Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA

Jian Zhao, Brendan M. Giles, Rhonda L. Taylor, Gabriel A. Yette, Kara M. Lough, Han Leng Ng, Lawrence J. Abraham, Hui Wu, Jennifer A. Kelly, Stuart B. Glenn, Adam J. Adler, Adrienne H. Williams, Mary E. Comeau, Julie T. Ziegler, Miranda Marion, Marta E. Alarcón-Riquelme, Graciela S. Alarcón, Juan Manuel Anaya, Sang Cheol Bae, Dam KimHye Soon Lee, Lindsey A. Criswell, Barry I. Freedman, Gary S. Gilkeson, Joel M. Guthridge, Chaim O. Jacob, Judith A. James, Diane L. Kamen, Joan T. Merrill, Kathy Moser Sivils, Timothy B. Niewold, Michelle A. Petri, Rosalind Ramsey-Goldman, John D. Reveille, R. Hal Scofield, Anne M. Stevens, Luis M. Vilá, Timothy J. Vyse, Kenneth M. Kaufman, John B. Harley, Carl D. Langefeld, Patrick M. Gaffney, Elizabeth E. Brown, Jeffrey C. Edberg, Robert P. Kimberly, Daniela Ulgiati, Betty P. Tsao, Susan A. Boackle, Johan Frostegård, Lennart Truedsson, Enrique De Ramón, José M. Sabio, María F. González-Escribano, Javier Martin, Norberto Ortego-Centeno, José Luis Callejas, Julio Sánchez-Román, Sandra D'Alfonso, Sergio Migliarese, Gian Domenico Sebastiani, Mauro Galeazzi, Torsten Witte, Bernard R. Lauwerys, Emoke Endreffy, László Kovács, Carlos Vasconcelos, Berta Martins Da Silva, R. Scherbarth, Pilar C. Marino, Estela L. Motta, Susana Gamron, Cristina Drenkard, Emilia Menso, Alberto Allievi, Guillermo A. Tate, Jose L. Presas, Simon A. Palatnik, Marcelo Abdala, Mariela Bearzotti, Alejandro Alvarellos, Francisco Caeiro, Ana Bertoli, Sergio Paira, Susana Roverano, Cesar E. Graf, Estela Bertero, Cesar Caprarulo, Griselda Buchanan, Carolina Guillerón, Sebastian Grimaudo, Jorge Manni, Luis J. Catoggio, Enrique R. Soriano, Carlos D. Santos, Cristina Prigione, Fernando A. Ramos, Sandra M. Navarro, Guillermo A. Berbotto, Marisa Jorfen, Elisa J. Romero, Mercedes A. Garcia, Juan C. Marcos, Ana I. Marcos, Carlos E. Perandones, Alicia Eimon, Sanatorio Parque, Cristina G. Battagliotti, Eduardo Acevedo, Mariano Cucho, Ignacio García De La Torre, Mario Cardiel Ríos, José Francisco Moctezuma, Marco Maradiaga Ceceña

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Objectives Systemic lupus erythematosus (SLE; OMIM 152700) is characterised by the production of antibodies to nuclear antigens. We previously identified variants in complement receptor 2 (CR2/CD21) that were associated with decreased risk of SLE. This study aimed to identify the causal variant for this association. Methods Genotyped and imputed genetic variants spanning CR2 were assessed for association with SLE in 15 750 case-control subjects from four ancestral groups. Allele-specific functional effects of associated variants were determined using quantitative real-time PCR, quantitative flow cytometry, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR. Results The strongest association signal was detected at rs1876453 in intron 1 of CR2 (pmeta=4.2×10-4, OR 0.85), specifically when subjects were stratified based on the presence of dsDNA autoantibodies (case-control pmeta=7.6×10-7, OR 0.71; case-only pmeta=1.9×10-4, OR 0.75). Although allele-specific effects on B cell CR2 mRNA or protein levels were not identified, levels of complement receptor 1 (CR1/CD35) mRNA and protein were significantly higher on B cells of subjects harbouring the minor allele (p=0.0248 and p=0.0006, respectively). The minor allele altered the formation of several DNA protein complexes by EMSA, including one containing CCCTC-binding factor (CTCF), an effect that was confirmed by ChIP-PCR. Conclusions These data suggest that rs1876453 in CR2 has long-range effects on gene regulation that decrease susceptibility to lupus. Since the minor allele at rs1876453 is preferentially associated with reduced risk of the highly specific dsDNA autoantibodies that are present in preclinical, active and severe lupus, understanding its mechanisms will have important therapeutic implications.

