| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
*
Graduate School of Biological Sciences, Mechanisms of Disease and Therapy Program, Mt. Sinai School of Medicine, New York, NY 10029;
Howard Hughes Medical Institute, Rheumatology Division, Washington University School of Medicine, St. Louis MO 63110; and
Department of Molecular Biology, Princeton University, Princeton, NJ 08544
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
, and TNF-
, as assessed in mice with targeted
mutations. Concurrent immunostaining for NK cells and viral Ags
revealed close association of NK cells and MCMV-infected cells in the
spleen and liver. Similar results were obtained in CD1-/-
and recombination activation gene-1-/- mice lacking NK T
or T and B cells, respectively, indicating specificity of staining for
NK cells. Thus, following MCMV infection, NK cells accumulate at sites
of viral replication in an IL-12-, IFN--, and TNF--independent
manner. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The spleen (in rodents) and peripheral blood (in humans) are the
primary sites from which NK cells are isolated for in vitro analysis,
but they have also been detected in the lymph node, thymus, and
nonlymphoid tissues including liver (2, 3). Unlike T and B
cells, whose specific organization within lymphoid tissues is well
characterized, NK cell distribution within lymphoid organs is less well
known, particularly during infection. Although there have been attempts
to define murine NK cell location in situ, these studies were
compromised by the coexpression of the available serologic markers such
as ASGM1 by other cell lineages, especially activated T cells
(9, 10, 11). In cell transfer experiments bone marrow NK cells
localize to areas within the spleen, where MCMV infected cells could
also be detected (12), suggesting that NK redistribution
to areas of infection may indeed occur. However, it is not clear
whether such colocalization actually occurs in situ during infection or
whether the observed effects reflect experimental cell trafficking. On
the other hand, NK cell-mediated antiviral effects in the liver have
been described as perforin-independent and IFN--dependent
(7), suggesting that NK cell localization in the liver may
be different from that in the spleen. Thus, the distribution of mouse
NK cells in situ, either normally or in the context of pathologic
processes, has not been clearly defined and may be helpful in
understanding NK cell control of viral infections.
The recent progress in understanding NK cell receptors has provided new serologic tools to address the issue of NK cell localization. Some of these receptors, such as gp49, belong to the Ig superfamily and are expressed by all activated NK cells (13). However, gp49 molecules are not expressed on resting NK cells and are constitutively expressed by a wide variety of other cell lineages (13, 14), thus precluding them as useful markers of NK cells in situ. Most NK cells also express receptors belonging to the C-type lectin superfamily (15) that are encoded in the NK gene complex (NKC) (16). Whereas some members of this superfamily, e.g., NKR-P1, in particular NKR-P1c (NK1.1), may be expressed by all NK cells (16, 17), others, such as the Ly-49 receptors, are expressed by subsets (18). The Ly49 family of molecules includes the inhibitory receptors for MHC class I, Ly49A, Ly49C/I, and Ly49G2, and the related activating receptors, Ly49D and Ly49H (3). Unlike gp49, Ly49 receptors are stably expressed once the adult NK cell receptor repertoire has been developed (19), and their expression does not appear to be significantly altered upon NK cell activation (20). All these receptors can be detected with specific mAbs by flow cytometry (3), suggesting that they may be useful for in situ localization studies.
In this study we examined the reactivity of all available NK cell receptor-specific mAbs by immunohistochemistry (IHC). Only one mAb, 4D11, specific for Ly49G2, which is expressed by approximately 50% of splenic NK cells (21, 22), was reactive with C57BL/6 tissue sections. Using this mAb we determined the normal in situ distribution of NK cells in spleen, lymph node, and thymus. This afforded us the opportunity to study changes that occurred during MCMV infection, and we show that the number of NK cells within various tissues as well as their specific location change during infection. Moreover, we show that NK cells are in close physical proximity to infected cells in both spleen and liver in situ. Thus, these studies provide significant new insight into NK cell responses in vivo.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The following mice were purchased from The Jackson Laboratory
(Bar Harbor, ME): C57BL/6 (B6), B6-IL-12-/-,
B6-IFN--/-, B6,129-TNF--/- (B6,
129-Tnftm1Gkl), B6,129SF2 (wild-type control for B6,
129-TNF--/-), and recombination activation
gene (Rag)-1-deficient C57BL/6
(C57BL/6J-Rag-1-/-).
