Abstract
B lymphocyte involvement in systemic lupus erythematosus has been recognized for several decades, mainly in the context of autoantibody production. Both mouse and human studies reveal that different types of antibody responses, as well as antibody-independent effector functions can be ascribed to distinct subpopulations (subsets) of circulating B cells. Characterizing human B cell subsets can advance the field of autoimmunity even further by establishing B cell signatures associated with disease severity, progression, and response-to-treatment. For this purpose, we have developed specialized B cell reagent panels for multiparameter flow cytometry, and combine their use with advanced bioinformatics strategies that together will likely be advantageous for improving the characterization, prognosis, and for possibly improving treatment regimens of chronic inflammatory diseases such as lupus.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Pons-Estel GJ et al (2008) Understanding the epidemiology and progression of systemic lupus erythematosis. Semin Arthritis Rheum 39:257–268
Borchers AT et al (2010) The geoepidemiology of systemic lupus erythematosus. Autoimmun Rev 9:A277–A287
Bertsias GK, Salmon JE, Boumpas DT (2010) Therapeutic opportunities in systemic lupus erythematosus: state of the art and prospects for the new decade. Ann Rheum Dis 69:1603–1611
Arbuckle MR et al (2003) Development of autoantibodies before the clinical onset of systemic lupus erythematosus. N Engl J Med 349:1526–1533
Anolik JH et al (2009) Insights into the heterogeneity of human B cells: diverse functions, roles in autoimmunity, and use as therapeutic targets. Immunol Res (2009) 45:144-158
Wardemann H et al (2003) Predominant autoantibody production by early human B cell precursors. Science 301:1374–1377
Yurasov S et al (2005) Defective B cell tolerance checkpoints in systemic lupus erythematosus. J Exp Med 201:703–711
Yurasov S et al (2006) Persistent expression of autoantibodies in SLE patients in remission. J Exp Med 203:2255–2261
Yurasov S et al (2005) B-cell tolerance checkpoints in healthy humans and patients with systemic lupus erythematosus. Ann N Y Acad Sci 1062:165–174
Yurasov S, Nussenzweig MC (2007) Regulation of autoreactive antibodies. Curr Opin Rheumatol 19:421–426
Tiller T et al (2007) Autoreactivity in human IgG+ memory B cells. Immunity 26:205–213
Sherer Y et al (2004) Autoantibody explosion in systemic lupus erythematosus: more than 100 different antibodies found in SLE patients. Semin Arthritis Rheum 34:501–537
Huang W et al (2002) The effect of anti-CD40 ligand antibody on B cells in human systemic lupus erythematosus. Arthritis Rheum 46:1554–1562
Erdei A et al (2009) Expression and role of CR1 and CR2 on B and T lymphocytes under physiological and autoimmune conditions. Mol Immunol 46:2767–2773
Casciola-Rosen LA, Anhalt G, Rosen A (1994) Autoantigens targeted in systemic lupus erythematosus are clustered in two populations of surface structures on apoptotic keratinocytes. J Exp Med 179:1317–1330
Schutters K, Reutelingsperger C (2010) Phosphatidylserine targeting for diagnosis and treatment of human diseases. Apoptosis 15:1072–1082
Pugh-Bernard AE et al (2001) Regulation of inherently autoreactive VH4-34 B cells in the maintenance of human B cell tolerance. J Clin Invest 108:1061–1070
Isenberg D et al (1993) Identification of the 9G4 idiotope in systemic lupus erythematosus. Br J Rheumatol 32:876–882
van Vollenhoven RF et al (1999) VH4-34 encoded antibodies in systemic lupus erythematosus: a specific diagnostic marker that correlates with clinical disease characteristics. J Rheumatol 26:1727–1733
Bhat NM et al (2002) VH4-34 encoded antibody in systemic lupus erythematosus: effect of isotype. J Rheumatol 29:2114–2121
Sanz I, Lee FE (2010) B cells as therapeutic targets in SLE. Nat Rev Rheumatol 6:326–337
Anolik JH et al (2004) Rituximab improves peripheral B cell abnormalities in human systemic lupus erythematosus. Arthritis Rheum 50:3580–3590