Original languageEnglish (US)
Pages (from-to)242-252
Number of pages11
JournalAnnals of the Rheumatic Diseases
Volume75
Issue number1
DOIs
StatePublished - Jan 1 2016

Fingerprint

Complement 3d Receptors
Electrophoretic mobility
Alleles
Association reactions
Autoantibodies
Chromatin
Antibodies
Assays
DNA
Cells
Chromatin Immunoprecipitation
Complement Receptors
Electrophoretic Mobility Shift Assay
Nuclear Antigens
Messenger RNA
Proteins
Flow cytometry
Gene expression
B-Lymphocytes
Introns

ASJC Scopus subject areas

  • Rheumatology
  • Immunology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Allergy

Cite this

Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA. / Zhao, Jian; Giles, Brendan M.; Taylor, Rhonda L.; Yette, Gabriel A.; Lough, Kara M.; Ng, Han Leng; Abraham, Lawrence J.; Wu, Hui; Kelly, Jennifer A.; Glenn, Stuart B.; Adler, Adam J.; Williams, Adrienne H.; Comeau, Mary E.; Ziegler, Julie T.; Marion, Miranda; Alarcón-Riquelme, Marta E.; Alarcón, Graciela S.; Anaya, Juan Manuel; Bae, Sang Cheol; Kim, Dam; Lee, Hye Soon; Criswell, Lindsey A.; Freedman, Barry I.; Gilkeson, Gary S.; Guthridge, Joel M.; Jacob, Chaim O.; James, Judith A.; Kamen, Diane L.; Merrill, Joan T.; Sivils, Kathy Moser; Niewold, Timothy B.; Petri, Michelle A.; Ramsey-Goldman, Rosalind; Reveille, John D.; Scofield, R. Hal; Stevens, Anne M.; Vilá, Luis M.; Vyse, Timothy J.; Kaufman, Kenneth M.; Harley, John B.; Langefeld, Carl D.; Gaffney, Patrick M.; Brown, Elizabeth E.; Edberg, Jeffrey C.; Kimberly, Robert P.; Ulgiati, Daniela; Tsao, Betty P.; Boackle, Susan A.; Frostegård, Johan; Truedsson, Lennart; De Ramón, Enrique; Sabio, José M.; González-Escribano, María F.; Martin, Javier; Ortego-Centeno, Norberto; Callejas, José Luis; Sánchez-Román, Julio; D'Alfonso, Sandra; Migliarese, Sergio; Sebastiani, Gian Domenico; Galeazzi, Mauro; Witte, Torsten; Lauwerys, Bernard R.; Endreffy, Emoke; Kovács, László; Vasconcelos, Carlos; Da Silva, Berta Martins; Scherbarth, R.; Marino, Pilar C.; Motta, Estela L.; Gamron, Susana; Drenkard, Cristina; Menso, Emilia; Allievi, Alberto; Tate, Guillermo A.; Presas, Jose L.; Palatnik, Simon A.; Abdala, Marcelo; Bearzotti, Mariela; Alvarellos, Alejandro; Caeiro, Francisco; Bertoli, Ana; Paira, Sergio; Roverano, Susana; Graf, Cesar E.; Bertero, Estela; Caprarulo, Cesar; Buchanan, Griselda; Guillerón, Carolina; Grimaudo, Sebastian; Manni, Jorge; Catoggio, Luis J.; Soriano, Enrique R.; Santos, Carlos D.; Prigione, Cristina; Ramos, Fernando A.; Navarro, Sandra M.; Berbotto, Guillermo A.; Jorfen, Marisa; Romero, Elisa J.; Garcia, Mercedes A.; Marcos, Juan C.; Marcos, Ana I.; Perandones, Carlos E.; Eimon, Alicia; Parque, Sanatorio; Battagliotti, Cristina G.; Acevedo, Eduardo; Cucho, Mariano; De La Torre, Ignacio García; Ríos, Mario Cardiel; Moctezuma, José Francisco; Ceceña, Marco Maradiaga.

