T cell senescence and contraction of T cell repertoire diversity in patients with chronic obstructive pulmonary disease

Pathogenetic mechanisms leading to chronic obstructive pulmonary disease (COPD) remain poorly understood. Because clonogenic T cells (CD4⁺CD28^null) were shown to be increased in autoimmune diseases we hypothesized that CD4⁺CD28^null T cells play a role in COPD. Here we describe that enhanced presence of CD4⁺CD28^null cells is associated with impaired lung function. Sixty‐four patients and controls were included. T cell phenotype was analysed using flow cytometry. Enzyme‐linked immunosorbent assays were utilized to determine cytokines. Statistical evaluations were performed using non‐parametric group comparisons and correlations. A logistic regression model was used to determine predictive values of CD4⁺CD28^null in the diagnosis of COPD. Populations of CD4⁺ T cells lacking surface co‐stimulatory CD28 were enlarged significantly in evaluated patients when compared with controls. Natural killer (NK)‐like T cell receptors (CD94, 158) and intracellular perforin, granzyme B were increased in CD4⁺CD28^null cells. Cytokine production after triggering of peripheral blood mononuclear cells (PBMCs) was elevated in patients at early disease stages. Receiver operating characteristic curve plotting revealed that presence of CD4⁺CD28^null T cells has a diagnostic value. These CD4⁺CD28^null T cells show increased expression of NK‐like T cell receptors (CD94, 158) and intracellular perforin and granzyme B. Furthermore, triggering of PBMCs obtained from patients with mild COPD led to increased interferon‐γ and tumour necrosis factor‐α production in vitro compared with controls. Our finding of increased CD4⁺CD28^null T cells in COPD indicates that chronic antigen exposure, e.g. through contents of smoke, leads to loss of CD28 and up‐regulation of NK cell receptors expression on T cells in susceptible patients.     

Characteristics of Expanded CD4+CD28null T Cells in Patients with Chronic Hepatitis B

Expanded CD4⁺CD28^null T cells have not been described in the circulation of patients with chronic hepatitis B (CHB). The aim of the present was to detect and characterize the surface phenotype and functional capacity of these cells in CHB patients. Expanded CD4⁺CD28^null T cells were detected in the circulation of CHB patients with high viral load and elevated aminotransferase levels. Most CD4⁺CD28^null T cells showed a CD27?CD45RA^?CD45RO⁺ surface phenotype. The markers CD56, CD57 and killer immunoglobulin-like receptor (KIR) were detected on CD4⁺CD28^null T cells, but the majority were positive for CD57. Functionally, CD4⁺CD28^null T cells were found to be potent cytotoxic T lymphocytes with perforin and granzyme B secretion profiles. These findings indicate that the expanded CD4⁺CD28^null T cells are cytotoxic memory T cells and display a distinct functional phenotype in comparison with CD4⁺CD28⁺ T cells. The presence of these cells appears to be associated with inflammatory conditions, suggesting that these elevated CD4⁺CD28^null T cells might be involved in the pathogenesis of CHB.     

Peripheral and Site‐Specific CD4+CD28null T Cells from Rheumatoid Arthritis Patients Show Distinct Characteristics

