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Division of Rheumatic Diseases, University of Connecticut Health Center, Farmington, CT 06030
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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after Ag stimulation.
Reactivation of memory cells by Ag feeding resulted in increased cell
size and up-regulation of CD28 and CD11c. CD8 mucosal memory cells
exhibited ex vivo lytic activity that was up-regulated dramatically
following Ag reencounter in vivo. Interestingly, reactivation of memory
cells did not require CD28-mediated costimulation. The ability of the
intestinal mucosa to maintain CD8 memory cells provides a potential
mechanism for effective mucosal vaccination. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The cellular and molecular basis of T cell memory remains poorly understood, and progress has been hampered by the inability to trace long-term Ag-specific memory T cells in vivo. Recently, the use of adoptive transfer of TCR transgenic T cells to normal mice (2, 8) and the production of tetrameric major histocompatibility Ag-peptide complexes (4, 9, 10) have allowed visualization of the fate of Ag-specific T cells in vivo. These systems provide the means for clarifying controversial issues regarding T cell activation and memory generation. In a few studies, Ag-specific memory T cells were defined and characterized based on their long-lived nature (2, 4, 10). The majority of those in vivo studies focused on memory T cells found in secondary lymphoid tissues, such as lymph nodes (LN) or spleen, but did not examine tertiary sites, such as the mucosa-associated lymphoid tissues. In one case, the presence of protective memory cells was not detected in the footpad of LCMV-immune mice after virus challenge at that site (11). This finding was used as evidence that persisting Ag is needed to maintain memory outside of secondary lymphoid tissues. However, the mucosae present unique anatomical and functional attributes as compared with other tissues. Because many pathogens gain access to the host through mucosal tissues, this could be an important site in which to focus immune memory responses. In fact, as previously suggested (12), in cases of mucosal infection, local clinical symptoms caused by infectious agents may occur rapidly such that reactivation of memory T cells in draining secondary lymphoid tissues and eventual recruitment of effector cells to the mucosa may be ineffective in controlling the initial infection. Thus, from the standpoint of protective immunity, memory T cell responses may be critical outside the secondary lymphoid tissues.
T lymphocytes in the intestinal mucosa exhibit functional and
phenotypic characteristics of activated or memory cells
(13, 14, 15, 16, 17). The phenotypic attributes of intestinal T cells
include low expression of CD62L (18) and high level
expression of activation markers such as CD11c (17).
Moreover, CD8 T cells, when freshly isolated from the intraepithelial
lymphocyte (IEL) or lamina propria (LP) compartments, show constitutive
cytolytic activity (16, 19). Although the precise origin
of these CTL remains unclear, recent data suggest that TCR
IEL
mature extrathymically, whereas the production of most TCR
ß IEL
requires the thymus (20, 21). Thus, although TCR IEL
may be activated in the intestinal epithelium, it is possible that
thymus-derived TCRß IEL are activated outside of the epithelium
and traffic to the intestinal mucosa. This was originally implied by
the pioneering studies of Sprent (22), in which activated,
alloreactive T cells were tracked in vivo. In recent studies, we have
used an adoptive transfer model in which OVA-specific CD8 T cells can
be tracked, and we demonstrated that activation of CD8 T cells outside
of the mucosa was required for entry of these cells into the LP and the
epithelium (23). OVA-specific CTL activity was detected in
the intestinal mucosa and was attributable to the migrating transgenic
donor cells. The latter was true whether immunization was via virus
infection or through injection of soluble OVA (sOVA) without adjuvant.
In contrast, CTL activity in secondary lymphoid tissue was generated
only in response to virus infection and not to sOVA. Thus, the
intestinal mucosa provides a potentiating environment for CTL
responses. In the present study, we have examined whether this
environment is conducive to maintenance of long-term CD8 T cell memory.
We find that memory CTL are retained in intestinal tissues and that
such cells can be clearly distinguished from peripheral memory T cells
by phenotype and function. These findings help explain the constitutive
lytic activity of TCRß IEL and also indicate that systemic
immunization could provide mucosal protection against certain
pathogens.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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C57BL/6J (Ly-5.1) mice were obtained from The Jackson Laboratory (Bar Harbor, ME). C57BL/6-Ly-5.2 mice were obtained from Charles River (Wilmington, MA) through the National Cancer Institute animal program. The OT-I mouse line was generously provided by W. R. Heath (Walter and Eliza Hall Institute, Parkville, Australia) and F. Carbone (Monash Medical School, Prahran, Victoria, Australia) (24), and was maintained as a C57BL/6-Ly-5.2 line on a RAG-/- background.
