Repertoire cloning of lupus anti-DNA autoantibodies.

Roben, Paul; Barbas, Shana M.; Sandoval, L; Lecerf, Jean‐Michel; Stollar, B D; Solomon, Andrew; Silverman, Gregg J.

doi:10.1172/jci119111

Cited by 77 publications

(70 citation statements)

References 64 publications

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“…Arginines have been shown to frequently enhance antibody affinity for DNA (27)(28)(29). Here, we show that this situation is clearly also the case for the human antibody 33.F12.…”

Section: Discussionsupporting

confidence: 60%

The evolution of human anti-double-stranded DNA autoantibodies

Wellmann

Letz

Herrmann

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

185

163

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It has been proposed that the anti-double-stranded DNA (dsDNA) response in patients with systemic lupus erythematosus (SLE) is antigen driven and that DNA or nucleosomes select anti-DNA reactive, somatically mutated B cells. We have used site-directed mutagenesis to systematically revert the somatic mutations of two human anti-dsDNA antibodies from SLE patients to analyze the resulting changes in DNA binding as well as binding to other autoantigens. Our data demonstrate that high-affinity binding to dsDNA and nucleosomes is acquired by somatic replacement mutations in a stepwise manner. Reactivity to surface structures of apoptotic cells is acquired by the same somatic mutations that generate high-affinity dsDNA binding. Importantly, revertant antibodies with germ-line V regions did not show any measurable DNA reactivity. We propose that anti-DNA autoantibodies are generated from nonautoreactive B cells during a normal immune response. B cells may acquire autoreactivity de novo during the process of somatic hypermutation. Nucleosomes, if available in lupus patients because of defects in clearing of apoptotic debris, might subsequently positively select high affinity anti-DNA B cells.somatic hypermutation ͉ systemic lupus erythematosus

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“…Arginines have been shown to frequently enhance antibody affinity for DNA (27)(28)(29). Here, we show that this situation is clearly also the case for the human antibody 33.F12.…”

Section: Discussionsupporting

confidence: 60%

The evolution of human anti-double-stranded DNA autoantibodies

Wellmann

Letz

Herrmann

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

185

163

View full text Add to dashboard Cite

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“…Mutation analysis performed with our phage Abs revealed that 40 -50% of the V H and V L genes coding for the Abs in both patients showed evidence of somatic hypermutation, supporting the idea that a significant part of the anti-␣ IIb ␤ 3 response developed in AITP or GT patients is driven by Ag. Our results confirm recent experiments where phage display was successfully used to isolate Abs from individuals with demonstrable serum Ab responses to a variety of Ags, including infectious agents such as HIV-1 (17), mutated protein in malignancy (18), and self Ags in autoimmune diseases (12)(13)(14)(15)(16)19). Isolated Abs have been shown to reflect the in vivo Ab response, and, even if theoretically, the combinatorial approach implies the possibility of pairings of L and H chains not present in the original source tissue.…”

Section: Discussionsupporting

confidence: 89%

“…Autoreactive B cells, even if present in naive blood samples, are certainly not proliferating, and, as a result, the RNA may not be in sufficient amounts to be amplified. Significantly, IgG anti-ds DNA were recovered from the library of a clinically active systemic lupus erythematosus patient, but not from the library of his healthy identical twin (12). Therefore, the minimally mutated Abs in our study may be either pathogenic by themselves or may be the template for the expansion of pathogenic Abs.…”

Section: Discussionmentioning

confidence: 86%

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Human IgG Monoclonal Anti-αIIbβ3-Binding Fragments Derived from Immunized Donors Using Phage Display

Jacobin

Laroche‐Traineau

Little

et al. 2002

The Journal of Immunology

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Previous studies of the immune response in polytransfused Glanzmann thrombasthenia (GT) patients and in autoimmune thrombocytopenic purpura (AITP) have relied on serum analysis and have shown the frequent development of Abs directed against the αIIbβ3 integrin. However, little is known about the molecular diversity of the humoral immune response to αIIbβ3 due to the paucity of mAbs issuing from these pathologies. We have isolated human IgG anti-αIIbβ3 binding fragments using combinatorial libraries of single-chain IgG created from the B cells of a GT and an AITP patient, both with serum Abs. Ab screening was performed using activated platelets or activated αIIbβ3-expressing Chinese hamster ovary cells. Sequencing of selected phage Abs showed that a broad selection of genes from virtually all V gene families had been used, indicating the diversity of the immune response. About one-half of the VH and VL segments of our IgG anti-αIIbβ3 fragments displayed extensive hypermutations in the complementarity-determining region, supporting the idea that an Ag-driven immune response was occurring in both patients. The H chain complementarity-determining region 3 analysis of phage Abs revealed motifs other than the well-known RGD and KQAGDV integrin-binding sequences. To our knowledge, our study is the first to illustrate multiple human IgG anti-αIIbβ3 reactivities and structural variations linked to the anti-platelet human immune response. Human αIIbβ3 Abs preferentially directed against the activated form of the integrin were further characterized because platelet αIIbβ3 inhibitors are potential therapeutic reagents for treating acute coronary syndromes. Currently available αIIbβ3 antagonists do not specifically recognize the activated form of the integrin.

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“…Moreover, the monoclonal anti-oxLDL Fabs reported in this study may be absent in patients during antibody selection from the semi-synthetic combinatorial antibody library. However, studies employing combinatorial libraries are valuable in the analysis of autoantibody structures (Farrar et al, 1997), and antibody repertoires produced from phage display libraries have identical immunologic properties to those present in donor sera (Barbas et al, 1992;Roben et al, 1996).…”

Section: Discussionmentioning

confidence: 99%