C57BL/6-CD1-/- mice were previously generated
in our laboratory (23). All mice contained the targeted
allele on the C57BL/6 background and were housed in a pathogen-free
barrier facility at Washington University. This facility is supervised
by veterinarians; animal protocols have been approved by the animal
studies committee at Washington University.
Virus and infections
Stocks of Smith strain MCMV (ATCC VR-194, lot 10; American Type Culture Collection, Manassas, VA) were generated from mouse salivary glands as previously described (24). Mice were infected on day 0 with 5 x 104 PFUs of MCMV via the i.p. route.
Abs and antiserum
The mAb 4D11 (rat IgG2a, anti-Ly-49G2) hybridoma is a kind gift from Dr. J. Ortaldo (National Cancer Institute, Frederick, MD), while mAb 270 (rat IgG2a anti-chicken neuronal receptor for acetylcholine) hybridoma was purchased from American Type Culture Collection. The following mAbs were purchased from BD PharMingen (San Diego, CA): APC-4D11, DX5 (anti-DX5), 2B4 (anti-2B4), 5E6 (anti-Ly49C/I), 4E5 (anti-Ly49D), PE-Gr-1 (anti-Ly6G), APC-2C11 (anti-CD3), and PE-PK136 (anti-NK1.1). FITC-4D11 was generated as previously described (25). Anti-MCMV antiserum was a gift from Dr. H. W. Virgin IV (Washington University School of Medicine, St. Louis, MO) (26).
Immunohistochemistry and immunocytochemistry
Spleens and livers were obtained from infected or uninfected mice at different days following infection, frozen in Tissue Freezing Medium (Triangle Biomedical Sciences, Durham, NC) on dry ice, and stored at -20 or -80°C until sectioning. Organs were sectioned at 5–10 µm using a Leica 1850 cryostat (Leica, Wetzlar, Germany) and stored at -20°C until used. Before staining, sections were allowed to equilibrate to room temperature, fixed in cold acetone, and rehydrated in PBS. Endogenous peroxidase was depleted by incubating sections for 10 min in 0.3% hydrogen peroxide in methanol. To reduce nonspecific staining 10% normal goat serum in PBS and avidin/biotin block (Vector Laboratories, Burlingame, CA) were applied to all sections before addition of Abs. For NK cell staining mAb 4D11 was incubated with sections for 1 h at room temperature or at 4°C overnight. Bound Abs were detected with HRP-conjugated goat anti-rat IgG (Jackson ImmunoResearch Laboratories, West Grove, PA). Staining was amplified using TSA Biotin System (New England Nuclear, Boston, MA) according to the manufacturer’s recommendations. This was followed by incubation with preformed complexes of avidin and biotin (Vector Laboratories) conjugated to HRP (Vectastain-ABC) or alkaline phosphatase (Vectastain-ABC-AP). Each incubation step was followed by three 5-min washes in PBS. The HRP substrate 3-amino-9-ethylcarbazole (AEC Substrate kit, Vector Laboratories) was used for deposition of red chromogen, while the alkaline phosphatase substrate, Vector substrate kit III (Vector Laboratories), was used for deposition of blue-purple chromogen. After sufficient chromogen deposition had been achieved, the reaction was stopped by washing the sections in running tap water. Sections were counterstained with hematoxylin, washed, and mounted with Crystal/Mount (Biomeda, Foster City, CA). In the experiments in which both NK cells and MCMV-infected cells were detected in the same section, NK cell staining was performed as described above, followed by blocking with 10% normal goat serum, then an overnight incubation with anti-MCMV immune sera (26). Bound Abs were detected with donkey anti-mouse IgG (Jackson ImmunoResearch Laboratories), and all other subsequent steps were performed as described above. mAb 270 was used as an isotype-matched control for 4D11 while nonimmune serum (26) was used as a control for the anti-MCMV immune serum. All mAbs and antiserum were diluted in 10% normal goat or donkey serum and used at concentrations previously determined to yield optimal staining.
