scholarly journals Lineages of human T-cell clones, including T helper 17/T helper 1 cells, isolated at different stages of anti–factor VIII immune responses

Blood ◽  
2009 ◽  
Vol 114 (7) ◽  
pp. 1423-1428 ◽  
Author(s):  
Ruth A. Ettinger ◽  
Eddie A. James ◽  
William W. Kwok ◽  
Arthur R. Thompson ◽  
Kathleen P. Pratt
Keyword(s):  
T Cells ◽  
T Cell ◽  
Immune Responses ◽  
Factor Viii ◽  
Hemophilia A ◽  
T Helper ◽  
T Cell Clones ◽  
Human T Cell ◽  
Cell Clones ◽  
Polarized Cells

AbstractThe development of neutralizing antibodies (inhibitors) after factor VIII (FVIII) infusions is a serious complication that affects approximately one-quarter of hemophilia A patients who have access to replacement therapy. To investigate the differentiation of naive T cells into FVIII-specific helper T cells that promote B-cell activation and antibody secretion, HLA-DRA-DRB1*0101-restricted T-cell clones that respond to a specific epitope in FVIII were isolated from a mild hemophilia A subject (the proband) 19 weeks and 21 months after his development of a high-titer inhibitor. Clones responding to the same epitope were also isolated from his multiply infused brother, who has not developed a clinically significant inhibitor. The 19-week proband clones were T helper (TH)17/TH1- or TH1/TH2-polarized, whereas all 8 clones isolated 21 months postinhibitor development were TH2-polarized cells. In contrast, all 6 clones from the brother who did not develop an inhibitor were TH1-polarized, indicating that tolerance to FVIII can be maintained even with circulating TH1-polarized cells that respond vigorously to in vitro FVIII stimulation. This is the first evidence that TH17/TH1-polarized cells play a role in hemophilic immune responses to FVIII. Furthermore, this is the first report of successful isolation and expansion of antigen-specific human TH17/TH1 clones using standard culture conditions.

Rupture of T Cell Tolerance to Self Factor VIII in Mild/Moderate Hemophilia A.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3076-3076 ◽  
Author(s):  
Marc G. Jacquemin ◽  
Renaud Lavend’homme ◽  
Benhida Abdellah ◽  
Kathelijne Peerlinck ◽  
Jos Vermylen ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Factor Viii ◽  
Cd4 T Cells ◽  
Hemophilia A ◽  
T Cell Tolerance ◽  
T Cell Clones ◽  
Cell Clones ◽  
Ifn Γ ◽  
Recombinant Fviii

Abstract Some patients with mild/moderate hemophilia A develop anti-Factor VIII (FVIII) antibodies following treatment with FVIII. In rare cases, the patient antibodies neutralize normal FVIII but do not recognize the patient FVIII. In most cases, the antibodies cross-react with the patient FVIII thereby changing the patient’s bleeding phenotype into that of a severe hemophilia A patient. We recently investigated the CD4+ T cells of a patient with mild hemophilia A, who produced antibodies recognizing only exogenous normal FVIII. Likewise, the patient CD4+ T cells recognized normal FVIII but not the patient FVIII. These observations raised the question of the specificity of FVIII-specific T cells occurring in mild/moderate hemophilia A patients and who developed an immune response towards both exogenous and self FVIII. We therefore investigated the specificity of CD4+ T lymphocytes in a mild hemophilia A patient (Ba), carrying a substitution Pro2292His in the FVIII gene, who produced high titer inhibitor antibodies recognizing both self and normal FVIII. The patient’s antibodies exclusively recognized the FVIII C2 domain, as determined in immunoprecipitation experiments using recombinant FVIII fragments produced in reticulocyte lysate. Eradication of the inhibitor was not successful despite using several therapeutic options (plasma exchanges, cyclophosphamide, low dose (25 U/kg) immune tolerance induction with plasma-derived Factor VIII and eventually anti-CD20 monoclonal antibody). Patient Ba FVIII specific T cells were expanded using dendritic cells. Out of 20 microcultures initiated with a total of 2 x 106 T cells, 3 cell lines specifically recognised FVIII. The frequency of FVIII-specific T cells in blood of this patient is therefore at least 1/700.000 CD4+ T cells. Patient Ba T cells were cloned using as antigen presenting cells an autologous lymphoblastoid cell line producing a non inhibitory anti-FVIII IgG4 antibody. Two FVIII-specific T cell clones were successfully derived. In control experiments, no FVIII-specific T cell clones could be derived from normal individuals. Both Ba T cell clones were activated by recombinant FVIII fragments encompassing the C1 and C2 domains. T cell activation was compared in presence of normal and His2292 recombinant FVIII. In the presence of 1 IU/ml normal or His2292 FVIII, clone 4E1 produced 0,793 ± 0,024 ng/ml and 0,277 ± 0,087 ng/ml IFN-γ, respectively, whereas clone 3F9 secreted 0,208 ± 0,07 and 0,238 ± 0,021 ng/ml, respectively. The concentration of normal FVIII inducing the same IFN-γ secretion as 1 IU/ml His2292 FVIII was only 0,45 IU/ml for clone 4E1 whereas it was 1,3 IU/ml for clone 3F9. Accordingly, one T cell clone recognizes patient His2292 FVIII at least as well as normal FVIII. These observations demonstrate that in a patient with mild/moderate hemophilia A who develops an immune response to his own FVIII, the T cell tolerance to self FVIII can also be broken. Such a cellular response to self FVIII may render restoration of tolerance to self and exogenous FVIII more difficult.

