ABSTRACT
We describe a unique case of fulminant infectious mononucleosis and recurrent Epstein-Barr virus reactivation presenting in an adolescent. Detailed assays of Epstein-Barr virus-specific T cell immunity revealed defects in the patient's T cell receptor signalling pathway characterized by a lack of interleukin-2 and CD25 expression, which may have contributed to her clinical course. Allogeneic stem cell transplantation reversed the clinical and laboratory phenotype.
Subject(s)
Herpesvirus 4, Human/immunology , Herpesvirus 4, Human/isolation & purification , Infectious Mononucleosis/diagnosis , Infectious Mononucleosis/immunology , Virus Activation , Adolescent , Child , Humans , Infectious Mononucleosis/therapy , Interleukin-2/deficiency , Interleukin-2 Receptor alpha Subunit/deficiency , Receptors, Antigen, T-Cell/deficiency , Recurrence , Stem Cell Transplantation , T-Lymphocytes/immunology , Young AdultABSTRACT
BACKGROUND: A 75-year-old man on methotrexate immunosuppression for myasthenia gravis presented with a 2-month history of lymphocytosis and bilateral inguinal adenopathy. There were no constitutional symptoms of fever, night sweats, or weight loss. INVESTIGATIONS: Physical examination, blood tests, flow cytometry, fluorescent in situ hybridization, immunoglobulin gene sequencing, viral load quantification by real-time polymerase chain reaction, excisional lymph-node biopsy, bone-marrow biopsy, tumor morphology and immunohistochemistry, sequential CT and PET scans. DIAGNOSIS: Methotrexate-associated mantle-cell lymphoma. MANAGEMENT: Cessation of methotrexate, anthracycline-based combination chemo-immunotherapy, and maintenance rituximab.
Subject(s)
Immunosuppressive Agents/adverse effects , Lymphoma, Mantle-Cell/chemically induced , Methotrexate/adverse effects , Myasthenia Gravis/drug therapy , Aged , Diagnosis, Differential , Humans , Immunosuppressive Agents/administration & dosage , Lymphocytes , Lymphocytosis/chemically induced , Lymphoma, Mantle-Cell/diagnosis , Lymphoma, Mantle-Cell/etiology , Male , Methotrexate/administration & dosage , Risk FactorsABSTRACT
In Hodgkin lymphoma (HL), the malignant Hodgkin Reed-Sternberg cells interact with the host microenvironment to create an immunosuppressive network that protects the lymphoma from immune attack. These mechanisms are not fully understood. We examined the role of the immunomodulatory protein galectin-1 (Gal-1) on Epstein-Barr virus-specific CD8(+) T cell responses in HL. Initial studies indicated Gal-1 expression in all in vitro established Hodgkin Reed-Sternberg cell lines. In situ analysis revealed Gal-1 expression in 26 of 42 classic HL, whereas Gal-1 was uniformly negative in nodular lymphocyte predominant HL. Gal-1(hi) expression was associated with male gender, older patients, reduced CD8(+) T cell infiltration at the tumor site, and most importantly, an impaired latent membrane protein 1 and 2-specific CD8(+) T-cell responses. In vitro exposure to recombinant Gal-1 inhibited proliferation and interferon-gamma expression by Epstein-Barr virus-specific T cells. These observations provide an important link between the Gal-1-mediated immunomodulatory networks and loss of antigen-specific T-cell function in classic HL.
Subject(s)
Galectin 1/metabolism , Herpesvirus 4, Human/metabolism , Hodgkin Disease/immunology , Lymphocytes, Tumor-Infiltrating/metabolism , Adult , CD8-Positive T-Lymphocytes/immunology , Cell Proliferation , Epstein-Barr Virus Infections/genetics , Epstein-Barr Virus Infections/metabolism , Epstein-Barr Virus Infections/virology , Female , Herpesvirus 4, Human/genetics , Hodgkin Disease/metabolism , Hodgkin Disease/virology , Humans , Interferon-gamma/metabolism , Male , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Reed-Sternberg Cells/immunology , Reed-Sternberg Cells/metabolism , T-LymphocytesABSTRACT
Immunosuppression following solid organ transplantation results in impaired T-cell immunity and risk of Epstein-Barr virus (EBV)-positive post-transplant lymphoproliferative disorders (PTLD). The B-cell targeting antibody rituximab has efficacy in PTLD. As B cells are the principle reservoir for EBV, we investigated the effect of rituximab on the persistence of EBV-specific CD8(+) T-cell immunity. To avoid the confounding factor of concurrent immunosuppression to prevent transplant rejection, immunity was analysed in non-transplanted lymphoma patients (i.e. a non-PTLD setting). Cytomegalovirus-specific T-cell immunity was assessed as an internal control. Our data demonstrated that circulating B cells were not critical for maintaining EBV-specific T-cell immunity.
