Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients
The microenvironment in the bone marrow — including lymphocytes — is part of the pathophysiology of multiple myeloma (MM). High dose chemotherapy followed by autologous stem cell transplantation is standard of care for younger patients. Aim: To determine the influence of reinfused lymphocyte subsets...
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| Veröffentlicht in: | Experimental Oncology |
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| Datum: | 2008 |
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2008
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| Zitieren: | Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients / R. Schmidmaier, N. Oversohl, B. Schnabel, C. Straka, B. Emmerich // Experimental Oncology. — 2008. — Т. 30, № 3. — С. 240–243. — Бібліогр.: 17 назв. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1859687196487843840 |
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| author | Schmidmaier, R. Oversohl, N. Schnabel, B. Straka, C. Emmerich, B. |
| author_facet | Schmidmaier, R. Oversohl, N. Schnabel, B. Straka, C. Emmerich, B. |
| citation_txt | Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients / R. Schmidmaier, N. Oversohl, B. Schnabel, C. Straka, B. Emmerich // Experimental Oncology. — 2008. — Т. 30, № 3. — С. 240–243. — Бібліогр.: 17 назв. — англ. |
| collection | DSpace DC |
| container_title | Experimental Oncology |
| description | The microenvironment in the bone marrow — including lymphocytes — is part of the pathophysiology of multiple myeloma (MM). High dose chemotherapy followed by autologous stem cell transplantation is standard of care for younger patients. Aim: To determine the influence of reinfused lymphocyte subsets on event free survival (EFS) and overall survival (OS). Methods: In peripheral blood (PB) and aphaeresis products (AP) of 41 MM patients lymphocyte subsets were determined by flow cytometry and were correlated with clinical outcome. Results: PB lymphocyte subsets did not influence EFS or OS. Residual plasma cells in the AP were not correlated with poor outcome, whereas a high percentage of B cells (CD19+) showed a trend towards reduced EFS (P = 0.051). A high amount of CD4 cells and an increased CD4/CD8 ratio were significantly associated with prolonged EFS. In contrast, high percentage of HLA-DR positive lymphocytes showed negative impact on EFS and OS (P = 0.03 and 0.02, respectively). Conclusion: Obtained data suggest the non-activated (HLA-DR negative) helper CD4+ T cells in the AP to be tumour protective.
Микроокружение в костном мозге, включая лимфоциты, оказывает влияние на патофизиологию множественной миеломы
(MM). Высокодозовая химиотерапия, за которой следует аутологическая трансплантация стволовых клеток, является
стандартным подходом при лечении более молодых пациентов. Цель: изучить влияние введенных субпопуляций лимфоцитов
на безрецидивную выживаемость (EFS) и общую выживаемость больных c ММ. Методы: методом проточной цитофлуориметрии
в периферической крови (PB) и продуктах афереза (АР) пациентов с ММ ( n = 41) изучали субпопуляции
лимфоцитов и возможную корреляцию с исходом болезни. Результаты: субпопуляции лимфоцитов РB не влияли на EFS
или OS. Остаточные клетки плазмы в АР не коррелировали с плохим прогнозом, в тоже время при высоком содержании
В-клеток (CD19+) отмечали тенденцию к снижению EFS (P = 0,051). Высокое содержание CD4-клеток и увеличение
соотношения CD4/CD8 были достоверно ассоциированы с повышением EFS. В отличие от этого, высокий процент HLADR-положительных
лимфоцитов имел отрицательное влияние на EFS и OS (P = 0,03 и 0,02 соответственно). Выводы:
полученные данные позволяют предположить, что неактивированные (HLA-DR-отрицательные) хелперные CD4+ T-клетки
в AP могут обладать антиопухолевыми свойствами.
|
| first_indexed | 2025-11-30T22:43:13Z |
| format | Article |
| fulltext |
240 Experimental Oncology 30, 240–243, 2008 (September)
High dose chemotherapy followed by autologous
blood stem cell transplantation (ASCT) is standard treat
ment for younger patients with symptomatic multiple
myeloma (MM). The lymphocytes in the microenviron
ment of MM cells are an essential part of the patho
physiology of this incurable disease [1]. It has been
previously shown that lymphocytes within the graft are
linked to the survival of MM patients [2]. However, only
little is known which lymphocyte subsets are responsible
for this tumor protective effect. In the presented study
we have analyzed the lymphocytes by immunopheno
typing in the peripheral blood (PB) and in the apheresis
product (AP) of 41 consecutive patients.
