Naïve T Cell Level and TCR Vβ Repertoire Usage in Patients with Chronic Myelogenous Leukemia.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4648-4648
Author(s):  
Yangqiu Li ◽  
Suxia Geng ◽  
Lijian Yang ◽  
Shaohua Chen ◽  
Qingsong Yin
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Receptor ◽  
Peripheral Blood ◽  
Clonal Expansion ◽  
Cell Receptor ◽  
Myelogenous Leukemia ◽  
Cd8 Cells ◽  
Normal Individuals ◽  
Excision Circles

Abstract During early T lymphopoiesis, rearrangement of the V, D and J segments of the TCR genes result in deletion of the intervening chromosomal DNA and formation of circular excision circles, the so-called signal joint T-cell receptor rearrangement excision circles (sjTRECs, TRECs). TRECs are assumed to have a high stability, and can not multiply and consequently are diluted during T cell proliferation. It has been used to evaluate thymic function in T-cell immune reconstruction after treatment in HTLV-I-infected or stem cell transplantation. Defects of cellular immunity, however, may also play a role in hematologic malignancies. On the other hand, examination of T cell receptor (TCR) gene repertoire is important to analysis the immune status of the patients with malignant neoplasms, because clonal expansion of T cells permit the identification of specific antigen response of T cells. Little is known about the feature of T-cell immune state in CML. In order to evaluate the thymic recent output function and the expansion feature of TCR V beta subfamily T cells in patients with chronic myelogenous leukemia (CML in chronic phase), TRECs level and TCR V beta repertoire usage and clonality were analyzed. Quantitative detection of TRECs in DNA of peripheral blood mononuclear cells from 20 cases with CML, purified CD4+ or CD8+ cells from 3 cases with CML were preformed by real-time PCR (TaqMan) analysis. And the TRECs-number was related to the number of T-cells by determination of the number of CD3-positive cells. The expression and cloanlity analysis of TCR V beta repertoire were detected by RT-PCR and genescan technique in PBMC from the 14 out of 20 patients. 9 normal individuals served as controls. A dramatic reduction of TRECs values in patients with CML was showed. The mean value of TRECs was 0.06±0.16 copies /1000 T cells in CML patients and 6.84±4.71 copies /1000 T cells in normal individuals, respectively (p<0.01). In 16 out of 20 CML cases no TRECs copies could be detected in peripheral blood, and lower value of TRECs could be identified in sorted CD4+ or CD8+ cells. The expression of 1–12 V beta subfamilies was found in samples from 14 patients. Clonal expanded T cells from 13 cases could be identified in some V beta subfamilies, which were preferentially used in V beta 3, V beta 10, V beta 19, V beta 21 and V beta 22 subfamilies. In conclusion, this is, to our knowledge, the first description of TRECs level in CML patients. These results showed a prominent reduction of TREC levels in CML. The predominant usage and clonal expansion of TCR Vβ subfamily T cells could be identified in CML patients, indicating the host could have the ability for specific immune response to leukemia associated antigen, in despite of T cell immunodeficiency was showed in patients.

The T-cell receptor Vβgene repertoire and clonal expansion from peripheral blood T cells in benzene-exposed workers in China

Hematology ◽  
2009 ◽  
Vol 14 (2) ◽  
pp. 106-110 ◽  
Author(s):  
Bo Li ◽  
Yangqiu Li ◽  
Shaohua Chen ◽  
Lijian Yang ◽  
Wei Yu ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Receptor ◽  
Peripheral Blood ◽  
Clonal Expansion ◽  
Cell Receptor ◽  
Exposed Workers

scholarly journals A Recurrent T-Cell Receptor γ Rearrangement of the Vγ9-JγP Type Represents the Major Clone of γδ T-Cells in Normal Individuals and Is Highly Suppressed in Lymphoproliferative Disorders

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 1119-1119 ◽  
Author(s):  
Philippe Szankasi ◽  
Jonathan Schumacher ◽  
Olga Efimova ◽  
Todd W. Kelley
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cell ◽  
T Cell Receptor ◽  
Peripheral Blood ◽  
Γδ T Cells ◽  
Cell Receptor ◽  
Lymphoproliferative Disorders ◽  
Normal Individuals ◽  
Tcr Repertoire