In: Annals of the Rheumatic Diseases, Vol. 75, No. 1, 01.01.2016, p. 242-252.

Research output: Contribution to journalArticle

Zhao, J, Giles, BM, Taylor, RL, Yette, GA, Lough, KM, Ng, HL, Abraham, LJ, Wu, H, Kelly, JA, Glenn, SB, Adler, AJ, Williams, AH, Comeau, ME, Ziegler, JT, Marion, M, Alarcón-Riquelme, ME, Alarcón, GS, Anaya, JM, Bae, SC, Kim, D, Lee, HS, Criswell, LA, Freedman, BI, Gilkeson, GS, Guthridge, JM, Jacob, CO, James, JA, Kamen, DL, Merrill, JT, Sivils, KM, Niewold, TB, Petri, MA, Ramsey-Goldman, R, Reveille, JD, Scofield, RH, Stevens, AM, Vilá, LM, Vyse, TJ, Kaufman, KM, Harley, JB, Langefeld, CD, Gaffney, PM, Brown, EE, Edberg, JC, Kimberly, RP, Ulgiati, D, Tsao, BP, Boackle, SA, Frostegård, J, Truedsson, L, De Ramón, E, Sabio, JM, González-Escribano, MF, Martin, J, Ortego-Centeno, N, Callejas, JL, Sánchez-Román, J, D'Alfonso, S, Migliarese, S, Sebastiani, GD, Galeazzi, M, Witte, T, Lauwerys, BR, Endreffy, E, Kovács, L, Vasconcelos, C, Da Silva, BM, Scherbarth, R, Marino, PC, Motta, EL, Gamron, S, Drenkard, C, Menso, E, Allievi, A, Tate, GA, Presas, JL, Palatnik, SA, Abdala, M, Bearzotti, M, Alvarellos, A, Caeiro, F, Bertoli, A, Paira, S, Roverano, S, Graf, CE, Bertero, E, Caprarulo, C, Buchanan, G, Guillerón, C, Grimaudo, S, Manni, J, Catoggio, LJ, Soriano, ER, Santos, CD, Prigione, C, Ramos, FA, Navarro, SM, Berbotto, GA, Jorfen, M, Romero, EJ, Garcia, MA, Marcos, JC, Marcos, AI, Perandones, CE, Eimon, A, Parque, S, Battagliotti, CG, Acevedo, E, Cucho, M, De La Torre, IG, Ríos, MC, Moctezuma, JF & Ceceña, MM 2016, 'Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA', Annals of the Rheumatic Diseases, vol. 75, no. 1, pp. 242-252. https://doi.org/10.1136/annrheumdis-2014-205584
Zhao, Jian ; Giles, Brendan M. ; Taylor, Rhonda L. ; Yette, Gabriel A. ; Lough, Kara M. ; Ng, Han Leng ; Abraham, Lawrence J. ; Wu, Hui ; Kelly, Jennifer A. ; Glenn, Stuart B. ; Adler, Adam J. ; Williams, Adrienne H. ; Comeau, Mary E. ; Ziegler, Julie T. ; Marion, Miranda ; Alarcón-Riquelme, Marta E. ; Alarcón, Graciela S. ; Anaya, Juan Manuel ; Bae, Sang Cheol ; Kim, Dam ; Lee, Hye Soon ; Criswell, Lindsey A. ; Freedman, Barry I. ; Gilkeson, Gary S. ; Guthridge, Joel M. ; Jacob, Chaim O. ; James, Judith A. ; Kamen, Diane L. ; Merrill, Joan T. ; Sivils, Kathy Moser ; Niewold, Timothy B. ; Petri, Michelle A. ; Ramsey-Goldman, Rosalind ; Reveille, John D. ; Scofield, R. Hal ; Stevens, Anne M. ; Vilá, Luis M. ; Vyse, Timothy J. ; Kaufman, Kenneth M. ; Harley, John B. ; Langefeld, Carl D. ; Gaffney, Patrick M. ; Brown, Elizabeth E. ; Edberg, Jeffrey C. ; Kimberly, Robert P. ; Ulgiati, Daniela ; Tsao, Betty P. ; Boackle, Susan A. ; Frostegård, Johan ; Truedsson, Lennart ; De Ramón, Enrique ; Sabio, José M. ; González-Escribano, María F. ; Martin, Javier ; Ortego-Centeno, Norberto ; Callejas, José Luis ; Sánchez-Román, Julio ; D'Alfonso, Sandra ; Migliarese, Sergio ; Sebastiani, Gian Domenico ; Galeazzi, Mauro ; Witte, Torsten ; Lauwerys, Bernard