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Blood, Vol. 92 No. 1 (July 1), 1998:
pp. 11-18
Intracellular Cytokine Profile of Cord and Adult Blood Lymphocytes
By
I.M.H. Chalmers,
G. Janossy,
M. Contreras, and
C. Navarrete
From the Department of Histocompatibility and Immunogenetics, North
London Centre (NBS), London, UK; and the Department of Immunology,
Royal Free Hospital, Pond Street, London, UK.
 |
ABSTRACT |
Umbilical cord blood (CB) transplantation is thought to be
associated with a reduced risk of severe graft-versus-host-disease (GVHD) compared with bone marrow transplantation (BMT). The cytokine cascade is known to be important in the pathogenesis of GVHD; however,
previous studies investigating the cytokine secretion pattern of CB
cells have been contradictory because of variations in experimental
techniques. In this study, the cytokine profile of cord and adult blood
lymphocytes and lymphocyte subsets has been assessed at the single-cell
level by flow cytometry, using CD4/CD8 and CD45RA/CD45RO markers. Cord
and adult blood mononuclear cells were stimulated with phorbol
12-myristate 13-acetate (PMA) and ionomycin in the presence of
monensin. After 4 to 24 hours of incubation, interleukin-2 (IL-2),
IL-4, interferon- (IFN-), and tumor necrosis factor- (TNF-)
production was measured by three-color flow cytometry. The results show
that cord blood lymphocytes (CBL) produce less IL-2, IL-4, IFN-, and
TNF- than adult peripheral blood lymphocytes (ABL). Further subset
analysis showed that in CBL the majority of cytokine producing cells
were CD4+CD45RA+, whereas in ABL the
cytokine-producing cells were both
CD4+CD45RO+ and
CD8+CD45RO+. These results suggest that the
reduced incidence of GVHD in CB transplantation may partly due to the
altered cytokine profile seen in CBL.
| |
INTRODUCTION |
ALLOGENEIC BONE MARROW transplantation
(BMT) is an accepted form of therapy for hematological malignancies, BM
failure syndromes, immunodeficiency states, and metabolic
disorders.1 However, its success is dependent on the prompt
identification of a suitable donor and on the avoidance of
opportunistic infections and severe graft-versus-host disease (GVHD).
To overcome some of these complications and to expand the available
donor pool, cord blood (CB) has been used as an alternative source of
hematopoietic stem cells.
CB is a rich source of primitive hematopoietic stem cells and
progenitor cells,3 with extensive proliferative and
self-renewal capacity ex vivo4,5 and possibly in
vivo.6 The first successful CB transplant was performed in
1988 on a patient with Fanconi anemia.6 Subsequently,
umbilical CB has been used in both related7 and unrelated
BMT,8 and in both settings there appears to be a reduced
incidence and severity of GVHD when compared with results obtained
using BM.9 Because GVHD is a major cause of morbidity and
mortality in allogeneic BMT, this feature of CB transplantation could
prove to be crucial for its use in stem cell transplantation.
The development of GVHD involves recognition of alloantigen(s) present
in the patient by donor T cells.10 However, the clinical manifestations largely result from cytokine
dysregulation.11 Therefore, it is important to investigate
whether altered cytokine production of CB could account for the reduced
incidence of GVHD observed in CB transplantation.
A variety of different experimental techniques, including enzyme-linked
immunosorbent assay (ELISA) or bioassays, have been used to measure
cytokine secretion by CB cells, leading to limited and inconclusive
results.12,13 In this study interleukin-2 (IL-2), IL-4,
tumor necrosis factor- (TNF-), and interferon- (IFN-)
production by phorbol 12-myristate 13-acetate
(PMA)-and-ionomycin-activated cord and adult lymphocytes has been
assessed at single-cell level by flow cytometry. CB contains primarily
unprimed T cells as documented by the expression of CD45RA marker on
the vast majority of both CD4 and CD8 subsets of T cells. Therefore, a
comprehensive phenotypic analysis of cytokine producing cells has been
performed using CD4/CD8 and CD45RA/CD45RO markers.
| |
MATERIALS AND METHODS |
Reagents.
PMA, ionomycin, and monensin were purchased from Sigma
(Poole, UK) and Calbiochem/Novachem (Nottingham, UK), respectively.
