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매독환자의 혈청 면역글로불린 G와 M항체에 반응하는 매독균 단백항원에 관한 연구

Other Titles
 Identification of protein antigens of treponema pallidum reacting with serum IgG and IgM antibodies of patients with syphilis 
Authors
 김동건 
Issue Date
1988
Description
의학과/박사
Abstract
[한글]

매독의 원인균인 T. pallidum을 구성하는 단백항원들을 알아내고, 그들의 생물학적, 생

화학적 특성을 규명하는 것은 매독의 병인론을 밝혀내는 기본요소가 되며, 나아가서는 백

신의 개발 및 새로운 면역학적 진단법 개발에 크게 기여할 수 있을 것이다. 이에 여러학

자들이 sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)와 immun

oblotting을 이용하여 매독균 단백항원의 정체 규명을 위한 연구를 해 왔으나, 연구자에

따라 매독환자 혈청에 출현하는 항체와 반응하는 매독균 항원의 수가 다르고 각 임상기별

로 항체와 반응하는 분자량에 따른 매독균 항원의 종류에도 차이가 있었다. 본 연구에서

는 매독 치료전에 출현하고 치료후에 소실되는 혈청 IgG 및 IgM 항체와 반응하는 매독균

단백항원을 규명하고 또한 T. pallidum과 T. phagedenis의 공유항원, T. pallidum 특이항

원을 규명하고자 다음과 같은 실험을 하였다.

가토의 고환에 계대접종하여 보존해 오던 T. pallidum, Nichols strain을 추출하여 Per

coll을 이용한 밀도구배 원심분리로 순수분리 하고 SDS-PAGE와 immunoblotting하였다. 이

를 각 임상기 매독환자 각기 3명으로부터 치료전과 치료후 3개월마다 채취하여 pooling한

혈청과 2기, 조기 잠복 또는 만기 잠복매독에서 치료후 2∼14년된 14명의 치료된 매독환

자 혈청과 반응시켜 autoradiography 혹은 immunoperoxidase 방법으로 IgG 및 IgM 항체에

반응하는 매독균 단백항원을 관찰하였다. 또한 thioglycollate broth media에 계대배양

해 오던 T. phagedenis, biotype Reiter를 SDS-PAGE와 immunoblotting한 후, 이를 각 임

상기 매독환자 각기 3명으로부터 치료전에 채취하여 pooling한 혈청과 반응시켜 autoradi

ography방법으로 혈청 IgG 항체와 반응하는 단백항원을 관찰하여 다음과 같은 성적을 얻

었다.

1. SDS-PAGE로 분리하여 염색한 결과, T. pallidum, Nichols strain과 T. phagedenis,

biotype Reiter에서 각각 45개 및 43개의 단백항원을 관찰하였다.

2. 치료전 매독환자 혈청 IgG 항체와 반응하는 분자량 47,000, 36,500, 15,500, 14,000

의 단백항원과 IgM 항체와 반응하는 분자량 47,000, 34,000, 29,500의 단백항원이 가장

강한 반응을 보여 이들이 매독균 주항원임을 관찰하였다.

3. 치료전, 후 매독환자에서 혈청 IgG 및 IgM 항체와 반응하는 매독균 단백항원을 관찰

한 결과, 1기, 2기 및 조기 잠복매독에서는 소수의 항원 소실과 반응도의 감소를 관찰하

였으나, 만기잠복 및 재감염된 매독에서는 변화를 볼 수 없었다.

4. 치료된 매독환자에서 혈청 항체와 매독균 주항원의 반응도를 관찰한 결과, IgM 항체

와 반응하는 분자량 47,700의 항원에서만 현저하게 반응도가 감소하였다.

5. T. pallidum과 T. phagedenis의 공유항원을 관찰한 결과 모두 11개였고, 분자량 86,

500, 68,500, 15,500, 14,000의 항원이 공유항원이 아닌 매독균 특이 항원임을 알 수 있

었다.

