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TRANSPLANTATION
From the Center for Hematologic Oncology and Department
of Biostatistics, Dana-Farber Cancer Institute; Department of Medicine,
Brigham and Women's Hospital; and Harvard Medical School; all of
Boston, MA.
Reconstitution of T-cell immunity after bone marrow transplantation
(BMT) is often delayed, resulting in a prolonged period of
immunodeficiency. Donor lymphocyte infusion (DLI) has been used to
enhance graft-versus-leukemia activity after BMT, but the effects of
DLI on immune reconstitution have not been established. We studied 9 patients with multiple myeloma who received myeloablative therapy and
T-cell-depleted allogeneic BMT followed 6 months later by infusion of
lymphocytes from the same donor. DLI consisted of
3 × 107 CD4+ donor T cells per kilogram
obtained after in vitro depletion of CD8+ cells. Cell
surface phenotype of peripheral lymphocytes, T-cell receptor (TCR) V Although allogeneic bone marrow transplantation
(BMT) provides potentially curative therapy for patients with a variety
of hematologic malignancies, previous studies have documented prolonged periods of cellular immunodeficiency following
transplantation.1-3 This prolonged period of
immunodeficiency places patients at high risk for infection with
opportunistic organisms and often results in significant morbidity and
mortality.4,5 Many aspects of T-cell function have been
examined after allogeneic BMT.4,6,7 These studies have
documented a variety of cellular defects in B- and T-cell function that
gradually improve after transplantation.8-13 These
functional deficiencies often persist for long periods after recovery
of phenotypically normal numbers of B and T cells in peripheral blood.
For example, several studies have demonstrated that 1 to 2 years are
required for the reconstitution of a normal T-cell receptor (TCR)
repertoire in patients who receive myeloablative therapy.14-16 Reconstitution of a normal T-cell repertoire
may occur more rapidly in children who have higher levels of thymic function, and this observation suggests that it may be possible to
develop methods for enhancing functional T-cell recovery after allogeneic BMT.8,17,18
In patients with hematologic malignancies, reconstitution of normal
allogeneic stem cells is also associated with the development of
immunity to residual tumor cells that have not been eliminated by the
transplantation preparative regimen.19,20 The
effectiveness of this graft-versus-tumor response has been demonstrated
by the high response rates observed in patients who receive donor
lymphocyte infusions (DLIs) for treatment of relapse after allogeneic
BMT. Responses occur most often in patients with chronic myelocytic leukemia (CML),21,22 but patients with multiple myeloma
and B-cell lymphoma also frequently respond to single infusions of donor lymphocytes without additional therapy.23-26 The
most significant toxicity associated with DLI is the development of
graft versus host (GVH) disease. Recent clinical trials have suggested
that the incidence and severity of GVH disease can be reduced by in vitro depletion of CD8+ T cells from the DLI
product.23,27,28 Importantly, graft-versus-leukemia activity appears to be maintained with infusion of defined numbers of
CD4+ donor cells. Although the immunologic targets of the
antileukemia response have not yet been well defined, previous studies
have demonstrated profound immunologic effects of CD4+ DLI
in patients who respond to this treatment.29-31 These
immunologic effects have included increased levels of T-cell
differentiation from hematopoietic stem cells, which results in
increased diversity of the TCR repertoire.30,32
The immunologic effects of DLI in patients with relapsed disease
suggested that DLI might provide a general method for enhancing cellular immune function following allogeneic hematopoietic stem cell
transplantation. We therefore examined reconstitution of cellular
immunity in a cohort of 9 patients with multiple myeloma who received
prophylactic infusion of CD8-depleted donor lymphocytes and compared
these results to a group of 7 similar patients who did not receive DLI.
All patients received myeloablative therapy followed by infusion of CD6
T-cell-depleted donor bone marrow (BM) from HLA-identical
siblings.33 Patients who received prophylactic DLI
received a single infusion of CD4+ T cells
(3 × 107/kg) from the same donor 6 months after BMT. In
patients with persistent disease after transplantation,
CD4+ DLI resulted in further reductions in tumor burden.