R. ; Endreffy, Emoke ; Kovács, László ; Vasconcelos, Carlos ; Da Silva, Berta Martins ; Scherbarth, R. ; Marino, Pilar C. ; Motta, Estela L. ; Gamron, Susana ; Drenkard, Cristina ; Menso, Emilia ; Allievi, Alberto ; Tate, Guillermo A. ; Presas, Jose L. ; Palatnik, Simon A. ; Abdala, Marcelo ; Bearzotti, Mariela ; Alvarellos, Alejandro ; Caeiro, Francisco ; Bertoli, Ana ; Paira, Sergio ; Roverano, Susana ; Graf, Cesar E. ; Bertero, Estela ; Caprarulo, Cesar ; Buchanan, Griselda ; Guillerón, Carolina ; Grimaudo, Sebastian ; Manni, Jorge ; Catoggio, Luis J. ; Soriano, Enrique R. ; Santos, Carlos D. ; Prigione, Cristina ; Ramos, Fernando A. ; Navarro, Sandra M. ; Berbotto, Guillermo A. ; Jorfen, Marisa ; Romero, Elisa J. ; Garcia, Mercedes A. ; Marcos, Juan C. ; Marcos, Ana I. ; Perandones, Carlos E. ; Eimon, Alicia ; Parque, Sanatorio ; Battagliotti, Cristina G. ; Acevedo, Eduardo ; Cucho, Mariano ; De La Torre, Ignacio García ; Ríos, Mario Cardiel ; Moctezuma, José Francisco ; Ceceña, Marco Maradiaga. / Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA. In: Annals of the Rheumatic Diseases. 2016 ; Vol. 75, No. 1. pp. 242-252.
@article{39d13ce320a94d8cb50754e29be9d92e,
title = "Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA",
abstract = "Objectives Systemic lupus erythematosus (SLE; OMIM 152700) is characterised by the production of antibodies to nuclear antigens. We previously identified variants in complement receptor 2 (CR2/CD21) that were associated with decreased risk of SLE. This study aimed to identify the causal variant for this association. Methods Genotyped and imputed genetic variants spanning CR2 were assessed for association with SLE in 15 750 case-control subjects from four ancestral groups. Allele-specific functional effects of associated variants were determined using quantitative real-time PCR, quantitative flow cytometry, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR. Results The strongest association signal was detected at rs1876453 in intron 1 of CR2 (pmeta=4.2×10-4, OR 0.85), specifically when subjects were stratified based on the presence of dsDNA autoantibodies (case-control pmeta=7.6×10-7, OR 0.71; case-only pmeta=1.9×10-4, OR 0.75). Although allele-specific effects on B cell CR2 mRNA or protein levels were not identified, levels of complement receptor 1 (CR1/CD35) mRNA and protein were significantly higher on B cells of subjects harbouring the minor allele (p=0.0248 and p=0.0006, respectively). The minor allele altered the formation of several DNA protein complexes by EMSA, including one containing CCCTC-binding factor (CTCF), an effect that was confirmed by ChIP-PCR. Conclusions These data suggest that rs1876453 in CR2 has long-range effects on gene regulation that decrease susceptibility to lupus. Since the minor allele at rs1876453 is preferentially associated with reduced risk of the highly specific dsDNA autoantibodies that are present in preclinical, active and severe lupus, understanding its mechanisms will have important therapeutic implications.",