Proinflammatory CD4⁺CD28^null T cells are frequently found in the circulation of patients with rheumatoid arthritis (RA), but are less common in the rheumatic joint. In the present study, we sought to identify functional differences between CD4⁺CD28^null T cells from blood and synovial fluid in comparison with conventional CD28‐expressing CD4⁺ T cells. Forty‐four patients with RA, displaying a distinct CD4⁺CD28^null T cell population in blood, were recruited for this study; the methylation status of the IFNG locus was examined in isolated T cell subsets, and intracellular cytokine production (IFN‐γ, TNF, IL‐17) and chemokine receptor expression (CXCR3, CCR6 and CCR7) were assessed by flow cytometry on T cells from the two compartments. Circulating CD4⁺CD28^null T cells were significantly more hypomethylated in the CNS‐1 region of the IFNG locus than conventional CD4⁺CD28⁺ T cells and produced higher levels of both IFN‐γ and TNF after TCR cross‐linking. CD4⁺CD28^null T cells from the site of inflammation expressed significantly more CXCR3 and CCR6 compared to their counterparts in blood. While IL‐17A production could hardly be detected in CD4⁺CD28^null cells from the blood, a significant production was observed in CD4⁺CD28^null T cells from synovial fluid. CD4⁺CD28^null T cells were not only found to differ from conventional CD4⁺CD28⁺ T cells in the circulation, but we could also demonstrate that synovial CD4⁺CD28^null T cells showed additional effector functions (IL‐17 coproduction) as compared to the same subset in peripheral blood, suggesting an active role for these cells in the perpetuation of inflammation in the subset of patients having a CD28^null population.     

Prevalence of circulating CD4CD28 T cells is associated with early atherosclerotic damage in patients with end-stage renal disease undergoing hemodialysis

CD4⁺ T-cell subsets lacking surface CD28 in peripheral blood have been suggested to predispose people to atherosclerosis. To determine if CD4⁺CD28^null T cells are involved in the immunopathological process of atherosclerotic damage in end-stage renal disease (ESRD) patients undergoing hemodialysis (HD), we characterized peripheral-blood CD4⁺CD28^null T cells from HD patients and investigated the association between these cells and early atherosclerotic damage. Four color flow cytometric analyses showed that HD patients had significantly higher percentages of CD4⁺CD28^null T cells in circulating blood than healthy subjects (HS). Most HD patient-derived CD4⁺CD28^null T cells expressed higher levels of CX3CR1 and produced more intracellular IFN-γ, perforin and granzyme B than their counterparts. Regression analyses demonstrated that the increased levels of CD4⁺CD28^null T cells were positively correlated to serum levels of C-reactive protein, suggesting systemic inflammation and atherosclerosis. Furthermore, phenotypic and functional studies of CD4⁺CD28^null T cells showed that these cells were closely correlated with impaired flow-mediated vasodilation and increased intima-media thickness in the carotid artery, which are markers of early atherosclerosis. These data suggested that CD4⁺CD28^null T cells are important effector cells in HD patients, and that these cells may have a critical role in mediating early atherosclerotic damage.     

Immunological role of CD4<Superscript>+</Superscript>CD28<Superscript>null</Superscript> T lymphocytes,natural killer cells,and interferon-gamma in pediatric patients with sickle cell disease: relation to disease severity and response to therapy

Sickle cell disease (SCD) is associated with alterations in immune phenotypes. CD4⁺CD28^null T lymphocytes have pro-inflammatory functions and are linked to vascular diseases. To assess the percentage of CD4⁺CD28^null T lymphocytes, natural killer cells (NK), and IFN-gamma levels, we compared 40 children and adolescents with SCD with 40 healthy controls and evaluated their relation to disease severity and response to therapy. Patients with SCD steady state were studied, focusing on history of frequent vaso-occlusive crisis, hydroxyurea therapy, and IFN-gamma levels. Analysis of CD4⁺CD28^null T lymphocytes and NK cells was done by flow cytometry. Liver and cardiac iron overload were assessed. CD4⁺CD28^null T lymphocytes, NK cells, and IFN-gamma levels were significantly higher in patients than controls. Patients with history of frequent vaso-occlusive crisis and those with vascular complications had higher percentage of CD4⁺CD28^null T lymphocytes and IFN-gamma while levels were significantly lower among hydroxyurea-treated patients. CD4⁺CD28^null T lymphocytes were positively correlated to transfusional iron input while these cells and IFN-gamma were negatively correlated to cardiac T2* and duration of hydroxyurea therapy. NK cells were correlated to HbS and indirect bilirubin. Increased expression of CD4⁺CD28^null T lymphocytes highlights their role in immune dysfunction and pathophysiology of SCD complications.     