Calero I, Sanz I (2010) Targeting B cells for the treatment of SLE: the beginning of the end or the end of the beginning? Discov Med 10:416–424
Shlomchik MJ et al (1994) The role of B cells in lpr/lpr-induced autoimmunity. J Exp Med 180:1295–1306
Chan OT et al (1999) A novel mouse with B cells but lacking serum antibody reveals an antibody-independent role for B cells in murine lupus. J Exp Med 189:1639–1648
Ahuja A et al (2007) Depletion of B cells in murine lupus: efficacy and resistance. J Immunol 179:3351–3361
Looney RJ et al (2004) B cell depletion as a novel treatment for systemic lupus erythematosus: a phase I/II dose-escalation trial of rituximab. Arthritis Rheum 50:2580–2589
Navarra SV et al (2001) Efficacy and safety of belimumab in patients with active systemic lupus erythematosus: a randomised, placebo-controlled, phase 3 trial. Lancet 377:721–731
Chan O, Shlomchik MJ (1998) A new role for B cells in systemic autoimmunity: B cells promote spontaneous T cell activation in MRL-lpr/lpr mice. J Immunol 160:51–59
Anolik JH et al (2007) Delayed memory B cell recovery in peripheral blood and lymphoid tissue in systemic lupus erythematosus after B cell depletion therapy. Arthritis Rheum 56:3044–3056
Sanz I et al (2008) Phenotypic and functional heterogeneity of human memory B cells. Semin Immunol 20:67–82
Wehr C et al (2004) A new CD21low B cell population in the peripheral blood of patients with SLE. Clin Immunol 113:161–171
Odendahl M et al (2000) Disturbed peripheral B lymphocyte homeostasis in systemic lupus erythematosus. J Immunol 165:5970–5979
Wirths S, Lanzavecchia A (2005) ABCB1 transporter discriminates human resting naive B cells from cycling transitional and memory B cells. Eur J Immunol 35:3433–3441
Palanichamy A et al (2009) Novel human transitional B cell populations revealed by B cell depletion therapy. J Immunol 182:5982–5993
Pascual V et al (1994) Analysis of somatic mutation in five B cell subsets of human tonsil. J Exp Med 180:329–339
Wellmann U et al (2005) The evolution of human anti-double-stranded DNA autoantibodies. Proc Natl Acad Sci USA 102:9258–9263
Tsuiji M et al (2006) A checkpoint for autoreactivity in human IgM+ memory B cell development. J Exp Med 203:393–400
Cappione A 3rd et al (2005) Germinal center exclusion of autoreactive B cells is defective in human systemic lupus erythematosus. J Clin Invest 115:3205–3216
Vinuesa CG, Sanz I, Cook MC (2009) Dysregulation of germinal centres in autoimmune disease. Nat Rev Immunol 9:845–857
Crotty S, Ahmed R (2004) Immunological memory in humans. Semin Immunol 16:197–203
Mamani-Matsuda M et al (2008) The human spleen is a major reservoir for long-lived vaccinia virus-specific memory B cells. Blood 111: 4653–4659
Anderson SM, Tomayko MM, Shlomchik MJ (2006) Intrinsic properties of human and murine memory B cells. Immunol Rev 211:280–294
Agematsu K et al (1997) B cell subpopulations separated by CD27 and crucial collaboration of CD27+ B cells and helper T cells in immunoglobulin production. Eur J Immunol 27:2073–2079
Tangye SG et al (2003) Intrinsic differences in the proliferation of naive and memory human B cells as a mechanism for enhanced secondary immune responses. J Immunol 170:686–694
Good KL, Avery DT, Tangye SG (2009) Resting human memory B cells are intrinsically programmed for enhanced survival and responsiveness to diverse stimuli compared to naive B cells. J Immunol 182:890–901
Tangye SG, Avery DT, Hodgkin PD (2003) A division-linked mechanism for the rapid generation of Ig-secreting cells from human memory B cells. J Immunol 170:261–269
Macallan DC et al (2005) B-cell kinetics in humans: rapid turnover of peripheral blood memory cells. Blood 105:3633–3640
van Gent R et al (2009) Refined characterization and reference values of the pediatric T- and B-cell compartments. Clin Immunol 133:95–107
Kruetzmann S et al (2003) Human immunoglobulin M memory B cells controlling Streptococcus pneumoniae infections are generated in the spleen. J Exp Med 197:939–945
Griffin DO, Holodick NE, Rothstein TL (2011) Human B1 cells in umbilical cord and adult peripheral blood express the novel phenotype CD20+CD27+CD43+CD70−RDDDDD. J Exp Med 208:67–80
