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*
Unité de Biologie Moléculaire du Gène, Institut National de la Santé et de la Recherche Médicale, Unité 277, Département d’Immunologie, Institut Pasteur, Paris, France;
Institut de Pharmacologie et de Biologie Structurale du Centre National de la Recherche Scientifique, Toulouse, France;
Howard Hughes Medical Institute, Department of Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, TN; and
Institut National de la Santé et de la Recherche Médicale, Unité 25, Hôpital Necker, Paris, France
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResults and DiscussionReferences |
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14invT cells (NKT cells), restricted by the
CD1d1 MHC 1b molecules, are a distinctive subset of T cells endowed
with pleiotropic functions. CD1d1-restricted NKT cells infiltrate the
granulomas induced by the s.c. injection of mycobacterial
phosphatidylinositoldimannoside (PIM2) but not of its
deacylated derivative. NKT cells are detectable as early as 6 hours
following the injection. Although the molecular structure of
PIM2 meets the requirements for presentation by CD1d1, Ab
blocking and adoptive transfer experiments of wild-type NKT cells into
CD1d1-/- mice show that CD1d1 expression is not required
for the early recruitment of NKT cells to the injection site. This
conclusion was confirmed by the finding that IL-12R
-/-
and CD40-/- mice were able to recruit NKT cells after
PIM2 challenge. Moreover, the injection of
-galactosylceramide, an NKT cell ligand that is recognized in the
context of CD1d1, promoted only a minor recruitment of NKT cells. By
contrast, injection of -galactosylceramide, a synthetic glycolipid
that binds to CD1d1 but does not activate the CD1d/TCR pathway,
resulted in the development of large granulomas rich in NKT cells.
Finally, local injection of TNF- mimics the effect of glycolipids.
It is concluded that NKT cells migrate to and accumulate at
inflammatory sites in the same way as other cells of the innate immune
system and that migration to and accumulation at inflammatory sites are
processes independent of the CD1d1 molecule. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResults and DiscussionReferences |
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14-J281 TCR -chain
predominantly associated with TCR V2, -7, and -8 -chains
in mice (5, 6, 7, 8) and V24-JQ/V 11 in humans
(9). Most of these cells also express NK markers, such as
NK1.1 in C57BL/6 mice (8). The murine
NK1.1+TCRint T cells (usually referred to
as NKT cells) are endowed with multiple, and sometimes conflicting,
markers, which have recently been reviewed (10). In the
mouse, they are predominantly found in the liver, spleen, thymus, and
bone marrow and are hardly detectable in blood. Thus far, a unique
ligand activates NKT cells through the CD1d-TCR axis: NKT cells respond
in a TCR-dependent manner to a particular glycolipid named
-galactosylceramide
(-GalCer)4 presented by CD1d1
molecules (11). However, -GalCer is not found in nature
except in some marine sponges, and the precise role of host and foreign
glycolipids in the physiology of V14inv NKT cells is
largely unknown (10).
The ability of the TCR of V14inv NKT cells to detect
bacterial glycolipids presented by CD1d molecules is a topic of
significant controversy. Mycobacterial glycolipids are assumed to play
a significant role in the host response to mycobacteria
(2). The injection of mycobacterial glycolipids,
particularly phosphatidylinositolmannosides (PIM), induces an acute
inflammatory reaction with, among other infiltrating lymphocytes, T
cells identified by the presence of the distinctive
V14inv TCR -chain, thus predominantly
CD1d1-restricted T cells (12). Because the structure of
PIMs conforms with the binding requirements of the hydrophobic groove
of CD1d1, it was assumed that the V14inv+ T cell
population had expanded in an Ag-driven manner with PIMs complexed with
CD1d1 molecules and presented to the TCR of V14inv T
cells. A primary role for NKT cells in the local inflammatory response
was also suggested.