Monoclonal antibodies (MoAbs) used were CD4-PerCP (Becton Dickinson,
Oxford, UK), CD8-PerCp (Becton Dickinson), CD45RA-FITC (Becton
Dickinson), CD45RO-FITC (Dako, High Wycombe, UK), IL-2-PE (Pharmingen,
Oxford, UK), IL-4-PE (Pharmingen), IFN--PE (Pharmingen), and
TNF--PE (Pharmingen). Isotype-specific anti-mouse IgG1 or IgG2a were
purchased from Becton Dickinson, while anti-mouse IgG1 and anti-rat
IgG2a were obtained from Pharmingen. The cells were fixed and
permeabilized with Fix and Perm solution (Calbiochem).
Cells.
Human CB was obtained from the placentas of normal full-term deliveries
at a local hospital, following ethical committee approval. Adult
peripheral blood (AB) mononuclear cells were isolated from buffy coats
obtained from local routine blood donations.
Mononuclear cells were obtained from all samples by centrifugation over
Ficoll-Hypaque gradients (Nycomed, Birmingham, UK). Before
separation CB was incubated with a glycophorin MoAb (Nycomed) at a
concentration of 100 µL/10 mL of blood for 5 minutes at room temperature to aid removal of nucleated red blood cells and to improve
separation. All cells were frozen in 10% dimethyl sulfoxide (DMSO)
using a controlled rate freezer, stored in liquid nitrogen, and then
thawed for use.
Intracellular cytokine analysis.
Cells were cultured in 24-well plates (1 × 106/mL) for
4 to 24 hours at 37°C and 5% CO2 in RPMI 1640 supplemented with 10% AB serum, 25 mmol/L HEPES, 100 nmol/L sodium
pyruvate, 2 mmol/L L-glutamine, and penicillin/streptomycin. Cells were
stimulated with PMA (5 ng/mL) and ionomycin (1 µmol/L). Monensin (3 µmol/L) was added 4 to 10 hours before the end of culture as
described by Jung et al.14 Cultured cells were washed twice
in phosphate-buffered saline (PBS)/1% bovine serum albumin (BSA) and
then stained with MoAb to the following cell-surface markers: CD4, CD8,
CD45RA, CD45RO. The cells were then fixed and permeabilized with Fix
and Perm solution (Calbiochem) according to manufacturer's
instructions. Finally, the cells were incubated with MoAb to the
following cytokines: IL-2, IL-4, IFN-, and TNF-.
Flow cytometry.
Phenotypic analyses of mononuclear preparations in CB and AB samples
were performed by two- and three-color flow cytometry using a Becton
Dickinson FACScan flow cytometer. A minimum of 10,000 lymphocyte-gated
events were acquired in list mode and analyzed with Lysis II (Becton
Dickinson), Cellnet (Becton Dickinson), and Flowmate (Dako) software.
Statistical analysis.
Student's t-tests were used to compare CB lymphocyte (CBL)
versus adult peripheral blood lymphocyte (ABL) cytokine production. P values <.05 were considered to be significant.
| |
RESULTS |
Kinetics of cytokine synthesis.
The kinetics of IL-2, IL-4, IFN-, and TNF- synthesis are shown in
Fig 1. After PMA and ionomycin stimulation
there is a rapid increase in IL-2 production in both CBL and ABL,
reaching a peak at 12 hours. In ABL there is a similar rapid increase
in TNF- and IFN-, maximal at 8 and 12 hours, respectively.
Although in CBL there was a much lower production of both TNF- and
IFN-, the maximal production was also at 8 and 12 hours,
respectively. Both cord and adult mononuclear cells (MNC)
produce low levels of IL-4, the peak production being at 8 hours.
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| Fig 1.
Kinetics of cytokine synthesis in cord and adult blood.
Cord and adult mononuclear cells were activated with PMA/ionomycin and
IL-2, IL-4, IFN-, and TNF- expression was measured every 4 hours
for 24 hours as described in Materials and Methods. The results are
expressed as a percentage of the lymphocyte gated cells (see Fig
2).
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Cytokine expression on cord and adult blood.