METHODS
Patients. 41 consecutive patients with multip
le mye loma received ASCT between 11/1995 and
07/2000. The study was approved by the Ethical
committee of the University Munich and informed
consent was obtained from each patient. Mobiliza
tion chemotherapy and GSCF was administered ac
cording to the IEV regimen [3]. Peripheral blood stem
cells were harvested when the postnadir absolute
leukocyte count rose above 5000/µl using a Cobe
Spectra (Cobe, Heimstetten, Germany) or an AS104
(Fresenius, St. Wendel, Germany) cell separator and
standard programs. The harvested blood stem cells
were mixed with an equal volume of a freezing solution
which was prepared with 5% HSA and 100% DMSO
(Cryoserv, Tera Pharmaceuticals, Midvale, Utah, USA)
(4 : 1). Relapse was defined according to the EBMT
criteria [4].
Flow cytometry. Cells in the AP and cells in the PB
at the day of apheresis were analyzed. Immunopheno
typing was performed with a threecolorfluorescence
using an EPICS XLMCL flow cytometer and the Sys
tem II software (Beckman Coulter, Krefeld, Germany).
100 µL of sample were incubated 15 min with saturating
concentrations of the fluorochromeconjungated an
tibodies [5]. IgG1FITC, IgG1PE, CD45FITC, CD3
FITC, CD38FITC, CD14PE, CD19PE, CD4PE, CD8
PE, CD56/CD16PE, HLADRPE, and CD138PR were
purchased from Immunotech (Marseille, France).
Statistics. SPSS 11.5 for Microsoft Windows
(SPSS Inc., Chicago, USA) was used for statistical
analysis. The median number of positive cells was
calculated for each marker and patients were assigned
to a group either above or below median. To compare
survival curves a logrank test was performed and
P < 0.05 was considered statistically significant.
RESULTS
Patients and stem cell harvest. The mean age
was 56 years (range: 38–68) and 25 of the 41 patients
were male (61%). Within the median follow up of
3.9 years 30 pts experienced relapse (73%) and 13 pts
died (32%). The median event free survival (EFS) was
858 days (range: 104–2141 days), the median overall
survival (OS) was 1406 days (range: 104–2557 days).
Immunophenotyping of AP lymphocytes.
A median of 78,000 leukocytes per µl (range: 12,000–
370,000/µl) was counted in the APs. 29% (7–63%)
were lymphocytes. The table shows the lymphocyte
pattern within the AP among the 41 tested patients. The
composition of lymphocyte subsets expectantly refers
rather well to values of the peripheral blood with 81% T
cells (CD3), 1% Bcells, and 4% NKcells (CD56/16).
A small but still significant MM cell contamination was
seen. The median CD4/CD8ratio was 1.6 (range 0.2–
7.0) and 5% of lymphocytes expressed the activation
HELPER T CELLS (CD3+/CD4+) WITHIN THE AUTOLOGOUS
PERIPHERAL BLOOD STEM CELL GRAFT POSITIVELY CORRELATE
WITH EVENT FREE SURVIVAL OF MULTIPLE MYELOMA PATIENTS
R. Schmidmaier*, N. Oversohl, B. Schnabel, C. Straka, B. Emmerich
Department of Hematology and Oncology, Medizinische Klinik Innenstadt,
Klinikum der Universität München, 80336 Munich, Germany
Background: The microenvironment in the bone marrow — including lymphocytes — is part of the pathophysiology of multiple myeloma
(MM). High dose chemotherapy followed by autologous stem cell transplantation is standard of care for younger patients. Aim: To
determine the influence of reinfused lymphocyte subsets on event free survival (EFS) and overall survival (OS). Methods: In periphe
ral blood (PB) and aphaeresis products (AP) of 41 MM patients lymphocyte subsets were determined by flow cytometry and were
correlated with clinical outcome. Results: PB lymphocyte subsets did not influence EFS or OS. Residual plasma cells in the AP were
not correlated with poor outcome, whereas a high percentage of B cells (CD19+) showed a trend towards reduced EFS (P = 0.051).
A high amount of CD4 cells and an increased CD4/CD8 ratio were significantly associated with prolonged EFS. In contrast, high
percentage of HLADR positive lymphocytes showed negative impact on EFS and OS (P = 0.03 and 0.02, respectively). Conclusion:
Obtained data suggest the nonactivated (HLADR negative) helper CD4+ T cells in the AP to be tumour protective.