However, these studies were conducted using partially purified
mycobacterial glycolipids. We have recently shown that
spectroscopically pure mycobacterial phosphatidylinositoldimannosides
(PIM2) induced an acute inflammatory reaction rich in
V14inv+ T cells (13). The availability
of highly purified reagents made it possible to investigate the precise
role of the CD1d1 and TCR molecules in the recruitment of
V14inv T cells in the acute inflammatory lesions that
these reagents induce. It could be concluded that neither a functional
CD1d1-TCR axis nor the expression of CD1d1 molecules was required for
NKT cell migration and that their local accumulation lacked Ag
specificity and was not Ag driven. In fact, the accumulation of the
cells appear to be determined by the ongoing acute inflammatory
process. V14inv+ T cells appear to be a novel
participant of the acute inflammatory response to bacterial
components.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResults and DiscussionReferences |
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C57BL/6 mice were purchased from IFFA CREDO (l’Arbresle,
France), CD40-/- mice (14) were purchased
from the Centre de l’Elevage, Centre National de Recherche
Scientifique (Orleans, France), IL12R-/- mice
(15) were obtained from The Jackson Laboratory (Bar
Harbor, ME), and V14tg mice on the C-/- background
were previously described (7, 16). All genetically
modified mice were backcrossed to the C57BL/6 background at least six
times, and animals were used between 6 and 12 wk of age. CD1d1 knockout
mice have been generated by one of us (17) and described
elsewhere (18).
Abs and reagents for FACS
For identification of NKT cells, we used FITC-anti-TCR
(H57-597) and PE-coupled anti-NK1.1 (PK136) (BD PharMingen, San
Diego, CA). To avoid nonspecific activation by engagement of the TCR
(19), we used a combination of NK1.1 and anti-CD5
(clone 53-7.3) (BD PharMingen) for cell sorting experiments.
Biotin-coupled anti-CD1d (1B1; BD PharMingen) was used for
evaluation of CD1d expression. For in vivo blocking and FACS staining
experiments, we used anti-CD1d, clone 20H2, kindly provided by A.
Bendelac (Princeton University, Princeton, NJ) (20). For
staining of granulocytes, dendritic cells, and B cells, we used the
respective Abs Gr-1 (RB6-8C5), CD11c (HL3), and CD19 (1D3) from BD
PharMingen. Phosphatidylinnositolmannosides bearing two mannosyl
residues (PIM2) were purified from Mycobacterium
tuberculosis strain H37Rv as described (13). TNF-
was purchased from R&D Systems (Wiesbaden, Germany).
RNA and cDNA preparation and PCR conditions
Specimens were disrupted in a Polytron homogenizer in 1 ml Trizol (Gibco, Cergy Pontoise, France). RNA extraction and cDNA preparation were conducted following standard procedures using (dT)17 primers and avian myeloblastosis virus reverse transcriptase from Roche (Meylan, France). For the detection of CD1d1-encoding mRNA, the following primers were used: 5'-CTCTAGGAGACCACGGACAAATA-3' and 5'-ACAC CTTCCGCTGCCTGCAGATG-3'.
Immunoscope analysis
The immunoscope technique for CDR3 spectratyping has been
described previously (21). For detection of the
V14-C rearrangement and the canonical V14-J281 transcript,
we used primers designed by Apostolou et al. (12).
Typically, a gaussian-like distribution of the peaks is observed in the
absence of antigenic stimulation, whereas clonal amplification results
either in a distortion of the gaussian distribution or by the
occurrence of isolated peaks.
Cloning and sequencing
PCR products were Pfu amplified, and blunted PCR products were ligated into the vector Blunt II-Topo (Invitrogen, Groningen, The Netherlands). Sequence analyses were conducted using a standard PerkinElmer (Wellesley, MA) protocol, and sequences were analyzed using ABI-Prism 373 sequencing equipment (Applied Biosystems, Foster City, CA).
Induction of granulomas
LPS-free mycobacterial glycolipids isolated from H37rv M.
tuberculosis were dissolved in chloroform-methanol-H2O
(60:35:5) at the indicated concentrations, an injection solution was
prepared by drying the solvent to the minimum, and the glycolipids were
conjugated to aluminum hydroxide (Alu-Gel-S; Serva, Heidelberg,
Germany) in 12.5% Alu-Gel-S-87.5% PBS. The latter solution served as
a negative control. One hundred microliters of the respective
glycolipid-alum suspension were injected s.c. into the lateral flank of
mice. At indicated time points, mice were sacrificed, and granulomas as
well as livers, spleens, and lymph nodes were recovered for further
analysis. For Ab-blocking experiments, mice were injected with 0.5 mg
anti-CD1d (clone 20H2) i.p. 12 h before induction of
granulomas. Synthetic - and -GalCer were kindly provided by Dr.
