Effects of Enterococcus faecalis sonicated extracts on IL-2, IL-4 and TGF-β1 production from human lymphocytes
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근관치료의 실패원인 중 중요한 세균으로 알려진 Enterococcus faecalis는 최근에 중요성이 더해지며 많은 연구들이 진행중이다. 여러가지 기전들이 보고되고 있으나 면역반응에 관한 연구는 거의 알려져 있지 않은 상태이다. 본 연구에서는 Enterococcus faecalis 의 초음파 분쇄 추출물을 성인의 말초혈액으로부터 얻은 임파구에 적용시켜서 여기서 분비되는 interleukin-2, interleukin-4, transforming growth factor-β1 의 농도를 Enzyme linked immunosorbent assay (ELISA) 로 측정하여 비교, 평가하는 것을 목적으로 한다. E. faecalis 를 적절한 조건에서 배양한 뒤 초음파 분쇄를 하여 추출물을 얻어냈다. 임파구는 건강한 성인의 말초혈액에서 추출하여 분리하였다. 임파구를 적절한 농도의 mitogen(Phytohemagglutinin: PHA)으로 자극시킨 뒤에 다양한 농도의 E. faecalis 초음파 추출물을 적용시키고 72시간 동안 배양하였다. ELISA 를 이용하여 IL-2, IL-4, TGF-β1 의 농도를 측정하였다. 실험결과는 Kruskal-Wallis test, Man-Whitney rank sum test (p < 0.05) 를 사용하여 통계처리 하였다. 실험결과 PHA 로 처리한 군은 아무것도 처리하지 않은 군에 비해서 IL-2, IL-4 의 수치가 유의성 있게 높았다 (p < 0.05). PHA 로 처리한 군중에서 고농도와 중농도의 sonic extract of E. faecalis (SEF) 로 처리한 군은 그렇지 않은 군에 비해서 IL-2, IL-4, 의 농도가 유의성 있게 낮았다 (p < 0.05). PHA 로 처리한 군중에서 저농도의 SEF 로 처리한 군은 그렇지 않은 군과 비교하여 유의할 만한 차이를 보이지 않았다. TGF-β1 의 농도는 모든 군에서 유의할 만한 차이를 보이지 않았다 (p > 0.05). 따라서, E. faecalis 의 추출물은 임파구의 IL-2, IL-4 의 분비능력을 저하시킨다고 할 수 있다.
Abstract
In order to examine the immunoresponse of host cells to Enterococcus faecalis, this in vitro study monitored the production of Interleukin-2(IL-2), Interleukin-4(IL-4) and Transforming growth factor-β1(TGF-β1) in human lymphocytes. Lymphocytes were activated with PHA in the presence or abscence of sonicated extracts of E. Faecalis(SEF) and further incubated for 72 hours. The level of each cytokine was measured by ELISA. Data were analyzed with Kruskal-Wallis test and Mann-Whitney U test (P<0.05). PHA-activated group did exhibit higher level of IL-2 and IL-4 than untreated control group. The levels of expression of both cytokines were significantly decreased following the treatment of high (25μg/ml) and medium concentration(12.5μg/ml) of SEF (P<0.05) than those of PHA activated group. But low concentration (5μg/ml) of SEF showed the similar level of IL-2 and IL-4 production as those of PHA activated group. TGF-β1 was unaffected by SEF treatment. These results suggested that E. faecalis may suppress IL-2 and IL-4 production by lymphocytes and this could be one of possible factors why E. faecalis are found frequently in the teeth with failed endodontic treatment.
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