Abnormal CD4 + T helper (Th) 1 cells and activated memory B cells are associated with type III asymptomatic mixed cryoglobulinemia in HCV infection

Virology Journal

Lai Wei¹  Xu Cong¹  Jianghua Wang¹  Huiying Rao¹  Henghui Zhang¹  Bo Feng¹  Wei Zhang¹  Fanyun Kong² 

[1] Peking University People’s Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, No.11 Xizhimen South Street, Beijing 100044, China;Department of Pathogenic biology and Laboratory of Infection and Immunology, Xuzhou Medical College, 84 West Huaihai Road, Xuzhou 221002, Jiangsu Province, China

关键词: Activated memory B cells;    CD4+ T helper cells;    Type III asymptomatic cryoglobulinemia;    Hepatitis C;   

Others  :  1224914
DOI  :  10.1186/s12985-015-0324-2

 received in 2015-02-02, accepted in 2015-06-12,  发布年份 2015

【 摘 要 】

Background

Mixed cryoglobulinemia (MC) in hepatitis C virus (HCV) infection is associated with abnormal immune responses mediated by T cells and B cells, while the relationships of different subsets of CD4 + T helper (Th) cells, B cells and associated cytokines with type III asymptomatic MC in HCV infection are poorly understood.

Methods

Fifty-four chronic hepatitis C (CHC) patients and 23 healthy controls (HCs) were enrolled in the study. Serum cryoglobulins were detected by cryoprecipitation. The types of cryoglobulin were determined by western blot. The phenotypes and frequencies of Th cell and B cell subsets were detected by flow cytometric analysis. The cytokines IFN-γ, IL-4, IL-17, IL-21, IL-22, and TGF-β were measured by enzyme-linked immunosorbent assay.

Results

Twenty-six CHC patients were detected with type III asymptomatic MC. The frequencies of Th2, Th17, follicular helper T (Tfh cells), Th22, and tissue-like B cells were significantly higher in CHC patients compared to HCs, while these cell subsets were not significantly different between CHC patients and HCV-related MC patients. The frequencies of Th1 and activated memory B cells increased in HCV-related MC patients compared to HCs, although the difference between the two cell subsets in CHC patients and HCs was not significant. The frequency of regulatory T cells (Treg cells) was higher in CHC patients than in HCV-related MC patients and HCs. Higher expressions of serum IFN-γ, IL-17, IL-21, and IL-22 were observed in CHC patients than in HCs, but the differences were not significantly different in CHC patients and HCV-related MC patients. The frequency of Th1 cells was associated with activated memory B cells in HCV-related MC patients, and the frequency of Th1 cells and activated memory B cells was closely related to HCV RNA in HCV-related MC patients.

Conclusions

The increased frequencies of Th17 cells, Tfh cells, Th22 cells, Treg cells, cytokines IL-17, IL-21, IL-22, and tissue-like B cells, were related to HCV infection but not type III asymptomatic MC. Higher frequencies of Th1 cells and activated memory B cells were associated with type III asymptomatic MC in HCV infection.

【 授权许可】

   
2015 Kong et al.

【 图 表 】

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 5.

Fig. 6.

