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* Laboratory for Experimental Immunology and
Department of Hematology, Academic Medical Center,
Department of Clinical Viro-Immunology, Sanquin Research at CLB, University of Amsterdam,
Division of Cellular Biochemistry, Netherlands Cancer Institute, and
¶ Department of Molecular Cell Biology, Faculty of Medicine, Vrije Universiteit, Amsterdam, The Netherlands; and
|| Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Interaction of CD27 with its transmembrane ligand, CD70, affects
Ag-driven T and B cell responses in several ways. Ligation of CD27 on
human activated B cells in vitro results in plasma cell differentiation
and enhanced Ig production (7, 13, 14, 15). On T cells,
triggering of CD27 enhances TCR-induced expansion and promotes the
generation of effector T cells with cytolytic function and
IFN-
-secreting capacity (16, 17, 18, 19, 20). In CD27-deficient
mice, reduced numbers of CD4+ and
CD8+ effector T cells were found in the lung
after primary and particularly secondary infection with influenza virus
(19). In line with this, persistent triggering of CD27 by
constitutive B cell-specific expression of mCD70 in CD70 transgenic
(TG)4 mice leads to increased
numbers of CD4+ and CD8+
effector T cells (20).
One way to control function of TNFR family members is the transient and tightly regulated expression of the ligand (4, 21, 22). In humans, CD70 is expressed on Ag receptor-activated, but not on resting, T and B cells. Although human (h) CD70 expression can be increased by CD28 or CD40 cross-linking, IL-4 down-regulates hCD70 expression (12, 23). Probably due to CD70’s transient expression, human tissue sections only show CD70 expression on some T cells and extrafollicular B cells. Occasionally, germinal centers containing a large number of CD70+ B cells are found (24). CD70 expression is also found in the medulla of human thymus (24). In the mouse, murine (m) CD70 has been detected on lymphocytes in vitro (25, 26). In accordance with studies on human lymphocytes, mCD70 was not detected on resting T cells. However, while in humans CD70 is expressed at significant levels at the plasma membrane of in vitro-activated T cells (23), in the mouse such cell surface expression was found to be very low (26). With regards to expression of mCD70 on resting and activated B cells, studies are partially conflicting (25, 26). Although CD70 has thus far not been detected on human DC, two reports have now documented membrane expression of CD70 on murine DC, one in infected mice and the other on in vitro-stimulated DC (27, 28).
In this study, newly developed anti-mCD70 mAb were used to document the expression pattern of mCD70 in vitro and in vivo. These findings are important to understand in which cell-cell interactions CD27-CD70 can participate. Moreover, this information is needed to establish to which extent the mouse can be used as a model to study the role of human CD27-CD70 interaction in vivo.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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C57BL/6 mice were bred in the facilities of The Netherlands Cancer Institute and CLB (Amsterdam, The Netherlands) under specific pathogen-free conditions. Mice used for experiments were between 6 and 8 wk of age at the start of the experiment. All animal experiments were conducted according to institutional and national guidelines and were approved by the Experimental Animal Committees of the respective institutes. Mice were infected intranasally with influenza virus strain A/NT/60/68 as described elsewhere (19).
Generation of anti-mCD70 mAb
Armenian hamsters (Cytogen, West Roxbury, MA) were injected three times i.p. with 107 irradiated (50 Gray) syngeneic fibroblasts transfected with the mCD70 cDNA, termed AR-mCD70 cells (29), at weekly intervals. Two weeks after the third injection, serum was tested for mCD70 reactivity. A booster injection (i.p., 107 cells) was given and 3 days thereafter, hamsters were sacrificed. Splenocytes were fused with SP2/0 murine myeloma cells as described (6, 29). Fourteen days after fusion, hybridoma supernatants were tested on AR-mCD70 cells and the 771 murine B lymphoma cell line (30) for the presence of anti-mCD70 Ab by flow cytometric analysis using PE-labeled goat anti-hamster Ig as second step reagent. After limiting dilution, two stable clones (3B9, 6D8) were obtained. Both mAbs were purified from culture supernatant using protein A bound to Sepharose CL-4B beads (Sigma-Aldrich, St. Louis, MO). F(ab')2 was generated and conjugated to biotin using standard procedures.