scholarly journals Public and private human T-cell clones respond differentially to HCMV antigen when boosted by CD3 copotentiation

2020 ◽  
Vol 4 (21) ◽  
pp. 5343-5356
Author(s):  
Laura R. E. Becher ◽  
Wendy K. Nevala ◽  
Shari Lee Sutor ◽  
Megan Abergel ◽  
Michele M. Hoffmann ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Clonal Expansion ◽  
Antigen Receptors ◽  
Fab Fragment ◽  
Public And Private ◽  
T Cell Clones ◽  
Human T Cell ◽  
Cell Clones ◽  
Human T Cells

Abstract Human cytomegalovirus (HCMV) induces long-lasting T-cell immune responses that control but do not clear infection. Typical responses involve private T-cell clones, expressing T-cell antigen receptors (TCRs) unique to a person, and public T-cell clones with identical TCRs active in different people. Here, we report the development of a pretherapeutic immunostimulation modality against HCMV for human T cells, CD3 copotentiation, and the clonal analysis of its effects in recall assays at single-cell resolution. CD3 copotentiation of human T cells required identification of an intrinsically inert anti-CD3 Fab fragment that conditionally augmented signaling only when TCR was coengaged with antigen. When applied in recall assays, CD3 copotentiation enhanced the expansion of both public and private T-cell clones responding to autologous HLA-A2(+) antigen-presenting cells and immunodominant NLVPMVATV (NLV) peptide from HCMV pp65 protein. Interestingly, public vs private TCR expression was associated with distinct clonal expansion signatures in response to recall stimulus. This implied that besides possible differences in their generation and selection in an immune response, public and private T cells may respond differently to pharmacoimmunomodulation. Furthermore, a third clonal expansion profile was observed upon CD3 copotentiation of T-cell clones from HLA-A2(−) donors and 1 HLA-A2(+) presumed-uninfected donor, where NLV was of low intrinsic potency. We conclude that human T-cell copotentiation can increase the expansion of different classes of T-cell clones responding to recall antigens of different strengths, and this may be exploitable for therapeutic development against chronic, persistent infections such as HCMV.

scholarly journals Variable Immortalizing Potential and Frequent Virus Latency in Blood-Derived T-Cell Clones Infected With Human T-Cell Leukemia Virus Type I

Blood ◽  
1997 ◽  
Vol 89 (9) ◽  
pp. 3303-3314 ◽  
Author(s):  
J.H. Richardson ◽  
P. Höllsberg ◽  
A. Windhagen ◽  
L.A. Child ◽  
D.A. Hafler ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Leukemia Virus ◽  
Type I ◽  
Cell Leukemia Virus Type ◽  
T Cell Clones ◽  
Human T Cell ◽  
Cell Clones ◽  
T Cell Leukemia Virus ◽  
Spontaneous Proliferation

Abstract Human T-cell leukemia virus type I (HTLV-I)-infected T cells expanded in vitro by single-cell cloning provide a unique system for investigating virus-cell interactions in nonimmortalized T cells. By analysis of clones generated randomly from the blood of virus carriers, we confirm that CD4 T cells are the major reservoir of HTLV-I in vivo and show that most infected cells contain a single integrated provirus. Contrary to the situation in HTLV-I immortalized cell lines, the HTLV-I provirus was found to be transcriptionally silent in a high proportion of randomly generated T-cell clones and could not be reactivated by mitogenic stimulation. The spontaneous proliferation previously documented in HTLV-I–infected T-cell clones was not observed in silently infected cells, and therefore correlates directly with the expression of tax and other viral genes. The only cytokine mRNA found to be significantly elevated in the virus-producing clones was interleukin-6; however, receptor-blocking experiments argue against a role for IL-6 in the virus-induced cell proliferation. We observed a striking variation in the ability of individual HTLV-I–producing clones to immortalize fresh peripheral blood lymphocytes. This ability did not correlate with the levels of viral mRNA expression, gag p24 production, spontaneous proliferation, or tax-transactivation, possibly suggesting a role for host cell factors as determinants of viral infectivity or immortalization. Studies to elucidate the basis of this phenotypic heterogeneity should enhance our understanding of viral spread and pathogenesis.

scholarly journals Lymphokine regulation of CD45R expression on human T cell clones.

