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Blood, Vol. 93 No. 2 (January 15), 1999:
pp. 737-745
Marrow Ablative and Immunosuppressive Effects of
131I-Anti-CD45 Antibody in Congenic and H2-Mismatched
Murine Transplant Models
By
Dana C. Matthews,
Paul J. Martin,
Cynthia Nourigat,
Frederick R. Appelbaum,
Darrell R. Fisher, and
Irwin D. Bernstein
From the Division of Clinical Research, Fred Hutchinson Cancer
Research Center, Seattle, WA; the Departments of Pediatrics
and Medicine, University of Washington, Seattle, WA; and the
Pacific Northwest National Laboratory, Richland, WA.
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ABSTRACT |
Targeted hematopoietic irradiation delivered by
131I-anti-CD45 antibody has been combined with conventional
marrow transplant preparative regimens in an effort to decrease
relapse. Before increasing the proportion of therapy delivered by
radiolabeled antibody, the myeloablative and immunosuppressive effects
of such low dose rate irradiation must be quantitated. We have examined the ability of 131I-anti-CD45 antibody to facilitate
engraftment in Ly5-congenic and H2-mismatched murine marrow transplant
models. Recipient B6-Ly5a mice were treated with
30F11 antibody labeled with 0.1 to 1.5 mCi 131I
and/or total body irradiation (TBI), followed by
T-cell-depleted marrow from Ly5b-congenic
(C57BL/6) or H2-mismatched (BALB/c) donors. Engraftment was achieved
readily in the Ly5-congenic setting, with greater than 80%
donor granulocytes and T cells after 0.5 mCi 131I
(estimated 17 Gy to marrow) or 8 Gy TBI. A higher TBI dose (14 Gy) was
required to achieve engraftment of H2-mismatched marrow, and
engraftment occurred in only 3 of 11 mice receiving 1.5 mCi 131I delivered by anti-CD45 antibody. Engraftment of
H2-mismatched marrow was achieved in 22 of 23 animals receiving 0.75 mCi 131I delivered by anti-CD45 antibody combined
with 8 Gy TBI. Thus, targeted radiation delivered via
131I-anti-CD45 antibody can enable engraftment of congenic
marrow and can partially replace TBI when transplanting
T-cell-depleted H2-mismatched marrow.
© 1999 by The American Society of Hematology.
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INTRODUCTION |
BONE MARROW transplantation has been used
for more than two decades to treat hematologic malignancies and
aplastic anemia and, more recently, genetic diseases such as
thalassemia and sickle cell anemia.1-6 However, the success
of marrow transplantation has been limited by the high relapse rates
seen in patients transplanted for advanced malignancies, by the
toxicity of preparative regimens, and by the lack of HLA-identical
family member donors for most patients. In principle, these limitations
could be addressed by the use of radiolabeled monoclonal antibodies
(MoAbs) to increase the irradiation to lymphohematopoietic tissues
while minimizing the exposure of normal organs. First, delivery of more
irradiation to sites of leukemic involvement should decrease the risk
of relapse. Second, targeted irradiation of lymphohematopoietic organs
should decrease the need for high-dose chemotherapy or total body
irradiation (TBI) to prevent rejection of the allogeneic graft.
Alternatively, adding targeted irradiation to full-dose systemic
therapy could enhance the level of immunosuppression, thereby
decreasing the risk of rejection of marrow from HLA-mismatched or
unrelated donors, especially when the marrow has been depleted of T
cells to prevent graft-versus-host disease (GVHD).
We have shown that infusion of radiolabeled antibodies against CD45
delivers irradiation selectively to lymphohematopoietic tissues.7-9 Significant supplemental doses of radiation
have been delivered to bone marrow and spleen in combination with
conventional marrow transplant preparative regimens without excessive
toxicity, and the relapse rates after these novel preparative regimens
have been low in preliminary studies. In the initial clinical studies, treatment with radiolabeled antibody was combined with standard preparative regimens containing cyclophosphamide and either
TBI9,10 or busulfan,11 because it is unknown
whether the relatively low dose rate radiation delivered by antibody
has sufficient antileukemic efficacy to prevent relapse and sufficient
immunosuppressive activity to prevent rejection of allogeneic marrow.