Analysis of peripheral blood lymphocytes revealed no significant
differences in the numbers of circulating CD3+ T cells, but
patients who received DLI developed increased numbers of
CD20+ B cells. CD4+ DLI was also associated
with increased levels of T-cell neogenesis and more rapid
reconstitution of the TCR repertoire. CD4+ DLI was also
associated with conversion of mixed chimerism to complete donor
hematopoiesis. Taken together, these results suggest that, in addition
to specifically enhancing antitumor immunity, prophylactic infusion of
CD4+ donor cells can provide a method for generally
improving reconstitution of T-cell immunity after allogeneic BMT.
Further studies can now be undertaken to examine the potential clinical
utility of this approach.
Patient treatment and preparation of patient samples
Flow cytometry analysis
RNA extraction, reverse transcription, and polymerase chain reaction RNA was extracted from 10 × 106 PBMCs using RNeasy Mini Kit (Quiagen, Valencia, CA) according to the manufacturer's protocol. First-strand complementary DNA (cDNA) was generated from 2 µg total RNA using random hexanucleotides (Pharmacia LKB Biotechnology, Piscataway, NJ) and reverse transcriptase (Superscript, GIBCO, Gaithersburg, MD). Each TCR V segment was
amplified with one of the 24 V subfamily-specific primers previously
described and a C primer recognizing both C1 and C2
regions.29,34 Polymerase chain reaction (PCR)
amplification of V5 and V13 required the use of 2 sets of primers
to identify the entire V subfamily. The C primer was conjugated
to fluorescent dye 6-FAM (Applied Biosystems, Foster City, CA) for CDR3
size analysis.
TCR V CDR3 fluorescent PCR
product was determined by electrophoresis on an automated 377 DNA sequencer (Applied Biosystems) using 4% polyacrylamide gels, and data
were analyzed by GeneScan software (Perkin Elmer Cetus Instruments, Emeryville, CA). A normal transcript size distribution, reflecting polyclonal cDNA, contains 8 to 10 peaks for each V
subfamily.35 The appearance of dominant peaks indicates
the presence of excess cDNA of identical size, suggesting the presence
of oligoclonal or clonal T-cell populations.
Analysis of hematopoietic chimerism Genomic DNA was extracted from 2 × 106 PBMCs using Wizard Genomic DNA Purification kit (Promega, Madison, WI) according to the manufacturer's recommendations. Prior to amplification, all DNA samples were quantified by UV spectrophotometry and diluted to working concentrations. Genomic DNA was extracted from samples of each donor-recipient pair before transplantation and amplified by PCR with a panel of 7 primer pairs specific for polymorphic microsatellite regions to identify an informative locus. The previously described primer sequences are designated as B7, H10, H12, H4, 3p2, pi, and CAR.15,36,37 The 3' primer of each pair was conjugated to fluorescent 6-FAM or Hex dye (Genosys Biotechnologies, The Woodlands, TX). PCR conditions included an initial denaturation of the DNA template at 94°C for 5 minutes, followed by denaturation at 94°C for 60 seconds, primer annealing at 55°C for 60 seconds, and primer extension at 72°C for 60 seconds for 40 cycles.30 A final 10-minute extension at 72°C followed the last cycle. Aliquots of the PCR products were electrophoresed on an automated 377 DNA sequencer (Applied Biosystems) using 4% polyacrylamide gel and analyzed by GeneScan software (Perkin Elmer Cetus Instruments). To quantify the donor-recipient ratio in patient samples, results were compared with standards derived from amplification of different mixtures of donor and recipient DNA (ranging from 90:10 to 10:90).TCR rearrangement excision circles To detect signal-joint TCR rearrangement excision circles (TRECs), a real-time quantitative PCR method was used.32,38 This method utilizes a fluoresceinated probe that hybridizes between the PCR primers. Each PCR reaction was performed in a 50 µL volume containing 0.09 µg genomic DNA, 1 × Taqman buffer A (Perkin Elmer Cetus Institute), 3 mM MgCl2, 300 nM each primer, 100 nM probe, 200 nM dATP, 200 nM DCTP, 200 nM DGTP, 400 nM dUTP, 17 units UNG, and 2 units AmpliTaq Gold DNA polymerase (Perkin Elmer). The PCR primer sequences were sense 5'-CGTGAGAACGGTGAATGAAGAGCAGACA-3', antisense 5'-CATCCCTTTCAACCATGCTGACACCTCT-3'. The probe sequence was 5'-VIC-TTTTTGTAAAGGTGCCCACTCCTGTGCACGGTGA-TAMRA-3'. A series of standard dilutions of plasmid containing the signal-joint breakpoint was used to quantitate TRECs in each patient and control DNA sample. By comparing the PCR cycle at which fluorescence was first significantly elevated above background (the CT or threshold cycle) in a patient sample relative to the standard curve of known concentration of the plasmid, it was possible to accurately quantitate the starting copy number of TRECs in the sample. Each patient and control DNA sample was run in duplicate on a 96-well plate along with the dilution series of the TREC plasmid. The same samples were also run on the same plate with established primers and probes for GAPDH. The