Up‐regulation of alternate co‐stimulatory molecules on proinflammatory CD28null T cells in bronchiolitis obliterans syndrome

Bronchiolitis obliterans syndrome (BOS) is associated with lack of immunosuppression of T cell proinflammatory cytokines and increased T cell granzyme B. Repeated antigen‐driven proliferation down‐regulates T cell CD28. We hypothesized that down‐regulation of CD28 and up‐regulation of alternate co‐stimulatory molecules (CD134, CD137, CD152 and CD154) on T cells may be associated with BOS. Co‐stimulatory molecules, granzyme B, perforin and intracellular cytokines were measured by flow cytometry on T cells from stable lung transplant patients (n = 38), patients with BOS (n = 20) and healthy controls (n = 10). There was a significant increase in the percentage of CD4/28^null and CD8/28^null T cells producing granzyme B, interferon (IFN)‐γ and tumour necrosis factor (TNF)‐α in BOS compared with stable patients. Down‐regulation of CD28 was associated with steroid resistance and up‐regulation of CD134, CD137, CD152 and CD154 on CD4⁺ T cells and CD137 and CD152 on CD8⁺ T cells. There was a significant correlation between increased CD28^null/CD137 T cells producing IFN‐γ, TNF‐α with BOS grade (r = 0·861, P < 0·001 for CD28^null/CD137 IFN‐γ/CD8) and time post‐transplant (r = 0·698, P < 0·001 for CD28^null/CD137 IFN‐γ/CD8). BOS is associated with down‐regulation of CD28 and up‐regulation of alternate co‐stimulatory molecules on steroid‐resistant peripheral blood proinflammatory CD4⁺ and CD8⁺ T cells. Therapeutic targeting of alternate co‐stimulatory molecules on peripheral blood CD28^null T cells and monitoring response using these assays may help in the management of patients with BOS.     

The life (and death) of CD4+CD28null T cells in inflammatory diseases

Inflammation contributes to the development and perpetuation of several disorders and T lymphocytes orchestrate the inflammatory immune response. Although the role of T cells in inflammation is widely recognized, specific therapies that tackle inflammatory networks in disease are yet to be developed. CD4⁺CD28^null T cells are a unique subset of helper T lymphocytes that recently shot back into the limelight as potential catalysts of inflammation in several inflammatory disorders such as autoimmunity, atherosclerosis and chronic viral infections. In contrast to conventional helper T cells, CD4⁺CD28^null T cells have an inbuilt ability to release inflammatory cytokines and cytotoxic molecules that can damage tissues and amplify inflammatory pathways. It comes as no surprise that patients who have high numbers of these cells have more severe disease and poor prognosis. In this review, I provide an overview on the latest advances in the biology of CD4⁺CD28^null T cells. Understanding the complex functions and dynamics of CD4⁺CD28^null T cells may open new avenues for therapeutic intervention to prevent progression of inflammatory diseases.     

High activation and skewed T cell differentiation are associated with low IL-17A levels in a hu-PBL-NSG-SGM3 mouse model of HIV infection

The humanized NOD/SCID/IL-2 receptor γ-chain^null (NSG) mouse model has been widely used for the study of HIV pathogenesis. Here, NSG mice with transgenic expression of human stem cell factor (SCF), granulocyte–macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-3 (NSG-SGM3) were injected with peripheral blood leukocytes (PBL mice) from two HIV-infected (HIV⁺) patients who were under anti-retroviral therapy (ART; referred as HIV⁺ mice) or one HIV-seronegative healthy volunteer (HIV⁻). Such mice are either hu-PBL-NSG-SGM3 HIV⁺ or HIV⁻ mice, depending on the source of PBL. The kinetics of HIV replication and T cell responses following engraftment were evaluated in peripheral blood and secondary lymphoid tissues. High HIV replication and low CD4 : CD8 ratios were observed in HIV⁺ mice in the absence of anti-retroviral therapy (ART). Consistent with high activation and skewed differentiation of T cells from the HIV-infected donor, HIV⁺ mice exhibited a higher T cell co-expression of human leukocyte antigen D-related (HLA-DR) and CD38 than HIV⁻ mice, as well as a shifted differentiation to a CCR7⁻CD45RA⁺ terminal effector profile, even in the presence of ART. In addition, HIV replication and the activation/differentiation disturbances of T cells were associated with decreased plasma levels of IL-17A. Thus, this hu-PBL-NSG-SGM3 mouse model recapitulates some immune disturbances occurring in HIV-infected patients, underlying its potential use for studying pathogenic events during this infection.     