1989 ◽  
Vol 170 (6) ◽  
pp. 2147-2152 ◽  
Author(s):  
S A Brod ◽  
C E Rudd ◽  
M Purvee ◽  
D A Hafler
Keyword(s):  
Molecular Weight ◽  
T Cells ◽  
T Cell ◽  
Activated T Cells ◽  
T Cell Clones ◽  
Human T Cell ◽  
Cell Clones ◽  
Human T Cells ◽  
Single Cell Cloning ◽  
Cell Expression

Whether the expression of higher molecular weight isoforms of the T-200 complex represents different lineages of T cells and/or a sequential stage of the differential pathway of T cells has been unclear. Understanding T cell expression of higher molecular weight isoforms of the T-200 complex (CD45R) may be important because of their association with regulation of immune responses. By direct single cell cloning, we observed a number of long-term T cell clones that expressed CD45RA (2H4). CD45RA expression could be further regulated by ionomycin or the cytokines IL-1 and IL-6, but not IL-2, IL-4, or IFN-gamma. These results indicate that CD45RA expression may define T cell lineages of activated T cells partially controlled by the cytokines IL-1 and IL-6. Further, these results may associate regulatory actions of IL-1 and IL-6 with their ability to increase CD45RA expression in subpopulations of human T cells.

Humanized E17 Hemophilic Mice Are a Major Breakthrough in the Design of New Preclinical Models for Developing Factor VIII Products with Reduced Immunogenicity.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 782-782 ◽  
Author(s):  
Birgit M. Reipert ◽  
Christina Hausl ◽  
Maria Sasgary ◽  
Maria Schuster ◽  
Rafi U. Ahmad ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Immune Responses ◽  
Factor Viii ◽  
Mhc Class Ii ◽  
Cd4 T Cells ◽  
Plasma Cells ◽  
Hemophilia A ◽  
Class Ii ◽  
Mhc Class Ii Molecules

Abstract MHC class II molecules are crucial for regulating adaptive immune responses against self and foreign protein antigens. They determine the antigenic peptides that are presented to CD4+ T cells and are essential for shaping the CD4+ T-cell repertoire in the thymus. Thus, the structure of MHC class II molecules is a major determinant for protein antigen immunogenicity. Structural differences between murine and human MHC class II complexes fundamentally limit the use of conventional murine hemophilia A models for dissecting immune responses to human factor VIII and developing new factor VIII products with reduced immunogenicity. To overcome this limitation, we humanized the murine E17 model of hemophilia A by introducing the human MHC class II haplotype HLA-DRB1*1501 on the background of a complete knockout of all murine MHC class II genes. Any anti-FVIII antibody response in this new humanized hemophilia A model is driven by CD4+ T cells that recognize FVIII-derived peptides that are presented by human HLA-DRB1*1501. The MHC class II haplotype HLA-DRB1*1501 is particularly relevant for the situation in hemophilia A patients because it is found in about 25% of Caucasians and 32% of Africans and has been shown to be associated with an increased risk that patients with severe hemophilia A have for developing FVIII inhibitors. We validated the relevance of this new model by asking the question whether HLA-DRB1*1501 hemophilic E17 mice develop FVIII inhibitors that are similar to those observed in patients with hemophilia A. Furthermore, we wanted to show that anti-FVIII antibody responses in these mice depend on the expression of the human DRB1*1501 molecule. Mice were treated with 8 intravenous doses of human FVIII and tested for anti-FVIII antibodies, anti-FVIII antibody-producing plasma cells and FVIII-specific T cells. About 90% of all humanized hemophilic E17 mice tested developed anti-FVIII antibodies that were similar to FVIII inhibitors found in patients. These antibodies were not restricted isotypically and contained mainly IgG1, IgG2a and IgG2b antibodies. Detection of antibodies in the circulation correlated with the presence of anti-FVIII antibody-producing plasma cells in the spleen. Development of anti-FVIII antibodies depended on the activation of FVIII-specific T cells and strictly depended on the expression of the HLA-DRB1*1501 molecule. Mice that did not express any MHC class II molecules did not develop anti-FVIII antibodies. We conclude that this new humanized E17 model for hemophilia A is a major advance towards developing suitable animal models needed to design future immunomodulatory strategies for patients with FVIII inhibitors and develop new FVIII products with reduced immunogenicity. Furthermore, it provides a tool for identifying T-cell epitopes of human FVIII restricted by MHC class II molecules that can be used for monitoring FVIII-specific T cells in patients who receive replacement therapy with FVIII products.