Radiolabeled antibody delivers radiation at an exposure rate that is
one to two orders of magnitude lower than the rate normally delivered
from an external beam source. Several studies have demonstrated the
influence of exposure rate on the amount of irradiation required to
achieve engraftment.12-15 van Os et
al14,15 compared exposure rates ranging from 0.5 to 40 cGy/min in a variety of murine transplant models. In a congenic donor
and recipient strain combination that differed by a single
nonimmunogenic marker, 5.5 Gy TBI delivered at 40 cGy/min was
sufficient to allow 80% engraftment in 50% of the recipients. In the
same model with TBI delivered at 2 cGy/min, a 7 Gy exposure was needed
to achieve the same degree of engraftment.15 In an
H2-compatible strain combination with disparity for multiple minor
histocompatibility antigens, 6 Gy TBI delivered at 40 cGy/min was
sufficient to allow engraftment. In the same model with TBI delivered
at 2 cGy/min, an 8 Gy exposure was needed.14 With T-cell-depleted marrow from H2-mismatched donors, the difference between low and high dose rate was at least 6 Gy, because full engraftment at the high-dose rate (40 cGy/min) occurred after 10 Gy,
but long-term engraftment did not occur after 16 Gy (the highest dose
tested) delivered at 2 cGy/min.14 These results suggest
that hematopoietic stem cells and precursors of cellular immunity have
a substantial ability to repair irradiation damage.
Little is known about the relative biologic effects of low dose rate
radiation resulting from the administration of radiolabeled antibody.
To address this issue, we initiated studies using
131I-labeled anti-CD45 antibody, with or without external
beam TBI, as a preparative regimen in two models of murine
transplantation. To measure marrow ablation,
B6-Ly5a recipients were transplanted with marrow
from congenic Ly5b donors.16,17 In this
model, the donor and recipient disparity is limited to CD45 allotype
and does not evoke a cellular immune response.18 In an
approach designed to test the immunosuppressive effects of the regimen,
the same recipients (H2b) were transplanted with
T-cell-depleted marrow from H2-mismatched (H2d) BALB/c
donors. Our results demonstrate that irradiation delivered solely by
131I-anti-CD45 antibody allows engraftment of marrow in
Ly5-congenic recipients but not in H2-mismatched recipients, where both
natural killer (NK) cells and T cells are known to cause
rejection. However, engraftment of T-depleted H2-mismatched marrow
could be accomplished by adding low-dose external beam TBI to the
radiation delivered by 131I-anti-CD45 antibody.
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MATERIALS AND METHODS |
Mice.
Male B6-Ly5a mice were bred at the Fred Hutchinson
Cancer Research Center (Seattle, WA) and housed under specific
pathogen-free conditions with acidified water and autoclaved chow.
C57BL/6 and BALB/c donors were purchased from Jackson Laboratory (Bar
Harbor, ME) and were 6 to 12 weeks old at the initiation of each
experiment.
MoAbs.
MoAbs GK1.5 (rat IgG2b anti-CD4), 30-H12 (rat
IgG2b anti-thy1.2), and 2.34 (rat IgG2b
anti-CD8) were prepared from culture supernatants of cells lines
obtained from ATCC (GK1.5 and 2.34) or Dr Jeffrey Ledbetter
(Bristol Meyers Squibb, Seattle, WA; 30-H12). A hybridoma cell line
secreting MoAb 30F11 (rat IgG2b), which recognizes all
murine CD45 isoforms, was the gift of Dr Ledbetter. Hybridoma cell
lines secreting MoAbs A20 (murine IgG2a), which recognizes
the Ly5.1 epitope encoded by the Ly5a allotype of
murine CD45, and 104 (murine IgG2a), which recognizes the
Ly5.2 epitope encoded by the Ly5b allotype of
murine CD45, were the gift of Dr Shoji Kimura (Sloan Kettering
Institute, New York, NY). Ascites containing each MoAb was produced in
BALB/c mice under specific pathogen-free conditions. Batch extraction
and purification of antibodies from ascites was performed using
Abx exchange resin (J.T. Baker, Phillipsburg, NJ) and
high-pressure liquid chromatography (Biosys 510 Beckman, Fullerton,
CA).
