Highly skewed T-cell receptor V-beta chain repertoire in the bone marrow is associated with response to immunosuppressive drug therapy in children with very severe aplastic anemia

Abstract Aplastic anemia is a bone-marrow-failure syndrome characterized by low blood-cell counts and a fatty bone marrow. In most cases, no obvious etiological factor can be identified, but clinical responses to immunosuppressive treatment (IST) strongly suggest an immune pathophysiology. Our previous study of T-cell receptor (TCR) Vβ (variable region of β-chain) repertoire usage by flow cytometry suggested that aplastic anemia results from antigen-specific lymphocyte attack on hematopoietic progenitors (Risitano et al. Lancet2004; 364:355). In the current work, 7 patients were investigated for Vβ pattern expression before first immunosuppresive treatment, at the remission, and again on relapse. The TCR Vβ repertoire was analyzed for CD4+ and CD8+ subsets, separately, by flow cytometry, using a monoclonal antibody set of 22 different Vβ chains. Most patients had very different patterns of Vβ usage from healthy individuals, and all but one showed expansion of at least one Vβ family before immunosuppressive treatment (Vβ family expansions were defined as 2 standard deviations (SD) from the means in controls). The median number of expanded Vβ families was 4 per patient among CD8CD28dim effector cells. At remission, almost all the initially expanded Vβ subfamilies decreased to less than 2SD of controls. At relapse, most of the expanded Vβ subsets were increased again. However, 5/7 patients showed new expanded Vβ subsets at recurrence of cytopenias, suggesting antigenic spread of new epitopes recognized by immune systems. Although no common pattern of specific expanded Vβ subsets could be identified among different patients, some Vβ subfamilies appeared to be more frequently involved (Vβ 5.1 and Vβ 5.2 were expanded in 4 of 7 patients both at initial presentation and relapse ). These data suggest that monitoring Vβ subsets in aplastic anemia, and potentially in other immune-mediated human diseases of a similar pathophysiology could be used to guide individual therapeutic decisions and in the development of new treatments.

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T-cell Receptor β Chain Variability in Bone Marrow and Peripheral Blood in Severe Acquired Aplastic Anemia

Blood Cells Molecules and Diseases ◽

10.1006/bcmd.1997.0127 ◽

1997 ◽

Vol 23 (1) ◽

pp. 110-122 ◽

Cited By ~ 23

Author(s):

Chantal Y. Manz ◽

Pierre-Yves Dietrich ◽

Valérie Schnuriger ◽

Catherine Nissen ◽

Aleksandra Wodnar-Filipowicz

Keyword(s):

Bone Marrow ◽

T Cell ◽

Aplastic Anemia ◽

T Cell Receptor ◽

Peripheral Blood ◽

Cell Receptor ◽

Β Chain

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Oligoclonal expansion of CD8+ CD57+ T cells with restricted T-cell receptor beta chain variability after bone marrow transplantation

Blood ◽

10.1182/blood.v83.2.587.bloodjournal832587 ◽

1994 ◽

Vol 83 (2) ◽

pp. 587-595 ◽

Cited By ~ 1

Author(s):

G Gorochov ◽

P Debre ◽

V Leblond ◽

B Sadat-Sowti ◽

F Sigaux ◽

...

Keyword(s):

Bone Marrow ◽

T Cell ◽

T Cell Receptor ◽

Gene Families ◽

Cell Receptor ◽

Pcr Analysis ◽

Gene Rearrangements ◽

Beta Chain ◽

Beta Gene ◽

Oligoclonal Expansion

A major expansion of CD8+57+ lymphocytes expressing an alpha-beta T- cell receptor (TCR) is frequent after bone marrow transplantation (BMT). We examined the clonality of the TCR beta gene repertoire in these expanded CD8+57+ cells after allogeneic or autologous BMT. We performed a polymerase chain reaction (PCR) analysis of the V beta chain usage in CD8+57+ cells purified from nine BMT recipients with a series of oligonucleotides specific for 20 V beta gene families. PCR products from selected TCR beta gene rearrangements were sequenced. The CD8+57+ cells from eight of nine patients used a restricted set of V beta families, with a marked predominance of two to three V beta gene families per patient, whereas the control autologous CD57- subset expressed the whole 20 V beta families. A direct sequencing analysis confirmed the V beta 16 and V beta 17 clonality in six patients, showing a striking homology in the CDR3 sequences of the V beta 16 products. The CD8+57+ cells, but not the CD57- cells, displayed an oligoclonal pattern of TCR rearrangements as shown by PCR analysis of TCR gamma gene rearrangements. Such an oligoclonal expansion of CD8+57+ cells, using a restricted set of the V beta gene families, may result from a specific TCR stimulation of a limited number of T-cell clones in BMT recipients.