Adoptive transfer and immunization
This method was adopted from Kearney et al. (8). A total of 2.5 x 106 pooled LN cells from OT-I-RAG-/- (Ly5.2+) mice was injected i.v. into C57BL/6J (Ly-5.1) mice. Two days later, 5 mg of OVA (grade VI; Sigma, St. Louis, MO) was administered by i.p. injection, or 1 x 106 PFU of VSV or VSV-OVA were injected i.v. The production of rVSV-OVA has been described previously (23). Lymphocytes were isolated at the indicated times and analyzed for the presence of transferred cells by flow-cytometric detection of Ly-5.2+ cells. Ab treatments were performed by i.p. injection of 100 µg of CTLA4-Ig or the CTLA4-Ig mutant 104 as control (25), which were generously provided by Philip Morton (G. D. Searle, St. Louis, MO). Mutant 104 does not bind to B7-1 or B7-2, but retains FcR binding. Injections were given daily starting on the day of immunization.
Isolation of lymphocyte populations
IEL and LP cells were isolated as described previously (26, 27). For cytotoxicity assays, panning of Percoll-fractionated IEL on anti-CD8 mAb-coated plates was performed to remove contaminating epithelial cells. LN and spleens were removed, and single cell suspensions were prepared using a tissue homogenizer. PLN included brachial, axillary, and superficial inguinal nodes. The resulting preparation was filtered through Nitex (Tetko Industries, Kansas City, MO), and the filtrate was centrifuged to pellet the cells.
Immunofluorescence analysis
Lymphocytes were resuspended in PBS/0.2% BSA/0.1% NaN3 (PBS/BSA/NaN3) at a concentration of 1 x 106–1 x 107 cells/ml, followed by incubation at 4°C for 30 min with 100 µl of properly diluted mAb. The mAbs either were directly labeled with FITC, PE, or CyChrome, or were biotinylated. For the latter, avidin-Red 670 (Av-R670; Life Technologies, Gaithersburg, MD) was used as a secondary reagent for detection. After staining, the cells were washed twice with PBS/BSA/NaN3 and fixed in 3% paraformaldehyde in PBS. Relative fluorescence intensities were then measured with a FACSCalibur (Becton Dickinson, San Jose, CA). Data were analyzed using Lysys II or WinMDI software.
Measurement of cytolytic activity
Cytolytic activity was measured using 51Cr sodium chromate-labeled EL4 cells (an H-2b thymoma) with or without the addition of 10 µg/ml of the OVA-derived peptide SIINFEKL. Serial dilutions of effector cells were incubated in 96-well round-bottom microtiter plates with 2.5 x 103 target cells for 6 h at 37°C. Percent specific lysis was calculated as: 100 x {[(cpm released with effectors) - (cpm released alone)]/[(cpm released by detergent) - (cpm released alone)]}.
Intracellular detection of IFN-
Lymphocyte populations were isolated from unimmunized OT-I mice
or from OT-I-transferred VSV-OVA-immunized mice 5 wk after infection.
Cells were cultured in DMEM/5% FCS/10% Nu Serum (Life Technologies)
with added HEPES, 2-ME, and antibiotics at a density of 1 x
106 cells/ml in a 24-well dish at 37°C. To
stimulate OT-I cells, cultures were treated with 1 µg/ml of SIINFEKL
peptide. Golgiplug (containing Brefeldin A; PharMingen, San Diego, CA)
was added to unstimulated and stimulated cultures at a concentration of
1 µl/ml. Cells were harvested after 5 h and stained for cell
surface Ags, as previously described. Cells were then fixed in 4%
paraformaldehyde/PBS for 20 min at 4°C, washed twice, and stored
overnight at 4°C. The next day, the cells were permeabilized by
incubating in Perm/Wash solution (PharMingen) for 20 min. The
permeabilized cells were incubated with anti-IFN- FITC (XMG1.2,
5 µg/ml; PharMingen) or control rat IgG1 FITC (R3-34, 5 µg/ml;
PharMingen) for 30 min at 4°C and washed twice in Perm/Wash solution.