Abs and Fc proteins
mAbs to mouse B220 (RA3.6B2), CD3
(145-2C11; 500A2), CD4
(GK1.5), CD8 (2.43), CD16/CD32 (2.4G2), MHC class II (M5/114.15.2),
CD27 (LG.3A10; Ref. 6), CD40 (FGK-40), Thy-1.1 (50 AD
22/15), and DEC-205 (NLDC-145) were purified from hybridoma supernatant
and conjugated to FITC and biotin according to standard procedures. mAb
to CD40 (HM-40.3) and CD28 (37.51), as well as B7-2-FITC (GL-1),
CD4-PerCP (RM4-5), CD8a-PerCP (53-6.7), CD11c-PE (HL3), CD19-FITC
(1D3), and CD3-allophycocyanin (145-2C11) were purchased from BD
PharMingen (San Diego, CA) and goat anti-mouse IgM
F(ab')2 was purchased from Jackson ImmunoResearch
Laboratories (West Grove, PA). PE-labeled goat anti-hamster Ig was
from Southern Biotechnology Associates (Birmingham, AL).
F(ab')2 of hamster-anti hCD97 mAb 1D2 (a kind
gift from Dr. J. Hamann, Academic Medical Center, Amsterdam, The
Netherlands) were labeled with biotin and served as control in flow
cytometry and immunohistology. Recombinant receptor Fc proteins were
kindly provided by Dr. R. Goodwin (Immunex, Seattle, WA).
Cells and cell culture
771 is a cell line derived from a B lymphoma induced by neonatal
inoculation of a C57BL/10 mouse with MCF 1233 murine leukemia virus
(30). Expression of mCD70 on this line originally
permitted the cDNA cloning (29). ARHO Armenian hamster
fibroblasts (6) and mCD70-transfected ARHO cells
(AR-mCD70) (29) have been described previously. Single
cell suspensions were obtained by grinding tissues through nylon
sieves, or flushing femurs and tibias with a needle. Erythrocytes were
removed by lysis with ammonium chloride solution. For T cell
purification, murine lymph node single cell suspensions were depleted
of MHC class II+, B220+,
IgM+ cells using anti-MHC class II,
anti-B220 mAb, goat anti-rat IgG, and rat anti-mouse IgM
MACS microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany),
and MACS (BS) deletion columns following the manufacturer’s
instructions. For B cell purification, CD19+
cells were positively selected from splenocytes using goat
anti-CD19 MACS microbeads beads and MACS
(MS+) separation columns following the
manufacturer’s instructions. Purity of the resulting populations was
tested by immunofluorescence with anti-CD19 and anti-CD3 mAbs
and exceeded 95%. DC were generated in vitro as described
(31). In short, bone marrow suspensions were depleted of
CD4+, CD8+,
B220+, and MHC class II+
cells using theindicated mAbs and goat anti-rat IgG MACS microbeads
and MACS (BS) depletion columns. Cells were seeded at
106/ml and grown in medium containing recombinant
mouse GM-CSF (1 ng/ml; BD PharMingen) and IL-4 (1 ng/ml; BD
PharMingen). DC were harvested at day 8. Cells were grown in IMDM
containing 10% heat inactivated FCS, penicillin, streptomycin, and LPS
(serotype 026:B; Sigma-Aldrich), recombinant IL-2, IL-4, IL-12, IFN-
(BD PharMingen), or IL-10 (Endogen, Woburn, MA). The D1 dendritic cell
(DC) line was cultured as described (32).
Biochemical analysis
771 cells (3 x 106/sample)
were labeled with 125I (Amersham International,
Amersham, U.K.) by the glucose/lactoperoxidase method and lysed in 1%
Nonidet P-40 buffer, containing 10 mM triethanolamine-HCl (pH 7.8), 150
mM NaCl, 5 mM EDTA, 1 mM PMSF, 20 µg/ml ovomucoid trypsin inhibitor,
1 mM N-
-p-tosyl-L-lysine
chloromethyl ketone, and 20 µg/ml leupeptin. After centrifugation,
the supernatant was divided. One part was precleared with pooled human
serum (5%) and protein G-Sepharose (Pharmacia, Uppsala, Sweden), the
other part was precleared with normal hamster serum (10%) and protein
A-Sepharose. mCD70 was subsequently precipitated with mCD27-Fc,
anti-mCD70 mAb, or an irrelevant control mAb and complexes were
adsorbed onto protein G- or A-Sepharose, eluted, and
electrophoretically separated by 5–15% SDS-PAGE, and visualized by
autoradiography.