To perform immunocytochemistry on sorted cells, cells were first applied to microscope slides by cytospin, then allowed to dry at room temperature for 10 min. Subsequently 4D11 staining was performed as described above for tissue sections. The nuclear morphology of sorted cells was determined by staining with KaryoMAX Giemsa (Invitrogen, Carlsbad, CA). All photomicrographs were acquired using a Nikon FDX-35 camera mounted on a Nikon Eclipse E400 microscope (Nikon, Tokyo, Japan).
Leukocyte isolation and determination of NK cell numbers
Leukocytes were isolated from liver, spleen, peripheral blood, and bone marrow as previously described (27, 28). The number of cells isolated from each tissue was determined by counting live cells using an hemocytometer. Dead cells were identified by trypan blue staining. The percentage of leukocytes that were CD3- NK1.1+ (NK cells) was determined by FACS analysis. The total number of NK cells was calculated by multiplying the percentage of NK cells by the number of leukocytes.
Flow cytometric analysis and sorting
Splenocytes were isolated from C57BL/6 mice as previously
described (27, 28). Cells were resuspended in 2.4G2
(anti-FcRII/III) supernatant to block nonspecific binding
through the FcR (29). Subsequently the desired
fluorochrome conjugated mAbs were added at 1
µg/106 cells and incubated on ice for 30 min,
then washed three times in wash solution. During FACS analysis, gates
were set to exclude propidium iodide-positive (dead) cells. In cell
sorting experiments cells were stained as described for flow cytometry,
except that an unconjugated isotype matched mAb was substituted for
2.4G2 for blocking. Cells positive for the desired Ag were then sorted
on a MoFlo cell sorter (Cytomation, Fort Collins, CO). The purity of
sorted cells was determined by FACS analysis after sorting.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
To determine the in situ distribution of NK cells, we analyzed the reactivity of mAbs specific for NK cell receptors by immunohistochemistry. These mAbs were either specific for receptors expressed by all NK cells, including DX5 (mAb DX5) (30), 2B4 (mAb 2B4) (31), and NK1.1 (mAb PK136) or receptors that are expressed by subsets of NK cells, e.g., Ly-49C/I (mAb 5E6), Ly-49D (mAb 4E5), Ly-49G2 (mAb 4D11), and Ly-49H (mAb 3D10). Preliminary experiments with a variety of immunohistochemical methods indicated that only one of the mAbs tested, 4D11, was reactive with C57BL/6 tissues (data not shown).
mAb 4D11 (anti-Ly-49G2) has been shown to react with approximately
50% of all splenic NK cells (NK1.1+
CD3-) by FACS analysis (20, 21).
Only a minor population of 4D11+ cells (<10%)
expresses the TCR/CD3 complex (21), and most of these are
NK T cells (32) (our unpublished observations).
However, less is known about mAb 4D11 reactivity with other leukocytes.
We therefore evaluated 4D11+ splenocytes for
expression of macrophage and neutrophil markers. As expected, these
cells did not express the macrophage marker F4/80 (data not shown).
Surprisingly, 4D11+ cells expressed Gr-1 (data
not shown), a marker generally described as being neutrophil specific
(33), suggesting that neutrophils may express Ly-49G2 or
that NK cells may express Gr-1 at low levels. To address these
possibilities, 4D11+ splenocytes were sorted and
stained with Giemsa (Fig. 1
B).