The fluorescence intensities were immediately measured on a
FACSCalibur.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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To visualize Ag-specific CD8 T cell activation in a defined
system, we used adoptive transfer of TCR transgenic CD8 T cells from
the mouse line, OT-I. The majority of OT-I T cells express a TCR
specific for an OVA-derived peptide in the context of
H-2Kb (24). Donor cells are tracked
by detection of differences in Ly-5.1 and Ly-5.2 expression between the
donor and host. We previously showed that immunization of
OT-I-transferred mice with either sOVA or VSV-OVA resulted in
substantial clonal expansion in peripheral and mucosal tissues during
primary responses (23). To determine the long-term outcome
of the OT-I immune response to VSV-OVA or sOVA, animals were analyzed
at late time points after adoptive transfer and immunization. Control
mice were infected with wild-type VSV. During the primary response (day
6 in the experiment shown), Ag-specific clonal expansion of donor
transgenic T cells occurs in PLN in response to VSV-OVA infection (Fig. 1
) or sOVA immunization
((23) and data not shown). Migration of donor OT-I T cells
into the intestinal epithelium was only detectable in OVA-immunized
mice and not in control mice. However, 35 days after immunization with
sOVA or wild-type VSV, donor cells were not detectable in PLN and were
barely detectable in IEL (Fig. 1). In contrast, OT-I cells were present
in PLN and the intestinal mucosa, including the epithelium (Fig. 1) and
LP (see below), of VSV-OVA-immunized mice. Phenotypic analysis
demonstrated that donor cells detected in the LN and the intestinal
mucosa expressed CD8ß and the transgenic TCRß (data not
shown). We defined these Ag-specific CD8 T cells present in the host
following immunogenic challenge as memory cells and specifically, those
that were found in the gut, as intestinal mucosa-specific memory
cells.
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ß IELs. After this time
point, the percentage of OT-I memory cells remained relatively stable
for at least 4 mo (Fig. 2), suggesting
that 
2 wk after immunization, the residual cells could be classified
as memory cells. Indeed, we have detected memory cells in the mucosa 1
yr after immunization (data not shown).
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Having defined memory CD8 populations in distinct anatomic
locations, we wished to determine whether each population could be
distinguished by phenotype. Therefore, we analyzed the expression of
activation and adhesion molecules by OT-I memory T cells from the LN
and the intestinal mucosa. Memory cells from LN or intestine expressed
comparable levels of CD8ß and TCR (data not shown). However,
striking differences were noted in expression of certain adhesion
molecules by LN vs intestinal mucosa-specific memory T cells (Fig. 3, , A and B). CD44
was rapidly up-regulated on OT-I cells in gut, LN (Fig. 3A),
and spleen (data not shown) after primary activation, and the level of
CD44 expression remained high in LN-specific memory T cells (Fig. 3B). In contrast, memory OT-I T cells found in the IEL
compartment exhibited more heterogenous expression of CD44 (Fig. 3B). OT-I memory cells in LP had CD44 levels only marginally
lower than that of splenic and LN memory OT-I cells. Similar to naive
host CD8 T cells, CD62L expression was heterogenous among naive PLN
OT-I T cells (Fig. 3A). Upon primary activation, two
distinct populations (CD62L high and CD62L low/intermediate) were
detectable in PLN (Fig. 3A) and in the spleen, although in
the latter CD62L low cells made up a larger portion of the cells as
compared with those in LN (data not shown). Multiparameter analysis
showed no difference in other activation Ag expression (CD44 and CD11a)
between these two populations. In contrast, primary activated as well
as memory mucosal OT-I cells for the most part lacked CD62L, although a
small population of CD62L+ cells was present
(Fig. 3). These phenotypes were identical with those of subsets of
endogenous host IEL and LP populations.
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Eß7 integrin is
expressed at high levels by most IEL and at lower levels by a subset of
peripheral CD8 T cells in normal mice (29). Similar to the
latter, naive OT-I LN cells expressed heterogenous levels of
Eß7, with many cells
lacking this integrin. Primary activated and memory LN OT-I cells
expressed low levels of
Eß7, indicating a loss
of
Eß7high
cells from LN (Fig. 3
, A and B). Interestingly,
after primary activation, most LP OT-I cells lacked
Eß7, while a distinct
population of OT-I IEL had heterogenous high levels of
Eß7. This result
suggested either that rapid up-regulation of
Eß7 had occurred on
OT-I IEL or that cells expressing
Eß7 preferentially
migrated to the epithelium. However, the majority of IEL-specific OT-I
memory cells expressed high levels of
Eß7 integrin, while LP
memory cells contained discernible
Eß7 high and
Eß7 low populations,
suggesting that up-regulation of
Eß7 occurred after
entry of cells into the mucosa. Overall, the memory T cells found in
each location resembled subsets of the resident CD8 populations with
regard to adhesion molecule expression. Memory CTL exhibit lytic activity that can be up-regulated by reexposure to Ag
Memory T cells from LCMV-immune mice exhibit low levels of
cytolytic activity (3). However, whether this is a
generalizable attribute of CD8 memory cells is unclear. This is an
important issue because more effective protection should be achieved if
memory CD8+ T cells can kill virus-infected cells
immediately without a lengthy reactivation process. Therefore, we
tested the lytic activity of mucosal and peripheral CD8 memory cells by
performing cytotoxicity assays on ex vivo LN, spleen, and IEL
populations containing OT-I memory T cells. Because the actual number
of potential effectors was determined by flow cytometry, lytic activity
on a per cell basis could be compared. At low E:T ratios (i.e., 1:1),
little OVA-specific killing activity was detected from MLN OT-I memory
T cells (Fig. 4A), whereas at
higher ratios (>10:1) MLN lytic activity could be detected (data not
shown). In contrast, lytic activity of splenic and IEL OT-I memory
cells was detectable at 1:1 E:T cell ratios, and higher ratios resulted
in substantial lytic activity (Fig. 4A).
