The importance of somatic mutations in the Vλ gene 2a2 in human monoclonal anti-DNA antibodies

Rahman, Anisur; Haley, Joanna; Radway-Bright, E L; Nagl, Sylvia; Low, D.G.; Latchman, David S.; Isenberg, David

doi:10.1006/jmbi.2000.4491

Cited by 28 publications

(53 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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.

190

163

View full text Add to dashboard Cite

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

show abstract

“…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.

190

163

View full text Add to dashboard Cite

show abstract

“…The loss of this light chain R has not previously been found to enhance binding to any antigen (20,31).…”

Section: Demonstrationmentioning

confidence: 89%

“…Human IgG was purified from the supernatant using a protein A column, and the product was analyzed for purity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and quantified by spectrophotometry. Purified human IgG was sent back to our institution, where the amount of whole IgG antibody was quantified by ELISA as previously described (20,25,30).…”

Section: Lambrianides Et Almentioning

confidence: 99%

“…In previous experiments we found that a computer model-based prediction that R27a in B3 V L was important in binding dsDNA/nucleosomes was confirmed by a reduction in binding when R27a was altered to serine (S) (20). In the present study we used similar techniques to investigate the importance of the R53 V H in binding to a range of antigens, in order to test the hypothesis that the introduction of this R plays a critical role in making B3 a high-affinity antinucleosome antibody.…”

mentioning

confidence: 90%

“…Antigen-driven somatic mutation in many of these highaffinity anti-dsDNA monoclonal antibodies results in accumulation of arginine (R), asparagine (N), and lysine (K) in the CDRs (13,15,16) In vitro expression experiments have shown that altering R residues in the CDRs of these antibodies can lead to significant alterations in binding to dsDNA (17)(18)(19)(20)(21), but few studies have investigated the influence of particular somatic mutations on binding to nucleosomes or ␣-actinin.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Arginine mutation alters binding of a human monoclonal antibody to antigens linked to systemic lupus erythematosus and the antiphospholipid syndrome

Lambrianides¹,

Giles²,

Ioannou³

et al. 2007

Arthritis & Rheumatism

Self Cite

View full text Add to dashboard Cite

Objective. Previous studies have shown the importance of somatic mutations and arginine residues in the complementarity-determining regions (CDRs) of pathogenic anti-double-stranded DNA (anti-dsDNA) antibodies in human and murine lupus, and in studies of murine antibodies, a role of mutations at position 53 in V H CDR2 has been demonstrated. We previously demonstrated in vitro expression and mutagenesis of the human IgG1 monoclonal antibody B3. The present study was undertaken to investigate, using this expression system, the importance of the arginine residue at position 53 (R53) in B3 V H .Methods. R53 was altered, by site-directed mutagenesis, to serine, asparagine, or lysine, to create 3 expressed variants of V H . In addition, the germline sequence of the V H 3-23 gene (from which B3 V H is derived) was expressed either with or without arginine at position 53. These 5 new heavy chains, as well as wild-type B3 V H , were expressed with 4 different light chains, and the resulting antibodies were assessed for their ability to bind to nucleosomes, ␣-actinin, cardiolipin, ovalbumin, ␤ 2 -glycoprotein I (␤ 2 GPI), and the N-terminal domain of ␤ 2 GPI (domain I), using direct binding assays.Results. The presence of R53 was essential but not sufficient for binding to dsDNA and nucleosomes. Conversely, the presence of R53 reduced binding to ␣-actinin, ovalbumin, ␤ 2 GPI, and domain I of ␤ 2 GPI. The combination B3 (R53S) V H /B3 V L bound human, but not bovine, ␤ 2 GPI.Conclusion. The fact that the R53S substitution significantly alters binding of B3 to different clinically relevant antigens, but that the alteration is in opposite directions depending on the antigen, implies that this arginine residue plays a critical role in the affinity maturation of antibody B3.

show abstract

Preparation and characterization of organic–inorganic hybrid composites based on multiepoxy silsesquioxane and cyanate resin

Liang

Guo

2006

J of Applied Polymer Sci

View full text Add to dashboard Cite

A multiepoxy cubic silsesquioxane was prepared by the hydrolysis and polycondensation of trifunctional monomer (␥-glycidoxypropyl)trimethoxysilane in a solvent mixture of methyl isobutyl ketone and anhydrous ethanol with a tetraethyl ammonium hydroxide aqueous solution acting as the catalyst, and it was successfully introduced into a cyanate resin and formed highly crosslinked organic-inorganic hybrid composites on a molecular level via a mutual cure reaction. The properties of the multiepoxy cubic silsesquioxane/bisphenol A dicyanate ester resin composites were investigated, and the results showed that introducing the cubic silsesquioxane unit into the cyanate resin successfully modified the local structure of the molecule, made the chain more rigid, restricted the chain mobility, and eventually improved the thermal stability and flame retardancy of the resin.

show abstract

The importance of somatic mutations in the Vλ gene 2a2 in human monoclonal anti-DNA antibodies

Cited by 28 publications

References 36 publications

The evolution of human anti-double-stranded DNA autoantibodies

The evolution of human anti-double-stranded DNA autoantibodies

Arginine mutation alters binding of a human monoclonal antibody to antigens linked to systemic lupus erythematosus and the antiphospholipid syndrome

Preparation and characterization of organic–inorganic hybrid composites based on multiepoxy silsesquioxane and cyanate resin

Contact Info

Product

Resources

About