author = "Jian Zhao and Giles, {Brendan M.} and Taylor, {Rhonda L.} and Yette, {Gabriel A.} and Lough, {Kara M.} and Ng, {Han Leng} and Abraham, {Lawrence J.} and Hui Wu and Kelly, {Jennifer A.} and Glenn, {Stuart B.} and Adler, {Adam J.} and Williams, {Adrienne H.} and Comeau, {Mary E.} and Ziegler, {Julie T.} and Miranda Marion and Alarc{\'o}n-Riquelme, {Marta E.} and Alarc{\'o}n, {Graciela S.} and Anaya, {Juan Manuel} and Bae, {Sang Cheol} and Dam Kim and Lee, {Hye Soon} and Criswell, {Lindsey A.} and Freedman, {Barry I.} and Gilkeson, {Gary S.} and Guthridge, {Joel M.} and Jacob, {Chaim O.} and James, {Judith A.} and Kamen, {Diane L.} and Merrill, {Joan T.} and Sivils, {Kathy Moser} and Niewold, {Timothy B.} and Petri, {Michelle A.} and Rosalind Ramsey-Goldman and Reveille, {John D.} and Scofield, {R. Hal} and Stevens, {Anne M.} and Vil{\'a}, {Luis M.} and Vyse, {Timothy J.} and Kaufman, {Kenneth M.} and Harley, {John B.} and Langefeld, {Carl D.} and Gaffney, {Patrick M.} and Brown, {Elizabeth E.} and Edberg, {Jeffrey C.} and Kimberly, {Robert P.} and Daniela Ulgiati and Tsao, {Betty P.} and Boackle, {Susan A.} and Johan Frosteg{\aa}rd and Lennart Truedsson and {De Ram{\'o}n}, Enrique and Sabio, {Jos{\'e} M.} and Gonz{\'a}lez-Escribano, {Mar{\'i}a F.} and Javier Martin and Norberto Ortego-Centeno and Callejas, {Jos{\'e} Luis} and Julio S{\'a}nchez-Rom{\'a}n and Sandra D'Alfonso and Sergio Migliarese and Sebastiani, {Gian Domenico} and Mauro Galeazzi and Torsten Witte and Lauwerys, {Bernard R.} and Emoke Endreffy and L{\'a}szl{\'o} Kov{\'a}cs and Carlos Vasconcelos and {Da Silva}, {Berta Martins} and R. Scherbarth and Marino, {Pilar C.} and Motta, {Estela L.} and Susana Gamron and Cristina Drenkard and Emilia Menso and Alberto Allievi and Tate, {Guillermo A.} and Presas, {Jose L.} and Palatnik, {Simon A.} and Marcelo Abdala and Mariela Bearzotti and Alejandro Alvarellos and Francisco Caeiro and Ana Bertoli and Sergio Paira and Susana Roverano and Graf, {Cesar E.} and Estela Bertero and Cesar Caprarulo and Griselda Buchanan and Carolina Guiller{\'o}n and Sebastian Grimaudo and Jorge Manni and Catoggio, {Luis J.} and Soriano, {Enrique R.} and Santos, {Carlos D.} and Cristina Prigione and Ramos, {Fernando A.} and Navarro, {Sandra M.} and Berbotto, {Guillermo A.} and Marisa Jorfen and Romero, {Elisa J.} and Garcia, {Mercedes A.} and Marcos, {Juan C.} and Marcos, {Ana I.} and Perandones, {Carlos E.} and Alicia Eimon and Sanatorio Parque and Battagliotti, {Cristina G.} and Eduardo Acevedo and Mariano Cucho and {De La Torre}, {Ignacio Garc{\'i}a} and R{\'i}os, {Mario Cardiel} and Moctezuma, {Jos{\'e} Francisco} and Cece{\~n}a, {Marco Maradiaga}",
year = "2016",
month = "1",
day = "1",
doi = "10.1136/annrheumdis-2014-205584",
language = "English (US)",
volume = "75",
pages = "242--252",
journal = "Annals of the Rheumatic Diseases",
issn = "0003-4967",
publisher = "BMJ Publishing Group",
number = "1",

