Sconocchia G, Titus J A, Segal D M
Experimental Immunology Branch, National Cancer Institute, Bethesda, MD 20892.
J Immunol. 1994 Dec 15;153(12):5473-81.
Previous studies have shown that target cells that bind to CD44 adhesion molecules on cloned cytotoxic T cells are lysed by the CTL. To determine whether CD44 is also a cytotoxic trigger molecule in human PBL, we tested a bispecific Ab consisting of anti-CD44 Fab cross-linked to a Fab against a target cell Ag, in cytotoxicity assays using PBL as effectors. We found that PBL mediated lysis in the presence of the anti-CD44 bispecific Ab provided that the effector cells were stimulated with either IL-2 or IL-12. Cell fractionation experiments showed that CD44-directed cytolysis was mediated exclusively by CD56+ low buoyant density cells, mainly by NK (CD16+) cells, but also to a lesser extent by CD56+ T cells. CD44-directed cytolysis appeared in these subsets 24 to 48 h after addition of IL-2 and paralleled the acquisition of Ab-independent (LAK) activity; in contrast, these cells mediated Ab-dependent cellular cytotoxicity and CD3 redirected lysis before stimulation with IL-2. Unstimulated CD56+ cells uniformly expressed high levels of CD44 that increased modestly after incubation with IL-2. No changes in isoform expression in the extracellular domain of CD44 could be detected upon activation with the use of isoform-specific mAbs. Thus, lymphokine stimulation caused CD44 to become a cytotoxic trigger in subsets of PBL that mediated other forms of cytotoxicity and expressed CD44 before activation, suggesting that activation of these cells was accompanied by a coupling of CD44 to their lytic machinery.
先前的研究表明,与克隆的细胞毒性T细胞上的CD44黏附分子结合的靶细胞会被细胞毒性T淋巴细胞(CTL)裂解。为了确定CD44在人外周血淋巴细胞(PBL)中是否也是一种细胞毒性触发分子,我们在以PBL作为效应细胞的细胞毒性试验中,测试了一种双特异性抗体,该抗体由与针对靶细胞抗原的Fab交联的抗CD44 Fab组成。我们发现,在存在抗CD44双特异性抗体的情况下,PBL介导了细胞裂解,前提是效应细胞用白细胞介素-2(IL-2)或白细胞介素-12(IL-12)进行刺激。细胞分级分离实验表明,针对CD44的细胞溶解完全由CD56 +低浮力密度细胞介导,主要由自然杀伤细胞(NK,CD16 +)介导,但也有较小程度由CD56 + T细胞介导。在添加IL-2后24至48小时,这些亚群中出现了针对CD44的细胞溶解,这与获得抗体非依赖性(LAK)活性平行;相反,这些细胞在被IL-2刺激之前介导了抗体依赖性细胞毒性和CD3重定向裂解。未刺激的CD56 +细胞均匀地表达高水平的CD44,在用IL-2孵育后适度增加。在用异构体特异性单克隆抗体激活后,未检测到CD44细胞外结构域异构体表达的变化。因此,淋巴因子刺激导致CD44在PBL亚群中成为细胞毒性触发因素,这些亚群介导其他形式的细胞毒性并在激活前表达CD44,这表明这些细胞的激活伴随着CD44与其裂解机制的偶联。