4D11+ cells displayed the nuclear morphology of
lymphocytes and not that of neutrophils (Fig. 1B). This
morphology was identical with that of sorted
NK1.1+ CD3- cells
representing NK cells (Fig. 1A). By contrast, sorted
Gr-1high cells displayed the characteristic
ringed polymorphonuclear (PMN) neutrophilic morphology (Fig. 1C). Thus, 4D11+ cells display the
morphology of lymphocytes and not that of PMN cells.
|
B). Approximately 50% of sorted
NK1.1+ CD3- cells showed
staining with mAb 4D11, consistent with results from flow cytometric
analysis of the same population (Fig. 1A). By contrast,
4D11-stained cells were rare in the Gr-1-sorted population and most
likely reflected rare contaminating NK cells from the sorting (Fig. 1C). Taken together with previous observations
(21), mAb 4D11 appears to recognize bonafide NK cells and
not other lineages. Importantly, it can be used in immunohistochemistry
for identification of NK cells. Normal distribution of NK cells in C57BL/6 mice
Although Ly-49G2 is expressed on approximately 50% of NK cells,
this proportion appears to be stable on bulk NK cell populations in all
tissues examined by flow cytometry (our unpublished
observation). The ability to perform immunohistochemistry with
mAb 4D11 therefore allowed us to assess the normal distribution of NK
cells in mouse lymphoid organs and liver. Chromogen deposition on
4D11+ cells formed a circular pattern (Fig. 2A, inset),
consistent with membrane expression of the Ly-49G2 receptor. In the
spleen these NK cells were distributed primarily in the red pulp;
however, an occasional NK cell could be found within lymphoid follicles
(Fig. 2A, arrow). In the lymph node, NK cells were found
primarily at the junction between adjacent lobes of the lymph node and
within the T cell area (Fig. 2C). The B cell areas of the
lymph node were generally devoid of NK cells (beyond the field of view
in Fig. 2C). Unlike the spleen and lymph node, NK cells in
the thymus were not localized to a particular region of the organ, but
were randomly distributed in both the cortex and medulla (Fig. 2E). Interestingly, no NK cells were detected in the liver
(data not shown), although NK1.1+
CD3- cells are easily detected by flow cytometry
of liver cell suspensions (34).
|
NK cell distribution is altered during infection
Having established the normal distribution of NK cells in various
organs, it was then possible to study alterations in these patterns
during pathologic processes. Since NK cells are required for early
control of MCMV replication, we sought to determine whether their
distribution is altered during MCMV infection. C57BL/6 (resistant
strain) mice were infected, and cryosections from spleen and livers
were stained with mAb 4D11 on successive days following infection. At
day 1 post infection the distribution of NK cells in spleen remained
unchanged (Fig. 3A). However,
by day 2 after infection the red pulp of the spleen became less
cellular, and clusters of NK cells were found to be associated with the
marginal zone and sinus area (Fig. 3C). An increase in the
number of NK cells associated with lymphoid follicles was also observed
(Fig. 3C). Analysis of day 3, 4, and 5 sections showed that
NK cells were no longer associated with the marginal zone and sinus,
with the pattern of distribution almost identical to that of an
uninfected spleen (Fig. 3E). It is unlikely that these
observed changes are due to effects of NK T cells, since similar
observations were made in CD1-/- mice (Fig. 3G).
|
, A and E).
However, some cells were associated with blood vessels and hepatic
sinusoids (Fig. 4A). On day 5 most NK cells in the liver
remained associated with inflammatory foci (Fig. 4C). This
is contrary to the spleen in which the NK cell distribution at this
time point appears similar to that of an uninfected organ (data not
shown). Analysis of Rag 1-/- and
CD1-/- livers showed results similar to those
of wild-type C57BL/6 mice, confirming that the stained cells were not T
or NK T lymphocytes (Fig. 4, F and G).