Flow cytometry
Cells (3 x 105) were seeded in 96-well round-bottom plates and incubated with anti-CD16/CD32 mAb (5 µg/ml) in FACS buffer (PBS, 0.5% BSA, 0.02% azide). After washing, cells were stained with the indicated mAbs and a subsequent incubation with allophycocyanin-labeled streptavidin (BD PharMingen) was performed to detect biotin-conjugated mAbs. All incubation steps were for 30 min at 4°C. Cell staining was analyzed on a FACSCalibur using CellQuest software (BD Biosciences, Mountain View, CA). Viable cell populations were gated on the basis of forward and side scatter and, where indicated, on the basis of a histogram profile.
Proliferation assay
For proliferation assays, purified murine T cells (105/well) were stimulated with Con A (Sigma-Aldrich) at the indicated concentrations and were cultured with mCD70-transfected or mock-transfected cells (5 x 104 cells/well). [3H]Thymidine incorporation was measured after 4 days of culture. [3H]Thymidine (0.4 µCi; Amersham International) was present for the last 16 h of culture.
Real-time PCR
Single cell suspensions of purified T cells (1 x 106/ml) were stimulated with immobilized anti-CD3 mAb (145-2C11, 10 µg/ml). Single cell suspensions of purified B cells (1 x 106/ml) were stimulated with anti-CD40 mAb (HM-40.3, 5 µg/ml) and LPS (10 µg/ml). After the indicated time of culture, cells (2 x 106) were collected and RNA was isolated with RNazol (Cinna/Biotecx Laboratories, Friendswood, TX). Single-strand cDNA was prepared in a 20-µl reaction volume with 500 ng of oligo(dT)12–18 and 100 U of Superscript II (Life Technologies, Rockville, MD). Murine CD70 cDNA was quantitated with the LightCycler System (Roche, Mannheim, Germany; www.lightcycler.com, technical note no. LC 10/2000). Using a FastStart DNA Master SYBR Green I kit and 1 µl of cDNA, a 20-µl real-time PCR was set-up amplifying mCD70 or hypoxanthine phosphoribosyltransferase (HPRT) from the cDNA derived from each sample. Real-time kinetic quantification allows measurement during the log-linear phase of a PCR. Analysis of this phase allows accurate determination of the starting concentration, using the equation: number of amplified molecules = initial number of molecules x efficiency constant (E)cycle number. In our approach, a standard curve of 771 cDNA was prepared to determine the amount of CD70 cDNA (E = 0.986) and HPRT cDNA (E = 0.982) in the T and B cell samples. The samples were normalized by dividing the amount of CD70 cDNA by the amount of cDNA of the housekeeping gene HPRT. The following primers were used: mCD70 (+ strand primer, 5'-GGATGCCGGAGGAAGGTCGCCC-3'; - strand primer, 5'-CAAGGGCATATCCACTGAACTC-3'); and HPRT (+ strand primer, 5'-TATGGACAGGACTGAACGTCTTGC-3'; - strand primer, 5'-GACACAAACATGATTCAAATCCCTGA-3'). Crossing points were calculated by the second derivative method using LightCycler System software. Real-time PCR was performed on a LightCycler PCR apparatus (Roche).
Immunohistology
For light microscopy, acetone-fixed cryosections of lung were incubated in methanol containing 0.02% H2O2, followed by incubation with 2% BSA in PBS. Next, they were incubated overnight at 4°C with biotinylated 3B9 anti-mCD70 F(ab')2 or biotinylated anti-Thy-1.1, diluted in PBS with 1% BSA. After thorough washing in cold PBS, sections were incubated with Vectastain ABC regent (Vector Laboratories, Burlingame, CA) for 30 min and developed with 0.05% 3,3'-diaminobenzidine with 0.01% H2O2 in 0.1 M phosphate buffer. After counterstaining with hematoxylin, sections were coverslipped for observation. For confocal microscopy, cryostat sections were fixed in acetone and incubated with anti-CD16/CD32 mAb (5 µg/ml). Sections were subsequently incubated with biotinylated 3B9 anti-mCD70 F(ab')2 at saturating concentrations for 2 h at room temperature. After thorough washing in cold PBS, sections were incubated with FITC-labeled mAb and streptavidin-conjugated Alexa Fluor 594 (Molecular Probes, Eugene, OR) overnight at 4°C. Stained sections were washed in cold PBS and coverslipped using antifading mounting medium (DAKO, Glostrup, Denmark). Omission of incubation with biotinylated anti-mCD70 mAb was used as negative control. Suspensions of cells isolated from lung-draining lymph nodes were analyzed for mCD70 expression as follows: Cells were pipetted onto cover slips, which had been coated with poly-L-lysine (50 µg/ml in PBS, 30 min, 37°C) and incubated for 30 min at 37°C. They were fixed for 5 min with methanol kept at -20°C. All subsequent incubations were at room temperature, as outlined for tissue sections. Coverslips were mounted in Vectashield (Vector Laboratories) and viewed under a Leica TCS NT confocal laser-scanning microscope (Deerfield, IL).