CD28 extinction in human T cells: altered functions and the program of T-cell senescence

Summary: The loss of CD28 expression on T cells is the most consistent biological indicator of aging in the human immune system, and the frequency of CD28^null T cells is a key predictor of immune incompetence in the elderly. There is also mounting evidence for the high frequency of these unusual T cells among patients with inflammatory syndromes or with chronic infections disproportionate with their age. In these pathological states, CD28^null T cells likely represent prematurely senescent lymphocytes due to persistent immune activation. Unlike the situation in CD28 gene knockout mice that have anergic CD28^0/0 T cells, human CD28^null T cells are functionally active, long‐lived, oligoclonal lymphocytes that lack or have limited proliferative capacity. Results of replicative senescence studies show that CD28^null T cells are derived from CD28⁺ precursors that have undergone repeated stimulation, indicating that CD28 silencing underlies the program of T‐cell aging. Dissection of the machinery regulating CD28 expression is paving the way in elucidating the molecular events leading to immune senescence as well as providing clues into the functional rejuvenation of senescent T cells.     

Expanded peripheral CD4+CD28null T cells and its association with atherosclerotic changes in patients with end stage renal disease on hemodialysis

End-stage renal disease (ESRD) patients, including those on hemodialysis, possess a high risk for cardiovascular diseases, as the first leading cause of death among them. Traditional risk factors do not utterly elucidate this. Throughout the last two decades, CD4⁺CD28^null T cells; an unusual subset of T lymphocytes, was detected high with excess cardiovascular (CV) mortality. We aimed to investigate the circulating CD4⁺CD28^null T cells frequency in ESRD patients on hemodialysis and to evaluate their relationship with atherosclerotic changes. High-resolution carotid ultrasonography was done to assess the common carotid artery intima media thickness in a number of ESRD patients, accordingly patients were selected and subdivided into two groups; 30 with atherosclerosis (mean [SD] age, 51.6 [6.3] years) and 30 without (mean [SD] age, 48.9 [5.5] years). Another 30 healthy individuals (mean [SD] age, 48.5 [6.8] years) were enrolled. Analysis of CD4⁺CD28^null T-cells frequency by flow-cytometry was performed in all studied subjects. CD4⁺CD28^null T cell percentage was significantly higher in ESRD patients, (mean [SD], 7.3 [2.7] %) compared to healthy individuals (mean [SD], 3.0 [0.8] %), (p < 0.001). Additionally, the expansion of these unusual T lymphocytes was significantly higher in ESRD patients with atherosclerotic changes (mean [SD], 9.47 [0.75] %) compared to those without atherosclerosis (mean [SD], 5.22 [2.14] %), (p < 0.001). In conclusion circulating CD4⁺CD28^null T lymphocyte population showed expansion in ESRD patients, and of interest in correlation to preclinical atherosclerotic changes.     