Key words: autologous graft, blood stem cell transplantation, correlation, eventfree survival, lymphocytes, multiple myeloma.
Received: June 3, 2008.
*Correspondence: Fax: +49 89 5160 4412
E-mail: ralf.schmidmaier@med.uni-muenchen.de
Abbreviations used: AP — apheresis products; EFS — event free
survival; MM — multiple myeloma; OS — overall survival; PB — pe-
ripheral blood.
Exp Oncol 2008
30, 3, 240–243
Experimental Oncology 30, 240–243, 2008 (September)30, 240–243, 2008 (September) (September) 241
marker HLADR. With respect to the clinical outcome
analysis below the displayed median values were used
for discrimination between high and low.
Table. Lymphocyte subsets within the peripheral blood stem cell grafts. After
IEV mobilization treatment and GCSF stimulation leukapheresis was performed
and lymphocyte subsets in the apheresis products of 41 consecutive multiple
myeloma patients were determined by flow cytometry as indicated. Median
and range are shown [% lymphocytes]. The median was used as cut off to
discriminate between high and low for further data analysis
Lymphocyte Subsets Median Range
CD3 81 7–63
CD19 1 0–15
CD4 45 14–76
CD8 29 11–63
CD56/16 4 1–19
HLA-DR 5 1–37
CD138/CD38 0.03 0.00–0.84
Survival according to lymphocyte subsets with-
in the graft. First, it was tested whether plasma cells
within the AP predict poor outcome. Figure a shows
that the median EFS was not significantly diffe rent in
patients with many plasma cells (CD138+CD38+ above
median) in the AP (1268 ± 258 vs 1066 ± 140 days).
This is in accordance to the results from PB. However,
the percentage of B cells (CD19+) within the AP had
a strong trend towards prediction of poor outcome
(P = 0.051, see Figure a).
Figure. Correlation of lymphocyte subsets within the peripheral
blood stem cell graft with clinical outcome (EFS event free sur
vival, OS overall survival) after high dose melphalan treatment.
Patients were grouped according to the amount of specific
lymphocyte subsets [% lymphocytes] within the stem cell graft
(high versus low defined as above or below median respectively).
EFS and OS were determined for the different groups and are
shown as Kaplan — Meier blots. a, EFS depending on B cells
(CD19) and plasma cells (CD138/CD38). b, EFS depending
on CD4 cells and CD4/CD8 ratio. c, EFS and OS depending on
HLADR positive cells
High leukocyte counts in the AP resulted in appa
rently longer EFS (1363 ± 164 vs 862 ± 111 days) and
slightly increased OS (1902 ± 138 vs 1663 ± 196 days),
although these results did not reach statistical signifi
cance (P = 0.074 and 0.288, respectively).
A high percentage of CD4 cells was associated with
a statistically significant prolongation of EFS (2179 ±
170 vs 1670 ± 212 days; P = 0.003), which was ad
ditionally reflected by the results obtained regarding
OS (Figure b). A low proportion of CD8 cells and con
sequently an increased T4/T8 ratio were significantly
associated with good EFS (see Figure b). In addition,
a small amount of activated lymphocytes (HLADR+)
almost doubled EFS: 1370 ± 173 vs 751 ± 104 days
(Figure c). This was also associated with a significantly
prolonged OS (P = 0.020).
Finally, none of the lymphocyte subsets in the pe
ripheral blood at the day of aphaeresis correlated with
any outcome parameter (OS, EFS).
DISCUSSION
The study was conducted between 1995 and 2000.
In the meantime several new drugs and treatment
regimens for relapsed patients have been developed.
Since probably all patients received these kind of
effective salvage treatment OS is most likely a poor
parameter to assess the influence of AP lymphocytes
on outcome. It seems reasonable to focus on EFS in
the discussion of the results.
There is a trend towards a better prognosis with
high leukocyte counts in the AP. This may be due to a
more pronounced GSCF responsiveness as an indica
tor for the quality of the bone marrow microenviron
ment [6]. However, peripheral blood leukocyte counts
on the day of apheresis did not predict for EFS (data not
shown). It has been shown previously that the absolute
number of infused lymphocytes during ASCT predicts
OS and time to progression in myeloma patients [2]. In
our study the relative numbers of lymphocytes (CD3+
and CD19+) were associated with a trend towards a
worse outcome, but this was due to the high num
bers of granulocytes that reflect good bone marrow
response to GCSF. The relative amount of CD4 cells
was associated with prolonged EFS in our study. The
EFS was the same as the relapse free survival. The
positive effect was therefore not due to prevention of
infectious complications but improved tumor control.