【 参考文献 】

• [1]Ko HM, Hernandez-Prera JC, Zhu H, Dikman SH, Sidhu HK, Ward SC et al.. Morphologic features of extrahepatic manifestations of hepatitis C virus infection. Clin Dev Immunol. 2012; 2012:740138.
• [2]Cacoub P, Gragnani L, Comarmond C, Zignego AL. Extrahepatic manifestations of chronic hepatitis C virus infection. Dig Liver Dis. 2014; 46 Suppl 5:S165-73.
• [3]Lauletta G, Russi S, Conteduca V, Sansonno L. Hepatitis C virus infection and mixed cryoglobulinemia. Clin Dev Immunol. 2012; 2012:502156.
• [4]Motyckova G, Murali M. Laboratory testing for cryoglobulins. Am J Hematol. 2011; 86:500-2.
• [5]Antonelli A, Ferri C, Ferrari SM, Ruffilli I, Colaci M, Frascerra S et al.. High serum levels of CXCL11 in mixed cryoglobulinemia are associated with increased circulating levels of interferon-gamma. J Rheumatol. 2011; 38:1947-52.
• [6]Loffreda S, Muratori P, Muratori L, Mele L, Bianchi FB, Lenzi M. Enhanced monocyte Th1 cytokine production in HCV-infected cryoglobulinemic patients. J Hepatol. 2003; 38:230-6.
• [7]Antonelli A, Ferri C, Ferrari SM, Ghiri E, Marchi S, Colaci M et al.. High interleukin-6 and tumor necrosis factor-alpha serum levels in hepatitis C infection associated or not with mixed cryoglobulinemia. Clin Rheumatol. 2009; 28:1179-85.
• [8]Saadoun D, Bieche I, Maisonobe T, Asselah T, Laurendeau I, Piette JC et al.. Involvement of chemokines and type 1 cytokines in the pathogenesis of hepatitis C virus-associated mixed cryoglobulinemia vasculitis neuropathy. Arthritis Rheum. 2005; 52:2917-25.
• [9]Antonelli A, Fallahi P, Ferrari SM, Colaci M, Giuggioli D, Saraceno G et al.. Increased CXCL9 serum levels in hepatitis C-related mixed cryoglobulinemia, with autoimmune thyroiditis, associated with high levels of CXCL10. J Interferon Cytokine Res. 2013; 33:739-45.
• [10]Antonelli A, Ferri C, Fallahi P, Ferrari SM, Frascerra S, Sebastiani M et al.. High values of CXCL10 serum levels in patients with hepatitis C associated mixed cryoglobulinemia in presence or absence of autoimmune thyroiditis. Cytokine. 2008; 42:137-43.
• [11]Antonelli A, Fallahi P, Ferrari SM, Sebastiani M, Manfredi A, Mazzi V et al.. Circulating CXCL11 and CXCL10 are increased in hepatitis C-associated cryoglobulinemia in the presence of autoimmune thyroiditis. Mod Rheumatol. 2012; 22:659-67.
• [12]Antonelli A, Ferri C, Fallahi P, Ferrari SM, Sebastiani M, Ferrari D et al.. High values of CXCL10 serum levels in mixed cryoglobulinemia associated with hepatitis C infection. Am J Gastroenterol. 2008; 103:2488-94.
• [13]Antonelli A, Fallahi P, Ferrari SM, Corrado A, Sebastiani M, Giuggioli D et al.. Parallel increase of circulating CXCL11 and CXCL10 in mixed cryoglobulinemia, while the proinflammatory cytokine IL-6 is associated with high serum Th2 chemokine CCL2. Clin Rheumatol. 2013; 32:1147-54.
• [14]Antonelli A, Ferrari SM, Ruffilli I, Fallahi P. Cytokines and HCV-related autoimmune disorders. Immunol Res. 2014; 60:311-9.
• [15]Boyer O, Saadoun D, Abriol J, Dodille M, Piette JC, Cacoub P et al.. CD4 + CD25+ regulatory T-cell deficiency in patients with hepatitis C-mixed cryoglobulinemia vasculitis. Blood. 2004; 103:3428-30.
• [16]Russi S, Lauletta G, Serviddio G, Sansonno S, Conteduca V, Sansonno L et al.. T cell receptor variable beta gene repertoire in liver and peripheral blood lymphocytes of chronically hepatitis C virus-infected patients with and without mixed cryoglobulinaemia. Clin Exp Immunol. 2013; 172:254-62.
• [17]Charles ED, Green RM, Marukian S, Talal AH, Lake-Bakaar GV, Jacobson IM et al.. Clonal expansion of immunoglobulin M + CD27+ B cells in HCV-associated mixed cryoglobulinemia. Blood. 2008; 111:1344-56.