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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To generate mAbs to characterize CD70 protein expression in the
mouse, Armenian hamsters were immunized with syngeneic fibroblasts
transfected with the mCD70 cDNA (29). Eight percent of the
supernatants of the resulting hybridomas were reactive in
immunofluorescence with the mCD70-expressing B cell line 771 and the
hamster fibroblast transfectant AR-mCD70. The selected mAbs, 3B9 and
6D8, could be used to isolate mCD70 from cell surface-iodinated 771 B
cells. As was shown for the murine CD27-Fc protein (29),
the anti-mCD70 mAbs not only precipitated monomeric mCD70, but also
the SDS-resistant dimeric and trimeric forms of mCD70 (Fig. 1
A). Flow cytometric analysis
showed that 3B9 and 6D8 blocked each other’s binding to 771 cells,
indicating that they react with the same epitope (data not shown). The
mAbs did not cross-react with human CD70 (data not shown).
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B, mCD70
transfectant AR-mCD70 incremented [3H]thymidine
incorporation of Con A-stimulated purified murine T cells. This was
inhibited by mAb 3B9 (Fig. 1B) as well as 6D8 (not shown).
Thus, addition of anti-mCD70 mAb 3B9 and 6D8 could block early and
late consequences of CD27 triggering, indicating that these
anti-mCD70 mAb prevent functional interaction between mCD70
and CD27. Analysis of mCD70 expression on in vitro-activated cells
In humans, CD70 is absent from resting lymphocytes, but can
readily be induced upon in vitro activation. Concordantly, bone marrow
cells, thymocytes, splenocytes, and lymph node cells, freshly isolated
from naive mice, showed no cell surface expression of mCD70, according
to flow cytometry (data not shown). Expression of mCD70 upon T and B
cell activation in vitro was analyzed by real-time PCR at the mRNA
level and by flow cytometry at the protein level. Purified T and B
cells were unstimulated, or incubated for 2 days with anti-CD3 mAb,
or anti-CD40 mAb with LPS, respectively. In unstimulated cells, no
PCR product was obtained with mCD70 primers, indicating a total lack of
mCD70 RNA expression in resting T and B cells (not shown). In
stimulated T cells, CD70 mRNA was detectable, but it was present at
much lower levels than in stimulated B cells (Fig. 2A). In accordance with the
difference in mCD70 mRNA expression between in vitro activated T and B
cells, cell surface mCD70 protein expression also differed. On
activated T cells, only a very low level of mCD70 was found, barely
detectable by flow cytometry, whereas mCD70 was easily detectable at
the cell surface of activated B cells (Fig. 2B). Expression
of mCD70 on activated T cells was not more pronounced at other time
points after activation, nor when T cells were additionally stimulated
with anti-CD28 mAb, IL-2, IL-4, IL-10, IL-12, or IFN- (data not
shown).
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50–70% positive cells at day 4 (Fig. 3A). These agents acted
synergistically in mCD70 up-regulation, allowing for expression of
mCD70 on almost all B cells, already at 2 days after activation. LPS
alone was much less potent in up-regulating mCD70 cell surface
expression, but acted in synergy with anti-CD40, allowing for
maximal expression at day 2 after stimulation on 70% of B cells
(Fig. 3B).
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consistently inhibited the induction of
mCD70 expression by anti-IgM Ab (Fig. 3C), but not by
LPS and anti-CD40 mAb (Fig. 3D). Addition of IL-10
slightly (1.5-fold), but consistently, reduced induction of mCD70
expression by LPS/anti-CD40 mAb (Fig. 3D), or LPS alone
(data not shown).
