近交系 BALB/c 및 Nude 마우스 腹腔渗出細胞와 血淸의 肝吸蟲 感染에 대한 免疫應答 = Immune Response to Clonorchis sinensis Infection by Peritoneal Exudate Cells and Serum in Nude and BALB/c Mice

국문 초록 (Abstract)

近交系 nude 마우스가 肝吸蟲 感染염에 대한 免疫의 移入에 關與하는지 究明하기 위하여 肝吸蟲 被襄幼蟲의 經口感染과 肝吸蟲 成蟲의 代射産物의 발바닥에 皮內注射로 感作한 肝吸蟲 및 nude 마우스의 腹腔渗出細胞와 血淸을 함께 recipient BALB/c 및 nude 마우스에 腹腔內注入하였다.
Donor BALB/c 및 nude 마우스는 各各 5마리씩 2群으로 나누어 第Ⅰ群에는 肝吸蟲 被襄幼蟲 20마리를 經口感染시켰고 第Ⅱ群에는 肝吸蟲 成蟲의 代謝産物과 Freund 不完全 adjuvant의 同量混合物 0.1㎖를 발바닥에 2週 間隔으로 2回 皮內注射하였다. 2週後에 痲醉下에 血淸과 腹腔渗出細胞를 採取하여 recipient BALB/c 및 nude 마우스의 腹腔內注入하였다.
Recipinet 마우스는 5마리씩 6群으로 나누어 第1群 BALB/c 마우스에는 第Ⅰ群 donor BALB/c 마우스의 腹腔渗出細胞 5×10 exp(5)과 血淸 0.1㎖를, 第2群 BALB/c 마우스에는 第Ⅱ群 donor BALB/c 마우스의 腹腔渗出細胞 5×10 exp(5)과 血淸 0.1㎖를, 第4群 nude 마우스에는 第Ⅱ群 nude 마우스의 腹腔渗出細胞 5×10 exp(5)과 血淸 0.1㎖를, 第5群 BALB/c 마우스와 第6群 nude 마우스에는 同系마우스의 非感作 腹腔渗出細胞 5×10 exp(5)과 血淸 0.1㎖를 各各 腹腔內注入하여 第1次 感作하였다.
7日 지난 後 6群의 recipient마우스에 肝吸蟲 被襄幼蟲을 20마리씩 challenge 經口感染시켜 第22日부터 EpG를 計算하였으며 challenge後 第50日에 全마우스를 屠殺하여 對照群마우스의 負荷蟲體數와 脾臟當 plaque 形成細胞數를 基準으로 하여 感作群마우스와 有意性을 檢定하여 免疫의 移入與否를 判定하였다.
Challenge 感作後 EpG의 變動에는 感作群과 非感作 對照群 마우스間 및 BALB/c와 nude 마우스間에 有意的 差를 認定할 수 없었다.
肝吸蟲 成蟲의 代謝産物을 발바닥에 皮內注射한 BALB/c 마우스의 腹腔渗出細胞와 血淸을 함께 腹腔內注入받은 BALB/c 마우스의 負荷 蟲體數는 非感作 對照群 同系마우스에 比하여 有意的으로 적었다. 그러나 nude 마우스에서는 非感作 對照群과의 사이에 有意的 差를 認定할 수 없었다.
脾臟에서 plaque 形成細胞는 感作 腹腔渗出細胞와 血淸을 함께 腹腔內注入받은 第2群 BALB/c 마우스에서만 少數 檢出할 수 있었다.

다국어 초록 (Multilingual Abstract)

In order to clarify the role of nude mice in the transfer of immunity against Clonorchis sinensis, the peritoneal exudate cells(PEC) and serum of the BALB/c and nude mice sensitized with the oral infection of C. sinensis metacercariae, and with the injection of the metabolic products of the adult flukes into footpads were injected intraperitoneally to the recipient mice.
The donor BALB/c and nude mice were divided into two groups. The BALB/c and nude mice of Group 1 were infected orally with the metacercariae of C. sinensis and those of Group Ⅱ were sensitized with two injections of the admixture of the metabolic products of adult C. sinensis and Freund's incomplete adjuvant into footpad at two weeks interval.
Two weeks after sensitization, the PEC and serum were collected from the mice under deep anesthesia.
Recipient mice(RM) were divided into six groups. The BALB/c mice of Group 1 were injected intraperitoneally(IP) with 5×10 exp(5) PEC and 0.1㎖ of serum from Group Ⅰdonor BALB/c mice, and those of Group 2 were injected IP with 5×10 exp(5) PEC and 0.1㎖ of serum from Group Ⅱ BALB/c mice. The nude mice of Group 3 were injected IP with 5×10 exp(5) PEC and 0.1㎖ of serum from group Ⅰ donor nude mice and those of Group 4 were injected IP with 5×10 exp(5) PEC and 0.1㎖ of serum from Group Ⅱ donor nude mice.
The control BALB/c and nude mice of Group 5 and Group 6 were injected IP with 5×10 exp(5) PEC and 0.1㎖ of serum from nonsensitized control mice, respectively.
Seven days after primary sensitization, the RM of six groups challenged orally with 20 metacercariae of C. sinensis and Eggs per Gram(EpG) of fecal samples were counted from 22 to 50 days after challenge. The RM were killed 50 days after challenge. The RM were killed 50 days after challenge, and the transfer of immunity to the RM was determined by significant differences in worm burdens and plague forming cells per spleen between sensitized and non-sensitized groups and between BALB/c and nude mice.
There were no significant differences in EpG pattern between the sensitized and non-sensitized groups and between BALB/c and nude mice.
The BALB/c mice given sensitized PEC and serum from Group Ⅱ BALB/c mice had fewer flukes than the non-sensitized group. The difference in worm burdens between sensitized and non-sensitized group were significant by the paired t-test, whereas no significant difference was found in worm burdens between BALB/c and nuce mice.
There were some plague forming cells in the BALB/c mice given immune PEC and serum. However, no plague forming cells were found in the nude mice.