이상의 결과로 매독균 주항원중 분자량 15,500, 14,400의 항원과 분자량 47,000의 항원

이 각각 향후 매독혈청검사 및 치료판정에 도움이 될 것으로 생각된다.







[영문]

Knowledge of the constituent protein antigens of T. pallidum, the causative

microorganism for syphilis, and their biological functions and biochemical

properties would not only serve to lay the groundwork in elucidating the

pathogenesis of syphilis, but would also further contribute significantly in the

development of both vaccinations and new immunodiagnostic methods for syphilis.

For these reasons, many researchers employing SDS-PAGE and immunoblotting

techniques have been attempting to characterize protein antigens of syphilis but

with differing results in terms of number of antigens reacting with antibodies in

the sera of patients with syphilis and with respect to antigens reacting with

antibodies in the sera of different stages of syphilis which were of different

types in terms of their molecular weights. This study was conducted to identify the

protein antigens reacting with IgG and IgM antibodies in the sera of patients with

syphilis, which appear before and disappear after treatment, the antigens common to

both T. pallidum and T. phagedenis, and the antigens specific only to T. pallidum.

T. pallidum, Nichols strain maintained by rabbit testicular passage was extracted

and purified by Percoll density gradient centrifugation, followed by SDS-PAGE and

immunoblotting. The protein antigens of T. pallidum reacting with IgG and IgM

antibodies in the sera were observed using autoradiography and immunoperoxidase

technique after the reactions between the prepared antigens of T. pallidum and the

sera of patients with syphilis in groups of 3 at each clinical stage before and

after treatment at 3 months intervals, where each group of 3 was pooled and the

sera of 14 patients with treated syphilis who had been treated for secondary, early

latent and late latent syphilis 2-14 years ago. T. phagedenis, biotype Reiter

maintained in the thioglycollate broth media was separated and transferred by

SDS-PAGE and immunoblotting. After the reaction between the prepared antigens of T.

phagedenis and the sera of patients with syphilis in groups of 3 at each clinical

stage before treatment, where each group of 3 was pooled, protein antigens of T.

phagedenis reacting with IgG antibodies in the sera were observed using

autoradiography.

The results obtained from the above observations were as follows;

1. After separation by SDS-PAGE and staining with Coomassie Blue dye, 45 protein

antigens of T. pallidum, Nichols strain and 43 protein antigens of T. phagedenis,

biotype Reiter were observed.

2. Before treatment, the most strongly reacting antigens of T. pallidum

precipitated by IgG antibodies in the sera of patients were polypeptides of

molecular weights 47,000, 36,500, 15,500 and 14,000 and those precipitated by IgM

antibodies were polypeptides of molecular weights 47,000, 34,000 and 29,500. So it

was observed that those were the major antigens of T. pallidum.

3. After observing protein antigens of T. pallidum reacting with IgG and IgM

antibodies in the sera of patients with syphilis before and after treatment, it was

seen that in primary, secondary and early latent syphilis there was a loss of

several antigens and a decrease in reactivity, but no changes occurred in late

latent and reinfected syphilis.

4. From the observation of the reaction between serum antibodies of patients with

treatedsyphilis and major antigens of T. pallidum, an evident decrease in

reactivity was observed only with protein antigen of molecular weight 47,000 which

reacts with IgM antibody.

5. The total number of antigens common to both T. pallidum and T. phagdenis was

observed to be 11, and antigens of molecular weights 86,500, 68,500, 15,500 and

14,000 showed to be non-common but antigens specific to T. pallidum.

From the above results, it could be concluded that of the major antigens of T.

pallidum, the antigens of the molecular weights 15,500 and 14,000 could serve to

develop newer serologic tests for syphilis, and that of molecular weight 47,000

could contribute to the assessment of the efficacy of treatment.
Full Text
https://ymlib.yonsei.ac.kr/catalog/search/book-detail/?cid=CAT000000005078
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1. College of Medicine (의과대학) > Others (기타) > 3. Dissertation
URI
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