Zuo L, Guo H, Zhang C
Department of Microbiology, Guiyang Medical College.
Zhonghua Yi Xue Za Zhi. 1996 Aug;76(8):597-9.
To study the measure for the reconstitution of severe combined immunodeficient disease mice (SCID mice) transplanted with human lymphocytes immunized with specific antigen in vitro.
Hu-SCID mice were constructed by intraperitoneal (i.p.) transfer of human tonsillar lymphocytes (TLC) first immunized with dengue virus-2 (DV2) in vitro, then boostered with different doses of DV2 antigen in vivo.
Human immunoglobulin (Ig) and specific human anti-DV2 antibodies could be detected in sera of the hu-SCID mice by ELISA. Two weeks after transplantion of the in vitro-immunized hu-TLC, the highest human IgG concentration was 412.86 micrograms/ml, two or four weeks later specific human anti-DV2 IgM or IgG and the neutralizing antibody could found in these sera, respectively, the highest titer was 1:6400. The levels of human IgG and human anti-DV2 antibodies were related to the dose of DV2 antigen. The presence of human B cells (CD20+) and T/cells (CD3+, CD4+, CD8+) were shown in the sleen, lung, intestine and mesentery of the hu-SCID mice as determined by immunoperoxidase staining, and the cells were distributed correspondingly with the lymphocyte cells in proper human tissue. Human anti-DV2 IgG could also be detected in the spleen cell culture obtained from the SCID mice.
By co-immunized in vitro and in vivo the SCID mice deduced stronger immune response for specific antigen, and the reconstitution of the human lymphocytes could be established successfully.
研究体外经特异性抗原免疫的人淋巴细胞移植的重症联合免疫缺陷病小鼠(SCID小鼠)的重建方法。
通过腹腔内(i.p.)注射体外先用登革病毒2(DV2)免疫,然后在体内用不同剂量DV2抗原加强免疫的人扁桃体淋巴细胞(TLC)构建人源化SCID小鼠(Hu-SCID小鼠)。
通过ELISA可在Hu-SCID小鼠血清中检测到人免疫球蛋白(Ig)和特异性人抗DV2抗体。体外免疫的人TLC移植后两周,人IgG最高浓度为412.86微克/毫升,两到四周后分别在这些血清中可检测到特异性人抗DV2 IgM或IgG以及中和抗体,最高滴度为1:6400。人IgG和人抗DV2抗体水平与DV2抗原剂量有关。通过免疫过氧化物酶染色在Hu-SCID小鼠的脾脏、肺、肠和肠系膜中显示有人B细胞(CD20+)和T细胞(CD3+、CD4+、CD8+),且这些细胞的分布与正常人体组织中的淋巴细胞相应。从SCID小鼠获得的脾细胞培养物中也可检测到人抗DV2 IgG。
通过体外和体内联合免疫,SCID小鼠对特异性抗原产生更强的免疫反应,可成功建立人淋巴细胞的重建。