2012/06/19 by Jia Yao Phuah, Joshua T. Mattila, Philana L. Lin +2 · 29 citations
Medicine · Immunology and Microbiology · #Tuberculosis Research and Epidemiology #Mycobacterium research and diagnosis #T-cell and B-cell Immunology
paper · doi:10.1016/j.ajpath.2012.05.009
In an attempt to contain Mycobacterium tuberculosis, host immune cells form a granuloma as a physical and immunological barrier. To date, the contribution of humoral immunity, including antibodies and specific functions of B cells, to M. tuberculosis infection in humans remains largely unknown. Recent studies in mice show that humoral immunity can alter M. tuberculosis infection outcomes. M. tuberculosis infection in cynomolgus macaques recapitulates essentially all aspects of human tuberculosis. As a first step toward understanding the importance of humoral immunity to control of M. tuberculosis infection in primates, we characterized the B-cell and plasma-cell populations in infected animals and found that B cells are present primarily in clusters within the granuloma. The B-cell clusters are in close proximity to peripheral node addressin-positive cells and contain cells positive for Ki-67, a proliferation marker. Granuloma B cells also express CXCR5 and have elevated HLA-DR expression. Tissues containing M. tuberculosis bacilli had higher levels of M. tuberculosis-specific IgG, compared with uninvolved tissue from the same monkeys. Plasma cells detected within the granuloma produced mycobacteria-specific antibodies. Together, these data demonstrate that B cells are present and actively secreting antibodies specific for M. tuberculosis antigens at the site of infection, including lung granulomas and thoracic lymph nodes. These antibodies likely have the capacity to modulate local control of infection in tissues. In an attempt to contain Mycobacterium tuberculosis, host immune cells form a granuloma as a physical and immunological barrier. To date, the contribution of humoral immunity, including antibodies and specific functions of B cells, to M. tuberculosis infection in humans remains largely unknown. Recent studies in mice show that humoral immunity can alter M. tuberculosis infection outcomes. M. tuberculosis infection in cynomolgus macaques recapitulates essentially all aspects of human tuberculosis. As a first step toward understanding the importance of humoral immunity to control of M. tuberculosis infection in primates, we characterized the B-cell and plasma-cell populations in infected animals and found that B cells are present primarily in clusters within the granuloma. The B-cell clusters are in close proximity to peripheral node addressin-positive cells and contain cells positive for Ki-67, a proliferation marker. Granuloma B cells also express CXCR5 and have elevated HLA-DR expression. Tissues containing M. tuberculosis bacilli had higher levels of M. tuberculosis-specific IgG, compared with uninvolved tissue from the same monkeys. Plasma cells detected within the granuloma produced mycobacteria-specific antibodies. Together, these data demonstrate that B cells are present and actively secreting antibodies specific for M. tuberculosis antigens at the site of infection, including lung granulomas and thoracic lymph nodes. These antibodies likely have the capacity to modulate local control of infection in tissues. Mycobacterium tuberculosis is an intracellular pathogen that is the causative agent of tuberculosis (TB), an infection that is estimated to affect a third of the world's population. Ten percent of infected individuals develop clinical symptoms of active TB; the remainder develop latent infection, which is clinically asymptomatic but can reactivate to cause active TB.1Wright A. Zignol M. WHO staffAnti-Tuberculosis Drug Resistance in the World. Fourth global report. The WHO/IUATLD Global Project on Anti-tuberculosis Drug Resistance Surveillance 2002–2007. WHO Press, Geneva2008Google Scholar, 2Brewer T.F. Heymann S.J. To control and beyond: moving towards eliminating the global tuberculosis threat.J Epidemiol Community Health. 2004; 58: 822-825Crossref PubMed Scopus (55) Google Scholar, 3Maartens G. Wilkinson R.J. Tuberculosis.Lancet. 