Y. Koezuka (Kirin Brewery, Gunma, Japan).
Adoptive transfer
To transfer highly purified NKT cell populations, spleen and
liver cells were prepared as described (7) from V14Tg
mice on a C-/- background and stained with
anti-CD5 and anti-NK1.1 mAbs. Double-positive, CD5-intermediate
cells were sorted using a MoFlow cell sorter (Cytomation, Geneva,
Switzerland) and incubated at 37°C for 2 h to allow
internalization of the bound Abs. The purity of sorted cells was
checked using anti-TCR and anti-NK1.1 mAbs or anti-CD1d
mAbs and was found to exceed 97% for purity of NKT cells and 99% for
the lack of CD1d expression in all experiments. Highly purified NKT
cells (2.5–3 x 106) were injected into the spleen of
CD1d-/- mice after short term anesthesia with
pentobarbital. Twelve hours after adoptive transfer, mice were injected
i.p. with 0.5 mg anti-CD1d clone 20H2 in 100 µl PBS to block
contaminating transferred CD1d1+ cells or
(auto)-up-regulation of this molecule on NKT cells during the transfer.
Twenty-four hours after the cell transfer, mice were injected s.c. for
granuloma induction as described above.
Collagenase treatment
Lesions were collected, cut, and stirred at 37°C for 90 min in 30 ml RPMI containing 20% FCS, 100 U/ml collagenase (C2139; Sigma-Aldrich, St. Louis, MO), and 5 U/ml DNase 1 (Sigma-Aldrich). At the middle and at the end of the incubation, the suspension was dissociated by multiple aspirations through a syringe for 2 min. The pellet was washed, and cells were purified on a Ficoll gradient and incubated with Abs.
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Results and Discussion |
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TopAbstractIntroductionMaterials and MethodsResults and DiscussionReferences |
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The nature of the cell populations recruited by injection of
PIM2 was first investigated in detail. The s.c. injection
of insolubilized, spectroscopically pure PIM2 induces an
acute inflammatory response with V14inv+ T cells as
one of the cellular components. The removal of the lipid moiety of
PIM2 abolished the inflammatory response (13).
In a first series of experiments, we re-examined the phenotype and the
proportion of the V14inv+ NKT cells present in the
day 7 lesions. Thus, mice were injected with PIM2, the
inflammatory lesions were excised at day 7, pooled, and dissociated
with collagenase; and the recovered cells were subjected to FACS. The
residual lesions left after injection of alum in PBS were used as
controls. The PIM2-induced lesions contained an average of
15% NK1.1+TCRint T cells in the
lymphocyte gate (Fig. 1
). The residual
lesions in the controls contained 1–3%
NK1.1+TCRint T cells, primarily as in
spleen. Thus, within the limits of the yield of recovery of living
cells, phenotypically defined
NK1.1+TCRint T cells accumulate in the
lesions a minimum of 6 times above their frequency in spleen. The
PIM2-induced lesions contained other cell types with the
following respective frequencies: granulocytes 20%; B cells 1%; and
CD11c+ cells 67%. The results of immunoscope analysis
confirmed an undetectable level of expression of the invariant TCR-
chain in PBS-induced lesions. In contrast, a peak centered around a
CDR3 length of 10 aa, which is typical of the invariant CD1d-restricted
TCR -chain, was detected in PIM2-induced lesions. This
peak was unique among all V14-C PCR products derived from
PIM2-induced lesions with no other V14+
rearrangements detectable, implying preferential recruitment of
CD1d-restricted T cells among all other rearranged V14+
T cells. Due to its sensitivity and consequently the possibility of
investigating individual lesions, Immunoscope analysis was used in all
subsequent studies. Thus V14inv+ T cells accumulate
within the acute inflammatory lesions caused by pure mycobacterial
glycolipids, confirming and extending earlier data using other reagents
and probes (12).
