Autoreactivity in Human IgG+ Memory B Cells

Slides:



Advertisements
Similar presentations
Comparison of A60 and three glycolipid antigens in an ELISA test for tuberculosis  Nancy Simonney, Jean Michel Molina, Mathieu Molimard, Eric Oksenhendler,
Advertisements

Influence of seasonal exposure to grass pollen on local and peripheral blood IgE repertoires in patients with allergic rhinitis  Yu-Chang B. Wu, PhD,
Peter D. Kwong, John R. Mascola  Immunity 
Volume 43, Issue 5, Pages (November 2015)
MicroRNAs Prevent the Generation of Autoreactive Antibodies
Human CD19 and CD40L deficiencies impair antibody selection and differentially affect somatic hypermutation  Menno C. van Zelm, PhD, Sophinus J.W. Bartol,
Volume 38, Issue 3, Pages (March 2013)
Christoph M. Hammers, Jing Chen, Chenyan Lin, Stephen Kacir, Don L
Volume 40, Issue 6, Pages (June 2014)
Volume 19, Issue 2, Pages (February 2016)
Volume 29, Issue 5, Pages (November 2008)
by Christian H. Ottensmeier, and Freda K. Stevenson
Volume 25, Issue 1, Pages (July 2006)
IgA and IgM VH repertoires in human colon: Evidence for clonally expanded B cells that are widely disseminated  Wolfgang Holtmeier, Andreas Hennemann,
Persistence of the IgE repertoire in birch pollen allergy
Dedicator of cytokinesis 8–deficient patients have a breakdown in peripheral B-cell tolerance and defective regulatory T cells  Erin Janssen, MD, PhD,
Volume 29, Issue 6, Pages (December 2008)
Antibodies and B Cell Memory in Viral Immunity
Volume 45, Issue 2, Pages (August 2016)
Volume 12, Issue 2, Pages (August 2003)
Autoreactivity in Human IgG+ Memory B Cells
Volume 47, Issue 6, Pages e4 (December 2017)
Lauren E. Higdon, Michael P. Cancro  Immunity 
Volume 29, Issue 6, Pages (December 2008)
Volume 38, Issue 2, Pages (February 2013)
Volume 28, Issue 2, Pages (February 2008)
TNF receptor superfamily member 13b (TNFRSF13B) hemizygosity reveals transmembrane activator and CAML interactor haploinsufficiency at later stages of.
Histone Modifications Associated with Somatic Hypermutation
IgG and IgM Autoantibody Differences in Discoid and Systemic Lupus Patients  Benjamin F. Chong, Lin-chiang Tseng, Thomas Lee, Rebecca Vasquez, Quan Z.
Volume 48, Issue 2, Pages e5 (February 2018)
Dedicator of cytokinesis 8–deficient patients have a breakdown in peripheral B-cell tolerance and defective regulatory T cells  Erin Janssen, MD, PhD,
Signaling lymphocytic activation molecule (SLAM)/SLAM-associated protein pathway regulates human B-cell tolerance  Laurence Menard, PhD, Tineke Cantaert,
Volume 25, Issue 3, Pages (September 2006)
Defective B-cell memory in patients with Down syndrome
De novo oligoclonal expansions of circulating plasmablasts in active and relapsing IgG4- related disease  Hamid Mattoo, PhD, Vinay S. Mahajan, MBBS, PhD,
Complex Antigens Drive Permissive Clonal Selection in Germinal Centers
Volume 161, Issue 7, Pages (June 2015)
Volume 11, Issue 3, Pages (September 1999)
Longitudinal analysis of ZIKV E–specific memory B cell responses in the DENV-experienced donors. Longitudinal analysis of ZIKV E–specific memory B cell.
Volume 28, Issue 6, Pages (December 2007)
Volume 29, Issue 2, Pages (August 2008)
Volume 12, Issue 5, Pages (May 2000)
Novel Role of the Ras Cascade in Memory B Cell Response
Volume 9, Issue 1, Pages (July 1998)
Volume 161, Issue 7, Pages (June 2015)
Volume 43, Issue 3, Pages (September 2015)
Volume 42, Issue 2, Pages (February 2015)
Volume 28, Issue 2, Pages (February 2008)
Fluorescent In Vivo Detection Reveals that IgE+ B Cells Are Restrained by an Intrinsic Cell Fate Predisposition  Zhiyong Yang, Brandon M. Sullivan, Christopher D.C.
Volume 44, Issue 4, Pages (April 2016)
Increased Junctional Diversity in Fetal B Cells Results in a Loss of Protective Anti- Phosphorylcholine Antibodies in Adult Mice  Cindy L Benedict, John.
Histone Modifications Associated with Somatic Hypermutation
Volume 25, Issue 9, Pages e4 (November 2018)
Volume 32, Issue 1, Pages (January 2010)
Volume 4, Issue 4, Pages (October 2008)
Volume 27, Issue 1, Pages (July 2007)
Vaccine Adjuvants Alter TCR-Based Selection Thresholds
VH usage of cross-reactive B cells induced by H5N1 or H7N9 vaccination
Figure 1 Central B cell tolerance is compromised in patients with NMOSD. Recombinant antibodies (rIgGs) derived from ... Figure 1 Central B cell tolerance.
Molecular and functional characterization of antibodies from PfCSP-reactive memory B cells and plasmablasts. Molecular and functional characterization.
Volume 17, Issue 6, Pages (November 2016)
Increased anti-DNA Abs in 2KO-Bcl6TC mice.
Antigen Selection of Anti-DSG1 Autoantibodies During and Before the Onset of Endemic Pemphigus Foliaceus  Ye Qian, Stephen H. Clarke, Valeria Aoki, Gunter.
Volume 13, Issue 1, Pages (July 2000)
Volume 31, Issue 5, Pages (November 2009)
Antibody deficiency in patients with ataxia telangiectasia is caused by disturbed B- and T-cell homeostasis and reduced immune repertoire diversity  Gertjan.
Affinity maturation of high-affinity human PfCSP NANP antibodies
Volume 28, Issue 5, Pages (May 2008)
Bcl-2 Obstructs Negative Selection of Autoreactive, Hypermutated Antibody V Regions during Memory B Cell Development  Shailaja Hande, Evangelia Notidis,
Volume 25, Issue 4, Pages e3 (October 2018)
Presentation transcript:

Autoreactivity in Human IgG+ Memory B Cells Thomas Tiller, Makoto Tsuiji, Sergey Yurasov, Klara Velinzon, Michel C. Nussenzweig, Hedda Wardemann  Immunity  Volume 26, Issue 2, Pages 205-213 (February 2007) DOI: 10.1016/j.immuni.2007.01.009 Copyright © 2007 Elsevier Inc. Terms and Conditions

Figure 1 IgH and IgL Chain Gene Features from IgG Memory B Cell Antibodies (A) IgG subclass distribution in single IgG+CD27+ memory B cells from three healthy donors. (B) The number of mutations in VH, Vκ, and Vλ genes in antibodies from IgG+ memory B cells. (C) The frequency of mutations in VH, Vκ, and Vλ genes in antibodies from IgG+ memory B cells calculated from the number of replacement (R; black bar) and silent (S; white bar) nucleotide exchanges per base pair in FWRs and CDRs. The R/S ratio for each region is indicated. (D) IgH V and J gene repertoire and IgH CDR3 positive charges and length in amino acids (aa) of antibodies from IgG+ memory B cells compared to mature naive B cells. Pie charts depict VH and JH family usage and the proportion of IgH CDR3s with 0, 1, 2, or ≥3 positive charges. p values for VH and JH usage and IgH CDR3-positive charges were calculated by 2 × 5 Fisher exact test. Bar graphs show frequencies of IgH CDR3s with 9 aa (white bars), 10–14 aa (light gray bars), 15–19 aa (dark gray bars), and ≥20 aa (black bars). p values for IgH CDR3 aa length were calculated by two-tailed nonparametric t test. The absolute number of sequences analyzed is indicated in the center of each pie chart. Values for mature naive B cells in this and other figures were published previously and are shown here for comparison (Tsuiji et al., 2006; Wardemann et al., 2003). Immunity 2007 26, 205-213DOI: (10.1016/j.immuni.2007.01.009) Copyright © 2007 Elsevier Inc. Terms and Conditions