2007; 370: 2030-2043Abstract Full Text Full Text PDF PubMed Scopus (257) Google Scholar In response to M. tuberculosis infection, the host immune system forms an organized conglomeration of cells known as a granuloma. Granulomas are crucial in control of mycobacterial pathogens, because they function as an immune and physical barrier to prevent widespread bacterial dissemination within the host.4Saunders B.M. Britton W.J. Life and death in the granuloma: immunopathology of tuberculosis.Immunol Cell Biol. 2007; 85: 103-111Crossref PubMed Scopus (250) Google Scholar Proper control of M. tuberculosis requires immune cells within the granuloma to kill internalized bacilli by activating macrophages while simultaneously balancing anti-inflammatory signals to reduce tissue damage. T cells in particular play a critical role in activating macrophages via the release of interferon gamma (IFN-γ) and tumor necrosis factor (TNF).5Scanga C.A. Mohan V.P. Yu K. Joseph H. Tanaka K. Chan J. Flynn J.L. Depletion of CD4(+) T cells causes reactivation of murine persistent tuberculosis despite continued expression of interferon gamma and nitric oxide synthase 2.J Exp Med. 2000; 192: 347-358Crossref PubMed Scopus (280) Google Scholar, 6Zhang S.Y. Boisson-Dupuis S. Chapgier A. Yang K. Bustamante J. Puel A. Picard C. Abel L. Jouanguy E. Casanova J.L. Inborn errors of interferon (IFN)-mediated immunity in humans: insights into the respective roles of IFN-alpha/beta, IFN-gamma, and IFN-lambda in host defense.Immunol Rev. 2008; 226: 29-40Crossref PubMed Scopus (252) Google Scholar The contribution of B cells to control of human M. tuberculosis infection and pathology remains unknown. Upon activation by antigen, mature B cells proliferate and differentiate into plasma cells for the sole purpose of generating antigen-specific antibodies. Antibodies can affect host-pathogen interactions by enhancing phagocytosis and antibody-dependent cell cytotoxicity, as well as by blocking pathogen-host receptor interactions.7Willcocks L.C. Smith K.G. Clatworthy M.R. Low-affinity Fcgamma receptors, autoimmunity and infection.Expert Rev Mol Med. 2009; 11: e24Crossref PubMed Scopus (63) Google Scholar, 8de Vallière S. Abate G. Blazevic A. Heuertz R.M. Hoft D.F. Enhancement of innate and cell-mediated immunity by antimycobacterial antibodies.Infect Immun. 2005; 73: 6711-6720Crossref PubMed Scopus (125) Google Scholar Antibody-mediated phagocytosis can modify macrophage behavior, depending on how the Fc portion of antibodies interacts with the Fc receptors expressed on macrophages.9Gallo P. Gonçalves R. Mosser D.M. The influence of IgG density and macrophage Fc (gamma) receptor cross-linking on phagocytosis and IL-10 production.Immunol Lett. 2010; 133: 70-77Crossref PubMed Scopus (68) Google Scholar, 10Halstead S.B. Mahalingam S. Marovich M.A. Ubol S. Mosser D.M. Intrinsic antibody-dependent enhancement of microbial infection in macrophages: disease regulation by immune complexes.Lancet Infect Dis. 2010; 10: 712-722Abstract Full Text Full Text PDF PubMed Scopus (327) Google Scholar B cells also present antigen to T cells and enhance CD4+ antigen-specific T-cell expansion.11Kleindienst P. Brocker T. Concerted antigen presentation by dendritic cells and B cells is necessary for optimal CD4 T-cell immunity in vivo.Immunology. 2005; 115: 556-564Crossref PubMed Scopus (42) Google Scholar, 12Dörner T. Crossroads of B cell activation in autoimmunity: rationale of targeting B cells.J Rheumatol Suppl. 2006; 77: 3-11PubMed Google Scholar B-cell depletion slows disease progression of what are predominantly T-cell-mediated autoimmune conditions, including multiple sclerosis13Monson N.L. Cravens P. Hussain R. Harp C.T. Cummings M. de Pilar Martin M. Ben L.H. Do J. Lyons J.A. Lovette-Racke A. Cross A.H. Racke M.K. Stüve O. Shlomchik M. Eagar T.N. Rituximab therapy reduces organ-specific T cell responses and ameliorates experimental autoimmune encephalomyelitis.PLoS One. 2011; 6: e17103Crossref PubMed Scopus (69) Google Scholar, 14Meinl E. Derfuss T. Krumbholz M. Pröbstel A.K. Hohlfeld R. Humoral autoimmunity in multiple sclerosis.J Neurol Sci. 2011; 306: 180-182Abstract Full Text Full Text PDF PubMed Scopus (48) Google Scholar and type 1 diabetes,15Mariño E. Silveira P.A. Stolp J. Grey S.T. B cell-directed therapies in type 1 diabetes.Trends Immunol. 