Although in the human system CD70 has only been found on cells of the
lymphoid lineage, in the mouse CD70 has been found on activated
CD11c+ DC (27, 28). The availability
of the murine DC line D1 (32) and culture procedures to
obtain primary murine DC from bone marrow cells in vitro permitted us
to test regulation of mCD70 expression on DC. As is shown in Fig. 4, D1 cells (Fig. 4A) and in
vitro-generated immature DC (CD11c+, MHC class
IIdull) (Fig. 4B) do not express
detectable mCD70 at the cell surface. However, after maturation, as
induced by LPS, anti-CD40 mAb or GM-CSF (34) in
particular primary DC, express high levels of mCD70.
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As mentioned above, we could not detect mCD70 by flow cytometry on
cell suspensions derived from primary and secondary lymphoid organs of
naive mice. To further document mCD70 expression in lymphoid organs of
nonimmunized mice, immunohistology was performed. In multiple
independent analyses of mice (10 animals of different strains), mCD70
expression in spleen and lymph nodes appeared to be extremely rare.
Occasionally, some scattered cells in the red pulp of the spleen and
dispersed cells in peripheral lymph nodes expressed mCD70 (data not
shown). However, consistently mCD70-expressing cells were found in
the medulla of the thymus (Fig. 5). These
cells also stained with an Ab to DEC-205, a carbohydrate receptor,
which is expressed by DC and thymic epithelial cells
(35).
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A). In these sections,
conspicuous fields of mCD70-positive cells were detected in
parabronchiolar regions. These were T cells, according to Thy-1.1
staining of serial sections (Fig. 6A). Such fields of
mCD70+ cells were never observed in noninfected
lung tissue (not shown). Lung tissue sections were also stained with
fluorescent Abs specific for T cells (CD3), B cells (B220), and DC
(CD11c) and examined by confocal microscopy. In this analysis,
lung-infiltrating T and B cells were detected in significant amounts.
However, DC were scarce. Double staining for mCD70 revealed that a
sizeable proportion of lung-infiltrating T cells expressed mCD70 (Fig. 6B). No mCD70-positive B cells or DC were found. Cells in
lung-draining lymph nodes were examined for mCD70 expression by
fluorescent staining of cytospins, followed by confocal microscopy
(Fig. 6C). This analysis revealed significant amounts of T
and B cells, identified by CD3 and B220 staining, while DC
(CD11c+) were very rare. Double staining for
mCD70 indicated that a significant proportion of T and B cells in these
lung-draining lymph nodes expressed mCD70 protein. The small number of
DC present permitted no reliable conclusion on mCD70 expression in DC.
Strikingly, both in T and B lymphocytes, the great majority of mCD70
staining appeared cytoplasmic, rather than at the cell membrane. This
can be seen particularly well in B cells, which show peripheral
staining for B220 which only rarely overlaps with the mCD70 staining
(Fig. 6C).
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). This experiment showed clearly
that mCD70 is detectable by flow cytometry at the plasma membrane of a
small proportion of lung-infiltrating T cells. Maximal expression was
found at the height of the response (day 8), as defined by the size of
the T cell infiltrate and the number of Ag-specific T cells
(19).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In line with the in vivo data, we could not detect mCD70 in vitro on
resting T cells or B cells or immature DC. Hartwig et al.
(25) found, in contrast to us and Oshima et al.
(26) that resting murine B cells express CD70. However,
this is unlikely because CD70 TG mice that constitutively express mCD70
on B cells have substantial disturbances in the composition of their
immune system, notably a progressive decline in B cell numbers
(20). With regard to regulation of mCD70 expression on
activated B cells, our findings partly confirm data presented by
Hartwig et al. (25) and Oshima et al. (26)
and is most compatible with CD70 expression data for human B cells. We
find induction of mCD70 expression after stimulation with anti-IgM
Ab, anti-CD40 mAb, or LPS, and a synergistic effect of these B cell
activators. In humans, the Th2 cytokine IL-4 negatively regulates CD70
expression on activated T and B lymphocytes (12, 23).
Strikingly, we found in this study that in the mouse, the Th1 cytokine
IFN- negatively regulates CD70 expression on IgM-activated B cells.
In CD70 TG mice, persistent CD27 triggering results in an increased
proportion of IFN--producing T cells, which inhibit B cell
maturation in the bone marrow (20). Therefore, the
down-regulation of mCD70 on activated B cells by IFN- may represent
a negative feedback mechanism.