Iodination and characterization.
Antibodies were iodinated with Na125I or Na131I
(ICN, Irvine, CA) using the Iodogen method19 for trace
labeling and the chloramine T-labeling method for high specific
activity labeling.20 The immunoreactivity (percentage of
counts able to bind at antigen excess) of each preparation was
determined by incubating antibody at low concentration (5 ng/mL) with
1.0 × 106 to 3 × 107
B6-Ly5a splenocytes for 1 hour at room temperature
and measuring unbound antibody as previously described.21
Preparations with less than 70% immunoreactivity were not used.
Antibody localization and estimation of radiation absorbed doses.
The double isotope labeling method of Pressman22 was used
to determine the biodistribution of 30F11 antibody as previously described.8 Animals were injected via tail vein with 200 µL volume containing 100 µg of antibody labeled with
131I (specific activity, 1 µCi/µg) and an equal amount
of rat IgG (Sigma Immunochemical, St Louis, MO) labeled with
125I. Groups of 5 mice were killed at multiple time points
from 1 to 96 hours after injection, and multiple tissues were sampled. The 125I and 131I contents of weighed samples
and standard samples of the injection mix were determined by
multichannel gamma counting (Packard 5000 Autogamma counter; Packard
Instrument Co, Downers Grove, IL). Counting data were adjusted for
cross-over from the 131I channel to the 125I
channel and were corrected for decay.
Time-activity curves were constructed for each organ from the data on
the percentage injected dose per gram (% ID/g) at each time point, and
the infinite-time integral was calculated for the area under the curve
to estimate the total number of disintegrations for dosimetry assuming
that both 30F11 antibody and Rat IgG were labeled with
131I. Absorbed fractions of  -particle energy for
131I were calculated by applying electron transport theory
(EGS423) to a dosimetric model for the laboratory
mouse.24,25 This model accounts for the size, shape,
position, and density of organs, as well as for the overlap and contact
from surrounding organs and tissues. The absorbed fractions included
contributions from same-organ irradiation as well as from cross-organ
irradiation. Contributions to the absorbed dose from 131I
penetrating gamma radiation in mouse organs were negligible (<1% of
the total dose) and were neglected. The femoral marrow was assumed to
be a cylinder of 0.5 mm in radius; therefore, only a proportion (46%)
of the radioiodine present in marrow was assumed to deposit its energy
inside the marrow. The -particle absorbed fraction for lymph nodes
was estimated to be 73%. Results were expressed as gray (Gy) per
millicurie (mCi) 131I.
Bone marrow transplantation.
Four days before marrow infusion, groups of 6 recipient
B6-Ly5a mice were treated with 100 µg of 30F11
antibody labeled with varying amounts of 131I at a maximum
specific activity of 1.5 mCi 131I/100 µg. Cages were
changed every other day for 1 week to decrease the irradiation from
excreted urine in the cage bedding. External beam TBI was administered
on the day of marrow infusion and was delivered at 0.2 Gy/min from dual
60Cobalt sources (J.L. Shepherd Co, San Fernando, CA).
Marrow cells were treated with hemolytic buffer and T cells were
depleted by complement-mediated lysis with antibodies against CD4, CD8,
and Thy-1, a method that routinely depleted 97% to 99% of T cells
(data not shown). A marrow cell dose of 1.0 × 107
nucleated cells (counted before T-cell depletion) was injected via tail
vein. This moderately high cell dose was selected to be certain that
cell dose was not a limiting factor for engraftment, especially
considering that some residual radiolabeled antibody remains in
hematopoietic tissues at 96 hours after antibody administration when
the marrow was infused. The marrow was depleted of T cells in all
experiments to allow comparison between Ly5-congenic and H2-mismatched
transplants. H2 disparity provides a rigorous test of
immunosuppression, and T-cell depletion allows recipients to survive
without lethal or debilitating GVHD.