Figure 2 shows the expression of IL-2,
IFN-, TNF-, and IL-4 on individual samples of cord and adult
lymphocytes, at the time of optimal production as determined by the
results in Fig 1 (12, 12, 8, and 8 hours, respectively). The forward
and side scatter (fsc/ssc) characteristics for cord and adult cells,
and the gates used for analysis of lymphocytes, are also shown in Fig
2. These results show that there is reduced production of IL-2,
IFN-, TNF-, and IL-4 in CBL compared with ABL. Further experiments were performed on six additional samples of cord and adult
blood to confirm these findings (Fig 3). In
these experiments, the mean percentage of cells secreting each cytokine
and the level of expression, indicated by the mean
fluorescence intensity (MFI), was analyzed (Fig 3A and B,
respectively). These results show that the percentage of positive cells
for each cytokine is significantly lower in CBL compared with ABL.
Likewise the level of expression of IFN- and TNF- was
significantly lower in CBL than in ABL. However, there was a similar
level of expression of IL-2 and IL-4, in both CBL and ABL positive
cells.
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| Fig 2.
Flow cytometric cytokine expression on cord and adult
lymphocytes. The method for cytokine analysis is described in Materials and Methods. The top two traces show the forward scatter/side scatter
distribution and the gate used to select lymphocytes for analysis. The
quadrants were set using isotype controls for each of the antibodies.
Percentages indicated in the right upper quadrant indicate the
percentage of the total lymphoid population (ie, the cells within the
region [R1] outlined in the dual scatter plot). For each plot cord
blood (left column) is compared with the adult blood (right column).
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| Fig 3.
(A) Cytokine expression on cord and adult blood. The
percentage positivity of six samples of cord and adult blood is shown following PMA/ionomycin activation. Results are expressed as a percentage of R1 (see Fig 2). (B) Cytokine expression on cord and adult
blood. The MFI for the cord and adult blood samples in (A) are shown.
The results are expressed as a mean of six samples of cord and adult
blood.
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CD45RA/CD45RO subset analysis.
Cytokine producing cells in CBL and ABL were further characterized, at
the time of peak production, according to expression of CD45RA/CD45RO.
The results are shown in Fig
4A. In CBL,
70% of IL-2-producing cells were CD45RA+ and 30% were
CD45RO+. Cells producing IFN- were 75%
CD45RA+ and 25% CD45RO+, while
TNF--producing cells were 90% CD45RA+ and 10%
CD45RO+. IL-4-producing cells were 75%
CD45RA+ and 25% CD45RO+. In contrast, in ABL,
the predominant IL-2-, IFN--, TNF--, and IL-4-producing cells
were CD45RO+, rather than CD45RA+ (83%
v 17%, 98% v 2%, 83% v 17%, and 94%
v 6%, respectively).
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| Fig 4.
(A) T-cell subset analysis. The cytokine producing cells
in cord and adult blood were characterized according to expression of
CD45RA and CD45RO. The results given were calculated using the ratio of
CD45RA to CD45RO, when these values were expressed as a percentage of
the total population analyzed. In each case the mean percentage
positivity as a percentage of the total lymphoid population is noted as
well as the number of samples tested (n). (B) T-cell subset analysis. The
cytokine-producing cells in cord and adult blood were further
characterized according to expression of CD4 and CD8. The values given
were calculated using the results expressed as a percentage of the
total population analyzed, as in (A).
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CD4/8 subset analysis.
Further phenotypic analysis of the cytokine-producing cells was also
performed according to CD4 and CD8 expression, and the results are
shown in Fig 4B. In CBL, IL-2 is produced predominantly by
CD4+ cells compared with CD8+ cells (80%
v 20%). Similarly, IFN-, TNF-, and IL-4 are produced mainly by CD4+ cells compared with CD8+ cells
(83% v 17%, 86% v 14%, and 61% v 39%,
respectively). In ABL, IL-2 is also produced predominantly by
CD4+ cells compared with CD8+ cells (75%
v 25%). However, IFN-, TNF-, and IL-4 are produced by
both CD4+ and CD8+ cells (49% v 51%,
44% v 56% and 42% v 58%).
The percentage and level of cytokine expression in different T-cell
subsets.