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14inv NKT cells was first probed by in vivo blocking
experiments. Injecting 0.5 mg anti-CD1d1 Abs can efficiently block
the in vivo availability of CD1d1 molecules in C57BL/6 mice. Similar to
the results of others (22), this concentration effectively
blocked CD1d1 function, because it prevented CD1d1-dependent
up-regulation of CD69 by -GalCer (Refs. 22 and
23 and data not shown). "CD1d1-blocked mice" were
then challenged with PIM2. The recruitment of
V14inv NKT cells in the normal-sized lesions induced by
injection of PIM2 was similar to that observed in untreated
wild-type mice (data not shown).
To rule out the possibility that the blocking effect of the Ab could
have been overcome during the ongoing inflammatory process and that the
skin compartment may not be freely accessible to injected Abs, we used
mice in which the CD1d1 gene has been inactivated and that are
subsequently devoid of all CD1d1-restricted T cells, including
V14inv NKT cells. Their liver contains 
2–3%
NK1.1+ TCRint T cells (18)
but only trace amounts of V14-J281 rearranged TCR -chains
(data not shown), although CD1d1 T cells used all CDR3 sizes of V14
rearrangements (Fig. 2A). To
test the ability of CD1d1-/- mice to mount an acute
inflammatory response, CD1d1-/- mice were challenged with
PIM2. Seven days later, a loosely organized cell
infiltrate, consisting predominantly of neutrophils and macrophages,
much in the same way as in 2m-/- mice
(12), had developed. No V14+ TCR -chains
and no NKT specific clonotypes were detected in the lesions (Fig. 2B). Thus, an inflammatory lesion can develop in the absence
of CD1d1 and CD1d1-restricted T cells, and no bias in the V14 or the
J281 usages was observed.
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14inv+NK1.1+TCRint
T cell compartment in CD1d1-/- mice. For this purpose and
to increase the number of NKT cells available, we used C57BL/6 mice
transgenic for a V14inv TCR -chain that had been
backcrossed onto a C-/- background as a source for
NK1.1+TCRint T cells. These mice lack all
other V rearrangements. Thus, 2.5–3 x 106
NK1.1+TCRint T cells sorted out from the
liver and the spleen of C57BL/6 V14-J281 Tg C-/-
mice (Fig. 3A) were injected
into the spleen of CD1d1-/- C+/+ recipient
mice. A search for NK1.1+TCRint cells in
the liver of reconstituted animals 7 days after the transfer showed the
presence of 9% NK1.1+TCRint cells among
total liver lymphocytes, a value lower than in +/+ animals (22%) but
significantly higher than in CD1d1-/- mice (2–3%) (Fig. 3B). The partial restoration in the number of
NK1.1+TCRint cells was due to the
transferred cells. Indeed, by Immunoscope (21) analysis, the
V14inv chain was detected in the spleen and liver of the
reconstituted CD1d1-/- mice as a peak migrating with a
CDR3 size of 10 aa, sufficient to distort the V14-C profiles,
whereas the gaussian pattern of V14+ T cells was
centered on the CDR3 size of 9 aa (Fig. 3C).
CD1d1-/- animals could thus be efficiently reconstituted
with CD1d1-restricted NKT cells. As an example, of 3 million cells
injected, 500,000 were recovered in the liver and 106
were recovered in the spleen, as deduced by FACS, suggesting that most
of the injected NKT cells have retained their original homing
properties in the liver and spleen. Because significant expression of
CD1d1 molecules by donor cells would interfere with the experiments,
the expression of CD1d1 was measured. DN
(CD4-CD8-) and CD4+ NKT cells, in
contrast to NK1.1-TCRhighCD1d-restricted T
cells using the invariant TCR -chain, do not express significant
levels of CD1d1 molecules on their surface (24). This was
confirmed by analysis of the sorted, wild-type
NK1.1+TCRint T cell population used in
transfer experiments, which did not significantly express CD1d1
molecules and CD1d1 mRNA before adoptive transfer (Fig. 4). Nonetheless, a possible up-regulation
of CD1d1 (24) on donor cells after the transfer was taken
into account by injecting of 0.5 mg anti-CD1d1 MAb (20H2; Ref.