scholarly journals Transformation of human T-cell clones by Herpesvirus saimiri: intact antigen recognition by autonomously growing myelin basic protein-specific T cells

1993 ◽  
Vol 90 (23) ◽  
pp. 11049-11053 ◽  
Author(s):  
F Weber ◽  
E Meinl ◽  
K Drexler ◽  
A Czlonkowska ◽  
S Huber ◽  
...  
Keyword(s):  
Signal Transduction ◽  
T Cells ◽  
T Cell ◽  
Myelin Basic Protein ◽  
Interferon Gamma ◽  
Basic Protein ◽  
Herpesvirus Saimiri ◽  
T Cell Clones ◽  
Human T Cell ◽  
Cell Clones

Herpesvirus saimiri has recently been shown to immortalize human T cells. It was unknown, however, whether Herpesvirus saimiri transformation affects T-cell receptor (TCR) expression and signal transduction. In the present study, we have transformed CD4+ human T-cell clones specific for human myelin basic protein. The transformed T cells were grown in interleukin 2 and divided in the absence of antigen and antigen-presenting cells. They retained the membrane phenotype of activated T cells and secreted the cytokines interferon gamma and lymphotoxin, but interleukin 4 was not detected. Further, the transformed T cells continued to express the original TCR as demonstrated by TCR variable-region-V beta-specific monoclonal antibodies and TCR sequencing. Antigen-specific recognition and signal transduction by the TCR were demonstrated by myelin-basic-protein-induced HLA-DR-restricted secretion of interferon gamma and lymphotoxin and by myelin-basic-protein-specific proliferation. Antigen specificity and reactivity have been maintained for > 1 year after transformation. Transformation with Herpesvirus saimiri now allows the production of virtually unlimited numbers of (auto)antigen-specific T cells expressing functional TCR and a stable membrane phenotype. This technology will facilitate studies of the pathogenesis of putative autoimmune diseases, such as multiple sclerosis, and may be of help in TCR-targeted immunotherapy.

scholarly journals Selection of a human T helper type 1-like T cell subset by mycobacteria.

1991 ◽  
Vol 174 (3) ◽  
pp. 583-592 ◽  
Author(s):  
J B Haanen ◽  
R de Waal Malefijt ◽  
P C Res ◽  
E M Kraakman ◽  
T H Ottenhoff ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cytokine Secretion ◽  
T Helper ◽  
Ifn Gamma ◽  
T Cell Clones ◽  
Cell Clones ◽  
Tuberculoid Leprosy ◽  
Helper Type

Mycobacteria elicit a cellular immune response in their hosts. This response usually leads to protective immunity, but may sometimes be accompanied by immunopathology due to delayed type hypersensitivity (DTH). A striking example in man is tuberculoid leprosy, which is characterized by high cellular immunity to Mycobacterium leprae and immunopathology due to DTH. Skin lesions of patients suffering from this disease have the characteristics of DTH reactions in which macrophages and CD4+ T lymphocytes predominate. In animal models, it has been shown that DTH responses are associated with the presence of a particular subset of CD4+ T cells (T helper type 1 [Th1]) that secrete only certain cytokines, such as interleukin 2 (IL-2), interferon gamma (IFN-gamma), and lymphotoxin, but no IL-4 or IL-5. We studied the cytokine release of activated M. leprae-reactive CD4+ T cell clones derived from tuberculoid leprosy patients. These T cell clones, which were reactive with mycobacterial heat shock proteins, exhibited a Th1-like cytokine secretion pattern with very high levels of IFN-gamma. Half of these clones secreted low levels of IL-4 and IL-5, but the ratio of IFN-gamma to IL-4 and IL-5 was much higher than that of T cell clones reactive with nonmycobacterial antigens. A Th1-like cytokine secretion pattern was also observed for T cell clones and polyclonal T cell lines from control individuals that recognized both heat shock and other mycobacterial antigens. The levels of IFN-gamma secreted by these clones were, however, significantly less than those of patient-derived T cell clones. This Th1-like pattern was not found with T cell clones from the same patients and healthy individuals generated in the same manner, but reactive with nonmycobacterial antigens. Our data thus indicate that mycobacteria selectively induce human T cells with a Th1-like cytokine secretion profile.

scholarly journals Structural requirements for binding of an immunodominant myelin basic protein peptide to DR2 isotypes and for its recognition by human T cell clones.