A comparison of the percentage of CD4+CD45RA+,
CD4+CD45RO+,
CD8+CD45RA+, and
CD8+CD45RO+ CBL and ABL producing IL-2,
IFN-, TNF-, and IFN- is shown in Fig
5. The level of expression as determined by
(MFI) of IL-2, IFN-, TNF-, and IL-4 in
CD4+CD45RA+,
CD4+CD45RO+,
CD8+CD45RA+, and
CD8+CD45RO+ subsets in CBL and ABL is also
shown in Fig 5.
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| Fig 5.
The percentage and level of expression of each cytokine
in different T-cell subsets. The percentage of IL-2-, IFN--,
TNF--, and IL-4-producing cells in
CD4+CD45RA+,
CD4+CD45RO+,
CD8+CD45RA+, and
CD8+CD45RO+ CBL and ABL is shown on
the left. The level of expression (MFI) of IL-2, IFN-, TNF-, and
IL-4 on CD4+CD45RA+,
CD4+CD45RO+,
CD8+CD45RA+, and
CD8+CD45RO+ CBL and ABL is shown on
the right. The results shown are the mean values of the samples shown
in Fig 4.
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There is a similar percentage of CD4CD45RA+ and
CD4+CD45RO+ CBL and ABL producing IL-2.
However, a lower number of CD8+CD45RA+ and
CD8+CD45RO+ CBL express IL-2 than ABL with a
similar phenotype. In CB, there is a reduced percentage of cells
producing IFN- and TNF- in all four T-cell subsets, compared with
AB. Although, a similar percentage of
CD4+CD45RA+ and
CD8+CD45RA+ CBL and ABL produce IL-4, there is
a lower percentage of CD4+CD45RO+ and
CD8+CD45RO+ CBL producing IL-4 compared to ABL
with the same phenotype.
In CBL and ABL there is a similar level of expression of IL-2 in all
four T-cell subsets. There is a similar level of expression of IL-4 in
CD4+CD45RA+,
CD8+CD45RA+, and
CD8+CD45RO+ CBL compared with ABL of similar
phenotype; however, there is a significant decrease in the level of
IL-4 in CD4+CD45RO CBL compared with CD4+CD45RO
ABL. Furthermore, there is a significant reduction in the level of
expression of IFN- and TNF- in all CB T-cell subsets, compared
with AB.
T-cell subsets in CBL and ABL.
To understand the differences in cytokine secretion described above, it
is important to consider the differences in T-cell subsets between CBL
and ABL. Figure 6 shows the mean percentage of CD4+CD45RA+,
CD4+CD45RO+,
CD8+CD45RA+, and
CD8+CD45RO+ subsets in CBL and ABL, expressed
as a percentage of the total lymphoid population. In CB, the majority
of T lymphocytes are either CD4+CD45RA+
(46% ± 6%) or CD8+CD45RA+
(19% ± 5%) with very few CD4+CD45RO+ and
CD8+CD45RO+ (6% ± 2% and 1% ± 2%,
respectively). In contrast, in AB there is a substantial proportion of
CD4+CD45RO+,
CD8+CD45RA+,
CD8+CD45RO+ with fewer
CD4+CD45RA+ (30% ± 12%, 21% ± 7%,
16% ± 7%, and 8% ± 4%, respectively).
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| Fig 6.
T-cell subsets in CBL ( ) and ABI ( ). The mean
percentage of CD4+CD45RA+,
CD4+CD45RO+,
CD8+CD45RA+, and
CD8+CD45RO+ subsets in six samples of CBL
and six samples of ABL is shown. The results are expressed as a
percentage of the total lymphoid population.
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| |
DISCUSSION |
Cytokine production has been extensively investigated in the past using
ELISA or bioassay techniques which measure cytokine secretion in
cell-culture supernatants. However, results have been variable and
inconclusive, largely because of the different experimental conditions
used.
In this study, the cytokine profile of CBL is reported, using a
modification of the technique described by Jung et al.14 This technique enables phenotypic characterization of
cytokine-producing cells, and therefore allows a more accurate and
detailed comparison of the cytokine profile of cord and adult
lymphocytes, at single-cell level.
Previous studies have shown that in stimulated CB, IL-2 secretion into
the supernatant is lower than15 or similar12,16 to activated AB. The results of this study show that although the
proportion of cells expressing IL-2 was lower in CBL, the level of
expression of IL-2 and the kinetics of IL-2 production are similar in
both CBL and ABL. Further subset analysis showed that the reduced
proportion of IL-2-producing cells in CB was predominantly caused by
the reduced percentage of IL-2-producing CD8+CD45RA+ and
CD8+CD45RO+ CBL compared to ABL with the same
phenotype.