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B). We then asked whether secondary challenge
resulted in an increase in the cytotoxicity of memory CD8 T cells. As
shown in Fig. 4A, oral OVA administration induced a
substantial increase in the lytic activity of memory cells in MLN,
spleen, and IEL. In the absence of OT-I T cell transfer, minimal lytic
activity was detected in this or in the primary response at the E:T
ratios used, presumably due to the low precursor frequency of
endogenous OVA-specific T cells (data not shown). Thus,
VSV-OVA-reactive OT-I memory cells in intestinal mucosa and in the
periphery can be reactivated by oral administration of sOVA.
In addition to cytotoxic activity, we analyzed the production of
IFN- following reactivation of memory OT-I cells in vitro (Fig. 5). After short-term culture (5 h) of
naive or memory splenic or LP OT-I cells in the presence or
absence of the antigenic peptide SIINFEKL, intracellular IFN- levels
were measured by flow cytometry. Without the addition of peptide,
neither naive nor memory OT-I cells produced detectable IFN-. Memory
cells isolated from the spleen or from LP, but not naive splenic OT-I
cells, rapidly up-regulated production of IFN- after restimulation
with antigenic peptide. Although naive OT-I cells challenged with Ag
did not produce detectable intracellular IFN-, >95% of memory OT-I
cells contained significant levels of IFN-. Similar results were
obtained using memory cells derived from LN or IEL (data not shown).
These results indicated that mucosal and systemic memory CD8 T cells
were functionally similar with regard to rapid up-regulation of lytic
activity and IFN- production following Ag reencounter.
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We previously reported that naive LN OT-I
CD8+ T cells are heavily dependent on the
costimulatory molecule B7-2 for primary in vivo activation
(23). Although it has been implied that reactivation of
memory cells may have less stringent requirements for CD28-mediated
costimulation, little is known about the role of costimulation during
in vivo reactivation of CD8 memory T cells. To test this, concomitant
with feeding of OVA, mice containing OT-I memory cells were treated
with CTLA4-Ig to block B7-1 and B7-2 interactions with CD28, or as a
control a CTLA4-Ig mutant that does not bind B7-1 or B7-2. In the MLN,
OT-I memory cells were reactivated after Ag feeding and CTLA4-Ig mutant
treatment, as evidenced by a major increase in cell size and by
up-regulation of CD28, CD11c (Fig. 6),
and CD44 (data not shown). However, CTLA4-Ig treatment had little
effect on the increase in cell size, indicating that CD28-mediated
costimulation was not required for optimal reactivation (as measured by
blastogenesis) of MLN CD8 memory cells by soluble Ag. In addition, CD28
up-regulation was not affected and CD11c induction was only partially
inhibited by CTLA4-Ig treatment. This result suggested that the
up-regulation of CD28 was TCR mediated and costimulatory molecule
independent. Similar results were obtained from analysis of splenic
OT-I memory cells (data not shown). Furthermore, intestinal mucosal
memory CD8 cell reactivation was largely unaffected by CTLA4-Ig
treatment (Fig. 7). The reactivation of
IEL memory cells was unaffected, and reactivation of LP memory CD8 T
cells was only marginally inhibited by CTLA4-Ig treatment, as measured
by cell size increase. CD28 and CD11c up-regulation on LP and IEL OT-I
memory cells was not inhibited by CTLA4-Ig treatment. The CTLA4-Ig
preparations employed were active because they effectively inhibited
primary OT-I activation and expansion in IEL as well as all other
tissues examined after feeding OVA (Fig. 7, E and
F, and data not shown). To determine whether the
reactivation of lytic activity was affected by B7 blockade, we tested
CTL activity in control and CTLA4-Ig-treated mice that had been fed Ag
(Fig. 8). Although there was substantial
up-regulation of lytic activity of splenic CD8 memory cells after Ag
reencounter, no inhibition of this reactivation by CTLA4-Ig treatment
was evident (Fig. 8). Similar results were obtained using cells from LN
or IEL (data not shown). Thus, costimulation was not required for
reexpression of phenotypic and functional modifications after secondary
Ag encounter. These results indicated that the stringency of
costimulatory requirements for memory CD8 T cell reactivation was
distinct from that of naive CD8 T cells and enforced the concept of a
rapid recall response by memory CTL.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The issue of maintaining memory has been of great interest because it is critical to vaccine efficacy. The relative importance of intrinsic and extrinsic signals in memory T cell survival has not been fully delineated. Accumulating data suggest that memory T cells are not dormant, but in constant cycle (1, 2, 36), although the signals that induce memory T cell division are not entirely clear. Persistence of cognate or cross-reactive Ags, continuous engagement of the TCR via MHC, and nonspecific inflammatory cytokines have been proposed to be involved in the maintenance of CD8 memory (reviewed in Refs. 37, 38). Current concepts suggest that long-term memory does not require persistence of Ag, but that continuous interaction with MHC class I molecules is essential to CD8 memory maintenance (38). Because CD8 memory cells are generally not found in nonmucosal tertiary tissues, except perhaps liver, our results suggest that the intestinal mucosa may provide organ-specific factors that aid in maintaining CD8 memory. This effect could be related to the inflammatory nature of the gut and the ability of the mucosa to induce CTL even when soluble Ag is used as immunogen. As we have previously shown, compared with peripheral T cell activation, costimulation via B7-1 is more important for CD8 T cell activation in the intestinal mucosa (23), and this and other constitutive costimulatory signals may be important in maintaining memory.