2011; 32: 287-294Abstract Full Text Full Text PDF PubMed Scopus (42) Google Scholar in mice and in humans. These studies reinforce the notion that B-cell antigen presentation is capable of mediating further effects on T cells to drive immune activation in the presence of antigen. Because M. tuberculosis is primarily an intracellular bacillus, the contribution of humoral immunity to protection was thought to be minimal. Studies in the late 19th and early 20th centuries on the protective effects of passive immunization yielded conflicting results, which have been attributed to variations in antisera preparation.16Abebe F. Bjune G. The protective role of antibody responses during Mycobacterium tuberculosis infection.Clin Exp Immunol. 2009; 157: 235-243Crossref PubMed Scopus (122) Google Scholar M. tuberculosis infection of B-cell-deficient mice have also yielded varied findings, ranging from increased pathology or bacterial burden to no apparent change in disease progression.17Maglione P.J. Xu J. Chan J. B cells moderate inflammatory progression and enhance bacterial containment upon pulmonary challenge with Mycobacterium tuberculosis.J Immunol. 2007; 178: 7222-7234Crossref PubMed Scopus (226) Google Scholar, 18Erber W.N. Asbahr H. Meyer B. Herrmann R.P. Davies J.M. Peanut agglutinin (lectin from Arachis hypogaea) binding to hemopoietic cells: an immunophenotypic study using a biotin streptavidin technique.Pathology. 1992; 24: 173-176Abstract Full Text PDF PubMed Scopus (4) Google Scholar The inconsistencies in these mouse studies make it difficult to ascertain the role of the humoral response against TB in humans. Several studies, however, have indicated that other components of the humoral response, including Fc receptors,19Maglione P.J. Xu J. Casadevall A. Chan J. Fc gamma receptors regulate immune activation and susceptibility during Mycobacterium tuberculosis infection.J Immunol. 2008; 180: 3329-3338Crossref PubMed Scopus (121) Google Scholar polymeric Ig receptors,20Tjärnlund A. Rodríguez A. Cardona P.J. Guirado E. Ivanyi J. Singh M. Troye-Blomberg M. Fernández C. Polymeric IgR knockout mice are more susceptible to mycobacterial infections in the respiratory tract than wild-type mice.Int Immunol. 2006; 18: 807-816Crossref PubMed Scopus (64) Google Scholar and intravenous immunoglobulin,21Roy E. Stavropoulos E. Brennan J. Coade S. Grigorieva E. Walker B. Dagg B. Tascon R.E. Lowrie D.B. Colston M.J. Jolles S. Therapeutic efficacy of high-dose intravenous immunoglobulin in Mycobacterium tuberculosis infection in mice.Infect Immun. 2005; 73: 6101-6109Crossref PubMed Scopus (63) Google Scholar can affect the outcome of M. tuberculosis infection in mice. These studies suggest that B-cell responses can confer protection against M. tuberculosis infection either directly or by modulating cellular immune responses (eg, macrophages and T-cell priming and activation). In the mouse model of M. tuberculosis infection, B cells are present in the lungs, often in aggregates that stain positive for peanut agglutinin (PNA), reminiscent of germinal centers in lymph nodes.17Maglione P.J. Xu J. Chan J. B cells moderate inflammatory progression and enhance bacterial containment upon pulmonary challenge with Mycobacterium tuberculosis.J Immunol. 2007; 178: 7222-7234Crossref PubMed Scopus (226) Google Scholar, 22Kahnert A. Höpken U.E. Stein M. Bandermann S. Lipp M. Kaufmann S.H. Mycobacterium tuberculosis triggers formation of lymphoid structure in murine lungs.J Infect Dis. 2007; 195: 46-54Crossref PubMed Scopus (114) Google Scholar Some studies have suggested that granulomas may also function as tertiary germinal centers, where the T-cell population is continuously activated via antigen presentation by B cells. Although B-cell aggregates have been identified in human lung tissue from TB patients,23Ulrichs T. Kosmiadi G.A. Trusov V. Jörg S. Pradl L. Titukhina M. Mishenko V. Gushina N. Kaufmann S.H. Human granulomas peripheral lymphoid to local host in the 2004; PubMed Scopus Google Scholar, S. G. Xu J. Tanaka K. C. J. Flynn