Figure 2 Self-Reactive Antibodies Are Enriched in the IgG+ Memory B Cell Pool (A) IgG+ memory B cell antibodies from healthy donors were tested for self-reactivity by HEp-2 cell lysate ELISA and IFA. Horizontal line shows cut-off OD405 for positive reactivity determined by comparison to low positive control serum (red line). Typical HEp-2 cell IFA staining patterns of antibodies cloned from IgG+ memory B cells are shown. Data shown are representative for at least two independent experiments. p values were calculated by 2 × 2 Fisher exact test. (B) Pie charts summarizing the frequency of HEp-2 self-reactive IgG+ memory B cell antibodies with nuclear (black), nuclear plus cytoplasmic (dark gray), and cytoplasmic (light gray) IFA staining patterns, and the frequency of nonreactive antibodies (white) in comparison to mature naive and IgM+ memory B cell clones (Tsuiji et al., 2006; Wardemann et al., 2003). The number of tested antibodies is indicated in each pie chart center. p values are in comparison to mature naive B cells and IgM+ memory B cells (Tsuiji et al., 2006; Wardemann et al., 2003) and were calculated by 2 × 5 Fisher exact test. (C) Serum IgG ANA amounts of three healthy donors and one patient with systemic lupus erythematosus (SLE101; Yurasov et al., 2005) were determined by HEp-2-ANA-ELISA. The manufacturer's instructions were followed to calculate relative units based on internal positive and negative control sera. Horizontal lines show positive, low positive, or negative cut-off titres as indicated. Immunity 2007 26, 205-213DOI: (10.1016/j.immuni.2007.01.009) Copyright © 2007 Elsevier Inc. Terms and Conditions

Figure 3 Polyreactive Antibodies Contribute to the IgG+ Memory B Cell Compartment IgG+ memory B cell antibodies from healthy donors were tested for polyreactivity by ELISA with dsDNA, ssDNA, LPS, and insulin. Dotted lines represent the high-positive control antibody ED38 (Meffre et al., 2004). Horizontal lines show cut-off OD405 for positive reactivity as determined by comparison to the negative control antibody mGO53 (green line) and low-positive control eiJB40 (red line; Wardemann et al., 2003). Data shown are representative for at least three independent experiments. Pie charts show the frequency of polyreactive clones from IgG+ memory B cells from all three donors compared to mature naive B cell and IgM+ memory B cell antibodies (Tsuiji et al., 2006; Wardemann et al., 2003). The number of tested antibodies is indicated in the center. p values are in comparison to mature naive B cells and IgM+ memory B cells (Tsuiji et al., 2006; Wardemann et al., 2003) and were calculated by 2 × 2 Fisher exact test. Immunity 2007 26, 205-213DOI: (10.1016/j.immuni.2007.01.009) Copyright © 2007 Elsevier Inc. Terms and Conditions

Figure 4 Somatic Hypermutation Contributes to Self-Reactivity in IgG Memory B Cell Antibodies (A) IgH and IgL chains from IgG+ memory B cell antibodies were reverted into their germline counterparts by PCR. Recombinant polyreactive (top left) and nonpolyreactive (bottom left) IgG+ memory B cell antibodies and their germline counterparts (right) were tested for polyreactivity by ELISA with dsDNA, ssDNA, insulin, and LPS. Representative graphs with dsDNA as antigen are shown. Dotted lines represent the high-positive control antibody ED38 (Meffre et al., 2004). Horizontal lines show cut-off OD405 for positive reactivity as determined by comparison to the previously published control antibodies mGO53 (negative control: green line; Wardemann et al., 2003) and eiJB40 (low-positive control: red line; Wardemann et al., 2003). Data shown are representative for ELISA reactivity measured with all four antigens. Reactivity with each antigen was tested in at least three independent experiments. (B) Typical HEp-2 cell IFA staining patterns of mutated IgG+ memory B cell antibodies (top) and their germline counterparts (bottom). Data shown are representative of two independent experiments. Immunity 2007 26, 205-213DOI: (10.1016/j.immuni.2007.01.009) Copyright © 2007 Elsevier Inc. Terms and Conditions

Figure 5 Monomeric IgM from Human Serum Is Less Self-Reactive than Serum IgG Polymeric IgM from pooled human plasma of healthy donors (open circles) and from serum of two healthy donors (PN, HW) was reduced under mild conditions to its monomeric IgM subunits (filled circles). Serum IgG antibodies from pooled human plasma of healthy donors and from three healthy donors (PN, HW, VB) were purified with Protein-G beads (squares). Equal molar quantities of the antibodies were tested by ELISA for reactivity against dsDNA, but data shown are also representative for ELISA reactivity with ssDNA, insulin, and LPS. Reactivity ELISAs with individual antigens were repeated at least three times. Control antibodies were the highly polyreactive ED38 (dotted line; Meffre et al., 2004) and negative mGO53 (triangles; Wardemann et al., 2003). Immunity 2007 26, 205-213DOI: (10.1016/j.immuni.2007.01.009) Copyright © 2007 Elsevier Inc. Terms and Conditions