Abstract INTRODUCTION γδ T-cells expressing T-cell receptor (TCR) type Vγ9Vδ2 are well-established mediators of anti-tumor immunity and have demonstrable HLA-independent cytotoxic activity against both B- and T-cell non-Hodgkin lymphoma cells. Initial clinical trials using ex vivo expanded, adoptively transferred γδ T-cells to enhance tumor immunity have shown some promise but overall results have been lackluster. This indicates that wholesale polyclonal expansion prior to transfer is not likely to be effective. Thus more refined approaches are necessary. This requires a better understanding of their TCR repertoire. We recently developed a next generation sequencing (NGS)-based method for evaluating the spectrum T-cell receptor gamma (TRG) gene rearrangements present in clinical samples. In an effort to better understand the TCR repertoire of γδ T-cells we used this strategy to evaluate a series of samples from normal individuals and from individuals with B and T-cell lymphoproliferative disorders (LPDs). METHODS DNA was isolated from samples from 11 normal individuals (all peripheral blood, PB), 11 patients with a T-cell LPD (6 PB, 3 bone marrow; BM, 2 FFPE tissues), and 5 patients with a B-cell LPD (Hairy cell leukemia; HCL, 4 PB, 1 BM). γδ T-cells were sorted from the PB of 4 of the healthy donors by FACS. TRG rearrangements were PCR amplified using consensus primers and NGS libraries were prepared and sequenced on the Ion Torrent PGM platform. The data was analyzed as follows. NGS typically yielded up to 400,000 sequencing reads which were grouped by identical V, J, and CDR3 sequences into unique rearrangements (typically 15-30,000). The prevalence of a particular TRG rearrangement (or CDR3 sequence) was determined by the number of individual NGS reads with this unique sequence per the entire data set (percent of total reads). All rearrangements were then ranked by their prevalence. RESULTS We sequenced the TRG repertoire of isolated γδ T-cells from the peripheral blood of normal individuals (n=4). We found that a recurrent Vγ9-JγP rearrangement with the CDR3 sequence CALWEVQELGKKIKVF was always (4 of 4 samples) the most prevalent rearrangement in normal γδ T-cells (3.2-11.7-fold more prevalent than the second most common rearrangement; representing 4-11.9% of total reads). Similarly, analysis of a larger set of unsorted normal peripheral blood samples demonstrated high prevalence of the same canonical CDR3 in 5 out of 7 samples, confirming that it is very common in most individuals relative to all TRG rearrangements (among the top 10 most prevalent CDR3s in 4 of 5 samples). We also sequenced the TRG repertoire in 11 samples from patients demonstrating evidence of involvement by a T-cell LPD. Unexpectedly, all Vγ9-JγP type rearrangements were strongly suppressed in these samples including the one with the canonical CDR3. The canonical rearrangement (present in 6 /11 cases), or the most abundant Vγ9-JγP rearrangement when the canonical rearrangement was absent (absent in 5/11 cases), represented on average 0.036 ±0.024 % of all NGS reads in the samples from patients with T-cell LPDs compared to 0.72 ±0.72 % of total NGS reads in the normal controls. This represents on average a 19.9 fold reduction in the T-cell LPD samples. The median rank by abundance of the top Vγ9-JγP rearrangement dropped from 9th (normals) to 291st (T-cell LPD cases) indicating that the suppression was not simply a consequence of the presence of an abundant malignant T-cell clone in the data. The overall distribution of TRG V-segment usage in the normal and neoplastic samples was comparable (Vγ9: 9.9 ±0.55 % and 7.2 ±2.79 %, respectively). In 2 samples from patients with HCL, Vγ9-JγP rearrangements were reduced to a similar extent to that seen in the T-cell LPD cases (9.5-fold reduced; average 0.06% of NGS reads; average rank order 451st). In the other 3 cases of HCL the findings were very similar to those seen in the normal samples. CONCLUSIONS We identified a recurrent Vγ9-JγP rearrangement by NGS representing the most abundant CDR3 in sorted γδ T-cells from normal individuals. This population, along with other clones with Vγ9 rearrangements, appeared specifically suppressed in all samples from patients with T-cell LPDs and in 2 or 5 samples from patients with B-cell LPDs (HCL), perhaps indicating a role in the disease process. Additional samples from a wider range of T- and B-cell LPDs are being analyzed. Disclosures No relevant conflicts of interest to declare.