• [18]Terrier B, Joly F, Vazquez T, Benech P, Rosenzwajg M, Carpentier W et al.. Expansion of functionally anergic CD21-/low marginal zone-like B cell clones in hepatitis C virus infection-related autoimmunity. J Immunol. 2011; 187:6550-63.
• [19]Saadoun D, Rosenzwajg M, Landau D, Piette JC, Klatzmann D, Cacoub P. Restoration of peripheral immune homeostasis after rituximab in mixed cryoglobulinemia vasculitis. Blood. 2008; 111:5334-41.
• [20]Santer DM, Ma MM, Hockman D, Landi A, Tyrrell DL, Houghton M. Enhanced activation of memory, but not naive, B cells in chronic hepatitis C virus-infected patients with cryoglobulinemia and advanced liver fibrosis. PLoS One. 2013; 8:e68308.
• [21]Holz LE, Yoon JC, Raghuraman S, Moir S, Sneller MC, Rehermann B. B cell homeostasis in chronic hepatitis C virus-related mixed cryoglobulinemia is maintained through naive B cell apoptosis. Hepatology. 2012; 56:1602-10.
• [22]Ishii K, Shinohara M, Kogame M, Shiratori M, Higami K, Kanayama K et al.. Effects of mutation number in interferon sensitivity determining region on peripheral blood CD4(+) T cell subsets (Th1, Th2) in chronic hepatitis C patients with hepatitis C virus genotype 1b and high viral load. Hepatol Int. 2012; 6:468-74.
• [23]Chang Q, Wang YK, Zhao Q, Wang CZ, Hu YZ, Wu BY. Th17 cells are increased with severity of liver inflammation in patients with chronic hepatitis C. J Gastroenterol Hepatol. 2012; 27:273-8.
• [24]Wu LY, Liu S, Liu Y, Guo C, Li H, Li W et al.. Up-regulation of interleukin-22 mediates liver fibrosis via activating hepatic stellate cells in patients with hepatitis C. Clin Immunol. 2015; 158:77-87.
• [25]Cabrera R, Tu Z, Xu Y, Firpi RJ, Rosen HR, Liu C et al.. An immunomodulatory role for CD4(+)CD25(+) regulatory T lymphocytes in hepatitis C virus infection. Hepatology. 2004; 40:1062-71.
• [26]Vogelzang A, McGuire HM, Yu D, Sprent J, Mackay CR, King C. A fundamental role for interleukin-21 in the generation of T follicular helper cells. Immunity. 2008; 29:127-37.
• [27]Pallikkuth S, Parmigiani A, Pahwa S. Role of IL-21 and IL-21 receptor on B cells in HIV infection. Crit Rev Immunol. 2012; 32:173-95.
• [28]Wang J, Shan Y, Jiang Z, Feng J, Li C, Ma L et al.. High frequencies of activated B cells and T follicular helper cells are correlated with disease activity in patients with new-onset rheumatoid arthritis. Clin Exp Immunol. 2013; 174:212-20.
• [29]Tangye SG, Ma CS, Brink R, Deenick EK. The good, the bad and the ugly - TFH cells in human health and disease. Nat Rev Immunol. 2013; 13:412-26.
• [30]Morita R, Schmitt N, Bentebibel SE, Ranganathan R, Bourdery L, Zurawski G et al.. Human blood CXCR5(+)CD4(+) T cells are counterparts of T follicular cells and contain specific subsets that differentially support antibody secretion. Immunity. 2011; 34:108-21.
• [31]Feng J, Hu X, Guo H, Sun X, Wang J, Xu L et al.. Patients with chronic hepatitis C express a high percentage of CD4(+)CXCR5(+) T follicular helper cells. J Gastroenterol. 2012; 47:1048-56.
• [32]Spaan M, Kreefft K, de Graav GN, Brouwer WP, de Knegt RJ, Ten Kate FJ et al.. CD4CXCR5 T cells in chronic HCV infection produce less IL-21, yet are efficient at supporting B cell responses. J Hepatol. 2014; 62:303-10.
• [33]Cacoub P, Musset L, Hausfater P, Ghillani P, Fabiani FL, Charlotte F et al.. No evidence for abnormal immune activation in peripheral blood T cells in patients with hepatitis C virus (HCV) infection with or without cryoglobulinaemia. Multivirc Group. Clin Exp Immunol. 1998; 113:48-54.