Assessment of engraftment.
Mice were examined daily for survival and for signs of GVHD or other
illness. At 4, 8, and 12 weeks posttransplant, orbital blood samples
were stained with 50 µL of biotinylated A20 (anti-Ly5.1) antibody or
biotinylated 104 (anti-Ly5.2) antibody at 100 µg/mL for 30 minutes
followed by phycoerythrin-streptavidin (PharMingen, San Diego, CA) and
fluorescein isothiocyanate (FITC)-conjugated CD3-specific antibody
(PharMingen). Control samples consisted of cells from unmanipulated
mice of both donor and host strains. Samples were analyzed by flow
cytometry using a FACScan (Becton Dickinson, San Jose, CA). Lymphocyte
and granulocyte populations were defined by forward and 90° scatter
characteristics. The lymphocyte population was analyzed separately
using two colors to distinguish CD3+ T cells and
CD3 B cells.
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RESULTS |
Biodistribution of 131I-anti-CD45 antibody and estimation
of radiation absorbed doses.
B6.Ly5a mice were injected with a 100 µg dose of
trace 131I-labeled anti-CD45 MoAb 30F11 or
125I-labeled rat IgG and the amount of isotope in major
organs was determined. The resulting time-activity curves demonstrated
greater uptake and retention of radiolabeled anti-CD45 antibody in
spleen, lymph nodes, and marrow as compared with the lung, the normal nontarget organ with the highest concentration
(Fig 1A). The corresponding time-activity
curves for 125I-labeled rat IgG (ie, IgG purified from sera
of normal rats), used as a nonspecific control, did not show any
specific retention in spleen, lymph nodes, or marrow (Fig 1B).
Anti-CD45 antibody was cleared from the blood far more rapidly than
control rat IgG, presumably by rapid binding of circulating antibody to
CD45 on readily accessible cells in spleen and marrow (Fig 1C).
Estimates of absorbed irradiation doses delivered to target and normal
organs by 30F11 antibody or polyclonal rat IgG, assuming they were
labeled with 131I, were calculated using the time-activity
curves for each tissue (Table 1). The
estimated absorbed irradiation delivered to spleen, lymph nodes, and
bone marrow by a 100 µg dose of antibody 30F11 labeled with 1 mCi
131I was, respectively, 6.5-, 4.0-, and 2.1-fold greater
than that delivered to lung. When 131I was delivered from
polyclonal rat IgG, the spleen, bone marrow, and lymph nodes all
absorbed less irradiation than the lung and absorbed approximately the
same amount of irradiation as the liver and kidney. The higher
estimated irradiation absorbed by nontarget organs after administration
of 131I-rat IgG as compared with 131I-30F11
antibody reflects the slower clearance of 131I-rat IgG from
the blood.
Estimated absorbed irradiation rates varied between tissues and over
time for a given tissue, as shown in Fig 2
for 0.5, 1.0, and 1.5 mCi of 131I. For antibody labeled
with 1.0 mCi of 131I, the highest irradiation rate for a
hematopoietic tissue was 3 cGy/min in the spleen between 4 and 24 hours
after infusion. Maximum lymph node dose rates ranged from 1.3 to 1.5 cGy/min. For brachial lymph nodes, 75% of the total estimated
irradiation was absorbed at a rate of at least 1 cGy/min. For marrow,
86% of the total dose was absorbed at an estimated rate of at least 0.6 cGy/min.
Engraftment of congenic marrow after 131I-30F11
antibody or TBI.
In the initial experiments with C57BL/6 donors and congenic
B6-Ly5a recipients, we administered a 100 µg dose
of 30F11 antibody labeled with 0 (ie, unlabeled antibody), 0.5, 1.0, 1.25, and 1.5 mCi of 131I, followed 4 days later by
injection of T-cell-depleted marrow or medium alone. All mice
receiving marrow survived, whereas 2 of 6 mice treated with 1.25 mCi
131I and 4 of 6 treated with 1.5 mCi 131I and
no marrow died between 11 and 15 days after radiolabeled antibody
injection (data not shown). Although postmortem examinations were not
performed, death at these time points after delivery of radiation is
consistent with death from marrow aplasia. All mice receiving at least
0.5 mCi 131I-anti-CD45 antibody demonstrated successful
engraftment (Table 2). Myeloid engraftment
occurred within 4 weeks posttransplant, but full T-cell engraftment did
not occur until 12 weeks, presumably reflecting delayed disappearance
of recipient T cells.