These results show that not only is there a lower proportion of
IFN-- and TNF--producing cells in CB, but there is also lower
expression of IFN- and TNF- on positive cells in each. Furthermore, in CBL the reduced percentage and level of expression of
these cytokines is seen in all four T-cell subsets. Some investigators have suggested that there may be a discrepancy between intracellular stores and secretion of TNF- in some clinical
settings.17 In our study we have not been able to compare
the intracellular cytokine production with secretion into the
supernatant, but previous data confirm that CBMNC secrete less TNF-
and IFN- into the supernatant than adult blood.18-20
With regard to IL-4, this study shows that both CBL and ABL display
similar kinetics and a similar level of expression of IL-4 on positive
cells. However, the proportion of IL-4-producing cells was
consistently lower in CBL compared with ABL, in keeping with previous
data.12,21 Subset analysis has shown that in CBL the
reduced proportion of IL-4-producing cells is related to the reduced
percentage of IL-4-producing-CD4+CD45RO+ and
CD8+CD45RO+ CBL. Overall, these results
indicate that although there is a reduction in intracellular IL-2 and
IL-4 production in CBL compared with ABL, the difference is less marked
than that seen in IFN- and TNF- production.
CB is known to contain primarily `unprimed' T cells expressing the
CD45RA phenotype, and this has been confirmed in this
study.22 Therefore, it has been suggested that the reason
for the altered cytokine profile seen in CBL is the low
proportion of CD45RO+ `memory' T cells
present.23,24 However, the results presented here show that
in CBL IL-2, IL-4, IFN-, and TNF- were produced predominantly by
CD45RA+ cells, in contrast to ABL where these cytokines are
predominantly produced by CD45RO+ cells. Furthermore, there
was a reduced level of expression of IFN- and TNF- in both
CD45RA+ and CD45RO+ CBL compared to ABL with
the same phenotype. The latter observation suggests that
CD45RA+ and CD45RO+. CBL are different to
CD45RA+ and CD45RO+ ABL. Thus, the reduction in
cytokine secretion by CBL may either be due to the reduced number of
CD45RO+ cells in CB and/or the reduced capacity for
CB CD45RA+ and CD45RO+ cells to produce
cytokines other than IL-2.
Further CD4/CD8 subset analysis showed that in CBL IL-2, IL-4, IFN-,
and TNF- are all predominantly produced by
CD4+CD45RA+ cells. On the other hand, in ABL
IL-2 is predominantly produced by CD4+CD45RO+
cells, while IL-4, IFN-, and TNF- are produced by both
CD8+CD45RO+ and
CD4+CD45RO+ cells. In keeping with previous
studies, these results have shown that there is a reduced proportion of
CD8+CD45RO+ cells in CBL, which again could in
part account for the marked reduction in IL-4, IFN-, and TNF-
secretion in CBL.25,26
PMA is a phorbol ester that acts on protein kinase C, while ionomycin
is a calcium ionophore that elevates intracellular calcium. These
agents act together to stimulate the T3-Ti-mediated and IL-2-mediated signal transduction pathways.27,28 In CB,
reduced cytokine production seen after PMA/ionomycin activation may
therefore be due to immaturity of the intracellular signaling
mechanism. The precise mechanism has yet to be defined. Kilpinen et
al29 have studied the production of NF- B, which has a
central role in the transcriptional activation of genes important in
production of cytokines, particularly IL-2. In their study, activation
with phorbol dibutyrate/calcium ionophore resulted in an increased activation of NF-B in CB compared with AB. Therefore, it seems unlikely that the defect in CB involves NF-B.29 On the
other hand, Pino-Otin et al30 have shown that CB T cells
have reduced expression of CD50 (ICAM-3), which has been shown to be
important in intracellular signaling.31,32 Therefore,
reduced CD50 expression may in part account for the altered cytokine
profile of CBL.