To the current knowledge more likely CD4 helper cells
than cytotoxic T cells are capable of tumor surveillance
and control [7–9]. Interestingly the most prominent
association was found in our study with respect to
activated lymphocytes (HLADR+). Two similar studies
did not reveal significant relations [10–11]. However in
the first peripheral blood was examined and in the latter
allogeneic transplantations were performed, making
both studies only poorly comparable. In the context
with the above mentioned results regarding CD4 cells
the negative impact of the activation marker HLADR
may be explained by the fact that activated T cells are
242 Experimental Oncology 30, 240–243, 2008 (September)
more differentiated and less potent to transfer regula
tory immune functions.
The number of plasma cells and as well the number
of B cells in the AP do not determine the clinical out
come of MM patients. This is in accordance with the
failure of purging strategies [12] and supports the hy
pothesis that the number of high dose chemotherapy
surviving cells within in the body are quite higher than
the number of plasma cells reinfused while ASCT. In
contrast, the relative B cell amount in the graft was as
sociated with an almost significant worsening of EFS.
This corresponds well to the results of B cell purging
in MM [13] and supports the hypothesis of precursor
B cells as origin of relapse [14–15]. However, clonali
ty was not assessed in the presented study and the
sample size were probably too small to reach statistical
significance. Further studies are needed to address
this topic.
Of note, a similar analysis was performed on
lymphocytes in the PB at the day of leukapheresis,
but none of the immunophenotyping parameters
correlated with the outcome of the patients (data not
shown). Most probably the highdose chemotherapy
diminishes the preexisting immune function. More
importantly, the lymphocytes within the graft signifi
cantly influence the immune function after ASCT and
herewith contribute to disease control [10, 16]. Indeed
it has been supposed that the lymphocytes within the
bone marrow influence the course of disease [17].
In summary, we suggest that a relative high pro
portion of nonactivated T helper cells within the
graft predicts proper tumor control in MM patients
after highdose chemotherapy. Targeted lymphocyte
preparation in the AP may further improve the results
of ASCT in MM.
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6. Straka C, Oduncu F, Hinke A, et al. Responsiveness to
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Experimental Oncology 30, 240–243, 2008 (September)30, 240–243, 2008 (September) (September) 243
Т-ХЕЛПЕРЫ (CD3+/CD4+) АУТОЛОГИЧЕСКОГО
ТРАНСПЛАНТАТА СТВОЛОВЫХ КЛЕТОК КРОВИ КОРРЕЛИРУЮТ
С БЕЗРЕЦИДИВНОЙ ВЫЖИВАЕМОСТЬЮ БОЛЬНЫХ
C МНОЖЕСТВЕННОЙ МИЕЛОМОЙ
Микроокружение в костном мозге, включая лимфоциты, оказывает влияние на патофизиологию множественной миеломы
(MM). Высокодозовая химиотерапия, за которой следует аутологическая трансплантация стволовых клеток, является
стандартным подходом при лечении более молодых пациентов. Цель: изучить влияние введенных субпопуляций лимфоци
тов на безрецидивную выживаемость (EFS) и общую выживаемость больных c ММ. Методы: методом проточной цито
флуориметрии в периферической крови (PB) и продуктах афереза (АР) пациентов с ММ ( n = 41) изучали субпопуляции
лимфоцитов и возможную корреляцию с исходом болезни. Результаты: субпопуляции лимфоцитов РB не влияли на EFS
или OS. Остаточные клетки плазмы в АР не коррелировали с плохим прогнозом, в тоже время при высоком содержании
Вклеток (CD19+) отмечали тенденцию к снижению EFS (P = 0,051). Высокое содержание CD4клеток и увеличение
соотношения CD4/CD8 были достоверно ассоциированы с повышением EFS. В отличие от этого, высокий процент HLA
DRположительных лимфоцитов имел отрицательное влияние на EFS и OS (P = 0,03 и 0,02 соответственно). Выводы:
полученные данные позволяют предположить, что неактивированные (HLADRотрицательные) хелперные CD4+ Tклетки
в AP могут обладать антиопухолевыми свойствами.