The lack of a requirement for B7 in reactivation of CD8 memory cells may be explained at the level of the APC or the T cell. Our results indicated that secondary lymphoid memory cells could mount a rapid recall response in the face of B7-1,2 blockade, even though presumably professional APCs had processed and presented intact OVA. In the case of IEL memory cells, reactivation by oral administration of Ag implied, but did not prove, that intestinal epithelial cells can act as APC in vivo, at least for presentation to memory cells. This hypothesis was supported by the costimulation independence of IEL memory cells, because normal IEC do not express B7 molecules (39) and there is scant evidence to indicate that APC reside in the mouse epithelium. There is little available information on costimulatory requirements for CD8 memory T cell reactivation in vivo. Reactivation of alloreactive CTL does not require costimulation in vitro (40), but whether this holds true in vivo is not known. The form of the reactivating Ag may also be important for the long-term outcome of the recall response in that reactivation during microbial infections rather than via administration of purified protein may provide additional signaling. Nonetheless, our results open the possibility that boosting of a T cell memory response could be induced using oral Ag, which would be undesirable when attempting to induce oral tolerance. We are currently testing the long-term effects of oral Ag dosing on memory T cells.
Although it has been known for many years that the phenotype of LP and
IEL CD4 and CD8 populations was suggestive of prior Ag exposure
(13, 14, 15, 16, 17), the origin, life span, and Ag reactivity of such
memory cells have not been defined. This has been particularly true for
IEL, whose origin has been a matter of considerable debate (20, 21). In addition, the poor proliferative response and in some
cases unique costimulatory requirements of these cells have
distinguished them from more traditional activated or memory cells
present in the secondary lymphoid organs (34, 35, 41, 42, 43).
Our current and previous results indicate that a specific subset of
CD8ß TCRß IEL is derived from the peripheral T cell pool
following activation (19, 23, 44). Thus, CD11a high,
Eß7 low, CD62L
high/intermediate cells appear to be recent arrivals in the intestinal
mucosa. Our results would also suggest that a subset of recent
immigrants generates long-lived memory cells that remain within the
mucosa and express high levels of
Eß7 integrin. Studies
with parabiotic mice show that a small population of TCRß cells of
unknown origin accumulates in the intestinal epithelium over time
(45), and these may represent memory cells and/or recently
activated T cells. However, the discrepancies in adhesion molecule
expression between the majority of memory OT-I
CD8+ T cells in the secondary lymphoid tissues
and those in the intestinal mucosa suggested that these populations
were distinct and were not part of a common pool. Comparison of the
percentage of OT-I memory T cells in the LN or the spleen and the
intestine in single mice supports this idea. That is, in some cases,
highly disparate percentages of donor memory OT-I T cells were detected
in the periphery as compared with the LP and IEL compartments,
suggesting that these populations were separately maintained or had
discordant life spans. Detailed trafficking studies will be needed to
determine the precise relationship between peripheral and mucosal
memory T cells.
The constitutive lytic activity of CD8 memory cells in the mucosa, as
shown in this study, helps explain the constitutive lytic activity of
IEL and LP CD8 T cells. Our original description of direct ex vivo
lytic activity of TCRß and TCR IEL utilized a redirected
lysis assay that bypasses TCR specificity (16, 46). This
assay is necessary to identify CTL in normal mucosa because the Ag
specificity of these cells is unknown. The lytic activity of TCRß
IEL is primarily contained within the Thy-1+
CD8ß subset (34). Mucosal memory CTL generated in the
OT-I transfer system retain Thy-1 and CD8ß, suggesting that this
population in normal mice contains substantial numbers of memory CTL.