20) 12 h after the transfer, a concentration
known to effectively block CD1d1 accessibility (22). The
liver lymphocytes of reconstituted animals did not express detectable
CD1d1 molecules 7 days after the transfer, upon FACS using each of the
two anti-CD1d1-mAb, namely, 20H2, used in the blocking on day 1,
and 1B1, which recognizes a different epitope (the labeling shown in
Fig. 3D is that due to 1B1).
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14-C rearrangements using a common C primer and
runoff experiments using the clonotypic J281 primer revealed the
presence of abundant V14inv TCR -chains but no other
rearranged V14 chains within the lesions (Fig. 4
A). The
expression of CD1d1 on donor cells was examined after completion of the
experiments. FACS of the cells present in the lesions could not be
performed, but RT-PCR analysis showed that no CD1d1-coding mRNA was
detectable in the infiltrating cell after 50 cycles of PCR and using
CD1d1-specific primers. By contrast, CD1d1 mRNA was routinely detected
in the infiltrates induced by PIM2 in wild-type mice
regardless of the cell type that produces it (Fig. 4B).
The V14inv C-/- transgenic mice used a
unique nucleotide TCR -chain sequence. The V14-J281 sequences
used by CD1d-restricted T cells of C57BL/6 mice are a mixture of
several slightly different nucleotide and amino acid sequences, which
may have a different origin and different roles. To test the possible
role of variants of the -chain and also the possible influence of
otherwise rearranged T cells on NKT cell populations, we transferred
into C57BL/6 CD1d1-/- mice,
NK1.1+TCRint T cells sorted out of
wild-type C57BL/6 mice, thus containing all variants of the TCR
-chain. Reconstitution of liver populations was as efficient as
above (data not shown). Reconstituted animals also reacted to
PIM2, and the invariant V14-J281 chains were
recovered from the normal-sized and organized infiltrates (data not
shown). Sequence analysis of the junctional CDR3 region of the
V14-J281-rearranged TCR -chains found in the lesions showed
the presence of two nucleotide sequences, GTGGTGGGCGAT and
GTGGTGGGGGAT, that have been previously described in NKT cells of
wild-type mice (Ref. 25 and data not shown).
Thus, NK1.1+TCRint T cells can migrate
into the inflammatory lesions induced by mycobacterial PIM2
in the absence of detectable CD1d1 expression and in a
CD1d1- environment.
Prior studies had shown a critical role for IL-12R and CD40 expression
in the CD1d1 dependent activation of mature NKT cells (26, 27). We therefore investigated the response to glycolipids of
IL-12R-/- and CD40-/- mice. C57BL/6
IL-12R-/- and CD40-/- mice possessed
similar numbers of liver and spleen
NK1.1+TCRint T cells as wild-type mice
(Fig. 5A). PIM2
injection in C57BL/6 IL-12R-/- and CD40-/-
animals induced a cellular infiltrate with a size, organization, and
recruitment of the specific TCR invariant -chain (Fig. 5, B and C) indistinguishable from wild-type animals
(data not shown).
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14inv NKT cells can migrate
to and accumulate in the inflammatory lesions induced by mycobacterial
PIM2 in the absence of CD1d1 expression and in a
CD1d1- context. Moreover, a functional CD1d1/TCR
activation pathway was not required.
The Ag-driven activation of V14inv+ T cells
limits the development of granulomatous lesions
These results do not exclude a role for the CD1d1/TCR axis in the
accumulation of NKT cells in lesions induced in wild-type mice. This
hypothesis could be probed by using glycolipids known to activate
V14inv+ T cells in a CD1d1/TCR-dependent manner and
thus the glycolipid purified out of a marine sponge, -GalCer, the
only well-defined ligand known thus far to activate NKT cells in a
CD1d1-TCR-dependent manner (11). A synthetic analog of
natural -GalCer, KRN7000, has been produced and used along
with its anomer, -GalCer, to probe the stereospecificity of the
interactions within the CD1d1-GalCer-TCR complex (11).