The two signal model dictates that productive activation of naive T cells requires one signal derived from the TCR (signal 1) and one costimulatory signal (signal 2). In the current concept of an adaptive immune response, these signals are provided by mature DC in secondary lymphoid organs. The constitutive expression of CD27 on naive T cells, the in vitro consequences of CD27-CD70 interaction and the observed CD70 expression on mature murine DC suggest that CD27-CD70 can provide signal 2. In our in vivo analyses, too few DC were present in the preparation to allow for conclusions on mCD70 expression. Further studies on ex vivo-purified DC preparations are necessary to investigate this issue.
In vitro, purified T cells from CD27-deficient mice show decreased
expansion in response to TCR stimulation, as compared with wild-type T
cells (19), strongly suggesting that mCD70 is present at
the surface of in vitro-activated murine T cells. In agreement with
Oshima et al. (26), we found that mCD70 was difficult to
detect by flow cytometry on such T cells. Cell surface expression was
not enhanced by anti-CD28 mAb, IL-2, IL-4, IL-10, IL-12, or IFN-
(data not shown). We addressed different posttranslational mechanisms
that might explain the lack of mCD70 membrane protein expression.
First, intracellular staining was done to examine whether mCD70 was
retained within the cell. Second, membrane staining after culture in
the presence of metalloprotease inhibitors or anti-mCD70 mAb was
performed to examine whether mCD70 was shed from the membrane. Third,
examined the possibility that mCD70 was shielded by soluble CD27
(17) by analyzing mCD70 expression on activated
CD27-deficient T cells. In none of these experiments, we could detect
substantial levels of mCD70 protein. Taken together, these data suggest
that induction of CD70 mRNA in in vitro-activated T cells is too low to
allow for easy detection of the protein by conventional flow
cytometry.
In contrast to the in vitro experiments, analyses of mice after influenza virus infection showed a strong expression of CD70 both in the lung and in the draining lymph nodes. These data indicate that factors, other than the cytokines that were tested in the in vitro culture experiments, contribute to the induction of mCD70. In draining lymph nodes, clear expression of mCD70 was found inside the cell, and flow cytometry of cell suspensions from these nodes also revealed very low plasma membrane expression (results not shown). Recent adoptive transfer experiments with CD27-deficient cells show that CD27 contributes to activated T cell expansion in lung-draining lymph nodes, arguing that mCD70 is indeed functionally expressed at this site (J. Hendriks, Y. Xiao, and J. Borst, manuscript in preparation). In the lung, mCD70 could clearly be detected at the cell surface of activated T cells. The percentage of mCD70-positive T cells in the lung according to flow cytometry was much lower than the percentage of mCD70-positive T cells seen in sections of lung tissue. Even though mCD70 could be induced at high levels on the surface of B cells activated in vitro, B cells in lung-draining lymph nodes were barely positive according to flow cytometry. However, intracellularly they expressed mCD70. Our data argue that mCD70 is induced at the protein level in T and B cells upon in vivo activation, but is to a large extent retained intracellularly.
In summary, our combined in vitro and in vivo data indicate that mCD70
plasma membrane expression is tightly controlled by both
transcriptional and posttranslational mechanisms. The consequences of
constitutive high-level mCD70 expression at the cell surface of B cells
in CD70 transgenic mice highlight why control of mCD70 expression
levels and therewith control of CD27-CD70 interaction is crucial. In
mCD70 transgenic mice that constitutively express mCD70 at a high level
on B cells under the CD19 promoter, T cells are stimulated to
differentiate into Th1-type effector cells, which produce IFN-.
IFN- production by these T cells inhibits B cell maturation in the
bone marrow, leading to progressive B cell depletion (20).
In addition, the T cell compartment in these mice declines in time,
most likely as a result of high turnover of the effector T cells (38).
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Footnotes |
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2 Address correspondence and reprint requests to Dr. René A. W. van Lier, Department of Experimental Immunology, Academic Medical Center, L1-158, P.O. Box 22600, 1100DD Amsterdam, The Netherlands. E-mail address: r.vanlier{at}amc.uva.nl
3 Abbreviations used in this paper: TG, transgenic; h, human; m, murine; DC, dendritic cell; HPRT, hypoxanthine phosphoribosyltransferase.