Induction of expansion of CD8ß IEL and up-regulation of lytic
activity are dependent on intestinal microbial flora (16, 47, 48). Thus, it appears likely that a subset of CD8ß IEL and
LP cells in healthy animals is comprised of bacterial Ag-specific
primary effector cells and memory cells. In the face of systemic or
mucosal infection or vaccination, Ag-specific effector cells would be
generated in MLN, Peyer’s patches (PP), or spleen, resulting in
migration to intestinal mucosa and generation of long-term memory. That
some IEL are derived from PP has been a longstanding theory
(49). Indeed, we observed activation of OT-I cells in PP
after immunization, but the population of activated OT-I cells was
always quantitatively larger in MLN than in PP (S.-K. Kim and L.
Lefrançois, unpublished results). It is perhaps likely that CD8
cells in both of these sites contribute to the activated T cell pool
following mucosal immunization. In any case, learning how to potentiate
mucosal CD8 effector and memory responses will provide tools for
improved vaccination against mucosal pathogens such as HIV.
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Leo Lefrançois, MC1310, Department of Medicine, UCONN Health Center, 263 Farmington Avenue, Farmington, CT 06030. E-mail address:
3 Abbreviations used in this paper: LCMV, lymphocytic choriomeningitis virus; IEL, intraepithelial lymphocyte; LP, lamina propria; MLN, mesenteric lymph node; PLN, peripheral lymph node; PP, Peyer’s patches; sOVA, soluble OVA; VSV, vesicular stomatitis virus.
Received for publication April 29, 1999. Accepted for publication July 28, 1999.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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+ intraepithelial lymphocytes. Science 243:1716.-T-cell receptor on intestinal CD8+ intraepithelial lymphocytes. Nature 333:855.[Medline]
IELß7 integrin on T cell receptor and T cell receptor and T cells. Eur. J. Immunol. 24:635.[Medline]
/ß T-cell receptor-positive intraepithelial lymphocytes and Peyer’s patch lymphocytes. Eur. J. Immunol. 23:1333.[Medline]
-T-cell antigen receptors. Proc. Natl. Acad. Sci. USA 90:8591.This article has been cited by other articles:
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|
  L. J. Ma, L. F. Acero, T. Zal, and K. S. Schluns Trans-Presentation of IL-15 by Intestinal Epithelial Cells Drives Development of CD8{alpha}{alpha} IELs J. Immunol., July 15, 2009; 183(2): 1044 - 1054. [Abstract] [Full Text] [PDF]
|
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P. K. Suvas, H. M. Dech, F. Sambira, J. Zeng, and T. M. Onami Systemic and Mucosal Infection Program Protective Memory CD8 T Cells in the Vaginal Mucosa J. Immunol., December 15, 2007; 179(12): 8122 - 8127. [Abstract] [Full Text] [PDF]
|
|
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A. B. Borowski, A. C. Boesteanu, Y. M. Mueller, C. Carafides, D. J. Topham, J. D. Altman, S. R. Jennings, and P. D. Katsikis Memory CD8+ T Cells Require CD28 Costimulation J. Immunol., November 15, 2007; 179(10): 6494 - 6503. [Abstract] [Full Text] [PDF]
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N. R. Roan and M. N. Starnbach Antigen-Specific CD8+ T Cells Respond to Chlamydia trachomatis in the Genital Mucosa J. Immunol., December 1, 2006; 177(11): 7974 - 7979. [Abstract] [Full Text] [PDF]
|
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C. Zaph, K. A. Rook, M. Goldschmidt, M. Mohrs, P. Scott, and D. Artis Persistence and Function of Central and Effector Memory CD4+ T Cells following Infection with a Gastrointestinal Helminth J. Immunol., July 1, 2006; 177(1): 511 - 518. [Abstract] [Full Text] [PDF]
|
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 |
|
 N. T. Joncker, J. Helft, A. Jacquet, V. Premel, and O. Lantz Intratumor CD4 T-cell accumulation requires stronger priming than for expansion and lymphokine secretion. Cancer Res., May 15, 2006; 66(10): 5443 - 5451. [Abstract] [Full Text] [PDF]
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H. Su, D. S. Chang, S. S. Gambhir, and J. Braun Monitoring the Antitumor Response of Naive and Memory CD8 T Cells in RAG1-/- Mice by Positron-Emission Tomography J. Immunol., April 1, 2006; 176(7): 4459 - 4467. [Abstract] [Full Text] [PDF]
