Cytotoxicity of lymphokine activated peritoneal macrophages against Trichomonas vaginalis

질트리코모나스에 대한 림포카인황성대식세포의 세포독성능

Trichomonas vaginalis is a parasitic nagellate in the urogenital tract of human. Innate cytotonicity of macrophages against T. vaginalis has been recognized, but any report on the cytotoxicity of Iymphokine-activated macrophages to T vaginalis is not yet available. The present study aimed to elucidate the Iymphokine-activated cell mediated cytotoxic effect against T. vaginalis by mouse peritoneal macrophages. Cytotoxicity was measured by counting the release of $^3H-thymidine$ from prelabeled protozoa, and tested in U-bottom microtiter plates. Nitrite concentration in culture supernatants was measured by standard Griess reaction. The results obtained are as follows: 1, The cytotoxicity of macrophages was increased by addition of rIL-2 or $rIFN-{\gamma}$$. 2, Cytotoxicity of macrophages was reduced by addition of rIL-4 to rOM-CSV, rIL-2 or $rIFN-{\gamma}$. 3. Crude Iymphokine mixed with anti-lL-2 decreased the cytotoxity of macrophages. 4. In case of macrophages cultured with $rIFN-{\gamma}$ or rIL-4, the concentration of nitrite was related with cytotokity of macrophages against T. vaginalis, but the cytotoxicity of macrophages cultured with rIL-2 and $rIFN-{\gamma}$ was decreased in spite of its high production of llitrite. From the results obtained, it is assumed that rIL-2 and $rIFN-{\gamma}$ enhance the cytotoxicity of macrophages while rIL-4 inhibits the cytotoxicity against T. vaginalis, and that the production of nitrite does not relate with the cytotoxicity of macrophages, but nitric oxide may play a role as an inhibitory factor on the proliferation of T. vaginalis.

정상 BALB/c 마우스의 복강에서 분리한 대식세포를 조제 림포카인 및 재조합 림포카인 (rGM-CSF, rIL-2, rIL-4, $rIFN-{\gamma}$)으로 18 시간 반응시켜 활성화시키고 대식세포 $1{\times}10^5$ 개와 $methyl-[^3H]-thymidine$으로 표지된 질트리코모나스 $1{\times}10^4$ 개를 10:1 비율로 넣고 18 시간 동안 $37^{\circ}C$, 5% $CO^2$ 항온항습기에서 반응시킨 다음 상청액 $100{\;}{\mu}l$씩을 scintillation cocktail 2ml에 넣어 방사능을 측정하여 세포독성을 관찰하고 nitrite 생산량을 측정하였다. rIL-2 40U/m1나 $rIFN-{\gamma}$ 100U/ml로 활성화시킨 대식세포는 대조군 보다 높은 세포독성을 보였으나 2개의 림포카인을 촌합시켰을때 낮은 세포독성을 보였다. rIL-4를 rGM-CSF, rIL-2 및 $rIFN-{\gamma}$와 각각 혼합시켰을 때 각 림포카인에 의한 대식세포의 세포독성은 감소되었다. 조제 림포 카인에서 IL-2에 대한 항체는 세포독성을 현저히 감소시켰다. Nitrite 생산량은 $rIFN-{\gamma}$ 및 rIL-4 각각으로 대식세포를 활성화시켰을 때 세포독성 정도와 비례되었고, rIL-2와 $rIFN-{\gamma}$을 혼합 시켰을 때 nitrite 생산량이 높고 증식이 억제되었으나 세포독성은 낮았다. 이러한 결과로 보아 rIL-2 및 $rIFN-{\gamma}$는 각각 단독으로도 대식세포를 활성화시켜 질트리모나스에 세포독성을 나타내며 rIL-4 는 세포독성을 억제시키는 것으로 생각되며, nitrite 생산량은 세포독성과 반드시 비례하지 않으나 nitric oxide가 질트리코모나스의 증식을 억제하는 것으로 생각된다.