J. Chan J. of the granuloma in murine and human cellular and tissue 2006; PubMed Scopus Google Scholar how these B-cell aggregates function in the of M. tuberculosis control is unknown. in the present study was to the of B cells and plasma cells within granulomas of M. cynomolgus model of M. tuberculosis infection been to all aspects of human in granuloma type and The study was of cynomolgus macaques from and from and infected with M. tuberculosis for other animals These studies all and all experimental and by the of of and macaques infected with to of the of M. tuberculosis via as S. A. A. S. C. E. Flynn J.L. Mycobacterium tuberculosis infection of cynomolgus macaques the of human M. tuberculosis Immun. PubMed Scopus Google Scholar, J.L. S. A. A. E. a model for tuberculosis Full Text Full Text PDF PubMed Scopus Google Scholar was by of to positive and by elevated peripheral cell responses to mycobacterial antigens to as via proliferation and J.L. S. A. A. E. a model for tuberculosis Full Text Full Text PDF PubMed Scopus Google Scholar, S. A. A. C. E. Flynn J.L. in Mycobacterium tuberculosis infection in cynomolgus Immun. 2006; PubMed Scopus Google Scholar for the present study from M. control in other TB using and and by a and as S. A. A. S. C. E. Flynn J.L. Mycobacterium tuberculosis infection of cynomolgus macaques the of human M. tuberculosis Immun. PubMed Scopus Google Scholar, J.L. S. A. A. E. a model for tuberculosis Full Text Full Text PDF PubMed Scopus Google Scholar of tissue in for or into for immunological studies, and bacterial as S. A. A. S. C. E. Flynn J.L. Mycobacterium tuberculosis infection of cynomolgus macaques the of human M. tuberculosis Immun. PubMed Scopus Google Scholar, J.L. S. A. A. E. a model for tuberculosis Full Text Full Text PDF PubMed Scopus Google Scholar, S. A. A. C. E. Flynn J.L. in Mycobacterium tuberculosis infection in cynomolgus Immun. 2006; PubMed Scopus Google Scholar was from the by with using a cells using to the cell from and on on with M. tuberculosis as tissue and with no as on with thoracic lymph as and other lymph as peripheral with and with These by a with specific on granuloma as S. A. A. C. E. Flynn J.L. in Mycobacterium tuberculosis infection in cynomolgus Immun. 2006; PubMed Scopus Google Scholar for of cell in antigen Antibodies for peripheral node CXCR5 and Cell with using with an model plasma cell in for at with and in at The tissue was with in The tissue was in optimal and using a at on and at The as from and from these tissue for T cells using and for B cells using and T-cell and HLA-DR identified on and B cells and T cells further identified on and for HLA-DR and within the population. B-cell within tissue by the of as from with cell from tissue and to tissue from tissue and to and M. tuberculosis-specific with of M. tuberculosis or mouse IgG in and at To the in was either at for or at in and at with in using cynomolgus mouse IgG antibody was as the agent at was as the to the for 1 at with with M. tuberculosis and mouse IgG and as from during and to 1 in with and and cells into well and for to at mouse IgG antibody was as the agent at and for 1 at well was using of to the with data with the data using granuloma are of to at and with using the using the was of lung granulomas characterized for to T cells, to B cells, and to cell cells and aggregates of cells within the of lung T cells a more within the compared with the of B cells, and to be within the B-cell T cells also present to the of the which was of cells that the B-cell clusters within the granulomas may be to germinal To the clusters have tertiary germinal T. Kosmiadi G.A. Trusov V. Jörg S. Pradl L. Titukhina M. Mishenko V. Gushina N. Kaufmann S.H. Human granulomas peripheral lymphoid to local host in the 2004; PubMed Scopus Google Scholar we for expression B P.J. Chan J. B cells the immune response against Mycobacterium Immunol. 2009; PubMed Scopus Google Scholar, F. R. in inflammatory Rev Immunol. 