scholarly journals T-cell receptor gene transfer exclusively to human CD8+ cells enhances tumor cell killing

Blood ◽  
2012 ◽  
Vol 120 (22) ◽  
pp. 4334-4342 ◽  
Author(s):  
Qi Zhou ◽  
Irene C. Schneider ◽  
Inan Edes ◽  
Annemarie Honegger ◽  
Patricia Bach ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Gene Transfer ◽  
Tumor Cell ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Cell Killing ◽  
Specific Gene ◽  
Cd8 Cells ◽  
Tumor Cell Killing

AbstractTransfer of tumor-specific T-cell receptor (TCR) genes into patient T cells is a promising strategy in cancer immunotherapy. We describe here a novel vector (CD8-LV) derived from lentivirus, which delivers genes exclusively and specifically to CD8+ cells. CD8-LV mediated stable in vitro and in vivo reporter gene transfer as well as efficient transfer of genes encoding TCRs recognizing the melanoma antigen tyrosinase. Strikingly, T cells genetically modified with CD8-LV killed melanoma cells reproducibly more efficiently than CD8+ cells transduced with a conventional lentiviral vector. Neither TCR expression levels, nor the rate of activation-induced death of transduced cells differed between both vector types. Instead, CD8-LV transduced cells showed increased granzyme B and perforin levels as well as an up-regulation of CD8 surface expression in a small subpopulation of cells. Thus, a possible mechanism for CD8-LV enhanced tumor cell killing may be based on activation of the effector functions of CD8+ T cells by the vector particle displaying OKT8-derived CD8-scFv and an increase of the surface density of CD8, which functions as coreceptor for tumor-cell recognition. CD8-LV represents a powerful novel vector for TCR gene therapy and other applications in immunotherapy and basic research requiring CD8+ cell-specific gene delivery.

In Utero Transplantation of Transduced Hematopoietic Stem Cells Results in Deletion of Transgene-Specific T Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3266-3266
Author(s):  
Pablo Laje ◽  
William H. Peranteau ◽  
Masayuki Endo ◽  
Philip W. Zoltick ◽  
Alan W. Flake
Keyword(s):  
Stem Cells ◽  
T Cells ◽  
T Cell ◽  
Hematopoietic Stem Cells ◽  
T Cell Receptor ◽  
Peripheral Blood ◽  
T Cell Development ◽  
Cell Receptor ◽  
In Utero ◽  
Hematopoietic Stem

Abstract The developing fetal immune system provides a unique opportunity to manipulate normal immunologic development for therapeutic prenatal and anticipated postnatal interventions. In previous studies we have shown that allogeneic in utero hematopoietic cell transplantation (IUHCT) results in donor specific tolerance that can subsequently facilitate non-myeloablative postnatal cellular or organ transplants. It follows that in utero injection of transduced hematopoietic stem cells (HSC) could potentially induce tolerance to a transgene encoded protein. We hypothesized that expression of a transduced antigenic protein by HSC and their progeny would alter thymic T cell development resulting in deletion of antigen specific T-cells. To test this hypothesis, we used the mammary tumor virus (MTV) superantigen system to evaluate the effect of IUHCT of transduced HSC on T cell development. In this system, expression of different MTV oncogenes by different I-E+ strains of mice results in deletion of T cells expressing the relevant Vβ T cell receptor. Specifically, mice which are Mtv7+ delete T cells expressing the Vβ6 T-cell receptor. In this study, CD150+CD48− enriched Balb/c (I-E+ Mtv7−) HSC were transduced with an HIV-based lentivirus expressing MTV7 under an MND promoter. 1.5E+05 transduced cells were injected intravascularly via the vitelline vein into E14 Balb/c fetuses. Non-injected age matched naive Balb/c mice served as the control group. The peripheral blood (PB) and thymuses of injected fetuses and control mice were harvested at day of life (DOL) 10, 20 and 60 and analyzed by flow cytometry for T lymphocyte Vβ6 expression. Additionally, the T cell composition of the thymus was assessed at DOL10 for CD4 and CD8 single positive (SP) and CD4/CD8 double positive (DP) cells. Thymic flow cytometric analysis at DOL10 revealed that greater than 98% of the T cells were CD4CD8 DP, a stage that has not yet undergone negative selection. No significant difference was noted in the percentage of thymic Vβ6+ DP T-cells at this time point or at DOL20 and DOL60. In contrast, there was a significant decrease in the percentage of Vβ6+ peripheral blood SP cells in those mice injected with MTV7 transduced HSC relative to control mice at DOL10, DOL20 and DOL60 (p<0.05) (Fig 1). The current study supports the ability of enriched transduced HSC to induce deletion of transgene specific T cells after IUHCT. In the future, this strategy may be useful to promote tolerance for pre or postnatal cellular or gene therapy. Figure Figure