• [34]Sansonno D, Lauletta G, Montrone M, Tucci FA, Nisi L, Dammacco F. Virological analysis and phenotypic characterization of peripheral blood lymphocytes of hepatitis C virus-infected patients with and without mixed cryoglobulinaemia. Clin Exp Immunol. 2006; 143:288-96.
• [35]Racanelli V, Frassanito MA, Leone P, Galiano M, De Re V, Silvestris F et al.. Antibody production and in vitro behavior of CD27-defined B-cell subsets: persistent hepatitis C virus infection changes the rules. J Virol. 2006; 80:3923-34.
• [36]Mizuochi T, Ito M, Saito K, Kasai M, Kunimura T, Morohoshi T et al.. Possible recruitment of peripheral blood CXCR3+ CD27+ CD19+ B cells to the liver of chronic hepatitis C patients. J Interferon Cytokine Res. 2010; 30:243-52.
• [37]Doi H, Tanoue S, Kaplan DE. Peripheral CD27(−)CD21(−) B-cells represent an exhausted lymphocyte population in hepatitis C cirrhosis. Clin Immunol. 2014; 150:184-91.
• [38]Diamant E, Melamed D. Class switch recombination in B lymphopoiesis: a potential pathway for B cell autoimmunity. Autoimmun Rev. 2004; 3:464-9.
• [39]Le Coz C, Joublin A, Pasquali JL, Korganow AS, Dumortier H, Monneaux F. Circulating TFH subset distribution is strongly affected in lupus patients with an active disease. PLoS One. 2013; 8:e75319.
• [40]Rawlings DJ, Schwartz MA, Jackson SW, Meyer-Bahlburg A. Integration of B cell responses through Toll-like receptors and antigen receptors. Nat Rev Immunol. 2012; 12:282-94.
• [41]Charles ED, Brunetti C, Marukian S, Ritola KD, Talal AH, Marks K et al.. Clonal B cells in patients with hepatitis C virus-associated mixed cryoglobulinemia contain an expanded anergic CD21low B-cell subset. Blood. 2011; 117:5425-37.
• [42]Schroder K, Hertzog PJ, Ravasi T, Hume DA. Interferon-gamma: an overview of signals, mechanisms and functions. J Leukoc Biol. 2004; 75:163-89.
• [43]Zhang H, Yang P, Zhou H, Meng Q, Huang X. Involvement of Foxp3-expressing CD4+ CD25+ regulatory T cells in the development of tolerance induced by transforming growth factor-beta2-treated antigen-presenting cells. Immunology. 2008; 124:304-14.
• [44]Zhang L, Hao CQ, Miao L, Dou XG. Role of Th1/Th2 cytokines in serum on the pathogenesis of chronic hepatitis C and the outcome of interferon therapy. Genet Mol Res. 2014; 13:9747-55.
• [45]Nelson DR, Gonzalez-Peralta RP, Qian K, Xu Y, Marousis CG, Davis GL et al.. Transforming growth factor-beta 1 in chronic hepatitis C. J Viral Hepat. 1997; 4:29-35.
• [46]Sousa GM, Oliveira IS, Andrade LJ, Sousa-Atta ML, Parana R, Atta AM. Serum levels of Th17 associated cytokines in chronic hepatitis C virus infection. Cytokine. 2012; 60:138-42.
• [47]Tran DQ. TGF-beta: the sword, the wand, and the shield of FOXP3(+) regulatory T cells. J Mol Cell Biol. 2012; 4:29-37.
• [48]Ramos-Casals M, Stone JH, Cid MC, Bosch X. The cryoglobulinaemias. Lancet. 2012; 379:348-60.
• [49]Rao HY, Sun DG, Yang RF, Liu F, Wang J, Feng B et al.. Outcome of hepatitis C virus infection in Chinese paid plasma donors: a 12-19-year cohort study. J Gastroenterol Hepatol. 2012; 27:526-32.
• [50]Rao H, Wei L, Lopez-Talavera JC, Shang J, Chen H, Li J et al.. Distribution and clinical correlates of viral and host genotypes in Chinese patients with chronic hepatitis C virus infection. J Gastroenterol Hepatol. 2014; 29:545-53.
• [51]Guo Z, Zhang H, Rao H, Jiang D, Cong X, Feng B et al.. DCs pulsed with novel HLA-A2-restricted CTL epitopes against hepatitis C virus induced a broadly reactive anti-HCV-specific T lymphocyte response. PLoS One. 2012; 7:e38390.
• [52]Musset L, Diemert MC, Taibi F, Du Thi Huong L, Cacoub P, Leger JM et al.. Characterization of cryoglobulins by immunoblotting. Clin Chem. 1992; 38:798-802.