4 This work was supported by The Netherlands Organization for Scientific Research Grant No. 901-07-095.
Received for publication November 27, 2001. Accepted for publication October 11, 2002.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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-mediated B cell depletion. Immunity 15:801.[Medline]
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D. V. Dolfi, A. C. Boesteanu, C. Petrovas, D. Xia, E. A. Butz, and P. D. Katsikis Late Signals from CD27 Prevent Fas-Dependent Apoptosis of Primary CD8+ T Cells J. Immunol., March 1, 2008; 180(5): 2912 - 2921. [Abstract] [Full Text] [PDF]
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H. R. Turnquist, G. Raimondi, A. F. Zahorchak, R. T. Fischer, Z. Wang, and A. W. Thomson Rapamycin-Conditioned Dendritic Cells Are Poor Stimulators of Allogeneic CD4+ T Cells, but Enrich for Antigen-Specific Foxp3+ T Regulatory Cells and Promote Organ Transplant Tolerance J. Immunol., June 1, 2007; 178(11): 7018 - 7031. [Abstract] [Full Text] [PDF]
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R. R. French, V. Y. Taraban, G. R. Crowther, T. F. Rowley, J. C. Gray, P. W. Johnson, A. L. Tutt, A. Al-Shamkhani, and M. J. Glennie Eradication of lymphoma by CD8 T cells following anti-CD40 monoclonal antibody therapy is critically dependent on CD27 costimulation Blood, June 1, 2007; 109(11): 4810 - 4815. [Abstract] [Full Text] [PDF]
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 A. M. Keller, T. A. Groothuis, E. A. M. Veraar, M. Marsman, L. M. de Buy Wenniger, H. Janssen, J. Neefjes, and J. Borst Costimulatory ligand CD70 is delivered to the immunological synapse by shared intracellular trafficking with MHC class II molecules PNAS, April 3, 2007; 104(14): 5989 - 5994. [Abstract] [Full Text] [PDF]
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P. J. Sanchez, J. A. McWilliams, C. Haluszczak, H. Yagita, and R. M. Kedl Combined TLR/CD40 Stimulation Mediates Potent Cellular Immunity by Regulating Dendritic Cell Expression of CD70 In Vivo J. Immunol., February 1, 2007; 178(3): 1564 - 1572. [Abstract] [Full Text] [PDF]
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M. A. Kapina, G. S. Shepelkova, V. V. Mischenko, P. Sayles, P. Bogacheva, G. Winslow, A. S. Apt, and I. V. Lyadova CD27low CD4 T Lymphocytes That Accumulate in the Mouse Lungs during Mycobacterial Infection Differentiate from CD27high Precursors In Situ, Produce IFN-{gamma}, and Protect the Host against Tuberculosis Infection J. Immunol., January 15, 2007; 178(2): 976 - 985. [Abstract] [Full Text] [PDF]
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C. W. Wieland, M. E. Kerver, S. Florquin, M. A. Nolte, J. Borst, R. van Lier, M. H. J. van Oers, and T. van der Poll CD27 contributes to the early systemic immune response to Mycobacterium tuberculosis infection but does not affect outcome Int. Immunol., November 1, 2006; 18(11): 1531 - 1539. [Abstract] [Full Text] [PDF]
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J. Huang, K. W. Kerstann, M. Ahmadzadeh, Y. F. Li, M. El-Gamil, S. A. Rosenberg, and P. F. Robbins Modulation by IL-2 of CD70 and CD27 Expression on CD8+ T Cells: Importance for the Therapeutic Effectiveness of Cell Transfer Immunotherapy. J. Immunol., June 15, 2006; 176(12): 7726 - 7735. [Abstract] [Full Text] [PDF]
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P. Soroosh, S. Ine, K. Sugamura, and N. Ishii OX40-OX40 Ligand Interaction through T Cell-T Cell Contact Contributes to CD4 T Cell Longevity J. Immunol., May 15, 2006; 176(10): 5975 - 5987. [Abstract] [Full Text] [PDF]
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 C.-L. Law, K. A. Gordon, B. E. Toki, A. K. Yamane, M. A. Hering, C. G. Cerveny, J. M. Petroziello, M. C. Ryan, L. Smith, R. Simon, et al. Lymphocyte Activation Antigen CD70 Expressed by Renal Cell Carcinoma Is a Potential Therapeutic Target for Anti-CD70 Antibody-Drug Conjugates Cancer Res., February 15, 2006; 66(4): 2328 - 2337. [Abstract] [Full Text] [PDF]