Ключевые слова: аутологический трансплантат, трансплантация стволовых клеток крови, корреляция, безрецидивная
выживаемость, лимфоциты, множественная миелома.
Copyright © Experimental Oncology, 2008
|
| id | nasplib_isofts_kiev_ua-123456789-139909 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1812-9269 |
| language | English |
| last_indexed | 2025-11-30T22:43:13Z |
| publishDate | 2008 |
| publisher | Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| record_format | dspace |
| spelling | Schmidmaier, R. Oversohl, N. Schnabel, B. Straka, C. Emmerich, B. 2018-06-21T13:36:30Z 2018-06-21T13:36:30Z 2008 Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients / R. Schmidmaier, N. Oversohl, B. Schnabel, C. Straka, B. Emmerich // Experimental Oncology. — 2008. — Т. 30, № 3. — С. 240–243. — Бібліогр.: 17 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/139909 The microenvironment in the bone marrow — including lymphocytes — is part of the pathophysiology of multiple myeloma (MM). High dose chemotherapy followed by autologous stem cell transplantation is standard of care for younger patients. Aim: To determine the influence of reinfused lymphocyte subsets on event free survival (EFS) and overall survival (OS). Methods: In peripheral blood (PB) and aphaeresis products (AP) of 41 MM patients lymphocyte subsets were determined by flow cytometry and were correlated with clinical outcome. Results: PB lymphocyte subsets did not influence EFS or OS. Residual plasma cells in the AP were not correlated with poor outcome, whereas a high percentage of B cells (CD19+) showed a trend towards reduced EFS (P = 0.051). A high amount of CD4 cells and an increased CD4/CD8 ratio were significantly associated with prolonged EFS. In contrast, high percentage of HLA-DR positive lymphocytes showed negative impact on EFS and OS (P = 0.03 and 0.02, respectively). Conclusion: Obtained data suggest the non-activated (HLA-DR negative) helper CD4+ T cells in the AP to be tumour protective. Микроокружение в костном мозге, включая лимфоциты, оказывает влияние на патофизиологию множественной миеломы (MM). Высокодозовая химиотерапия, за которой следует аутологическая трансплантация стволовых клеток, является стандартным подходом при лечении более молодых пациентов. Цель: изучить влияние введенных субпопуляций лимфоцитов на безрецидивную выживаемость (EFS) и общую выживаемость больных c ММ. Методы: методом проточной цитофлуориметрии в периферической крови (PB) и продуктах афереза (АР) пациентов с ММ ( n = 41) изучали субпопуляции лимфоцитов и возможную корреляцию с исходом болезни. Результаты: субпопуляции лимфоцитов РB не влияли на EFS или OS. Остаточные клетки плазмы в АР не коррелировали с плохим прогнозом, в тоже время при высоком содержании В-клеток (CD19+) отмечали тенденцию к снижению EFS (P = 0,051). Высокое содержание CD4-клеток и увеличение соотношения CD4/CD8 были достоверно ассоциированы с повышением EFS. В отличие от этого, высокий процент HLADR-положительных лимфоцитов имел отрицательное влияние на EFS и OS (P = 0,03 и 0,02 соответственно). Выводы: полученные данные позволяют предположить, что неактивированные (HLA-DR-отрицательные) хелперные CD4+ T-клетки в AP могут обладать антиопухолевыми свойствами. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Uncategorized Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients Т-хелперы (CD3+/CD4+) аутологического трансплантата стволовых клеток крови коррелируют с безрецидивной выживаемостью больных c множественной миеломой Article published earlier |
| spellingShingle | Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients Schmidmaier, R. Oversohl, N. Schnabel, B. Straka, C. Emmerich, B. Uncategorized |
| title | Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients |
| title_alt | Т-хелперы (CD3+/CD4+) аутологического трансплантата стволовых клеток крови коррелируют с безрецидивной выживаемостью больных c множественной миеломой |
| title_full | Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients |
| title_fullStr | Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients |
| title_full_unstemmed | Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients |
| title_short | Helper T cells (CD3+/CD4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients |
| title_sort | helper t cells (cd3+/cd4+) within the autologous peripheral blood stem cell graft positively correlate with event free survival of multiple myeloma patients |
| topic | Uncategorized |
| topic_facet | Uncategorized |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/139909 |
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