|
Inasmuch as 4D11-reactive cells were undetectable by IHC in
uninfected liver, their appearance after infection could be due to an
increase in the total number of NK cells associated with the liver or a
selective change in the distribution of Ly49G2+
cells. To address these possibilities, we evaluated the total number of
liver-associated NK cells in uninfected and infected mice by flow
cytometry and cell count. On day 3 postinfection the percentage (Fig. 5A) and total number (Fig. 5B) of liver NK cells in infected mice were significantly
increased compared with uninfected controls, whereas the percentage of
Ly49G2+ cells was not markedly altered (Fig. 5C and data not shown). Thus, the capacity to detect
previously undetectable 4D11+ NK cells in liver
following infection may be due to an increase in the total number of NK
cells within this tissue.
|
, A and B) at day 3 postinfection. To
better understand these results we analyzed the kinetics of the change
in total NK and 4D11+ cell number in liver,
spleen, and bone marrow NK cells. The results show a continuous
increase in both bulk liver NK cells and 4D11+
cells throughout the duration of the study (6 days; Fig. 5C). The magnitude of increase in liver NK cells was nearly
5-fold by day 3 and 10-fold by day 6 (Fig. 5C). By contrast,
only small changes were noted in the bone marrow, but the number of
bone marrow NK cells remained at or below preinfection levels
throughout the study (Fig. 5C). In the spleen the number of
NK cells decreased initially and then increased as the infection
progressed, reaching about a 2-fold increase by day 6 (Fig. 5C). The changes in NK cell number in the spleen were modest
compared with those observed in the liver. While these changes could be
due to trafficking, recruitment, or proliferation, these results
nevertheless suggest that during MCMV infection, the distribution of NK
cells favors their accumulation within the liver and spleen. Role of cytokines in NK cell distribution
To address the mechanisms involved in the observed changes in NK
cell distribution during MCMV infection, we investigated the role of
certain cytokines that have been shown to affect NK cells. IL-12
administration has been shown to induce an IFN--dependent NK cell
recruitment to liver in C57BL/6 mice (35). This is similar
to the effect of poly(I:C) administration (36) that is
blocked by anti-TNF- antiserum (37). We therefore
hypothesized that our observed changes during MCMV infection may be
IL-12, IFN-, or TNF- dependent. Using gene-targeted mice we
assessed the potential role of these cytokines in NK cell recruitment
during infection. Following MCMV infection, livers from mutant and
wild-type mice were analyzed to determine the number of NK cells
associated with the tissue and compared with that in uninfected mice.
To our surprise MCMV infection induced an increase in liver NK cells in
all mutant and wild-type mice (Fig. 6).
Therefore, the liver accumulation of NK cells during MCMV infection is
independent of IL-12, IFN-, or TNF- alone.
|
Perforin-mediated cytotoxicity requires physical interaction
between effector and target cells (8). Since NK cell
control of MCMV in the spleen is perforin dependent (7),
close association between NK cells and virally infected cells should
occur. Staining of sequential sections of the spleen for either MCMV or
NK cells show that the distribution pattern of NK cells is similar to
that of infected cells (Fig. 7, A and B), indicating that NK cells may colocalize
with infected cells. Indeed, staining for both NK cells and
MCMV-infected cells in the same section revealed that these cells were
in close proximity, although not all NK cells were found in close
proximity of virally infected cells (Fig. 7C). In the liver
MCMV-infected cells were detected as clusters of two or more cells
randomly distributed throughout the tissue (Fig. 7D). Some
infected cells were associated with inflammatory foci (data not shown).