GAPDH copy number served as a control for both the quality and amount of genomic DNA in the sample.Statistical analysis Univariate analyses were done using the Wilcoxon Mann-Whitney rank sum test to compare median cell counts, change in cell counts, and complexity scores between those patients who received DLI and those who did not receive DLI at each time point. The Fisher exact test was used to compare the percent of patients who returned to the normal complexity score and the percent of patients who converted to complete donor chimerism over the first 18 months following BMT. The normal complexity score was defined as 137.6, the 95% lower confidence interval of scores for the 7 healthy donors.15 The median time to complete donor chimerism was estimated using the Kaplan-Meier method and compared between the 2 groups using the Gehan Wilcoxon statistic. A mixed model using an autoregressive correlation structure and the method of restricted maximum likelihood39 was fit to account for repeated measures over time for the absolute cell counts, change in cell counts, complexity score, and change from baseline in the log (base 10)-transformed number of TREC copies per 105 CD3+ T cells. Those samples with TREC values lower than the limit of detection were set equal to the limit of detection (100 copies per 105 CD3+ cells)
Patient characteristics Nine patients with multiple myeloma received myeloablative therapy followed by infusion of BM from HLA-identical siblings and infusion of CD4+ lymphocytes from the same donor 6 months later (Table 1). The median age was 47 years (range, 43-54 years). Eight patients received total body irradiation (14 Gy) and cyclophosphamide (120 mg/kg), and 1 patient received cyclophosphamide (120 mg/kg) plus busulfan (16 mg/kg). In each case, donor marrow was depleted of CD6+ T cells as previously described, and patients did not receive prophylactic immunosuppressive therapy after BMT.40 None of these patients developed greater than grade 1 acute GVH disease after BMT. At the time of transplantation, 7 patients were in partial response (PR) and 2 were in complete remission (CR). Prior to DLI 6 months after BMT, 2 patients had evidence of progressive disease, 3 were in PR, 2 had stable disease, and 2 were in CR. After DLI, all 7 patients with evidence of disease demonstrated evidence of a further response.33 Five patients achieved CR, and 2 demonstrated a further PR. Two patients in CR before DLI remained in CR.
Seven patients (5 with multiple myeloma and 2 with non-Hodgkin
lymphoma) received similar myeloablative therapy followed by infusion
of CD6 T-cell-depleted marrow from HLA-identical sibling donors. These
patients did not receive prophylactic DLI after BMT. Clinical
characteristics of these patients are summarized in Table
2. The total number of CD3+
cells per kilogram infused was similar in both groups. However, 2 patients in the no-DLI group developed significant GVH disease after
BMT. Patient no. 10 relapsed 9 months after BMT and was excluded from
further immunologic analysis after this time.
Lymphocyte reconstitution following BMT and CD4+ DLI To identify the immunologic effects of prophylactic CD4+ DLI, we first examined well-defined lymphocyte subsets in peripheral blood during the first 6 months after BMT and for 1 year after DLI. The absolute number of CD3+, CD4+, and CD8+ T cells, CD20+ B cells, and CD56+ natural killer (NK) cells present at various times is summarized in Figure 1A. Each of these subsets had recovered by 3 months after BMT, but CD4+ T cells and CD20+ B cells were only present at low levels. There was little change in these subsets 6 to 9 months after BMT, but the number of CD3+CD8+ cells decreased at 12 and 18 months after BMT. There was no significant change in the number of NK cells throughout this period. The number of CD20+ B cells increased from a median of 144 (range, 99-243 cells) 3 months after BMT to 730 (range, 160-1257 cells) 12 months after BMT and 631 (range, 159-3167 cells) at 18 months after BMT. These differences were statistically significant at 9 and 12 months (P = .03 and P = .05, respectively). The increase in CD20+ B cells occurred in patients who achieved CR as well as in patients who continued to demonstrate only PR after DLI. The number of CD3+CD4+ T cells was low and stable in the first 6 months after BMT and gradually increased at later time points. No significant differences were found in lymphocyte subpopulations between patients who developed GVH disease (grade 2-4) and patients without GVH disease after DLI. A similar analysis of lymphocyte subsets in patients who did not receive prophylactic DLI is shown in Figure 1B. When compared with patients who received CD4+ DLI, the only statistically significant difference was the lower absolute number of CD20+ B cells at 9 (P = .001) and 12 months (P = .04) after BMT. No significant differences between the 2 groups at any time point were detected in the absolute number of CD3+ (P = .23), CD4+ (P = .60), CD8+ (P = .18), or CD56+ (P = .10) cells.
