Lang A B, Fürer E, Schürch U, Cross A S, Larrick J W, Cryz S J
Swiss Serum and Vaccine Institute, Berne.
Dev Biol Stand. 1990;71:121-6.
A number of human monoclonal antibodies (HmAb) recognizing type-specific determinants expressed by the lipopolysaccharide (LPS) of Pseudomonas aeruginosa and by the capsular polysaccharide (CPS) of Klebsiella were generated for potential treatment of nosocomial infections. The goal is to administer these type-specific HmAb prophylactically as a "cocktail" providing broad coverage. Lymphoblastoid cell lines (LCL) secreting HmAb recognizing P. aeruginosa LPS, toxin A or Klebsiella CPS were obtained by Epstein Barr Virus (EBV) transformation of peripheral blood lymphocytes (PBL) from donors immunized with either a polyvalent Klebsiella CPS or P. aeruginosa O-polysaccharide-toxin A conjugate vaccine. LCL secreting antibodies of the desired specificities were fused to a heteromyeloma cell line. Stable clones were selected by limiting dilution. Hybridomas secreting IgM HmAb which recognized P. aeruginosa Habs serotype 3 and 4 and all 7 Fisher immunotypes were isolated. All were able to prevent fatal experimental P. aeruginosa sepsis in mice when passively transferred. In addition, 4 lines secreting IgG HmAb which neutralize the cytotoxic activity of toxin A were characterized. IgM and IgA secreting hybridoma cells with specificity for Klebsiella CPS of 22 different serotypes were also isolated. Preliminary studies indicate that these HmAb are opsonic.
为了潜在治疗医院感染，制备了多种识别铜绿假单胞菌脂多糖（LPS）和肺炎克雷伯菌荚膜多糖（CPS）所表达的型特异性决定簇的人单克隆抗体（HmAb）。目标是将这些型特异性HmAb作为“鸡尾酒”预防性给药，以提供广泛的覆盖范围。通过用多价肺炎克雷伯菌CPS或铜绿假单胞菌O-多糖-毒素A结合疫苗免疫供体的外周血淋巴细胞（PBL）进行爱泼斯坦-巴尔病毒（EBV）转化，获得了分泌识别铜绿假单胞菌LPS、毒素A或肺炎克雷伯菌CPS的HmAb的淋巴母细胞系（LCL）。将分泌所需特异性抗体的LCL与异源骨髓瘤细胞系融合。通过有限稀释选择稳定克隆。分离出分泌识别铜绿假单胞菌Habs血清型3和4以及所有7种费舍尔免疫型的IgM HmAb的杂交瘤。当被动转移时，所有这些抗体都能够预防小鼠致命的实验性铜绿假单胞菌败血症。此外，还鉴定了4个分泌中和毒素A细胞毒活性的IgG HmAb的细胞系。还分离出了对22种不同血清型的肺炎克雷伯菌CPS具有特异性的分泌IgM和IgA的杂交瘤细胞。初步研究表明，这些HmAb具有调理作用。