As in the spleen, some NK cells could be found in the proximity of
infected cells, although not all infected cells were found associated
with NK cells (Fig. 7D). Thus, following MCMV infection NK
cells show physical proximity with infected cells, especially in the
spleen.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The role of NK cells in the control of acute MCMV infection is now well established (38, 39). However, the details of NK cellular responses during infection have remained unclear because of the technical difficulties in identifying and tracking them in situ. The major effector functions of NK cells are cytokine secretion and direct cytotoxicity (3, 5); NK cells may therefore need to be located at sites of viral replication to perform these functions. The liver is a major site of viral replication during MCMV infection; accumulation of NK cells within this organ is consistent with the hypothesis that NK cells need to home to such a site to achieve their effector functions. Previous in situ analysis of cells associated with liver inflammation during MCMV infection show colocalization of ASGM1+ and MCMV-infected cells (40). However, the physical relationship between NK cells and MCMV-infected cells remained inconclusive, since cells of other lineages also express ASGM1, especially activated T cells (9, 10, 11). While this manuscript was in preparation, Andrews et al. (11) showed that NK cells in the liver do associate with MCMV-infected cells by direct immunofluorescent staining using PE-conjugated mAb PK136 (anti-NK1.1). However, this study was not able to determine histologic architecture and did not address changes in NK cell numbers in various compartments during infection. Here, we not only demonstrate that NK cells are found in close proximity of infected cells, but in addition we show that their numbers increase significantly in the liver, but not in spleen or bone marrow.
Surprisingly, we were unable to detect NK cells with 4D11 reactivity in uninfected liver tissues. Yet, NK cells can be easily isolated from single cell suspensions of leukocytes from this organ. It is possible that in uninfected livers, NK cells are resident within the blood vasculature and are washed away during the sample preparation, whereas they are retained in infected livers due to infiltration of the parenchyma. Alternatively, the density of NK cells in the liver may be too low to detect, since there are approximately 103-fold more NK cells per gram of uninfected tissue in spleen compared with liver (our unpublished observation), and subsequently, reflecting at least a 5-fold increase in liver NK cells, they can be visualized by immunohistochemistry. Interestingly the number of NK cells in the spleen decreases initially during infection. Although this could be due to other mechanisms, including cell death, the subsequent increase in liver NK cells suggests that recruitment to the liver may have occurred. Alternatively, it is possible that 4D11 staining is present in infected livers due to up-regulation of Ly49G2 expression. This seems unlikely, since the percentage of 4D11+ cells remains relatively constant during infection, and the level of Ly49G2 expression is comparable in liver and spleen by FACS. Taken together, while we cannot rule out the possibility that other factors may have contributed to our ready detection of NK cells in infected livers, the observed increase in the total number of NK cells associated with the liver as well as an increase in the number of NK cells in the parenchyma may have contributed to the immunohistochemical detection of NK cells in infected liver.
NK cell responses are differentially regulated among organs. For
instance, during MCMV infection perforin-dependent mechanisms are
required for control of viral replication in the spleen, but not in the
liver (7), suggesting a critical role for granule-mediated
cytotoxicity. Conversely, IFN- appears to be critical for MCMV
clearance in the liver (5, 7). Furthermore, the effect of
Cmv1, a previously described autosomal dominant genetic
locus controlling MCMV replication, is primarily mediated in the spleen
(41). Our analyses of uninfected spleens show that NK
cells were located primarily in the red pulp. Inasmuch as the blood
circulation in the red pulp is open (42), NK cells are
positioned to directly interact with contents of the blood or to
respond to products secreted by red pulp macrophages that are part of
the reticulo-endothelial system (42). Following MCMV
infection there was an alteration in the normal NK cell distribution
pattern. In marked contrast to uninfected spleens, the red pulp was
practically devoid of NK cells. Furthermore, the lymphoid follicle and
marginal zone areas, which rarely contained NK cells in uninfected
spleen, had an accumulation of these cells. This was not surprising,
since previous studies as well as our own observations revealed that
cells in the marginal zone are the predominant MCMV-infected cells in
the spleen (11, 43). Here we clearly show that some
splenic NK cells are in physical proximity to infected cells,
consistent with a role for cytotoxicity in NK cell-mediated resistance
to MCMV. As mentioned earlier, not all detected NK cells were
associated with virally infected cells, suggesting that NK cells may be
heterogeneous in their response to infection with only specific subsets
involved in viral clearance. Interestingly, we recently identified
Ly-49H as an NK cell activation receptor that mediates resistance to
MCMV (44). NK cell subsets expressing this receptor may
therefore respond differently than others. While the mAb to Ly49H
(3D10) has not been useful in immunohistochemistry (our unpublished
observation), it is interesting to note that approximately 60%
of the 4D11-reactive cells express Ly49H (20). In
conclusion, our current data establish the normal distribution of NK
cells in lymphoid organs and clarify their physical relationship to
infected cells in the spleen.