2006; 6: PubMed Scopus Google Scholar and a mature germinal cells to at the of the B-cell actively cells detected within the that active cellular proliferation was with within the B-cell clusters in the but as cells than a compared with mature B-cell germinal is for and positive expression is with the presence of of germinal centers cells detected in close proximity to cells within the of the granuloma is in germinal centers in mouse but in the present study of granulomas no was within the however, was within the macrophage of the granuloma. was present within the B-cell of lymph also human and plasma cells W.N. Asbahr H. Meyer B. Herrmann R.P. Davies J.M. Peanut agglutinin (lectin from Arachis hypogaea) binding to hemopoietic cells: an immunophenotypic study using a biotin streptavidin technique.Pathology. 1992; 24: 173-176Abstract Full Text PDF PubMed Scopus (4) Google Scholar and These suggest that expression is in primates, compared with and we as a germinal marker. these data that that the clusters in granulomas of germinal of the B cells within the granuloma was to cellular was on of lung and lymph node to B-cell within infected identified on and and B cells further on expression cells found to be present at within M. compared with lung on a in of cells was and HLA-DR expression on B cells was to activated B cells within infected on the that activated B cells have antigen presentation The of cells from or HLA-DR was HLA-DR expression was higher on cells from compared with within animals and Although HLA-DR was on cells from lung it was to data from uninvolved because B cells present within to in expression be within the cells from lung compared with uninvolved lung tissue The receptor CXCR5 was to activated that to signals into lymphoid CXCR5 expression was on B-cell clusters within the however, of and CXCR5 was within the of the granuloma that B-cell clusters within the granuloma germinal the presence of plasma cells further the within the B cells are in Plasma cells identified within the granuloma on higher to from granuloma for plasma cell in was on granuloma but it to plasma cells lymph node contain cells positive with within the B-cell As of plasma cells, a IgG was to and plasma cells within lymph and from infected plasma cells for mycobacterial antigens and within lung and for within present in on compared with uninvolved lung and the of plasma cells in compared with of plasma cells specific for and within To antibody levels within tissue lung and lymph node from by for IgG antibodies. of and uninvolved lung from the same also an in IgG within lung The same was within compared with IgG levels within lung and lymph node The importance of B cells in control of human M. tuberculosis infection remains despite studies on antibody responses in F. Bjune G. The protective role of antibody responses during Mycobacterium tuberculosis infection.Clin Exp Immunol. 2009; 157: 235-243Crossref PubMed Scopus (122) Google Scholar, R. M.A. S. G. P. K. A. P.A. Plasma antibody as for Immunol. 2011; 18: PubMed Scopus Google Scholar, of antibodies to in pulmonary tuberculosis with of active Infect Dis. 2011; PubMed Google Scholar In the present we the presence of B cells and antibodies in the granulomas of M. using a model that recapitulates the pathology granuloma and infection in humans infected with M. tuberculosis. B cells have been to to control of M. tuberculosis in mouse P.J. Xu J. Chan J. B cells moderate inflammatory progression and enhance bacterial containment upon pulmonary challenge with Mycobacterium tuberculosis.J Immunol. 2007; 178: 7222-7234Crossref PubMed Scopus (226) Google Scholar and are reminiscent of germinal centers of lymphoid on cell and A. Höpken U.E. Stein M. Bandermann S. Lipp M. Kaufmann S.H. Mycobacterium tuberculosis triggers formation of lymphoid structure in murine lungs.J Infect Dis. 2007; 195: 46-54Crossref PubMed Scopus (114) Google Scholar the structure of murine granulomas is to granulomas in humans. we demonstrate that B cells are present as a population within lung granulomas and are organized into clusters with germinal and plasma cells and activated B cells. Several studies have suggested that B cells within the granuloma are reminiscent of germinal centers, on T. Kosmiadi G.A. Trusov V. Jörg S. Pradl L. Titukhina M. Mishenko V. Gushina N. Kaufmann S.H. Human granulomas peripheral lymphoid to local host in the 2004; PubMed Scopus Google Scholar, M. Yang L. M. Yang J. C. H. B. B cell is with the increased and expression in the of with Immunol. 