In subsequent experiments, mice received varying doses of TBI
or a broader range of 131I doses before infusion of
T-cell-depleted congenic marrow. A near-linear relationship between
radiation dose and engraftment of donor marrow was found for
131I doses of up to 0.5 mCi
(Fig 3) and TBI doses of up to 8 Gy
(Fig 4). Fifty percent engraftment of
granulocytes, B cells, and T cells was observed after treatment with
0.2 to 0.3 mCi of 131I-labeled antibody, which corresponded
to an estimated marrow radiation dose of 7 to 10 Gy. Engraftment after
8 Gy TBI (dose rate, 20 cGy/min) was equivalent to engraftment after
0.5 mCi of 131I delivered via 30F11 antibody, with 80%
donor cells. The estimated irradiation absorbed by marrow after
treatment with 0.5 mCi of 131I on 100 µg 30F11 was
approximately 17 Gy, more than twice the 8 Gy required when the
irradiation was delivered by external beam.
Engraftment of T-cell-depleted H2-mismatched marrow after
131I-anti-CD45 antibody alone or with external beam TBI.
In the initial experiments with BALB/c donors and MHC-mismatched
B6-Ly5a recipients, we administered either a 100 µg dose of antibody 30F11 labeled with 0.5 to 1.5 mCi of
131I on day  4 or 2 to 16 Gy TBI on day 0, followed
by infusion of T-cell-depleted marrow or medium alone. With external
beam TBI and no marrow infusion, the LD50 was approximately
10 Gy, and no recipients survived exposures of  14 Gy (data not
shown). With infusion of T-cell-depleted H2-incompatible marrow,
engraftment (>80% donor T cells at 3 months posttransplant) was
observed in all recipients prepared with 14 Gy TBI delivered by
external beam (Fig 5). With
131I-labeled antibody and no marrow infusion, the
LD50 was between 0.5 and 1.0 mCi, and no recipients
survived after administration of 1.5 mCi (data not shown). With
infusion of T-cell-depleted H2-incompatible marrow, 1.5 mCi of
131I-labeled antibody was not sufficient to permit
engraftment. Only 1 of 6 mice receiving the highest isotope dose had
more than 80% T cells of donor origin (Fig
6A).
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| Fig 5.
Engraftment of T-cell-depleted BALB/c marrow in
B6-Ly5a recipients treated with 6 to 16 Gy TBI.
Proportion of T cells of donor origin (mean ± SD) 3 months after
transplantation. Data point labels indicate the number of mice with
80% donor T cells of total surviving mice (of 6 treated mice per
group).
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| Fig 6.
Engraftment of T-cell-depleted BALB/c marrow in mice
treated with 100 µg 30F11 antibody labeled with 0.5 to 1.5 mCi
131I alone (A) or combined with 4 Gy TBI (B). Proportion of
T cells of donor origin (mean ± SD) 3 months after transplantation.
Data point labels indicate the number of mice with 80% donor T
cells of total surviving mice (of 6 treated mice per group).
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It was not possible to deliver more irradiation with the use of
131I-30F11 antibody alone, because labeling the antibody to
a specific activity higher than 15 mCi/mg impaired immunoreactivity of
the antibody, and antibody doses higher than 100 µg delayed clearance from both blood and marrow, causing the infused donor marrow to be
damaged by isotope persisting for more than 4 days (data not shown). To
circumvent this problem, we tested the immunosuppressive effects of
radiation from 131I-30F11 antibody combined with 4 Gy
external beam TBI. This combined preparative regimen resulted in
engraftment in most recipients treated at 131I doses of at
least 1.0 mCi 131I (Fig 6B). In two experiments,
5 of 12 mice receiving 1.5 mCi survived and demonstrated donor
engraftment, but the remaining 7 mice treated with 1.5 mCi died between
9 and 14 days after marrow transplant. These results suggest that the
margin between immunosuppressive and toxic doses of radiation is small
when high doses of radioisotope are combined with 4 Gy TBI.