IL-12, IFN-, and TNF- were previously implicated in induction of
NK cell accumulation in liver (35, 36). Since these
cytokines are secreted during MCMV infection (45, 46), we
assessed their role in mediating our observed increase in liver NK cell
numbers during infection. To our surprise the increase in the number of
liver NK cells was independent of each individual cytokine. This may be
due to a redundancy in the mechanisms involved in NK cell recruitment.
In MCMV infection a multitude of cytokines is produced. Any one alone
may be able to induce an increase in the number of liver NK cells.
Alternatively, the increase in NK cell number may be chemokine
mediated, since NK cells can respond to several chemokines in vitro,
including RANTES, macrophage inflammatory protein-1 (MIP-1),
MIP-1
, macrophage chemoattractant protein-1 (MCP-1), and MCP-2
(47). However, the physiological role of these chemokines
in NK cell recruitment is not well understood. Recently, it was shown
that the recruitment of NK cells to the liver during MCMV infection was
abrogated in MIP-1-/- mice
(40), suggesting that MIP-1-/-
may be required for NK cell recruitment and that NK cell recruitment is
needed for protection. It is important to note, however, that no
inflammation could be detected in the liver of the
MIP-1-/- mice. This suggests that MIP-1
may not be specifically required for NK cell recruitment, but may
instead be a more general proinflammatory chemokine; the lack of NK
cells may therefore be an indirect effect.
Thus, the current studies indicate the in situ localization of NK cells and provide valuable insight into understanding the role of NK cells during infection.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Wayne M. Yokoyama, Rheumatology Division, Box 8045, Department of Medicine, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110. E-mail address: yokoyama{at}imgate.wustl.edu
3 Abbreviations used in this paper used in this paper: MCMV, murine CMV; ASGM1, asialo ganglio-N-tetraoglyceramide; NKC, NK gene complex; MIP-1, macrophage inflammatory protein-1; MCP-1, macrophage chemoattractant protein-1; MCP-2, macrophage chemoattractant protein-2; PMN, polymorphonuclear; Rag, recombination activation gene; IHC, immunohistochemistry.
Received for publication May 14, 2001. Accepted for publication August 21, 2001.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
interferon and tumor necrosis factor in macrophage activation during murine cytomegalovirus and herpes simplex virus infections. J. Virol. 69:904.[Abstract]
regulates acute and latent murine cytomegalovirus infection and chronic disease of the great vessels. J. Exp. Med. 188:577. T cells with potent cytotoxicity in the liver of mice via production of IL-12 from Kupffer cells. J. Immunol. 156:2436.[Abstract]
and VCAM-1 and is rapidly down-regulated by a mechanism involving T cells and expression of Fas. J. Immunol. 161:6014. is a principal cytokine involved in the recruitment of NK cells to liver parenchyma. J. Immunol. 153:333.[Abstract]
(MIP-1)-dependent pathways. J. Exp. Med. 187:1.This article has been cited by other articles:
|
|
 |
|
  P. Jennings and D. Yuan NK Cell Enhancement of Antigen Presentation by B Lymphocytes J. Immunol., March 1, 2009; 182(5): 2879 - 2887. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Giroux, E. Yurchenko, J. St.-Pierre, C. A. Piccirillo, and C. Perreault T Regulatory Cells Control Numbers of NK Cells and CD8{alpha}+ Immature Dendritic Cells in the Lymph Node Paracortex J. Immunol., October 1, 2007; 179(7): 4492 - 4502. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.-R. Chang, K.-J. Wang, Y.-F. Lu, L.-J. Yang, W.-J. Chen, Y.-H. Lin, H.-H. Chang, and S.