}

TY - JOUR

T1 - Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA

AU - Zhao, Jian

AU - Giles, Brendan M.

AU - Taylor, Rhonda L.

AU - Yette, Gabriel A.

AU - Lough, Kara M.

AU - Ng, Han Leng

AU - Abraham, Lawrence J.

AU - Wu, Hui

AU - Kelly, Jennifer A.

AU - Glenn, Stuart B.

AU - Adler, Adam J.

AU - Williams, Adrienne H.

AU - Comeau, Mary E.

AU - Ziegler, Julie T.

AU - Marion, Miranda

AU - Alarcón-Riquelme, Marta E.

AU - Alarcón, Graciela S.

AU - Anaya, Juan Manuel

AU - Bae, Sang Cheol

AU - Kim, Dam

AU - Lee, Hye Soon

AU - Criswell, Lindsey A.

AU - Freedman, Barry I.

AU - Gilkeson, Gary S.

AU - Guthridge, Joel M.

AU - Jacob, Chaim O.

AU - James, Judith A.

AU - Kamen, Diane L.

AU - Merrill, Joan T.

AU - Sivils, Kathy Moser

AU - Niewold, Timothy B.

AU - Petri, Michelle A.

AU - Ramsey-Goldman, Rosalind

AU - Reveille, John D.

AU - Scofield, R. Hal

AU - Stevens, Anne M.

AU - Vilá, Luis M.

AU - Vyse, Timothy J.

AU - Kaufman, Kenneth M.

AU - Harley, John B.

AU - Langefeld, Carl D.

AU - Gaffney, Patrick M.

AU - Brown, Elizabeth E.

AU - Edberg, Jeffrey C.

AU - Kimberly, Robert P.

AU - Ulgiati, Daniela

AU - Tsao, Betty P.

AU - Boackle, Susan A.

AU - Frostegård, Johan

AU - Truedsson, Lennart

AU - De Ramón, Enrique

AU - Sabio, José M.

AU - González-Escribano, María F.

AU - Martin, Javier

AU - Ortego-Centeno, Norberto

AU - Callejas, José Luis

AU - Sánchez-Román, Julio

AU - D'Alfonso, Sandra

AU - Migliarese, Sergio

AU - Sebastiani, Gian Domenico

AU - Galeazzi, Mauro

AU - Witte, Torsten

AU - Lauwerys, Bernard R.

AU - Endreffy, Emoke

AU - Kovács, László

AU - Vasconcelos, Carlos

AU - Da Silva, Berta Martins

AU - Scherbarth, R.

AU - Marino, Pilar C.

AU - Motta, Estela L.

AU - Gamron, Susana

AU - Drenkard, Cristina

AU - Menso, Emilia

AU - Allievi, Alberto

AU - Tate, Guillermo A.

AU - Presas, Jose L.

AU - Palatnik, Simon A.

AU - Abdala, Marcelo

AU - Bearzotti, Mariela

AU - Alvarellos, Alejandro

AU - Caeiro, Francisco

AU - Bertoli, Ana

AU - Paira, Sergio

AU - Roverano, Susana

AU - Graf, Cesar E.

AU - Bertero, Estela

AU - Caprarulo, Cesar

AU - Buchanan, Griselda

AU - Guillerón, Carolina

AU - Grimaudo, Sebastian

AU - Manni, Jorge

AU - Catoggio, Luis J.