In two experiments we combined 0.75 mCi of 131I-anti-CD45
antibody with TBI doses varying between 2 and 10 Gy
(Fig 7). With 6 Gy TBI, engraftment was
observed in 9 of 11 recipients (experiment no. 1) and 4 of 11 recipients (experiment no. 2), and with 8 Gy TBI, engraftment was
observed in 12 of 12 recipients (experiment no. 1) and 10 of 11 recipients (experiment no. 2). These results suggest that the
immunosuppressive effect of 131I-anti-CD45 antibody labeled
with 0.75 mCi 131I can replace 6 to 8 Gy TBI in producing
engraftment equivalent to that seen with 14 Gy TBI alone. A dose of
0.75 mCi 131I is estimated to deliver approximately 25 Gy
to marrow, 50 Gy to lymph nodes, and 80 Gy to spleen.
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| Fig 7.
Engraftment of T-cell-depleted BALB/c marrow in mice
treated with 100 µg 30F11 antibody labeled with 0.75 mCi
131I combined with 2 to 10 Gy TBI. Proportion of T cells of
donor origin (mean ± SD) 3 months after transplantation. Data point
labels indicate the number of mice with 80% donor T cells of total
surviving mice (of 12 treated mice per group).
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DISCUSSION |
We asked if radiation delivered by 131I-labeled anti-CD45
antibody could enable engraftment of congenic marrow and of
H2-incompatible marrow. Using congenic marrow,
131I-anti-CD45 antibody alone enabled engraftment, with
80% engraftment when antibody was labeled with 0.5 mCi
131I, and thus could replace TBI. When donor and recipient
were H2-incompatible, 131I-anti-CD45 antibody could
partially replace TBI, demonstrating its potential to permit reduction
of external beam TBI and possibly provide a less toxic marrow
transplant preparative regimen.
Several factors influence the relationship between radiation dose and
successful engraftment, including recipient variables such as strain,
health status, and presensitization to donor
alloantigens26,27 and graft variables such as
histocompatibility, stem cell source, cell dose, and number of T
cells.12,28-36 Engraftment is also influenced by the
exposure rate of irradiation and by the fractionation schedule.12-15,37-39 Delivering radiation with
131I-anti-CD45 antibody enabled engraftment of donor cells
in congenic mice. All B6-Ly5a recipients treated
with 0.75 mCi 131I and 3 of 6 treated with 1.0 mCi
131I conjugated to anti-CD45 antibody survived without
infusion of donor marrow, demonstrating that the radiation delivered by
these doses of isotope was not myeloablative and did not have fatal nonhematopoietic toxicity. Nonetheless, recipients treated with a much
lower dose of 131I delivered by anti-CD45 antibody (0.3 mCi) had 60% T-cell, 70% B-cell, and 45% myeloid engraftment after
infusion of Ly5-congenic marrow. Eighty percent engraftment required 8 Gy TBI or 0.5 mCi of 131I. The 17 Gy estimated irradiation
absorbed by marrow delivered by antibody labeled with 0.5 mCi
131I supports the concept that radiation delivered at the
low continuous dose rate and low linear energy transfer of
-particles such as 131I has a lower relative biological
efficacy (RBE) than radiation delivered at the much more rapid rate
used for TBI.