Wei C et al (2007) A new population of cells lacking expression of CD27 represents a notable component of the B cell memory compartment in systemic lupus erythematosus. J Immunol 178:6624–6633
Fecteau JF, Cote G, Neron S (2006) A new memory CD27-IgG+ B cell population in peripheral blood expressing VH genes with low frequency of somatic mutation. J Immunol 177:3728–3736
Klein U, Rajewsky K, Kuppers R (1998) Human immunoglobulin (Ig)M+ IgD+ peripheral blood B cells expressing the CD27 cell surface antigen carry somatically mutated variable region genes: CD27 as a general marker for somatically mutated (memory) B cells. J Exp Med 188:1679–1689
Qian Y et al (2010) Elucidation of seventeen human peripheral blood B-cell subsets and quantification of the tetanus response using a density-based method for the automated identification of cell populations in multidimensional flow cytometry data. Cytometry B Clin Cytom 78(Suppl 1):S69–S82
Dunn-Walters DK, Isaacson PG, Spencer J (1995) Analysis of mutations in immunoglobulin heavy chain variable region genes of microdissected marginal zone (MGZ) B cells suggests that the MGZ of human spleen is a reservoir of memory B cells. J Exp Med 182:559–566
Weller S et al (2008) Somatic diversification in the absence of antigen-driven responses is the hallmark of the IgM+ IgD+ CD27+ B cell repertoire in infants. J Exp Med 205:1331–1342
Wu YC et al (2010) High-throughput immunoglobulin repertoire analysis distinguishes between human IgM memory and switched memory B-cell populations. Blood 116:1070–1078
Seifert M, Kuppers R (2009) Molecular footprints of a germinal center derivation of human IgM+(IgD+)CD27+ B cells and the dynamics of memory B cell generation. J Exp Med 206:2659–2669
Wasserstrom H et al (2008) Memory B cells and pneumococcal antibody after splenectomy. J Immunol 181:3684–3689
Iwata Y et al (2011) Characterization of a rare IL-10-competent B-cell subset in humans that parallels mouse regulatory B10 cells. Blood 117:530–541
Dogan I et al (2009) Multiple layers of B cell memory with different effector functions. Nat Immunol 10:1292–1299
Pape KA et al (2011) Different B cell populations mediate early and late memory during an endogenous immune response. Science 331:1203–1207
Weller S et al (2001) CD40-CD40L independent Ig gene hypermutation suggests a second B cell diversification pathway in humans. Proc Natl Acad Sci USA 98:1166–1170
Mietzner B et al (2008) Autoreactive IgG memory antibodies in patients with systemic lupus erythematosus arise from nonreactive and polyreactive precursors. Proc Natl Acad Sci USA 105:9727–9732
Snapper CM, Mond JJ (1993) Towards a comprehensive view of immunoglobulin class switching. Immunol Today 14:15–17
Rodriguez-Bayona B et al (2010) Decreased frequency and activated phenotype of blood CD27 IgD IgM B lymphocytes is a permanent abnormality in systemic lupus erythematosus patients. Arthritis Res Ther 12:R108
Korganow AS et al (2010) Peripheral B cell abnormalities in patients with systemic lupus erythematosus in quiescent phase: decreased memory B cells and membrane CD19 expression. J Autoimmun 34:426–434
Houtkamp MA et al (2001) Adventitial infiltrates associated with advanced atherosclerotic plaques: structural organization suggests generation of local humoral immune responses. J Pathol 193:263–269
Kyaw T et al (2010) Conventional B2 B cell depletion ameliorates whereas its adoptive transfer aggravates atherosclerosis. J Immunol 185:4410–4419
Sjoberg BG et al (2009) Low levels of IgM antibodies against phosphorylcholine-A potential risk marker for ischemic stroke in men. Atherosclerosis 203:528–532
Zhou X, Hansson GK (1999) Detection of B cells and proinflammatory cytokines in atherosclerotic plaques of hypercholesterolaemic apolipoprotein E knockout mice. Scand J Immunol 50:25–30
Ait-Oufella H et al (2010) B cell depletion reduces the development of atherosclerosis in mice. J Exp Med 207:1579–1587