AU - Soriano, Enrique R.

AU - Santos, Carlos D.

AU - Prigione, Cristina

AU - Ramos, Fernando A.

AU - Navarro, Sandra M.

AU - Berbotto, Guillermo A.

AU - Jorfen, Marisa

AU - Romero, Elisa J.

AU - Garcia, Mercedes A.

AU - Marcos, Juan C.

AU - Marcos, Ana I.

AU - Perandones, Carlos E.

AU - Eimon, Alicia

AU - Parque, Sanatorio

AU - Battagliotti, Cristina G.

AU - Acevedo, Eduardo

AU - Cucho, Mariano

AU - De La Torre, Ignacio García

AU - Ríos, Mario Cardiel

AU - Moctezuma, José Francisco

AU - Ceceña, Marco Maradiaga

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Objectives Systemic lupus erythematosus (SLE; OMIM 152700) is characterised by the production of antibodies to nuclear antigens. We previously identified variants in complement receptor 2 (CR2/CD21) that were associated with decreased risk of SLE. This study aimed to identify the causal variant for this association. Methods Genotyped and imputed genetic variants spanning CR2 were assessed for association with SLE in 15 750 case-control subjects from four ancestral groups. Allele-specific functional effects of associated variants were determined using quantitative real-time PCR, quantitative flow cytometry, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR. Results The strongest association signal was detected at rs1876453 in intron 1 of CR2 (pmeta=4.2×10-4, OR 0.85), specifically when subjects were stratified based on the presence of dsDNA autoantibodies (case-control pmeta=7.6×10-7, OR 0.71; case-only pmeta=1.9×10-4, OR 0.75). Although allele-specific effects on B cell CR2 mRNA or protein levels were not identified, levels of complement receptor 1 (CR1/CD35) mRNA and protein were significantly higher on B cells of subjects harbouring the minor allele (p=0.0248 and p=0.0006, respectively). The minor allele altered the formation of several DNA protein complexes by EMSA, including one containing CCCTC-binding factor (CTCF), an effect that was confirmed by ChIP-PCR. Conclusions These data suggest that rs1876453 in CR2 has long-range effects on gene regulation that decrease susceptibility to lupus. Since the minor allele at rs1876453 is preferentially associated with reduced risk of the highly specific dsDNA autoantibodies that are present in preclinical, active and severe lupus, understanding its mechanisms will have important therapeutic implications.

AB - Objectives Systemic lupus erythematosus (SLE; OMIM 152700) is characterised by the production of antibodies to nuclear antigens. We previously identified variants in complement receptor 2 (CR2/CD21) that were associated with decreased risk of SLE. This study aimed to identify the causal variant for this association. Methods Genotyped and imputed genetic variants spanning CR2 were assessed for association with SLE in 15 750 case-control subjects from four ancestral groups. Allele-specific functional effects of associated variants were determined using quantitative real-time PCR, quantitative flow cytometry, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR. Results The strongest association signal was detected at rs1876453 in intron 1 of CR2 (pmeta=4.2×10-4, OR 0.85), specifically when subjects were stratified based on the presence of dsDNA autoantibodies (case-control pmeta=7.6×10-7, OR 0.71; case-only pmeta=1.9×10-4, OR 0.75). Although allele-specific effects on B cell CR2 mRNA or protein levels were not identified, levels of complement receptor 1 (CR1/CD35) mRNA and protein were significantly higher on B cells of subjects harbouring the minor allele (p=0.0248 and p=0.0006, respectively). The minor allele altered the formation of several DNA protein complexes by EMSA, including one containing CCCTC-binding factor (CTCF), an effect that was confirmed by ChIP-PCR. Conclusions These data suggest that rs1876453 in CR2 has long-range effects on gene regulation that decrease susceptibility to lupus. Since the minor allele at rs1876453 is preferentially associated with reduced risk of the highly specific dsDNA autoantibodies that are present in preclinical, active and severe lupus, understanding its mechanisms will have important therapeutic implications.

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