1994 ◽  
Vol 179 (1) ◽  
pp. 279-290 ◽  
Author(s):  
K W Wucherpfennig ◽  
A Sette ◽  
S Southwood ◽  
C Oseroff ◽  
M Matsui ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Myelin Basic Protein ◽  
Basic Protein ◽  
Peptide Binding ◽  
Peptide Epitope ◽  
T Cell Clones ◽  
High Affinity ◽  
Human T Cell ◽  
Cell Clones

Immunodominant T cell epitopes of myelin basic protein (MBP) may be target antigens for major histocompatibility complex class II-restricted, autoreactive T cells in multiple sclerosis (MS). Since susceptibility to MS is associated with the DR2 haplotype, the binding and presentation of the immunodominant MBP(84-102) peptide by DR2 antigens were examined. The immunodominant MBP(84-102) peptide was found to bind with high affinity to DRB1*1501 and DRB5*0101 molecules of the disease-associated DR2 haplotype. Overlapping but distinct peptide segments were critical for binding to these molecules; hydrophobic residues (Val189 and Phe92) in the MBP(88-95) segment were critical for peptide binding to DRB1*1501 molecules, whereas hydrophobic and charged residues (Phe92, Lys93) in the MBP(89-101/102) sequence contributed to DRB5*0101 binding. The different registers for peptide binding made different peptide side chains available for interaction with the T cell receptor. Although the peptide was bound with high affinity by both DRB1 and DRB5 molecules, only DRB1 (DRB1*1501 and 1602) but not DRB5 molecules served as restriction elements for a panel of T cell clones generated from two MS patients suggesting that the complex of MBP(84-102) and DRB1 molecules is more immunogenic for MBP reactive T cells. The minimal MBP peptide epitope for several T cell clones and the residues important for binding to DRB1*1501 molecules and for T cell stimulation have been defined.

CD4+ T-cell clones specific for wild-type factor VIII: a molecular mechanism responsible for a higher incidence of inhibitor formation in mild/moderate hemophilia A

Blood ◽  
2003 ◽  
Vol 101 (4) ◽  
pp. 1351-1358 ◽  
Author(s):  
Marc Jacquemin ◽  
Valérie Vantomme ◽  
Cécile Buhot ◽  
Renaud Lavend'homme ◽  
Wivine Burny ◽  
...  
Keyword(s):  
T Cell ◽  
Factor Viii ◽  
Mhc Class Ii ◽  
Hemophilia A ◽  
Cd4 T Cell ◽  
Wild Type ◽  
T Cell Clones ◽  
Mhc Molecules ◽  
Cell Clones ◽  
C1 Domain

Mild/moderate hemophilia A patients carrying certain mutations in the C1 domain of factor VIII (FVIII) have a higher risk of inhibitor occurrence. To analyze the mechanisms responsible for inhibitor development in such patients, we characterized FVIII-specific CD4+ T-cell clones derived from a mild hemophilia A patient carrying an Arg2150His substitution in the C1 domain and who presented with a high titer inhibitor toward normal but not self-FVIII. All T-cell clones recognized synthetic peptides encompassing Arg2150. The peptides were presented to the T-cell clones by DRB1*0401/DRB4*01 or DRB1*1501/DRB5*01. Interestingly, the latter haplotype was previously reported as being associated with an increased incidence of inhibitor formation. Peptide I2144-T2161 also bound to other DR molecules such as DRB1*0101 and DRB1*0701, indicating that the peptide binds to major histocompatibility complex (MHC) class II molecules expressed in more than 60% of the population. None of the T-cell clones recognized recombinant FVIII carrying the substitution Arg2150His, even when FVIII was presented by an FVIII-specific B-cell line. The mutation likely alters T-cell recognition of the mutated peptide associated to MHC molecules, because the mutated peptide bound to immunopurified DR molecules nearly as effectively as the native peptide. These observations demonstrate that T cells of this patient with mutation Arg2150His distinguish between self- and wild-type FVIII and provide a plausible mechanism for the frequent occurrence of an inhibitor in patients carrying this substitution. A similar phenomenon may occur with other mutations associated to an increased incidence of inhibitor formation.

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