While both - and -GalCer molecules bind CD1d1 molecules
(28, 29), only the anomer can activate
V14inv NKT cells through the TCR (11),
leading to rapid cytokine production (30) and
activation-induced cell death (31). The in vivo activation
by KRN7000 of V14inv NKT cells through their TCR
is also well documented (11). The availability of these
reagents made it possible to probe directly in vivo the hypothesis of
an Ag-driven accumulation of NKT cells by injecting each of the two
anomers as insoluble complexes. Thus, C57BL/6 mice were injected with 5
µg -GalCer or -GalCer. Spectroscopically pure mycobacterial
PIM2 was used as a positive control of NKT recruitment by
bacterial glycolipids. The injected animals were sacrificed at day 7
after the injection. Minor signs of inflammation and no recruitment of
V14inv TCR -chain were observed in PBS-injected mice
(Fig. 6). All other mice showed signs of
an ongoing inflammatory process. Mice injected with the anomer
showed small cell infiltrates, primarily necrotic and markedly
diminished in size (<1 mm diameter), and analysis of the cellular
infiltrate showed no V14-J281 transcripts in three of six animals
and barely detectable V14-J281 transcripts in the three other
mice (Fig. 6). By contrast, mice injected with the anomer developed
a strong inflammatory reaction, and well-structured lesions with a core
of neutrophils surrounded by a dense rim of macrophages and lymphocytes
wrapped into a fibroblast layer. These lesions were markedly enriched
primarily in invariant V14-J281 transcripts, as did wild-type
mice injected with PIM2 (Fig. 6). Systemic injection of
-GalCer leads to prompt apoptosis of NKT cells (31).
Results obtained using a systemic injection of -GalCer and a short
period of time are difficult to compare with those obtained using the
same reagent in an immobilized form and 7 days of exposure to it.
However, it appears likely that the lack of significant in situ
V14inv NKT cell infiltration in the animals injected
with -GalCer is due to cell death, a phenomenon not observed after
injection of -GalCer, with no prejudice of -GalCer acting locally
or at the periphery.
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14inv NKT cells in the acute
inflammatory lesions.
V14inv NKT cells are attracted precociously by
inflammatory cytokines
Because the accumulation of NKT cells is not Ag driven, the cells
could migrate into the lesions either due to chemoattraction or because
of a facilitated passage through the endothelium. The injected
glycolipids induce an acute inflammatory response. Because the early
release of TNF- is a hallmark of inflammatory processes, and TNF
is among the first cytokines detectable at the site of injection of
PIM2 a few hours after the injection (C. Ronet, manuscript
in preparation), we injected C57BL/6 mice s.c. with 0.1 µg TNF-
adsorbed on alum, using PBS-alum as a control, to mimic the local
production of an inflammatory cytokine. The cell infiltrates were
collected 3, 6, and 24 h after the injection. The presence of
V14+ T cells at the site of injection was studied by
Immunoscope analysis (Fig. 7). Cells
using the V14inv TCR -chain were detected in
TNF--induced lesions as early as 3–6 h after the injection. The
recruitment was selective given that no V14+
rearrangements other than V14-J281 with a CDR3 length of 10 aa
were detected. The early recruitment of T cells using the invariant TCR
-chain led us to examine the early response to pure
PIM2. Indeed, the invariant TCR -chain was occasionally
detected 6 h after the injection and unambiguously detected as the
only V14+ rearranged TCR -chain 12 h after the
injection, whereas it was absent from the controls (Fig. 7). There is
presently no proof that TNF- is the only proinflammatory cytokine
released at the time of injection. These findings strongly suggest that
V14inv NKT cells could be recruited to the site of
glycolipid injection through a cascade of events induced by an early
release of TNF-, among other cytokines that may be involved.
