단클론항체를 이용한 B세포 자가성장인자의 특징
Characterization of B Cell Autocrine Growth Factor using Monoclonal Antibody
소속 상세정보
이헌구/Hern Ku Lee
박재승/김성광/하대유/장용석/Jae Seung Park/Sung Kwang Kim/Tai You Ha/Yong Suk Jang
KMID : 0357119950170040317
Abstract
정상 B 세포유래 자가성 장인자 (B cell-derived B cell growth factor; B-BCGF)는 존재
만 확인되었을 뿐 그 분자적 특성이나 생물학적 기능에 관해서는 전혀 알려져 있지 않다.
따라서 본 연구에서는 이 물질의 본체를 연구하려는 목적의 일환으로 B-BCGF에 대한 단
클론 항체의 개발을 시도하였다. B-BCGF로는 Staphylococcus aureus Cowan I Strain
(SAC, 0.002%) 으로 24시간 자극한 배양액(SAC-sup)을 사용하였다. 농축한 SAC-sup을 마
우스 비장에 주사 면역하여 얻은 단클론 항체는 SAC 으로 자극한 B 세포 증식을 억제하고
SAC-sup으로 부터 BCGF활성을 가진 64Kda 인 물질을 침전 시켰다. 이 항체를 이용한
affinity 크로마토그라피로 정제된 BCGF 는 그 양이 너무 작아 염기서열을 결정하는데 실
패하였다. 이들 항체 또는 항-IgM 항체 +IL2로 자극한 B 세포의 증식은 억제하지 못하여
SAC특이적인 기능을 나타내었으며 IsM 항체 생산으로 알아본 B 세포분화 역시 40-50%
유의하게 억제하였으나 활성화의 지표인 CD23및 CD25의 발현에는 별다른 영향을 미치지
못하였다.
#초록#
It has been reported that malignant B cell line or Epstein Barr virus transformed as
well as normal B cells are capable of producing an autocrine B cell growth factor
(B-BCGF). However, the molecular nature and biological roles of B-BCGF were poorly
understood. To inverstigate this issue, we have tried to develop monoclonal
antibodies(mAbs) to B-BCGF. The purified tonsillar B cells were stimulated with
Staphylococcus aureus Cowan I strain (SAC). The 24h culture supematant(SAC-sup)
was used as the source of B-BCGF. The mice were immunized by injection of
concentrated SAC-sup via spleen. Five hybridomas which inhibited SAC-stimulated B
cell proliferation and removed B-BCGF activity from SAC-sup were selected, cloned and
mAbs were prepared as ascites from. SAC-sup which eluated from affinity column
conjugated with the mAbs(HJ 93 and HI 205) were found to have BCGF
activity,indicating that these mAbs are specific for B-BCGF of SAC-sup. The mAb,HJ
93, precipitated the molecule with M.W of 64KDa which had BCGF activity.The mAb
did not affect the expression of CD23 or CD25 which are known as activation markers
of B cells, but significantly inhibited SAC-or SAC+IL2-stimulated B cell proliferation as
well as IgM secretion from SAC+IL-2-stimulated B cells.Taken together. these data
indicated that a mAb capable of binding to 64KDa B-BCGF and inhibiting
SAC-stimulated B-cell proliferation as well as differentiation was developed.
키워드
Autocrine B cell growth factor; monoclonal antibody.;
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