Distinct contributions of CD4+ and CD8+naive and memory T-cell subsets to overall T-cell–receptor repertoire complexity following transplantation of T-cell–depleted CD34-selected hematopoietic progenitor cells from unrelated donors

Blood ◽  
2002 ◽  
Vol 100 (5) ◽  
pp. 1915-1918 ◽  
Author(s):  
Matthias Eyrich ◽  
Tanja Croner ◽  
Christine Leiler ◽  
Peter Lang ◽  
Peter Bader ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Progenitor Cells ◽  
T Cell Receptor ◽  
Peripheral Blood ◽  
Cell Receptor ◽  
T Cell Subsets ◽  
Memory T Cell ◽  
Tcr Repertoire ◽  
Unrelated Donors

Normalization of restricted T-cell–receptor (TCR) repertoire is critical following T-cell–depleted (TCD) stem cell transplantation. We present a prospective study analyzing respective contributions of naive and memory T-cell subsets within the CD4+ and CD8+ compartments to the evolution of overall TCR-repertoire complexity following transplantation of CD34-selected peripheral blood progenitor cells from unrelated donors. During the first year after transplantation, sorted CD4/45RA, CD4/45R0, CD8/45RA, and CD8/45R0 subsets were analyzed at 3-month intervals for TCR-repertoire complexity by CDR3 size spectratyping. Skew in TCR-repertoire was observed only in early memory-type T cells. CD4+ and CD8+ subsets differed in clonal distribution of CDR3 sizes, with rapid Gaussian normalization of bands in CD4/45R0+ T cells. Naive T cells displayed normal repertoire complexity and contributed significantly to skew correction. Our data provide direct evidence for an important role of de novo maturation of naive T cells in normalization of an initially restricted TCR-repertoire following transplantation of CD34-selected, TCD-depleted peripheral blood progenitors from unrelated donors.

T cells containing T cell receptor excision circles are inversely related to HIV replication and are selectively and rapidly released into circulation with antiretroviral treatment

AIDS ◽  
2003 ◽  
Vol 17 (8) ◽  
pp. 1145-1149 ◽  
Author(s):  
Mireya Diaz ◽  
Daniel C Douek ◽  
Hernan Valdez ◽  
Brenna J Hill ◽  
Dolores Peterson ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Receptor ◽  
Antiretroviral Treatment ◽  
Cell Receptor ◽  
Hiv Replication ◽  
Excision Circles

Increased Frequency of CD4 - 8 - T Cells Bearing T-Cell Receptor ab Chains in Peripheral Blood of Atomic Bomb Survivors Exposed to High Doses

1994 ◽  
Vol 139 (1) ◽  
pp. 67 ◽  
Author(s):  
Yoichiro Kusunoki ◽  
Seishi Kyoizumi ◽  
Yuko Hirai ◽  
Shoichiro Fujita ◽  
Mitoshi Akiyama
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Receptor ◽  
Peripheral Blood ◽  
Atomic Bomb ◽  
Cell Receptor ◽  
Atomic Bomb Survivors ◽  
High Doses
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