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D. Masopust, V. Vezys, E. J. Wherry, D. L. Barber, and R. Ahmed Cutting Edge: Gut Microenvironment Promotes Differentiation of a Unique Memory CD8 T Cell Population J. Immunol., February 15, 2006; 176(4): 2079 - 2083. [Abstract] [Full Text] [PDF]
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 E. Ramsburg, J. Publicover, L. Buonocore, A. Poholek, M. Robek, A. Palin, and J. K. Rose A Vesicular Stomatitis Virus Recombinant Expressing Granulocyte-Macrophage Colony-Stimulating Factor Induces Enhanced T-Cell Responses and Is Highly Attenuated for Replication in Animals J. Virol., December 15, 2005; 79(24): 15043 - 15053. [Abstract] [Full Text] [PDF]
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M. S. Bharhani, J. S. Grewal, M. J. Pilgrim, C. Enocksen, R. Peppler, L. London, and S. D. London Reovirus Serotype 1/Strain Lang-Stimulated Activation of Antigen-Specific T Lymphocytes in Peyer's Patches and Distal Gut-Mucosal Sites: Activation Status and Cytotoxic Mechanisms J. Immunol., March 15, 2005; 174(6): 3580 - 3589. [Abstract] [Full Text] [PDF]
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J. W. Peacock, S. K. Nordone, S. S. Jackson, H.-X. Liao, N. L. Letvin, A. G. Yafal, L. Gritz, G. P. Mazzara, B. F. Haynes, and H. F. Staats Gender Differences in Human Immunodeficiency Virus Type 1-Specific CD8 Responses in the Reproductive Tract and Colon following Nasal Peptide Priming and Modified Vaccinia Virus Ankara Boosting J. Virol., December 1, 2004; 78(23): 13163 - 13172. [Abstract] [Full Text] [PDF]
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 A. C. Kirby, M. Sundquist, and M. J. Wick In Vivo Compartmentalization of Functionally Distinct, Rapidly Responsive Antigen-Specific T-Cell Populations in DNA-Immunized or Salmonella enterica Serovar Typhimurium-Infected Mice Infect. Immun., November 1, 2004; 72(11): 6390 - 6400. [Abstract] [Full Text] [PDF]
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Y. Lou, G. Wang, G. Lizee, G. J. Kim, S. E. Finkelstein, C. Feng, N. P. Restifo, and P. Hwu Dendritic Cells Strongly Boost the Antitumor Activity of Adoptively Transferred T Cells In vivo Cancer Res., September 15, 2004; 64(18): 6783 - 6790. [Abstract] [Full Text] [PDF]
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A. L. Marzo, V. Vezys, K. D. Klonowski, S.-J. Lee, G. Muralimohan, M. Moore, D. F. Tough, and L. Lefrancois Fully Functional Memory CD8 T Cells in the Absence of CD4 T Cells J. Immunol., July 15, 2004; 173(2): 969 - 975. [Abstract] [Full Text] [PDF]
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M. C. Gold, M. W. Munks, M. Wagner, C. W. McMahon, A. Kelly, D. G. Kavanagh, M. K. Slifka, U. H. Koszinowski, D. H. Raulet, and A. B. Hill Murine Cytomegalovirus Interference with Antigen Presentation Has Little Effect on the Size or the Effector Memory Phenotype of the CD8 T Cell Response J. Immunol., June 1, 2004; 172(11): 6944 - 6953. [Abstract] [Full Text] [PDF]
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D. Masopust, V. Vezys, E. J. Usherwood, L. S. Cauley, S. Olson, A. L. Marzo, R. L. Ward, D. L. Woodland, and L. Lefrancois Activated Primary and Memory CD8 T Cells Migrate to Nonlymphoid Tissues Regardless of Site of Activation or Tissue of Origin J. Immunol., April 15, 2004; 172(8): 4875 - 4882. [Abstract] [Full Text] [PDF]
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 K. S. Schluns, E. C. Nowak, A. Cabrera-Hernandez, L. Puddington, L. Lefrancois, and H. L. Aguila Distinct cell types control lymphoid subset development by means of IL-15 and IL-15 receptor {alpha} expression PNAS, April 13, 2004; 101(15): 5616 - 5621. [Abstract] [Full Text] [PDF]
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S.-K. Kim and R. M. Welsh Comprehensive Early and Lasting Loss of Memory CD8 T Cells and Functional Memory during Acute and Persistent Viral Infections J. Immunol., March 1, 2004; 172(5): 3139 - 3150. [Abstract] [Full Text] [PDF]
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 F. G. Lakkis and M. H. Sayegh Memory T Cells: A Hurdle to Immunologic Tolerance J. Am. Soc. Nephrol., September 1, 2003; 14(9): 2402 - 2410. [Full Text] [PDF]
