Anti-nuclear antibody reactivity in lupus may be partly hard-wired into the primary B-cell repertoire.
When monoclonal ANAs and non-ANAs generated from a genetically simplified mouse model of lupus, B6.Sle1, were recently compared, the ANAs exhibited three sequence motifs in their immunoglobulin heavy chains, including increased cationicity in CDR3 ("motif A"), reduced anionicity in CDR2 ("motif B") and increased aspartate at H50 ("motif C"). The present study was designed to elucidate the extent to which these ANA-associated sequence motifs might be hard-wired into the primary B-cell repertoire in lupus. The immunoglobulin heavy chain sequence of total splenic B-cells, follicular B-cells and marginal zone B-cells from B6.Sle1 congenic mice and C57BL/6 controls were amplified by single-cell PCR and compared. Analysis of the primary immunoglobulin heavy chain repertoire indicated that the first two sequence motifs "A" and "B" were already encoded in the naïve repertoire of B6.Sle1(z) mice, whereas the third motif "C" was introduced in part by somatic mutation. Site-directed mutagenesis confirmed that non-anionic CDR2 and cationic CDR3 residues in the immunoglobulin heavy chain facilitated nuclear antigen binding in concert, whereas aspartate at H50 strongly vetoed DNA-binding, while preserving nucleosome reactivity. Hence, anti-nuclear antibodies appear to arise as a consequence of two distinct processes-genetically programmed selection of specific CDR charge motifs into the primary immunoglobulin repertoire, with secondary contribution from somatic mutation. Polymorphisms in the lupus susceptibility gene Ly108 that impair central B-cell tolerance may be mechanistically responsible for these early repertoire differences in lupus.
Chang
Sooghee
S
Department of Internal Medicine, University of Texas Southwestern Medical School, Dallas, TX 75390, USA.
Yang
Liu
L
Moon
Young Mee
YM
Cho
Young Gyu
YG
Min
So Youn
SY
Kim
Tae Joo
TJ
Kim
Young Joo
YJ
Patrick
Wilson
W
Kim
Ho-Youn
HY
Mohan
Chandra
C
D020816:Amino Acid Motifs / Q000235:genetics / Q000276:immunology; D000818:Animals; D000974:Antibodies, Antinuclear / Q000235:genetics* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D000950:Antigens, Ly / Q000235:genetics* / Q000276:immunology; D034961:Antigens, Nuclear / Q000235:genetics / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D004195:Disease Models, Animal; D007143:Immunoglobulin Heavy Chains / Q000235:genetics* / Q000276:immunology; D008180:Lupus Erythematosus, Systemic / Q000235:genetics* / Q000276:immunology; D051379:Mice; D011110:Polymorphism, Genetic / Q000235:genetics / Q000276:immunology; D000071176:Signaling Lymphocytic Activation Molecule Family
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D000950:Antigens, Ly; D034961:Antigens, Nuclear; D007143:Immunoglobulin Heavy Chains; C000604063:Slamf6 protein, mouse; D000071176:Signaling Lymphocytic Activation Molecule Family
Scand J Immunol. 1998 Sep;48(3):223-32
9743205
Proteins. 1991;11(3):159-75
1749770
Science. 2003 Sep 5;301(5638):1374-7
12920303
J Clin Invest. 2007 Jun;117(6):1558-65
17510706
Methods. 1997 Jan;11(1):20-6
8990085
Arthritis Rheum. 2007 Sep;56(9):3057-69
17763445
Ann N Y Acad Sci. 1997 Apr 5;815:327-30
9186671
J Exp Med. 2004 Jan 19;199(2):255-64
14718514
Arthritis Rheum. 1996 Jun;39(6):894-903
8651982
J Immunol. 1996 Sep 15;157(6):2430-9
8805642
J Exp Med. 1990 Jan 1;171(1):265-92
2104919
J Mol Biol. 1994 Oct 21;243(2):283-97
7523684
Eur J Immunol. 1991 Jul;21(7):1725-31
2060580
Mol Immunol. 1994 Dec;31(18):1377-90
7529877
J Biol Chem. 1987 Jan 15;262(2):643-8
3805000
Science. 2006 Jun 16;312(5780):1665-9
16778059
Mol Immunol. 1993 Aug;30(12):1069-75
8366857
J Exp Med. 1991 Jun 1;173(6):1357-71
1903427
J Immunol. 1997 Dec 15;159(12):6083-90
9550408
J Exp Med. 2004 Feb 2;199(3):381-98
14757744
Curr Biol. 1994 Jul 1;4(7):573-83
7953531
J Immunol. 1993 Jun 1;150(11):4966-77
8496598
Int Immunol. 1995 Jun;7(6):1013-9
7577795
Proc Natl Acad Sci U S A. 1976 Mar;73(3):804-8
1062791
J Exp Med. 1992 Sep 1;176(3):761-79
1512540
Cell. 1996 May 3;85(3):303-6
8616885
J Exp Med. 1994 Sep 1;180(3):925-32
8064241
J Exp Med. 2005 Mar 7;201(5):703-11
15738055
Int Immunol. 2004 Feb;16(2):377-84
14734623
J Biol Chem. 1994 Feb 4;269(5):3615-22
8106406
Nature. 1995 May 25;375(6529):334-8
7753200
Annu Rev Immunol. 1994;12:487-520
8011289
J Biol Chem. 1994 Feb 4;269(5):3623-32
8106407
Immunol Rev. 1992 Aug;128:123-49
1427921
Immunity. 2001 Dec;15(6):947-57
11754816
Immunogenetics. 2000 Jan;51(1):20-9
10663558
J Immunol. 1995 Sep 15;155(6):3223-33
7673735
J Clin Invest. 2005 Nov;115(11):3205-16
16211091
J Exp Med. 1997 Apr 7;185(7):1317-26
9104818
Mol Immunol. 2001 Jan;38(1):9-18
11483206
J Immunol. 1997 Jun 15;158(12):6019-28
9190957
J Immunol. 1994 Jun 15;152(12):5988-96
8207223
Mol Immunol. 2003 Dec;40(8):491-9
14563368
J Exp Med. 1999 May 17;189(10):1639-48
10330443
Mol Immunol. 2009 Aug;46(13):2671-81
19556006
Curr Opin Rheumatol. 2000 Sep;12(5):364-8
10990170
Mol Immunol. 2002 Oct;39(5-6):333-47
12220891
N Engl J Med. 1998 May 7;338(19):1359-68
9571257
J Clin Invest. 1998 Mar 15;101(6):1362-72
9502778
J Immunol. 2003 Oct 1;171(7):3886-94
14500691
Immunol Res. 1994;13(2-3):172-85
7775808