B-cell reconstitution was also analyzed in a cohort of 87 patients with other hematologic malignancies who received the same myeloablative conditioning regimen followed by CD6 T-cell-depleted BMT (Figure 1C).6,40 The median absolute number of B cells in this additional control group of patients was 111.5 (range, 12.5-678.2 cells) (n = 41), 139.5 (range, 17.6-563 cells) (n = 31), 211.1 (range, 57.6-1242 cells) (n = 25), 260.2 (range, 28-1164 cells) (n = 39), and 221.1 (range, 67.2-678.7 cells) (n = 15) at 3, 6, 9, 12, and 18 months, respectively. As shown in Figure 1C, there was no difference between the 2 groups who received only T-cell-depleted BMT; P = .43, P = .44, P = .99, P = .73 at 3, 9, 12, and 18 months after BMT, respectively. Because we did not find any difference between these 2 groups, they were combined and compared with the group of patients who received DLI. In this analysis, B-cell reconstitution was significantly greater in patients who received DLI (P = .0012). Reconstitution of TCR V
spectratyping. As shown for patient no. 6 in Figure
2B, V spectratype profiles 3 months
after BMT were markedly abnormal compared with normal donor spectratype
shown in Figure 2A. Most patient profiles contained only clonal or
oligoclonal peaks, indicating marked limitation of the TCR repertoire.
In contrast, spectratype profiles 3 months after CD4+ DLI
(Figure 2C) demonstrated marked improvement with few oligoclonal profiles. To compare TCR repertoires in different patients, we utilized
a complexity score to quantify changes in repertoire after BMT and
DLI.15 Based on the analysis of 7 healthy donors, we calculated a score of 137 to be the lower limit of the 95% confidence interval for the mean of the healthy donors. As
shown in Figure 3, all patients in both
groups had low complexity scores 3 months after BMT compared with the
healthy donors median score of 82 (range, 75-111) in the DLI
group and 84 (range, 80-98) in the no-DLI group. In patients who
received DLI, complexity scores pre-DLI (6 months after BMT) were not
significantly higher than at 3 months after BMT (data not shown). TCR
complexity scores improved at 9 months after BMT in patients who
received CD4+ DLI. In contrast, complexity scores remained
low in patients who did not receive DLI. In comparison with patients
who received DLI, these values were significantly lower at all
subsequent time points; P = .02, P = .05, and
P = .04 at 9, 12, and 18 months, respectively. Comparison
of the rate of recovery confirmed that complexity scores improved more
rapidly after BMT in patients who received DLI
(P = .01).
Effect of CD4+ DLI on hematopoietic chimerism Analysis of informative polymorphic microsatellite regions was used to distinguish between donor and recipient cells and to quantify the extent of hematopoietic chimerism after BMT. All patients had evidence of engraftment with donor cells, but only 1 patient in the DLI group had established complete donor hematopoiesis at 3 and 6 months after BMT (Figure 4). All other patients in the DLI group were found to have stable mixed hematopoietic chimerism prior to DLI, with 25% to 80% donor cells in peripheral blood. By 3 months after DLI, 5 additional patients had established complete donor chimerism. By 1 year after DLI, 8 of 9 patients had complete donor hematopoiesis and only 1 patient had persistent mixed chimerism. All patients who did not receive DLI also had evidence of mixed hematopoietic chimerism 3 months after BMT. With further follow-up, a gradual increase in the fraction of patients who converted to complete donor hematopoiesis was also noted in this group. However, at 9 months after BMT, only 1 of 7 patients had established complete donor hematopoiesis, and only 1 additional patient developed complete donor hematopoiesis 1 year after BMT without DLI. Although the fractions of patients who eventually converted to complete donor chimerism in the 2 groups were not significantly different (P = .26), the median time to achieve complete donor chimerism was 9 months with DLI and 18 months without DLI. This difference is statistically significant (P = .05, Wilcoxon).
