Dziarski R
J Immunol. 1984 Nov;133(5):2537-44.
Using a plaque assay with immunoglobulin (Ig)-coated SRBC, we and others have previously reported that the majority of polyclonally activated mouse lymphocytes secreted antibodies that appeared to be IgM anti-IgG autoantibodies. Careful reexamination of this assay, with application of several highly purified mouse serum and myeloma IgG and IgM preparations, revealed that IgM, which was a minor contaminant of Ig preparations, rather than IgG, was responsible for the formation of these plaques. High numbers of plaques could also be detected in assays with polyclonally activated human lymphocytes, Ig-coated SRBC, and anti-Ig developing sera. Of all IgG-, IgM- or IgA-secreting cells, 40 to 100% were detected with SRBC coated with gamma-globulin or Ig of the same isotype as the isotype to which the developing serum was specific; in general, low proportions of all PFC were detected with SRBC coated with Ig of a different isotype. Studies on the sequence of events leading to the formation of plaques with Ig-sensitized SRBC (both in humans and mice) revealed that antibodies detected in these assays were not able to bind to the Ig-coated SRBC (without the presence of developing serum), and therefore were not anti-Ig autoantibodies. It is our conclusion that the plaque assays with Ig-coated SRBC represent another type of a reverse hemolytic PFC assay that detects cells secreting antibodies regardless of their specificity, and these plaques are formed due to the cross-linking by the anti-Ig developing serum of the Ig coated on SRBC and the Ig secreted by lymphocytes. Our results confirmed preferential induction of anti-DNA antibody secreting cells in mice by showing that these antibodies indeed bind to DNA coated on SRBC. In cultures of polyclonally activated human lymphocytes, anti-DNA and anti-erythrocyte autoantibody-secreting cells were over 10 to 100 times less frequent than in mice. These results, therefore, disprove the concept of preferential induction of anti-Ig autoantibodies in the polyclonal activation of mouse and human lymphocytes, and show that anti-DNA and anti-erythrocyte autoantibodies are easily induced in the polyclonal activation of mouse, but not human, lymphocytes.
通过使用免疫球蛋白（Ig）包被的绵羊红细胞（SRBC）进行空斑试验，我们和其他人之前曾报道，大多数多克隆激活的小鼠淋巴细胞分泌的抗体似乎是IgM抗IgG自身抗体。在应用几种高度纯化的小鼠血清以及骨髓瘤IgG和IgM制剂对该试验进行仔细重新检查后发现，是作为Ig制剂中微量污染物的IgM而非IgG导致了这些空斑的形成。在多克隆激活的人淋巴细胞、Ig包被的SRBC和抗Ig显色血清的试验中也能检测到大量空斑。在所有分泌IgG、IgM或IgA的细胞中，40%至100%可通过包被有与显色血清特异性针对的同种型相同的γ球蛋白或Ig的SRBC检测到；一般来说，用包被有不同同种型Ig的SRBC检测到的所有空斑形成细胞（PFC）比例较低。对导致用Ig致敏的SRBC形成空斑的事件顺序的研究（在人和小鼠中均有）表明，在这些试验中检测到的抗体无法结合到Ig包被的SRBC上（在没有显色血清的情况下），因此不是抗Ig自身抗体。我们的结论是，用Ig包被的SRBC进行的空斑试验代表了另一种类型的反向溶血空斑形成细胞试验，该试验可检测分泌抗体的细胞，而不考虑其特异性，并且这些空斑是由于抗Ig显色血清使包被在SRBC上的Ig与淋巴细胞分泌的Ig交联而形成的。我们的结果通过证明这些抗体确实能结合到包被在SRBC上的DNA，证实了小鼠中抗DNA抗体分泌细胞的优先诱导。在多克隆激活的人淋巴细胞培养物中，抗DNA和抗红细胞自身抗体分泌细胞的频率比小鼠低10至100倍以上。因此，这些结果反驳了在小鼠和人淋巴细胞的多克隆激活中优先诱导抗Ig自身抗体的概念，并表明抗DNA和抗红细胞自身抗体在小鼠而非人淋巴细胞的多克隆激活中容易被诱导。