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 B. F. Israel, M. Gulley, S. Elmore, S. Ferrini, W.-h. Feng, and S. C. Kenney Anti-CD70 antibodies: a potential treatment for EBV+ CD70-expressing lymphomas Mol. Cancer Ther., December 1, 2005; 4(12): 2037 - 2044. [Abstract] [Full Text] [PDF]
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 G. J. de Bree, E. M.M. van Leeuwen, T. A. Out, H. M. Jansen, R. E. Jonkers, and R. A.W. van Lier Selective accumulation of differentiated CD8+ T cells specific for respiratory viruses in the human lung J. Exp. Med., November 21, 2005; 202(10): 1433 - 1442. [Abstract] [Full Text] [PDF]
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J. Hendriks, Y. Xiao, J. W. A. Rossen, K. F. van der Sluijs, K. Sugamura, N. Ishii, and J. Borst During Viral Infection of the Respiratory Tract, CD27, 4-1BB, and OX40 Collectively Determine Formation of CD8+ Memory T Cells and Their Capacity for Secondary Expansion J. Immunol., August 1, 2005; 175(3): 1665 - 1676. [Abstract] [Full Text] [PDF]
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T. N. J. Bullock and H. Yagita Induction of CD70 on Dendritic Cells through CD40 or TLR Stimulation Contributes to the Development of CD8+ T Cell Responses in the Absence of CD4+ T Cells J. Immunol., January 15, 2005; 174(2): 710 - 717. [Abstract] [Full Text] [PDF]
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A. F. Ochsenbein, S. R. Riddell, M. Brown, L. Corey, G. M. Baerlocher, P. M. Lansdorp, and P. D. Greenberg CD27 Expression Promotes Long-Term Survival of Functional Effector-Memory CD8+ Cytotoxic T Lymphocytes in HIV-infected Patients J. Exp. Med., December 6, 2004; 200(11): 1407 - 1417. [Abstract] [Full Text] [PDF]
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V. Y. Taraban, T. F. Rowley, and A. Al-Shamkhani Cutting Edge: A Critical Role for CD70 in CD8 T Cell Priming by CD40-Licensed APCs J. Immunol., December 1, 2004; 173(11): 6542 - 6546. [Abstract] [Full Text] [PDF]
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R. Arens, M. A. Nolte, K. Tesselaar, B. Heemskerk, K. A. Reedquist, R. A. W. van Lier, and M. H. J. van Oers Signaling through CD70 Regulates B Cell Activation and IgG Production J. Immunol., September 15, 2004; 173(6): 3901 - 3908. [Abstract] [Full Text] [PDF]
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R. van Doorn, R. Dijkman, M. H. Vermeer, J. J. Out-Luiting, E. M. H. van der Raaij-Helmer, R. Willemze, and C. P. Tensen Aberrant Expression of the Tyrosine Kinase Receptor EphA4 and the Transcription Factor Twist in Sezary Syndrome Identified by Gene Expression Analysis Cancer Res., August 15, 2004; 64(16): 5578 - 5586. [Abstract] [Full Text] [PDF]
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 P. Garcia, A. B. de Heredia, T. Bellon, E. Carpio, M. Llano, E. Caparros, P. Aparicio, and M. Lopez-Botet Signalling via CD70, a member of the TNF family, regulates T cell functions J. Leukoc. Biol., July 1, 2004; 76(1): 263 - 270. [Abstract] [Full Text] [PDF]
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Y. Xiao, J. Hendriks, P. Langerak, H. Jacobs, and J. Borst CD27 Is Acquired by Primed B Cells at the Centroblast Stage and Promotes Germinal Center Formation J. Immunol., June 15, 2004; 172(12): 7432 - 7441. [Abstract] [Full Text] [PDF]
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R. Arens, K. Schepers, M. A. Nolte, M. F. van Oosterwijk, R. A.W. van Lier, T. N.M. Schumacher, and M. H.J. van Oers Tumor Rejection Induced by CD70-mediated Quantitative and Qualitative Effects on Effector CD8+ T Cell Formation J. Exp. Med., June 7, 2004; 199(11): 1595 - 1605. [Abstract] [Full Text] [PDF]
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T. F. Rowley and A. Al-Shamkhani Stimulation by Soluble CD70 Promotes Strong Primary and Secondary CD8+ Cytotoxic T Cell Responses In Vivo J. Immunol., May 15, 2004; 172(10): 6039 - 6046. [Abstract] [Full Text] [PDF]
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