Quantitative analysis of TCR excision circles Quantitative measurement of TRECs was also used to examine reconstitution of T cells after BMT and DLI. TRECs are produced in individual T cells as a byproduct of TCR gene rearrangement. Previous studies have demonstrated that these episomal DNA fragments can therefore be used to identify recent thymic emigrants and to measure recovery of T-cell neogenesis after BMT.32,38 Using a quantitative PCR assay, TRECs were measured in the same samples used to evaluate TCR V repertoire, lymphocyte phenotype, and chimerism.
Because only CD3+ cells contain TRECs, results of assays
summarized in Figure 5 show the number of
TREC copies measured per 105 CD3+ T cells.
Prior to transplantation, the median TREC count was 6.1 × 103 copies. This value was not statistically
different from the TREC counts of 10 healthy donors of similar
ages (data not shown). The median value of TREC counts 3 months after
BMT was significantly lower than pretransplantation values
(P = .04), and TREC counts were below the limit of
detection for this assay (< 100 copies per 105
CD3+ cells) in 4 patients. TREC values increased 6 months
after BMT but remained significantly lower than pre-BMT values
(P = .01). At 6 months after BMT, only 3 patients still
had undetectable TRECs in circulating T cells. By 9 months after BMT,
TREC values had returned to pretransplantation levels and were
significantly higher than pre-DLI values (P = .022). TREC
levels remained normal throughout the 1-year follow-up period after
DLI. Also shown in Figure 5, 1 patient had relatively low TREC values
at all the time points evaluated. This patient had relatively low TREC
copies before transplantation and was also the single individual who demonstrated persistent mixed hematopoietic chimerism after DLI. TREC
values were compared in patients who developed grade 2-4 GVH disease
(n = 5) and patients without GVH disease (n = 4) after DLI and were
not significantly different (P > .06 Mann-Whitney rank
sum test).
TRECs were also measured in patients who did not receive DLI. As shown
in Figure 6, median TREC counts in
peripheral blood CD3+ cells were similar in both groups
prior to BMT and at 3 months after transplantation. TREC values
returned to normal levels 9 months after BMT in patients who received
DLI, but TREC values remained significantly low in patients who did not
receive DLI. TREC values increased in patients who did not receive DLI
at 12 and 18 months after BMT, but recovery of normal TREC counts was significantly slower in patients who did not receive DLI
(P < .0001).
Previous reports from several centers have documented high response rates following DLI in patients with multiple myeloma who relapse after allogeneic BMT. Responses are maintained when CD8+ cells are depleted from the donor lymphocytes prior to infusion, and previous clinical trials have demonstrated that approximately 50% of patients with relapsed multiple myeloma have clinical responses after infusion of 3 × 107 to 15 × 107 CD4+ cells per kilogram.23 To determine whether earlier infusion of donor lymphocytes would provide more effective control of myeloma, we undertook a clinical trial to examine the antitumor effects of prophylactic DLI in patients who received myeloablative therapy followed by infusion of CD6 T-cell-depleted BM from HLA-identical sibling donors.33 No prophylactic immune suppressive therapy was administered, and patients without GVH disease received a single infusion of 3 × 107 CD4+ from the same donor 6 months after BMT. Despite administration of myeloablative therapy, most patients continued to have evidence of persistent myeloma 6 months after BMT. Importantly, all patients with persistent disease had evidence of further disease response following CD4+ DLI. These observations confirmed previous studies indicating that allogeneic immune responses contribute to the control of multiple myeloma following BMT and support the further use of donor immune responses to improve outcomes of allogeneic BMT in this disease. The prophylactic infusion of donor lymphocytes in a defined cohort of patients also provided us with an opportunity to examine the immunologic effects of this treatment. Although the antitumor activity of DLI has been well documented, the immunologic mechanisms responsible for tumor rejection have not been defined, and the effects of DLI on cellular immune function in the recipient have not been well characterized. To examine these immunologic effects, serial samples of PBMCs were obtained over an 18-month period from a cohort of 9 patients enrolled in this clinical trial who received prophylactic DLI. The immunologic studies carried out with these samples were designed to provide a quantitative assessment of reconstitution of T-cell immunity rather than the recovery of T-cell responsiveness to specific antigens. For comparison, similar