Dersimonian H, Schwartz R S, Barrett K J, Stollar B D
Department of Biochemistry, Tufts University Health Science Campus, Boston, MA.
J Immunol. 1987 Oct 1;139(7):2496-501.
In order to identify the V region genes encoding systemic lupus erythematosus (SLE)-derived anti-DNA autoantibodies, we have determined the nucleotide sequence of heavy chain mRNA from several DNA-binding immunoglobulins secreted by human hybridomas. We used the technique of cDNA primer extension for determining sequences of the VH, D, and JH gene segments of anti-DNA autoantibodies from three different primary hybridoma growths from an SLE patient and one hybridoma from a leprosy patient. Immunoglobulins from two of the SLE hybridomas expressed the same idiotype, Id-16/6, which is also expressed on immunoglobulins in sera of patients with active SLE. Their mRNA sequences showed complete homology to each other in the V, D, and J genes and more than 99% homology to the VH26 germ-line gene sequence, a member of the human VHIII gene family. The VH mRNA sequence of the third SLE hybridoma, 21/28, which was idiotypically unrelated to the other two, was 93% homologous to a different VH germ-line gene sequence, HA2, a member of the human VHI gene family. The fourth anti-DNA-producing hybridoma, 8E10, was derived from a leprosy patient of different ethnic origin than the SLE patient. It was idiotypically related to 21/28 and expressed a VH segment gene identical to that of 21/28. Hybridomas 21/28 and 8E10 shared sequence homology with the VH26 anti-DNA antibodies in the first complementarity-determining region. In addition, 21/28 shared sequence homology with the Id-16/6+ group in the region encoded by the D and J gene segments. Our findings indicate that some SLE autoantibodies are encoded by unmodified or scarcely modified VH germ-line genes that are conserved in the human population and identify two distinct VH germ-line genes that can encode segments of anti-DNA immunoglobulins.
为了鉴定编码系统性红斑狼疮(SLE)来源的抗DNA自身抗体的V区基因,我们测定了人杂交瘤分泌的几种DNA结合免疫球蛋白重链mRNA的核苷酸序列。我们使用cDNA引物延伸技术来确定来自一名SLE患者的三种不同原代杂交瘤生长物以及一名麻风病患者的一种杂交瘤的抗DNA自身抗体的VH、D和JH基因片段的序列。来自两个SLE杂交瘤的免疫球蛋白表达相同的独特型,即Id-16/6,活跃SLE患者血清中的免疫球蛋白也表达该独特型。它们的mRNA序列在V、D和J基因中彼此完全同源,与人类VHIII基因家族成员VH26种系基因序列的同源性超过99%。第三个SLE杂交瘤21/28的VH mRNA序列与其他两个杂交瘤在独特型上不相关,与另一种不同的VH种系基因序列HA2同源性为93%,HA2是人类VHI基因家族的成员。第四个产生抗DNA的杂交瘤8E10来自一名与SLE患者不同种族的麻风病患者。它在独特型上与21/28相关,并且表达与21/28相同的VH片段基因。杂交瘤21/28和8E10在第一个互补决定区与VH26抗DNA抗体具有序列同源性。此外,21/28在由D和J基因片段编码的区域与Id-16/6+组具有序列同源性。我们的研究结果表明,一些SLE自身抗体由在人类群体中保守的未修饰或几乎未修饰的VH种系基因编码,并鉴定出两个不同的VH种系基因,它们可以编码抗DNA免疫球蛋白的片段。