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 H. Beacock-Sharp, A. M. Donachie, N. C. Robson, and A. M. Mowat A role for dendritic cells in the priming of antigen-specific CD4+ and CD8+ T lymphocytes by immune-stimulating complexes in vivo Int. Immunol., June 1, 2003; 15(6): 711 - 720. [Abstract] [Full Text] [PDF]
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H.-C. Wang, Q. Zhou, J. Dragoo, and J. R. Klein Most Murine CD8+ Intestinal Intraepithelial Lymphocytes Are Partially But Not Fully Activated T Cells J. Immunol., November 1, 2002; 169(9): 4717 - 4722. [Abstract] [Full Text] [PDF]
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M. P. Rubinstein, A. N. Kadima, M. L. Salem, C. L. Nguyen, W. E. Gillanders, and D. J. Cole Systemic Administration of IL-15 Augments the Antigen-Specific Primary CD8+ T Cell Response Following Vaccination with Peptide-Pulsed Dendritic Cells J. Immunol., November 1, 2002; 169(9): 4928 - 4935. [Abstract] [Full Text] [PDF]
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W. N. D'Souza, K. S. Schluns, D. Masopust, and L. Lefrancois Essential Role for IL-2 in the Regulation of Antiviral Extralymphoid CD8 T Cell Responses J. Immunol., June 1, 2002; 168(11): 5566 - 5572. [Abstract] [Full Text] [PDF]
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K. Haglund, I. Leiner, K. Kerksiek, L. Buonocore, E. Pamer, and J. K. Rose High-Level Primary CD8+ T-Cell Response to Human Immunodeficiency Virus Type 1 Gag and Env Generated by Vaccination with Recombinant Vesicular Stomatitis Viruses J. Virol., February 22, 2002; 76(6): 2730 - 2738. [Abstract] [Full Text] [PDF]
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W. Z. Mehal, F. Azzaroli, and I. N. Crispe Antigen Presentation by Liver Cells Controls Intrahepatic T Cell Trapping, Whereas Bone Marrow-Derived Cells Preferentially Promote Intrahepatic T Cell Apoptosis J. Immunol., July 15, 2001; 167(2): 667 - 673. [Abstract] [Full Text] [PDF]
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E. Y. Choi, Y. Yoshimura, G. J. Christianson, T. J. Sproule, S. Malarkannan, N. Shastri, S. Joyce, and D. C. Roopenian Quantitative Analysis of the Immune Response to Mouse Non-MHC Transplantation Antigens In Vivo: The H60 Histocompatibility Antigen Dominates Over All Others J. Immunol., April 1, 2001; 166(7): 4370 - 4379. [Abstract] [Full Text] [PDF]
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J. W. Huleatt, I. Pilip, K. Kerksiek, and E. G. Pamer Intestinal and Splenic T Cell Responses to Enteric Listeria monocytogenes Infection: Distinct Repertoires of Responding CD8 T Lymphocytes J. Immunol., March 15, 2001; 166(6): 4065 - 4073. [Abstract] [Full Text] [PDF]
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D. Masopust, J. Jiang, H. Shen, and L. Lefrancois Direct Analysis of the Dynamics of the Intestinal Mucosa CD8 T Cell Response to Systemic Virus Infection J. Immunol., February 15, 2001; 166(4): 2348 - 2356. [Abstract] [Full Text] [PDF]
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R. J. Hogan, E. J. Usherwood, W. Zhong, A. D. Roberts, R. W. Dutton, A. G. Harmsen, and D. L. Woodland Activated Antigen-Specific CD8+ T Cells Persist in the Lungs Following Recovery from Respiratory Virus Infections J. Immunol., February 1, 2001; 166(3): 1813 - 1822. [Abstract] [Full Text] [PDF]
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 D. Masopust, V. Vezys, A. L. Marzo, and L. Lefrançois Preferential Localization of Effector Memory Cells in Nonlymphoid Tissue Science, March 23, 2001; 291(5512): 2413 - 2417. [Abstract] [Full Text]
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, lytic activity
of memory cells 2 days after sOVA feeding. Lytic activity at the E:T
used in the absence of transferred OT-I cells was <5%. Lytic activity
against nonpeptide-coated targets was <5%. B, The size
of memory cells before and after Ag feeding was assessed by analysis of
forward light scatter properties in conjunction with Ly-5.2 staining.
This experiment has been performed four times with comparable
results.
, Ag fed treated with CTLA4-Ig mutant; •, no treatment. Lytic
activity against nonpeptide-coated targets was <5%. This experiment
has been performed three times with similar results.