Previous studies have shown that when the external beam TBI dose rate
is decreased from 25 to 1 cGy/min, the LD50 for early gastrointestinal toxicity is increased from 12 to 21 Gy and the LD50 for late nonhematopoietic deaths occurring by 1 year
is increased from 10 to 21 Gy in BALB/c mice supported with syngeneic
marrow transplants.40 In that study, no mice treated with
20 Gy at 1 cGy/min had detectable histologic changes in the lung or
kidney. In CBA mice treated with thoracic irradiation at high dose rate (180 cGy/min), the minimum dose required to produce a detectable elevation in breathing rate at 28 weeks after treatment increased from
13.8 Gy rad when the irradiation was delivered as a single fraction to
30.4 Gy when the irradiation was delivered in 7 fractions.41 In a study with -emitting radionuclide
90Y administered as inhaled fused aluminosilicate
particles, estimated lung radiation doses up to 27 Gy did not decrease
survival as compared with untreated controls.42 These
results are consistent with our observations of lower RBE of radiation
delivered by 131I-labeled antibody compared with equivalent
doses delivered as conventional TBI.
In a transplant model demanding profound immunosuppression, engraftment
of T-cell-depleted H2-mismatched marrow could not be achieved using
131I-CD45 antibody alone. Engraftment occurred in only a
minority of mice receiving the highest 131I dose delivered
(1.5 mCi). Several factors might explain the inability to achieve
engraftment after treatment with radiolabeled antibody, despite the
delivery of estimated radiation doses of almost 100 Gy to lymph nodes,
50 Gy to marrow, and 160 Gy to spleen. First, a blood-thymus barrier
impeded antibody localization in the thymus, with a dose of
approximately 19 Gy delivered by 1.5 mCi 131I. Second, the
absorbed radiation actually delivered to cells that cause graft
rejection might be less than estimated from the biodistribution
studies. The estimates of absorbed dose in hematopoietic tissues
represent the average for cells that are assumed to reside in that
tissue for at least the first 96 hours after radiolabeled antibody
infusion and do not account for heterogeneity within the tissue. T
cells or NK effectors that circulate during some or all of the period
of radiation delivery may thus receive a lower radiation dose. A cell
that circulates during the entire period of radiation delivery will
receive at least the radiation dose estimated for total body (5.9 Gy/mCi 131I). It is not possible to quantitate the
additional radiation received by a circulating cell because it involves
not only radiation delivered by antibody bound to the cell and to
neighboring cells in circulation as well as that present in plasma
(which depends on geometry of blood vessels in relationship to the path
length of the isotope), but also radiation delivered during the time the cell traffics in tissues such as the spleen where greater radiation
effects are present. Within lymph nodes, radiation delivery may be
heterogeneous, and the amount of irradiation delivered to individual
nodes could vary. Finally, cellular repair mechanisms might be able to
keep pace with the rate of damage inflicted by the low radiation dose
rate delivered from 131I, thereby avoiding cell death.
Our results with CD45-congenic donor/recipient pairs suggest that
treatment with 131I-anti-CD45 antibody alone is sufficient
to allow engraftment of donor cells at 131I doses that are
well tolerated. This raises the possibility that 131I-labeled anti-CD45 antibody might provide a less toxic
method for selective ablation of marrow and might enable reconstitution with autologous hematopoietic cells modified by gene therapy. Furthermore, the finding that engraftment of T-cell-depleted
H2-mismatched marrow reliably occurred when 0.75 mCi of
131I-anti-CD45 antibody was combined with 8 Gy TBI suggests
that targeted radiation may be able to replace nearly half of the TBI ordinarily administered in such transplants. Such an approach holds the
potential for enhanced antileukemic efficacy while inducing less
systemic toxicity, thereby improving the outcome after marrow transplantation for acute leukemia.
| |
ACKNOWLEDGMENT |
The authors are indebted to Minna Zheng, Jennifer Smith, and Carol Dean
for their expert technical assistance.
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FOOTNOTES |
Submitted April 30, 1998;
accepted September 22, 1998.
Supported by National Institutes of Health Grant No. CA01690, the Adler
Foundation, and the Parker-Hughes Trust.
The publication costs of this
article were defrayed in part by
page charge payment. This article
must therefore be hereby marked
"advertisement"
in accordance with 18 U.S.C. section
1734 solely to indicate this fact.
Address reprint requests to Dana C. Matthews, MD, Fred Hutchinson
Cancer Research Center D1-100, 1100 Fairview Ave N, PO Box 19024, Seattle, WA 98109; e-mail: dmatthew{at}fhcrc.org.
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Abstract
Introduction