Halliley JL et al (2010) Peak frequencies of circulating human influenza-specific antibody secreting cells correlate with serum antibody response after immunization. Vaccine 28:3582–3587
Gonzalez-Garcia I et al (2006) Immunization-induced perturbation of human blood plasma cell pool: progressive maturation, IL-6 responsiveness, and high PRDI-BF1/BLIMP1 expression are critical distinctions between antigen-specific and nonspecific plasma cells. J Immunol 176:4042–4050
Odendahl M et al (2005) Generation of migratory antigen-specific plasma blasts and mobilization of resident plasma cells in a secondary immune response. Blood 105:1614–1621
Lee FE et al (2010) Circulating antibody-secreting cells during acute respiratory syncytial virus infection in adults. J Infect Dis 202:1659–1666
Fairfax KA et al (2008) Plasma cell development: from B-cell subsets to long-term survival niches. Semin Immunol 20:49–58
Oracki SA et al (2010) Plasma cell development and survival. Immunol Rev 237:140–159
Caraux A et al (2010) Circulating human B and plasma cells. Age-associated changes in counts and detailed characterization of circulating normal CD138− and CD138+ plasma cells. Haematologica 95:1016–1020
Bernasconi NL, Traggiai E, Lanzavecchia A (2002) Maintenance of serological memory by polyclonal activation of human memory B cells. Science 298:2199–2202
Jacobi AM et al (2010) Effect of long-term belimumab treatment on B cells in systemic lupus erythematosus: extension of a phase II, double-blind, placebo-controlled, dose-ranging study. Arthritis Rheum 62:201–210
Moir S et al (2008) Normalization of B cell counts and subpopulations after antiretroviral therapy in chronic HIV disease. J Infect Dis 197:572–579
Jacobi AM et al (2008) Activated memory B cell subsets correlate with disease activity in systemic lupus erythematosus: delineation by expression of CD27, IgD, and CD95. Arthritis Rheum 58:1762–1773
Habib LK, Finn WG (2006) Unsupervised immunophenotypic profiling of chronic lymphocytic leukemia. Cytometry B Clin Cytom 70:124–135
Diaz-Romero J et al (2010) Hierarchical clustering of flow cytometry data for the study of conventional central chondrosarcoma. J Cell Physiol 225:601–611
Cappione AJ et al (2004) Lupus IgG VH4.34 antibodies bind to a 220-kDa glycoform of CD45/B220 on the surface of human B lymphocytes. J Immunol 172:4298–4307
Herzenberg LA et al (2006) Interpreting flow cytometry data: a guide for the perplexed. Nat Immunol 7:681–685
Eisen MB et al (1998) Cluster analysis and display of genome-wide expression patterns. Proc Natl Acad Sci USA 95:14863–14868
D’haeseleer (2005) How does gene expression clustering work? Nat Biotechnol 23:1499–1501
Quackenbush J (2001) Computational analysis of microarray data. Nat Rev Genet 2:418–427
Misra J et al (2002) Interactive exploration of microarray gene expression patterns in a reduced dimensional space. Genome Res 12:1112–1120
Lugli E et al (2007) Subject classification obtained by cluster analysis and principal component analysis applied to flow cytometric data. Cytometry A 71:334–344
Lee FE et al (2011) Circulating human antibody-secreting cells during vaccinations and respiratory viral infections are characterized by high specificity and lack of bystander effect. J Immunol 186:5514–5521
Wei et al (2011) OMIP-003: Phenotypic analysis of human memory B cells. Cytometry A 79:894–896
Acknowledgments
We thank John Jung and Ravi Misra for reading the manuscript and members of the Sanz lab for help and advice. Supported by NIH R01 AI049660-01A1 and U19 Autoimmunity Center of Excellence AI56390 to I.S.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kaminski, D.A., Wei, C., Rosenberg, A.F., Lee, F.EH., Sanz, I. (2012). Multiparameter Flow Cytometry and Bioanalytics for B Cell Profiling in Systemic Lupus Erythematosus. In: Perl, A. (eds) Autoimmunity. Methods in Molecular Biology, vol 900. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-720-4_6
Download citation
DOI: https://doi.org/10.1007/978-1-60761-720-4_6
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-719-8
Online ISBN: 978-1-60761-720-4
eBook Packages: Springer Protocols