Subject(s)
Antibodies, Monoclonal/pharmacology , Antineoplastic Agents/pharmacology , CD8-Positive T-Lymphocytes/drug effects , Epstein-Barr Virus Infections/immunology , Lymphoma, Non-Hodgkin/drug therapy , Aged , Antibodies, Monoclonal, Murine-Derived , CD8-Positive T-Lymphocytes/immunology , Cells, Cultured , Female , Humans , Immunity, Cellular/drug effects , Lymphoma, Non-Hodgkin/immunology , Male , Middle Aged , Rituximab , T-Lymphocytes, Cytotoxic/drug effects , T-Lymphocytes, Cytotoxic/immunologyABSTRACT
In Hodgkin lymphoma (HL), the malignant Hodgkin Reed-Sternberg (HRS) cells constitute only 0.5% of 10% of the diseased tissue. The surrounding cellular infiltrate is enriched with T cells that are hypothesized to modulate antitumor immunity. We show that a marker of regulatory T cells, LAG-3, is strongly expressed on infiltrating lymphocytes present in proximity to HRS cells. Circulating regulatory T cells (CD4+ CD25hi CD45 ROhi, CD4+ CTLA4hi, and CD4+ LAG-3hi were elevated in HL patients with active disease when compared with remission. Longitudinal profiling of EBV-specific CD8+ T-cell responses in 94 HL patients revealed a selective loss of interferon-gamma expression by CD8+ T cells specific for latent membrane proteins 1 and 2 (LMP1/2), irrespective of EBV tissue status. Intratumoral LAG-3 expression was associated with EBV tissue positivity, whereas FOXP3 was linked with neither LAG-3 nor EBV tissue status. The level of LAG-3 and FOXP3 expression on the tumor-infiltrating lymphocytes was coincident with impairment of LMP1/2-specific T-cell function. In vitro pre-exposure of peripheral blood mononuclear cells to HRS cell line supernatant significantly increased the expansion of regulatory T cells and suppressed LMP-specific T-cell responses. Deletion of CD4+ LAG-3+ T cells enhanced LMP-specific reactivity. These findings indicate a pivotal role for regulatory T cells and LAG-3 in the suppression of EBV-specific cell-mediated immunity in HL.
Subject(s)
Antigens, CD/biosynthesis , CD8-Positive T-Lymphocytes/physiology , Gene Expression Regulation, Neoplastic , Hodgkin Disease/blood , Hodgkin Disease/metabolism , Lymphocytes, Tumor-Infiltrating/metabolism , Viral Matrix Proteins/metabolism , Adolescent , Adult , Aged , CD8-Positive T-Lymphocytes/metabolism , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Remission Induction , Lymphocyte Activation Gene 3 ProteinABSTRACT
PURPOSE: Latent Epstein-Barr virus (EBV) genomes are found in the malignant cells of approximately one-third of Hodgkin's lymphoma (HL) cases. Detection and quantitation of EBV viral DNA could potentially be used as a biomarker of disease activity. EXPERIMENTAL DESIGN: Initially, EBV-DNA viral load was prospectively monitored from peripheral blood mononuclear cells (PBMC) in patients with HL. Subsequently, we analyzed viral load in plasma from a second cohort of patients. A total of 58 patients with HL (31 newly diagnosed, 6 relapsed, and 21 in long-term remission) were tested. Using real-time PCR, 43 PBMC and 52 plasma samples were analyzed. RESULTS: EBV-DNA was detectable in the plasma of all EBV-positive patients with HL prior to therapy. However, viral DNA was undetectable following therapy in responding patients (P = 0.0156), EBV-positive HL patients in long-term remission (P = 0.0011), and in all patients with EBV-negative HL (P = 0.0238). Conversely, there was no association seen for the EBV-DNA load measured from PBMC in patients with active EBV-positive HL patients as compared with EBV-negative HL, or patients in long-term remission. EBV-DNA load in matched plasma/PBMC samples were not correlated. CONCLUSIONS: We show that free plasma EBV-DNA has excellent sensitivity and specificity, and can be used as a noninvasive biomarker for EBV-positive HL and that serial monitoring could predict response to therapy. Additional prospective studies are required to further evaluate the use of free plasma EBV-DNA as a biomarker for monitoring response to treatment in patients with EBV-positive HL.