It has been suggested that the production of IFN- and IL-2 by donor
T cells and that of IL-1 and TNF- by donor mononuclear cells play a
central role in the pathogenesis of GVHD.33 The involvement
of TNF- and IFN- has also been described in the skin explant
model.34 Other investigators have found raised levels of
IFN- and TNF- in the serum of patients with
GVHD.35,36 Previous studies have supported the early role
of IL-2 in GVHD, probably through stimulation of the production of
other cytokines such as IL-1 and TNF-.37 However,
further studies have shown that IL-2 mRNA levels in peripheral blood
were not found to be increased in patients with GVHD.38
In considering the reduced incidence of GVHD in CB transplantation it
may be important to investigate the cytokine profile of CB T cells
posttransplant, because the cytokine profile of CB T cells may be
modified in vivo. Indeed, recent studies by Sornasse et
al39 have shown that the cytokine profile of naïve CB CD4+ T cells can be modified by the addition of
differentiation-inducing cytokines.
Nevertheless, the results presented here show that CBL have an altered
intracellular cytokine profile compared with ABL, particularly with
respect to IFN- and TNF-, which might account for the reduced capacity of transplanted CB T cells to induce severe GVHD. Furthermore, the reduction of cytokine production seen in CBL may be due to the
predominance of CD4+CD45RA+ cells. Defining the
underlying mechanisms of reduced cytokine production is of crucial
importance in understanding the apparent reduced incidence and severity
of GVHD in CB transplantation.
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FOOTNOTES |
Submitted September 8, 1997;
accepted April 1, 1998.
Address reprint requests to C. Navarrete, PhD, Department of
Histocompatibility and Immunogenetics, North London Centre (NBS), Colindale Ave, London NW9 5BG, UK.
The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" is accordance with 18 U.S.C. section 1734 solely to indicate this fact.
| |
REFERENCES |
1.
Tabbara IA:
Allogeneic bone marrow transplantation.
South Med J
89:857,
1996[Medline]
[Order article via Infotrieve]
2.
Tabbara IA:
Allogeneic bone marrow transplantation: Acute and late complications.
Anticancer Res
16:1019,
1996[Medline]
[Order article via Infotrieve]
3.
Broxmeyer HE,
Douglas GW,
Hangoc G,
Cooper S,
Bard J,
English D,
Arny M,
Thomas L,
Boyse EA:
Human Umbilical cord blood as a potential source of transplantable haemopoietic stem/progenitor cells.
Proc Natl Acad Sci USA
86:3828,
1989[Abstract/Free Full Text]
4.
Broxmeyer HE,
Hangoc G,
Cooper S,
Ribeiro RC,
Graves V,
Yoder M,
Wagner J,
Vadhan-Raj S,
Rubinstein P,
Broun ER:
Growth characteristics and expansion of human umbilical cord blood and estimation of its potential for transplantation of adults.
Proc Natl Acad Sci USA
89:4109,
1992[Abstract/Free Full Text]
5.
Lu L,
Xiao M,
Shen RN,
Grigsby S,
Broxmeyer HE:
Enrichment, characterisation and responsiveness of single primitive CD34+++ human cord blood hemopoietic progenitor with high proliferative and replating potential.
Blood
81:41,
1993[Abstract/Free Full Text]
6.
Gluckmann E,
Broxmeyer HE,
Auerbach AD:
Hemopoietic reconstitution in a patient with Fanconi's anemia by means of umbilical cord blood from an HLA-identical sibling.
N Engl J Med
1989:1174,
1989
7.
Wagner JE,
Kernan N,
Steinbuch M,
Broxmeyer H,
Gluckman E:
Allogeneic sibling umbilical-cord blood transplantation in children with malignant and non-malignant disease.
Lancet
346:214,
1995[Medline]
[Order article via Infotrieve]
8.
Kurtzberg J,
Laughlin M,
Graham ML,
Smith C,
Olspn J,
Halperin EC,
Ciocci G,
Carrier C,
Stevens CE,
Rubinstein P:
Placental blood as a source of hemopoietic stem cells for transplantation into unrelated recipients.
N Engl J Med
335:157,
1996[Abstract/Free Full Text]
9.
Wagner JE:
Umbilical cord blood transplantation: Overview of the clinical experience.
Blood Cells
20:227,
1994[Medline]
[Order article via Infotrieve]
10.
Korngold R,
Sprent J:
T cell subsets and graft-versus-host disease.