-L. Wang Characterization of Early Gamma Interferon (IFN-{gamma}) Expression during Murine Listeriosis: Identification of NK1.1+ CD11c+ Cells as the Primary IFN-{gamma}-Expressing Cells Infect. Immun., March 1, 2007; 75(3): 1167 - 1176. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Walzer, M. Blery, J. Chaix, N. Fuseri, L. Chasson, S. H. Robbins, S. Jaeger, P. Andre, L. Gauthier, L. Daniel, et al. Identification, activation, and selective in vivo ablation of mouse NK cells via NKp46 PNAS, February 27, 2007; 104(9): 3384 - 3389. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Qian, X. Jiang, H. An, Y. Yu, Z. Guo, S. Liu, H. Xu, and X. Cao TLR agonists promote ERK-mediated preferential IL-10 production of regulatory dendritic cells (diffDCs), leading to NK-cell activation Blood, October 1, 2006; 108(7): 2307 - 2315. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Bajenoff, B. Breart, A. Y.C. Huang, H. Qi, J. Cazareth, V. M. Braud, R. N. Germain, and N. Glaichenhaus Natural killer cell behavior in lymph nodes revealed by static and real-time imaging J. Exp. Med., March 20, 2006; 203(3): 619 - 631. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A.-C. Iversen, P. S. Norris, C. F. Ware, and C. A. Benedict Human NK Cells Inhibit Cytomegalovirus Replication through a Noncytolytic Mechanism Involving Lymphotoxin-Dependent Induction of IFN-{beta} J. Immunol., December 1, 2005; 175(11): 7568 - 7574. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Baratin, S. Roetynck, C. Lepolard, C. Falk, S. Sawadogo, S. Uematsu, S. Akira, B. Ryffel, J.-G. Tiraby, L. Alexopoulou, et al. Natural killer cell and macrophage cooperation in MyD88-dependent innate responses to Plasmodium falciparum PNAS, October 11, 2005; 102(41): 14747 - 14752. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. E. Berg, E. Crossley, S. Murray, and J. Forman Relative Contributions of NK and CD8 T Cells to IFN-{gamma} Mediated Innate Immune Protection against Listeria monocytogenes J. Immunol., August 1, 2005; 175(3): 1751 - 1757. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. H. Robbins, M. S. Tessmer, T. Mikayama, and L. Brossay Expansion and Contraction of the NK Cell Compartment in Response to Murine Cytomegalovirus Infection J. Immunol., July 1, 2004; 173(1): 259 - 266. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. J. Roberts, Y. Lin, P. M. Spence, L. Van Kaer, and R. R. Brutkiewicz CD1d1-Dependent Control of the Magnitude of an Acute Antiviral Immune Response J. Immunol., March 15, 2004; 172(6): 3454 - 3461. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. P. Makrigiannis, E. Rousselle, and S. K. Anderson Independent Control of Ly49g Alleles: Implications for NK Cell Repertoire Selection and Tumor Cell Killing J. Immunol., February 1, 2004; 172(3): 1414 - 1425. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Friese, M. Platten, S. Z. Lutz, U. Naumann, S. Aulwurm, F. Bischof, H.-J. Buhring, J. Dichgans, H.-G. Rammensee, A. Steinle, et al. MICA/NKG2D-Mediated Immunogene Therapy of Experimental Gliomas Cancer Res., December 15, 2003; 63(24): 8996 - 9006. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. A. Rabinovich, J. Li, J. Shannon, R. Hurren, J. Chalupny, D. Cosman, and R. G. Miller Activated, But Not Resting, T Cells Can Be Recognized and Killed by Syngeneic NK Cells J. Immunol., April 1, 2003; 170(7): 3572 - 3576. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. S. Falk, M. Mach, D. J. Schendel, E. H. Weiss, I. Hilgert, and G. Hahn NK Cell Activity During Human Cytomegalovirus Infection Is Dominated by US2-11-Mediated HLA Class I Down-Regulation J. Immunol., September 15, 2002; 169(6): 3257 - 3266. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