2011; PubMed Scopus Google Scholar germinal formation or lymphoid been with in autoimmune and disease F. R. in inflammatory Rev Immunol. 2006; 6: PubMed Scopus Google Scholar and was a more of antigen to drive cellular activation at In the of the the B-cell clusters may be within the the on a may drive antigen presentation for T-cell the of granuloma B-cell clusters to lymph node germinal centers and the increased HLA-DR expression present on lung B cells. may the of B cells from granulomas to present mycobacterial antigens and T-cell as well as the of granuloma B cells for these compared with peripheral B cells. The increased of antigen-specific IgG at the site of infection to the notion that antibodies may modulate the host-pathogen interactions in the granuloma. plasma cells present within lung granulomas suggest that antibody is however, a of antigen-specific IgG and plasma cells was within which that M. tuberculosis-specific plasma cells are and within These lymph are often and contain M. tuberculosis-specific T S. A. A. S. C. E. Flynn J.L. Mycobacterium tuberculosis infection of cynomolgus macaques the of human M. tuberculosis Immun. PubMed Scopus Google Scholar, J.L. S. A. A. E. a model for tuberculosis Full Text Full Text PDF PubMed Scopus Google Scholar is likely that the plasma cells in lung are from activated B cells that are to the Upon into plasma cells, these B cells likely expression and to the the of cells within the granuloma. Although B-cell clusters within the granuloma with lymph node germinal centers, the granuloma B-cell clusters contain B cells, of mature cells The role of antibody remains in passive immunization and therapy have yielded conflicting in human Studies in humans have been largely to using M. tuberculosis-specific antibodies as of disease by IgG specific to M. tuberculosis R. M.A. S. G. P. K. A. P.A. Plasma antibody as for Immunol. 2011; 18: PubMed Scopus Google Scholar, of antibodies to in pulmonary tuberculosis with of active Infect Dis. 2011; PubMed Google Scholar, R. J. M. D.M. Flynn J.L. K. P.A. antibodies to Mycobacterium tuberculosis by for of Immunol. 2008; PubMed Scopus Google Scholar may be a role for antibodies at the site of is that M. tuberculosis bacilli receptors than and receptors, in increased macrophage Vallière S. Abate G. Blazevic A. Heuertz R.M. Hoft D.F. Enhancement of innate and cell-mediated immunity by antimycobacterial antibodies.Infect Immun. 2005; 73: 6711-6720Crossref PubMed Scopus (125) Google Scholar, K. H. M.A. A. G.A. in pulmonary tuberculosis.J Infect Dis. 2005; 192: PubMed Scopus Google Scholar These are likely by cross-linking of receptors expressed on macrophages and by immune of IgG and M. tuberculosis from M. receptor knockout mice also suggest that are in the P.J. Xu J. Casadevall A. Chan J. Fc gamma receptors regulate immune activation and susceptibility during Mycobacterium tuberculosis infection.J Immunol. 2008; 180: 3329-3338Crossref PubMed Scopus (121) Google Scholar that macrophage activation is for control of M. tuberculosis, antibodies may play a role in macrophage function within the granuloma. phagocytosis may be as in or B S.B. Mahalingam S. Marovich M.A. Ubol S. Mosser D.M. Intrinsic antibody-dependent enhancement of microbial infection in macrophages: disease regulation by immune complexes.Lancet Infect Dis. 2010; 10: 712-722Abstract Full Text Full Text PDF PubMed Scopus (327) Google Scholar In infected with M. tuberculosis, increased B-cell activation was to be with to contain the S. L. Yang G. P. S. B. L. G. pulmonary tuberculosis in a host immune Biol. 2011; PubMed Scopus Google Scholar enhancement of infection may also play a role in the of infection for M. tuberculosis. present demonstrate that activated B cells, plasma cells, and antibodies are within the and have of germinal studies are to on the or immune roles for these cells in and for and and for Chan of for for specific and of for peanut agglutinin in of that are in the of The of PDF of the of in Cell with and Cell and of the of The of the The and contribution have as PDF