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感染白色念珠菌B血清型的家兔中的抗原血症:通过酶免疫测定法进行检测及抗原的初步特性分析

Antigenemia in rabbits infected with Candida albicans serotype B: detection by enzyme immunoassay and preliminary characterization of the antigen.

作者信息

Reiss E, Kuykendall R J, Kaufman L

出版信息

J Med Vet Mycol. 1986 Aug;24(4):259-69.

PMID:3528447
Abstract

Peroxidase-labeled IgG against Candida albicans serotype A cell walls failed to detect antigenemia in rabbits infected with C. albicans type B. Therefore type B antisera were produced in rabbits immunized with either type B cell walls; whole blastoconidia heated and killed at 60 degrees C (HK); a sublethal intravenous dosage of 10(6) colony forming units (c.f.u.); or a normally lethal dose of 10(7) c.f.u. injected into subcutaneously implanted plastic chambers. The four antisera were conjugated to peroxidase for a double antibody sandwich enzyme immunoassay. Sixteen rabbits, immunosuppressed with cortisone, received 10(7) type B blastoconidia intravenously. On day 4 their kidneys contained between 3.4 X 10(8) and 1.4 X 10(11) c.f.u. Antigenemia was detected only with the conjugated anti-HK-IgG. A mean concentration of 246 ng ml-1 of circulating antigen was detected only when serum-antigen complexes were dissociated by boiling in EDTA. The properties of the circulating antigen were as follows: relative molecular weight between 50 000 and 100 000 da; stable to boiling and proteinases, labile to sodium periodate oxidation, and bound by concanavalin A. These properties are consistent with a polysaccharide or glycoprotein, probably the mannoprotein of the cell wall. Antigenic specificity was probed further using IgG produced against heat-killed C. albicans serotype A. Antigenemia resulting from infection with serotype B was detected in 5 of 5 rabbits tested with the anti-HK-serotype B IgG but only with 3 of 5 tested with the anti-HK-serotype A IgG. In addition, the concentrations detected with serotype B reagents were 31 times higher.

摘要

抗白色念珠菌 A 血清型细胞壁的过氧化物酶标记 IgG 未能在感染 B 型白色念珠菌的兔中检测到菌血症。因此,用 B 型细胞壁、在 60℃加热并杀死的完整芽生孢子(HK)、10⁶ 个菌落形成单位(c.f.u.)的亚致死静脉剂量或注入皮下植入塑料腔室的 10⁷ c.f.u. 的正常致死剂量对兔进行免疫,制备 B 型抗血清。将这四种抗血清与过氧化物酶偶联,用于双抗体夹心酶免疫测定。16 只用可的松免疫抑制的兔静脉注射 10⁷ 个 B 型芽生孢子。在第 4 天,它们的肾脏中含有 3.4×10⁸ 至 1.4×10¹¹ 个 c.f.u.。仅用偶联的抗 HK-IgG 检测到菌血症。仅在通过在 EDTA 中煮沸使血清 - 抗原复合物解离时,才检测到循环抗原的平均浓度为 246 ng/ml。循环抗原的特性如下:相对分子量在 50000 至 100000 道尔顿之间;对煮沸和蛋白酶稳定,对高碘酸钠氧化不稳定,并与伴刀豆球蛋白 A 结合。这些特性与多糖或糖蛋白一致,可能是细胞壁的甘露糖蛋白。使用针对热杀死的白色念珠菌 A 血清型产生的 IgG 进一步探究抗原特异性。在用抗 HK - B 血清型 IgG 检测的 5 只兔中,有 5 只检测到由 B 血清型感染引起的菌血症,但在用抗 HK - A 血清型 IgG 检测的 5 只兔中,只有 3 只检测到。此外,用 B 血清型试剂检测到的浓度高 31 倍。

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