Transplantation
44:335,
1987[Medline]
[Order article via Infotrieve]
11.
Antin JH,
Ferrara JLM:
Cytokine dysregulation and acute graft-versus-host disease.
Blood
80:2964,
1992[Abstract/Free Full Text]
12.
Roncarolo MG,
Bigler M,
Ciuti E,
Martino S,
Tovo PA:
Immune responses by cord blood cells.
Blood Cells
20:573,
1994[Medline]
[Order article via Infotrieve]
13.
von Freeden U,
Zessack N,
van Valen F,
Burdach S:
Defective Interferon gamma production in neonatal T cells is independent of interleukin-2 receptor binding.
Pediatr Res
30:270,
1991[Medline]
[Order article via Infotrieve]
14.
Jung T,
Schauer U,
Heusser C,
Neumann C,
Reiger C:
Detection of intracellular cytokines by flow cytometry.
J Immunol Methods
159:197,
1993[Medline]
[Order article via Infotrieve]
15.
Watson W,
Ramdahin R,
Harman C:
Immunoglobulin and cytokine production by neonatal lymphocytes.
Clin Exp Immunol
83:169,
1991[Medline]
[Order article via Infotrieve]
16.
Andersson U,
Andersson J,
Lindfors A,
Wagner K,
Moller G,
Heusser C:
Simultaneous production of interleukin 2, interleukin 4, and interferon- by activated human blood lymphocytes.
Eur J Immunol
20:1591,
1990[Medline]
[Order article via Infotrieve]
17.
Hoffmann MW,
Wonigeit K,
Steinhoff G,
Herzbeck H,
Flad HD,
Pichlmayr R:
Production of cytokines (TNF-alpha, IL-1-beta) and endothelial cell activation in human liver allograft rejection.
Transplantation
55:329,
1993[Medline]
[Order article via Infotrieve]
18.
Sautois B,
Fillet G,
Beguim Y:
Comparative cytokine production by in vitro stimulated mononucleated cells from cord and adult blood.
Exp Hematol
25:103,
1997[Medline]
[Order article via Infotrieve]
19.
Wilson C,
Westall J,
Johnston L,
Lewis DB,
Dower SK,
Alpert AR:
Decreased production of interferon-gamma by human neonatal cells.
J Clin Immunol
77:860,
1986
20.
Wakasugi N,
Virelizier J-L,
Arezana-Seisdedos F,
Rothhut B,
Huerta J-MM,
Russo-Marie F,
Fiers W:
Defective IFN- production in the human neonate.
J Immunol
134:172,
1985[Abstract]
21.
Lewis DB,
Prickett KS,
Larsen A,
Grabstein K,
Weaver M,
Wilson CB:
Restricted production of interleukin 4 by activated human T cells.
Proc Natl Acad Sci USA
85:9743,
1988[Abstract/Free Full Text]
22. (suppl 1)
Chalmers IMH,
Aird A,
Green T,
Janossy G,
Contreras M,
Navarrete C:
Cellular and cytokine responses in cord and adult peripheral blood mononuclear cells.
Br J Haematol
97:2,
1997
23.
Kruse A,
Neustock P,
Reuter M,
Kirchner H:
Interferon and lymphokine production by human placental and cord blood cells.
Pediatr Res
30:270,
1993
24.
Akbar AN,
Terry L,
Timms A,
Beverley PC,
Janossy G:
Loss of CD45R and gain of UCHL1 reactivity is a feature of primed T cells.
J Immunol
140:2171,
1988[Abstract]
25.
Harris DT,
Schumacher MJ,
Locascio J,
Besencon FJ,
Olson GB,
Deluca D,
Shenker L,
Bard J:
Phenotypic and functional immaturity of umbilical cord blood T lymphocytes.
Proc Natl Acad Sci USA
89:10006,
1992[Abstract/Free Full Text]
26.
Flomenberg N,
Keever CA:
Cord blood transplants: Potential utility and potential limitations.
Bone Marrow Transplant
10:115,
1992
27.
Scheinman RI,
Cogswell PC,
Lofquist AK,
Baldwin AS:
Role of Transcriptional activation of IB synthesis in mediation of immunosupression by glucocorticoids.
Science
270:283,
1995[Abstract/Free Full Text]
28.