MHC class II-deficient mice allow functional human CD4+ T-cell development

Humanized mouse models have been developed to study cell-mediated immune responses to human pathogens in vivo. How immunocompetent human T cells are selected in a murine thymus in such humanized mice remains poorly explored. To gain insights into this mechanism, we investigated the differentiation of human immune compartments in mouse MHC class II-deficient immune-compromised mice (humanized Ab0 mice). We observed a strong reduction in human CD4⁺ T-cell development but despite this reduction Ab0 mice had no disadvantage during Epstein–Barr virus (EBV) infection. Viral loads were equally well controlled in humanized Ab0 mice compared to humanized NSG mice, and improved T-cell recognition of autologous EBV-transformed B cells was observed, especially with respect to cytotoxicity. MHC class II blocking experiments with CD4⁺ T cells from humanized Ab0 mice demonstrated MHC class II restriction of lymphoblastoid cell line recognition. These findings suggest that a small number of CD4⁺ T cells in humanized mice can be solely selected on human MHC class II molecules, presumably expressed by reconstituted human immune cells, leading to improved effector functions.     

Soluble PD-1 rescues the proliferative response of simian immunodeficiency virus-specific CD4 and CD8 T cells during chronic infection

Phenotypic and functional studies of the programmed death-1 (PD-1) molecule on CD4⁺ and CD8⁺ T cells were performed on peripheral blood mononuclear cells from uninfected and simian immunodeficiency virus (SIV)-infected rhesus macaques. These data demonstrated a rapid upregulation of PD-1 expression on tetramer-positive CD8⁺ T cells from MamuA.01⁺ SIV-infected macaques upon infection. Upregulation of PD-1 on total CD8⁺ T cells was not detectable. In contrast, CD4⁺ T-cell PD-1 expression was markedly higher in total CD4⁺ T cells during chronic, but not acute, infection and there was a correlation between the level of PD-1 expression on naive and central memory CD4⁺ T cells and the levels of viral loads. Such association was emphasized further by a marked decrease of PD-1 expression on tetramer-positive CD8 T cells as well as on CD4⁺ T cells on longitudinal samples collected before and after the initiation of antiretroviral therapy and downregulation of viral replication in vivo. Cloning of PD-1 and its two ligands from several non-human primate species demonstrated > 95% conservation for PD-1 and PD-L2 and only about 91% homology for PD-L1. Functional studies using soluble recombinant PD-1 protein or PD-1–immunoglobulin G fusion proteins induced marked increases in the SIV-specific proliferative responses of both CD4⁺ and CD8⁺ T cells from rhesus macaques. The results of these studies serve as a foundation for future in vivo trials of the use of rMamu-PD-1 to potentially enhance and/or restore antiviral immune responses in vivo.     

Evidence that CD4+, but not CD8+ T cells are responsible for murine interleukin-2-deficient colitis

Mice deficient in interleukin-2 production (IL-2^null mice) develop colonic inflammation closely resembling ulcerative colitis in humans. Although this disease is marked by substantial infiltration of the colon by CD8⁺ and CD4⁺ T lymphocytes, no function has yet been assigned to these T cell subsets in the development of colitis in the IL-2^null mouse. For the present study, we investigated the involvement of T lymphocytes in the onset of colitis in IL-2^null mice, and examined the possible role played by cytotoxic T cells. Both lamina propria lymphocytes (LPL) and intraepithelial lymphocytes (IEL) of the colon of IL-2^null mice were potently cytotoxic ex vivo in short-term redirected cytotoxic lymphocyte (CTL) assays. In contrast, colonic T cells of wild-type animals showed little or no constitutive cytotoxic T cell activity. Colonic CTL were detectable prior to the appearance of disease in IL-2^null animals and CTL activity was confined to the TcRαβ, rather than to the TcRγδ IEL subset. IL-2^null animals crossed with major histocompatibility complex class I-deficient mice [IL-2^null × β₂ microglobulin (β₂m^null] mice also developed colitis, which appeared even earlier than in most IL-2^null mice. These findings suggest that neither CD8⁺ IEL nor LPL were causal in the onset of colitis in IL-2^null animals. In IL-2^null × β₂m^null mice, an ulcerative colitis-like disease was evident from histological studies and immunohistological staining which showed very large numbers of CD4⁺ lymphocytes within the intestinal mucosa. Significant ex vivo killing by CD4⁺ T cells was observed in IL-2^null × β₂m^null animals, although this required an extended incubation time compared to colonic CD8⁺ T cells. Peripheral as well as colonic CD4⁺ T cells in IL-2^null and IL-2^null × β₂m^null animals, were activated as judged by their cell surface phenotype (CD45RB^lo, L-selectin^lo and CD69⁺). In light of these findings, we propose that infiltrating CD4⁺, but not CD8⁺ T cells are central to the inflammation observed in the intestinal mucosa in IL-2^null colitis.     