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In this paper, we show that V14inv NKT cells
accumulate in the inflammatory lesions caused by spectroscopically pure
or synthetic glycolipids, in the absence of the CD1d1 molecules and
CD1d1 activation pathway, rather than activated and expanded in situ in
an Ag-driven manner. The present data show that V14inv
NKT cells, presumably because they already possess an activated
phenotype, are attracted within hours to these inflammatory sites
primarily, along with neutrophils. V14inv NKT cells thus
participate in the first wave of leukocytes attracted to inflammatory
sites. In this respect, they behave like early cells of the innate
immune system. A similar conclusion has been reached after studies on
the T cells that infiltrate the skin during psoriasis; NKT cells were
recruited at inflammatory sites and were associated with the ongoing
inflammatory process (32). However, not all inflammatory
processes are associated with the recruitment of NKT cells, because we
did not detect the latter in the lesions of skin sarcoidosis
(33). The strict dependency on the presence of a lipidic
moiety (13) does not imply a role for the sole CD1d1
molecule. Indeed, blood molecules like CD14 are known to be carriers of
bacterial glycolipids like LPS (34). Would it be proved
that PIM2 can be loaded into CD14 molecules and that NKT
cells be activated at the periphery through CD14/TLR or other pathways
rather than through the CD1d/TCR pathway, the emerging picture of NKT
cells would be that of cells educated through their contact with
CD1d1+ cells in the thymus but that play part of their
roles at the periphery in the absence of the restriction element. Then,
in contrast to the physiology of conventional T cells, NKT cells are
selected in a manner that is distinct from the way the cells act at the
periphery.
Their role during the early stages on inflammation is not known. They
may contribute to the inflammatory response due to their immediate
ability to release locally IFN
in the absence of primary
stimulation, as already described in an animal model of psoriasis
(35). Finally, the absence of a dominant role of the
CD1d1/TCR axis in the accumulation of NKT cells does not rule out the
possibility that the infiltrating V14inv NKT cells may
control CD1d1-positive APCs cells by suppressing ongoing inflammation.
This mechanism has recently been proposed for human V24-JQ cells
(36). Nonetheless, it implies a function for
V14inv T cells in the immunosurveillance of injured
tissues, perhaps through a control of inflammatory processes rather
than exclusively in the defense against invading pathogens. This may be
a common feature underlying the pleiotropic function of NKT cells.
As an extension of our previous studies (12), we have
analyzed of the response to mycobacterial PIMs, presently
PIM2, in view of correlating the expression of CD1d
molecules with the accumulation of NKT cells. The response to other
inflammatory molecules like lipidoarabinomannan (37) or
granuloma-causing mycobacterial glycolipids like trehalose-mycolate
(38) was not studied. However, the finding that synthetic
-GalCer molecules induce very similar lesions and cause the local
accumulation of NKT cells strongly suggests that the response to
PIM2 is simply an example of a more general phenomenon
concerning the innate response to inflammatory glycolipids for which
LPS and lipoteichoic acids are the best studied representatives.
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Acknowledgments |
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Footnotes |
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2 M.M. and C.R. contributed equally to the work.
3 Address correspondence and reprint requests to Dr. Gabriel Gachelin, Unité de Biologie Moléculaire du Gène, Institut National de la Santé et de la Recherche Médicale Unite 277, Departement d’Immunologie, Institut Pasteur, 25 Rue du Dr. Roux, 75015 Paris, France. E-mail address: ggachel{at}pasteur.fr
4 Abbreviations used in this paper: GalCer, galactosylceramide; PIM, phosphatidylinositolmannoside; PIM2, phosphatidylinositoldimannoside.
Received for publication July 31, 2001. Accepted for publication October 29, 2001.
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References |
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TopAbstractIntroductionMaterials and MethodsResults and DiscussionReferences |
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repertoires and differential distribution in lymphoid tissues. J. Immunol. 165:2481. chain is used by a unique subset of major histocompatibility complex class I-specific CD4+ and CD4-8- T cells in mice and humans. J. Exp. Med. 190:1097.
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production by NK-T cells. J. Immunol. 165:4076.24-invariant natural killer T cells in T-cell-reactive leprosy together with a highly biased T cell receptor V repertoire. Am. J. Pathol. 157:509.24JQ T cells: a feedback mechanism for immune regulation. J. Immunol. 165:3756.This article has been cited by other articles:
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