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T-cell receptor Vβ CDR3 oligoclonality frequently occurs in childhood refractory cytopenia (MDS-RC) and severe aplastic anemia

Leukemia ◽

10.1038/leu.2008.23 ◽

2008 ◽

Vol 22 (6) ◽

pp. 1170-1174 ◽

Cited By ~ 18

Author(s):

A C H de Vries ◽

A W Langerak ◽

B Verhaaf ◽

C M Niemeyer ◽

J Stary ◽

...

Keyword(s):

T Cell ◽

Aplastic Anemia ◽

T Cell Receptor ◽

Cell Receptor ◽

Severe Aplastic Anemia ◽

T Cell Receptor Vβ

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Sublethal gamma-radiation induces differentiation of CD4-/CD8- into CD4+/CD8+ thymocytes without T cell receptor beta rearrangement in recombinase activation gene 2-/- mice.

Journal of Experimental Medicine ◽

10.1084/jem.180.4.1517 ◽

1994 ◽

Vol 180 (4) ◽

pp. 1517-1521 ◽

Cited By ~ 38

Author(s):

J C Zúñiga-Pflücker ◽

D Jiang ◽

P L Schwartzberg ◽

M J Lenardo

Keyword(s):

Bone Marrow ◽

T Cell ◽

Gamma Radiation ◽

B Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Cell Development ◽

Beta Chain ◽

Stage Of Development ◽

Heat Stable Antigen

DNA recombination of the immunoglobulin (Ig) or T cell receptor (TCR) gene loci is an essential step in the production of lymphocytes bearing antigen-specific receptors. Mice that lack the ability to rearrange their Ig and TCR gene loci are devoid of mature B and T cells. Complete rearrangement and expression of the TCR-beta chain has been suggested to allow immature thymocytes to switch from the CD4-/CD8- to the CD4+/CD8+ stage of thymic development. Thus, thymocytes from severe combined immune deficient (SCID) mice or mice deficient in recombinase activation genes (RAG), which do not undergo proper DNA rearrangement, are arrested at the early CD4-/CD8- stage of development. B cell precursors in SCID or RAG mice do not progress from the B220+/sIgM-/heat stable antigen (HSA)+/CD43+ to the B220+/sIgM-/HSA+/CD43- stage. In an attempt to reconstitute RAG-2-/- mice with bone marrow- or fetal liver-derived progenitor cells, we subjected these mice to sublethal doses of gamma-radiation. It is surprising that in the absence of donor cells, irradiated RAG-2-/- mice revealed a dramatic change in their lymphoid phenotype. 14 d after irradiation, the majority of thymocytes had advanced to the CD4+/CD8+ stage of T cell development and a small number of bone marrow precursors had progressed to the CD43-, HSAhi stage of B cell development. Analysis of the resulting CD4+/CD8+ thymocytes revealed no surface expression of the TCR/CD3 complex and no V-D-J rearrangement of the TCR-beta gene locus. Our findings provide evidence for a novel pathway that allows the transition of thymocytes from the CD4-/CD8- to the CD4+/CD8+ stage and that does not appear to require TCR-beta chain rearrangement.

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Oligoclonal expansion of CD8+ CD57+ T cells with restricted T-cell receptor beta chain variability after bone marrow transplantation

Blood ◽

10.1182/blood.v83.2.587.587 ◽

1994 ◽

Vol 83 (2) ◽

pp. 587-595 ◽

Cited By ~ 105

Author(s):

G Gorochov ◽

P Debre ◽

V Leblond ◽

B Sadat-Sowti ◽

F Sigaux ◽

...

Keyword(s):

Bone Marrow ◽

T Cell ◽

T Cell Receptor ◽

Gene Families ◽

Cell Receptor ◽

Pcr Analysis ◽

Gene Rearrangements ◽

Beta Chain ◽

Beta Gene ◽

Oligoclonal Expansion

Abstract A major expansion of CD8+57+ lymphocytes expressing an alpha-beta T- cell receptor (TCR) is frequent after bone marrow transplantation (BMT). We examined the clonality of the TCR beta gene repertoire in these expanded CD8+57+ cells after allogeneic or autologous BMT. We performed a polymerase chain reaction (PCR) analysis of the V beta chain usage in CD8+57+ cells purified from nine BMT recipients with a series of oligonucleotides specific for 20 V beta gene families. PCR products from selected TCR beta gene rearrangements were sequenced. The CD8+57+ cells from eight of nine patients used a restricted set of V beta families, with a marked predominance of two to three V beta gene families per patient, whereas the control autologous CD57- subset expressed the whole 20 V beta families. A direct sequencing analysis confirmed the V beta 16 and V beta 17 clonality in six patients, showing a striking homology in the CDR3 sequences of the V beta 16 products. The CD8+57+ cells, but not the CD57- cells, displayed an oligoclonal pattern of TCR rearrangements as shown by PCR analysis of TCR gamma gene rearrangements. Such an oligoclonal expansion of CD8+57+ cells, using a restricted set of the V beta gene families, may result from a specific TCR stimulation of a limited number of T-cell clones in BMT recipients.

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