studies were carried out with cryopreserved samples from 7 patients who underwent similar therapy except that they did not receive prophylactic DLI. We first examined the reconstitution of well-defined lymphocyte subsets of PBMCs using flow cytometry. Although all patients received T-cell-depleted donor marrow, CD3+ cells had recovered by 3 months after BMT. Consistent with the results of previous studies, these cells were predominately CD8+, and the number of CD4+ T cells recovered very slowly after BMT.6 NK cells recovered rapidly after BMT, but B cells recovered slowly. Infusion of CD4+ donor lymphocytes at 6 months after BMT had no discernable effect on the number of CD4+ or CD8+ CD3+ T cells or NK cells. However, CD20+ B cells were significantly increased in patients who received CD4+ DLI. The increase in circulating B cells was most evident at 3 and 6 months after DLI, and this observation was confirmed in further comparison with a large cohort of 87 patients with other hematologic malignancies who had undergone T-cell-depleted BMT without DLI. The demonstration of increased B-cell reconstitution after CD4+ DLI is consistent with previous studies indicating that tumor rejection following DLI results from a coordinated immune response that includes B-cell as well as T-cell responses.41 Thus, patients who respond to DLI often have marrow infiltration with polyclonal plasma cells, and high-titer antibody responses to a variety of leukemia-associated antigens have been documented in these patients.41 The effect of DLI on T-cell reconstitution was also examined through
analysis of TCR repertoire. Healthy individuals maintain a
highly complex T-cell repertoire, and this can be quantified through
examination of TCR V In conjunction with assessment of TCR repertoire, we also examined the
number of TRECs in CD3+ PBLs. TRECs are generated as a
byproduct of TCR Although the mechanism by which infusion of a relatively small number of mature CD4+ donor T cells can enhance thymic function and T-cell neogenesis is not known, we have previously observed that reconstitution of a normal T-cell repertoire was also associated with the establishment of complete donor hematopoiesis.15 We therefore included an examination of hematopoietic chimerism in our assessment of T-cell reconstitution and found that CD4+ DLI also appeared to enhance the conversion of mixed chimerism to complete donor hematopoiesis. Five patients converted to complete donor hematopoiesis 3 months after DLI, and 8 of 9 patients had complete donor hematopoiesis 1 year after DLI. Interestingly, the single individual who maintained mixed hematopoietic chimerism after DLI was also the only individual who had minimal recovery of TRECs after DLI. These observations suggest that reconstitution of cellular immunity after transplantation is facilitated by the establishment of full donor hematopoiesis, but further studies will be necessary to determine the mechanism whereby this occurs in these patients. In summary, our examination of cellular immunity after prophylactic DLI provides consistent evidence that CD4+ DLI initiates a profound and long-lasting effect on T-cell immunity after allogeneic stem cell transplantation. Interestingly, these immunologic effects reflect changes in global reconstitution of T-cell immunity and appear to occur independently of any specific response to residual recipient tumor cells or tumor-associated antigens. Further studies in animal model systems as well as in patients who receive DLI will be necessary to better define the mechanism underlying this immunologic effect. Nevertheless, these studies suggest that infusion of CD4+ donor T cells may provide a clinically simple and straightforward method for enhancing reconstitution of T-cell immunity as well as promoting antitumor immunity after allogeneic BMT. This hypothesis can be examined in future clinical trials to determine whether prophylactic CD4+ DLI might improve clinical outcomes by reducing the incidence of opportunistic infections after allogeneic stem cell transplantation
Submitted September 17, 2001; accepted February 5, 2002.
Supported by NIH grants AI29530, HL04293, and CA78378. R.J.S. is a Clinical Research Scholar of the Leukemia and Lymphoma Society. K.C.A. is a Doris Duke Distinguished Clinical Scientist.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Jerome Ritz, Dana-Farber Cancer Institute, 44 Binney St
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Top
Abstract
Introduction
gene rearrangement and therefore provide an
independent measure of T-cell neogenesis from undifferentiated
hematopoietic stem cells. Although the thymus undergoes involution in
adults, analysis of TRECs in healthy individuals has
demonstrated that T-cell neogenesis continues despite thymic involution, and this contributes to the maintenance of a diverse T-cell
repertoire.