Minocycline attenuates HIV‐1 infection and suppresses chronic immune activation in humanized NOD/LtsZ‐scidIL‐2Rγnull mice

More than a quarter of a century of research has established chronic immune activation and dysfunctional T cells as central features of chronic HIV infection and subsequent immunodeficiency. Consequently, the search for a new immunomodulatory therapy that could reduce immune activation and improve T‐cell function has been increased. However, the lack of small animal models for in vivo HIV study has hampered progress. In the current study, we have investigated a model of cord blood haematopoietic progenitor cells (CB‐HPCs) ‐transplanted humanized NOD/LtsZ‐scidIL‐2Rγ^null mice in which progression of HIV infection is associated with widespread chronic immune activation and inflammation. Indeed, HIV infection in humanized NSG mice caused up‐regulation of several T‐cell immune activation markers such as CD38, HLA‐DR, CD69 and co‐receptor CCR5. T‐cell exhaustion markers PD‐1 and CTLA‐4 were found to be significantly up‐regulated on T cells. Moreover, increased plasmatic levels of lipopolysaccharide, sCD14 and interleukin‐10 were also observed in infected mice. Treatment with minocycline resulted in a significant decrease of expression of cellular and plasma immune activation markers, inhibition of HIV replication and improved T‐cell counts in HIV‐infected humanized NSG mice. The study demonstrates that minocycline could be an effective, low‐cost adjunctive treatment to regulate chronic immune activation and replication of HIV.     

Lymph node and circulating T cell characteristics are strongly correlated in end‐stage renal disease patients,but highly differentiated T cells reside within the circulation

Ageing is associated with changes in the peripheral T cell immune system, which can be influenced significantly by latent cytomegalovirus (CMV) infection. To what extent changes in circulating T cell populations correlate with T cell composition of the lymph node (LN) is unclear, but is crucial for a comprehensive understanding of the T cell system. T cells from peripheral blood (PB) and LN of end‐stage renal disease patients were analysed for frequency of recent thymic emigrants using CD31 expression and T cell receptor excision circle content, relative telomere length and expression of differentiation markers. Compared with PB, LN contained relatively more CD4⁺ than CD8⁺ T cells (P < 0·001). The percentage of naive and central memory CD4⁺ and CD8⁺ T cells and thymic output parameters showed a strong linear correlation between PB and LN. Highly differentiated CD28^null T cells, being CD27^–, CD57⁺ or programmed death 1 (PD‐1⁺), were found almost exclusively in the circulation but not in LN. An age‐related decline in naive CD4⁺ and CD8⁺ T cell frequency was observed (P = 0·035 and P = 0·002, respectively) within LN, concomitant with an increase in central memory CD8⁺ T cells (P = 0·033). Latent CMV infection increased dramatically the frequency of circulating terminally differentiated T cells, but did not alter T cell composition and ageing parameters of LN significantly. Overall T cell composition and measures of thymic function in PB and LN are correlated strongly. However, highly differentiated CD28^null T cells, which may comprise a large part of circulating T cells in CMV‐seropositive individuals, are found almost exclusively within the circulation.     