Auphan N,
Didonato JA,
Rosette C,
Helmberg A,
Karin M:
Immunosupression by glucocorticoids: inhibition of NF-B activity through induction of IB synthesis.
Science
270:286,
1995[Abstract/Free Full Text]
29.
Kilpinen S,
Henttinnen T,
Lahdenpohja N,
Hulkkonen J,
Hurme M:
Signals leading to the activation of NF-B transcription factor are stronger in neonatal than adult T lymphocytes.
Scand J Immunol
44:85,
1996[Medline]
[Order article via Infotrieve]
30.
Pino-Otin MR,
Juan M,
de la Fuente MA,
Vinas O,
Martinez-Caceres E,
Fernandez MD,
Miralles A,
Vilella R,
Yague J,
Vives J,
Gaya A:
CD50 (intercellular adhesion molecule-3) is expressed at higher levels on memory than on naive human T cells but induces a similar calcium mobilization on both subsets.
Tissue Antigens
46:32,
1995[Medline]
[Order article via Infotrieve]
31.
Cid MC,
Esparza J,
Juan M,
Miralles A,
Ordi J,
Vilella R,
Urbano MA,
Gayà A,
Vives J,
Yagüe J:
Signalling through CD50 (ICAM-3) stimulates T lymphocyte binding to human umbilical vein endothelial cells and extracellular matrix proteins via an increase in beta 1 and beta 2 integrin function.
Eur J Immunol
24:1377,
1994[Medline]
[Order article via Infotrieve]
32.
Vilella R,
Mila J,
Lozano F,
Alberola-Ila J,
Places L,
Vives J:
Involvement of the CDw50 molecule in allorecognition.
Tissue Antigens
36:203,
1990[Medline]
[Order article via Infotrieve]
33.
Ferrara JLM,
Deeg HJ:
Mechanisms of disease: Graft-versus-host disease.
N Engl J Med
324:667,
1991[Medline]
[Order article via Infotrieve]
34.
Dickinson AM,
Sviland L,
Hamilton PJ,
Usher P,
Taylor P,
Jackson G,
Dunn J,
Proctor SJ:
Cytokine involvement in predicting clinical graft-versus-host disease in allogeneic bone marrow transplant recipients.
Bone Marrow Transplant
13:65,
1994[Medline]
[Order article via Infotrieve]
35.
Niederweiser D,
Herold M,
Woloszczuk W,
Aulitzky W,
Meister B:
Endogenous IFN-gamma during human bone marrow transplantation.
Transplantation
50:620,
1990[Medline]
[Order article via Infotrieve]
36.
Holler E,
Kolb HJ,
Hintermeier-Knabe R:
Role of tumour necrosis factor alpha in graft-versus-host disease.
Transplant Proc
25:1234,
1993[Medline]
[Order article via Infotrieve]
37.
Theobald M,
Nierle T,
Bunjes D,
Arnold R,
Heimpel H:
Hosp-specific interleukin-2-secreting donor T-cell precursors as predictors of graft-versus-host disease in bone marrow transplantation between HLA-identical siblings.
N Engl J Med
327:1613,
1992[Abstract]
38.
Tanaka J,
Imamura M,
Kasai M,
Sakurada K,
Miyazaki T:
Cytokine gene expression after allogeneic bone marrow transplantation.
Leuk Lymphoma
16:413,
1995[Medline]
[Order article via Infotrieve]
39.
Sornasse T,
Larenas PV,
Davis KA,
de Vries JE,
Yssel H:
Differentiation and stability of T helper 1 and 2 cells derived from naive human neonatal CD4+ T cells analysed at the single-cell level.
J Exp Med
184:473,
1996[Abstract/Free Full Text]
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|
|
|
|
|
|
|
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3662 - 3671.
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[PDF]
|
|
|
|
|
|
|
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|
|
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|
|
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[PDF]
|
|
|
|
|
|
|
K. Sato, H. Nagayama, K. Tadokoro, T. Juji, and T. A. Takahashi
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162(7):
3865 - 3872.
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[Full Text]
[PDF]
|
|
|
|
|
|
|
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Blood,
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93(3):
1120 - 1123.
[Full Text]
[PDF]
|
|
|
|
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Abstract
Introduction
Abstract