female mice, the alveoli stained with anti-p28 or anti-p8 serum were found to react also with anti-gp47 serum. However, during lactation, the presence of p28 and p8 could not be detected in the alveoli stained with anti-gp47 serum. Thus, infected alveoli express virus antigens differently. In the mammary glands of all mice studied, some alveoli did not display any staining when the latter three antisera were used. The specific antigenic pattern, maintained in differentiated mammary tumours, became a diffuse and unlocalized staining after transformation into the undifferentiated state. With the exception of the kidney, where gp47 was found in the glomeruli, all other organs of the mouse did not stain with any of the antisera. The presence of gp47 in the glomeruli, probably related to immune-complex deposits, increases with the age of the mouse and is most noticeable in tumour-bearing females. In mice free of milk-borne MMTV, examined during the first week of lactation, antigen gp47 could not be detected. Internal antigens p28 and p8 were detected in nearly all mammary cells as numerous small cytoplasmic dots. This suggests a limited expression of an endogenous virus genome; this expression would be modified when milk-borne virus is produced.

摘要

采用间接免疫荧光技术研究小鼠乳腺肿瘤病毒(MMTV)感染的瑞士白化小鼠品系中病毒抗原的表达。所用抗血清对以下物质具有单特异性:糖多肽gp160和gp47,以及内部多肽p28和p8。在经乳汁传播的MMTV感染的小鼠中,在首次怀孕期间以及之后的整个生命过程中,在乳腺和乳腺肿瘤中均检测到了所有这四种抗原。抗原gp160位于所有腺细胞的分泌边界,无论乳腺细胞是否产生病毒,其质膜以及病毒包膜均含有该抗原,这通过使用吸收后的抗血清得以证明。其他三种抗原具有病毒特异性。抗p47血清使细胞的分泌边界染色,而与p28和p8相关的荧光由一些大的胞质内斑点组成。p28和p8的特异性抗体仅被破碎的病毒粒子吸收,证实这些多肽是内部抗原。对怀孕和老年雌性小鼠的腺体进行连续切片观察发现,用抗p28或抗p8血清染色的肺泡也与抗gp47血清发生反应。然而,在哺乳期,在用抗gp47血清染色的肺泡中未检测到p28和p8的存在。因此,受感染的肺泡以不同方式表达病毒抗原。在所有研究的小鼠乳腺中,当使用后三种抗血清时,一些肺泡未显示任何染色。在分化的乳腺肿瘤中维持的特异性抗原模式,在转化为未分化状态后变为弥漫性且无定位的染色。除了在肾小球中发现gp47的肾脏外,小鼠的所有其他器官均未被任何抗血清染色。肾小球中gp47的存在可能与免疫复合物沉积有关,其随小鼠年龄增长而增加,在荷瘤雌性小鼠中最为明显。在哺乳期第一周检查的未感染经乳汁传播的MMTV的小鼠中,未检测到抗原gp47。在几乎所有乳腺细胞中均检测到内部抗原p28和p8,表现为众多小的胞质内小点。这表明内源性病毒基因组的表达有限;当产生经乳汁传播的病毒时,这种表达会发生改变。

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