Human CXCR5+PD-1+ CD8 T cells in healthy individuals and patients with hematologic malignancies

Immune checkpoint blockade (ICB) has revolutionized cancer therapy, but varying response rates illustrate the need for biomarkers of response. Studies in mice have identified a subset of CD8 T cells that is essential for response to PD-1 ICB. These CD8 T cells co-express CXCR5, PD-1 and Tcf1, and provide effector T cells upon PD-1 ICB. It is unknown whether similar T cells play a role in PD-1 ICB in humans. We studied human peripheral blood and lymph nodes (LNs) for the frequency, phenotype, and functionality of CXCR5⁺PD-1⁺ CD8 T cells. We find that CXCR5⁺PD-1⁺ CD8 T cells are memory-like cells, express Tcf1, and lack expression of effector molecules. CXCR5⁺PD-1⁺ CD8 T cells produce cytokines upon stimulation, but have limited proliferative capacity. We studied patients with hematologic malignancies with varying response rates to PD-1 ICB. Specifically in chronic lymphocytic leukemia, in which PD-1 ICB does not induce clinical responses, CXCR5⁺PD-1⁺ CD8 T cells show loss of the memory phenotype and increased effector differentiation. In conclusion, we identified CXCR5⁺PD-1⁺ CD8 T cells in human peripheral blood and LN, which could play a similar role during PD-1 ICB. Future studies should analyze CXCR5⁺PD-1⁺ CD8 T cells during PD-1 ICB and their importance for therapeutic response.     

Accumulation of CD5<Superscript>+</Superscript>CD19<Superscript>+</Superscript> B lymphocytes expressing PD-1 and PD-1L in hypertrophied pharyngeal tonsils

Programmed death-1 (PD-1) is one of the most important inhibitory co-receptors expressed predominantly on activated T and B lymphocytes whose expression could be sustained by permanent antigenic stimulation accompanying chronic or recurrent tonsillitis. The expression of PD-1 and PD-1L was analyzed using flow cytometry on hypertrophied tonsils collected from 57 children. We observed high expression of PD-1 and PD-1L on certain lymphocytes subpopulations of hypertrophied tonsils; among T cells, the expression of PD-1 on protein level was higher on CD4⁺ cells (70.3 %) than on CD8⁺ cells (35 %). Interestingly, a limited expression of PD-1 was observed on CD19⁺ B lymphocytes (6.5 %), while CD5⁺CD19⁺ B cells overexpressed PD-1 (52.5 %). Moreover, the expression of PD-1L was also higher on CD5⁺CD19⁺ B cells (16.5 %) than on CD19⁺ B cells (3.5 %) and on CD4⁺ T cells (20 %) than on CD8⁺ T cells (10 %). PD-1 and PD-1L expressions correlated only on CD5⁺CD19⁺ cells. In conclusion, high expression of PD-1 and PD-1L on T and B cells could represent hallmark of immune system adaptation to chronic antigenic exposition in patients with tonsillitis.     

Functional engraftment of human peripheral T and B cells and sustained production of autoantibodies in NOD/LtSzscid/IL‐2Rγ−/− mice

NOD/LtSzscid/IL‐2Rγ^?/? (NSG) mice have advantages in establishing humanized mouse models. However, transferring human PBMCs into these mice often causes lethal GVH disease. In this study, we discovered an improved method for the engraftment of normal or pathological human PBMCs into NSG mice and examined the subsequent induction of specific immune responses. We sequentially transferred human CD4⁺ memory T (Tm) and B cells obtained from PBMCs of healthy adults or patients with autoimmune diseases into NSG mice. Removing naïve CD4⁺ T cells from the transferred PBMCs allowed successful engraftment without lethal GVH disease. The transferred Tm cells were found to reside mainly in the spleen and the lymphoid nodules, where they expressed MHC class II molecules and produced cytokines, including IL‐21. Surprisingly, the transferred B cells were also well maintained in the lymphoid organs, underwent de novo class‐switch recombination, and secreted all isotypes of human Igs at significant levels. Moreover, transferring patient‐derived Tm and B cells resulted in sustained production of IgM‐rheumatoid factor and antiaminoacyl transfer RNA synthetase Abs in these mice. These results suggest that transfer of Tm and B cells derived from human PBMCs into NSG mice could be a useful method for the study of human autoimmune mechanisms.