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Department of Microbiology and Center for Immunology, University of Minnesota, Minneapolis, MN 55455
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The capacity of T cells to mediate immunological memory is associated
with several changes in function (2). Naive T cells
express high levels of molecules that facilitate migration from blood
into lymph nodes, for example CD62 ligand (CD62L), and low levels of
molecules that facilitate migration into nonlymphoid tissues, for
example CD44. In contrast, Ag-experienced T cells express low levels of
CD62L and high levels of CD44, at least at early times after initial
activation. These patterns suggest that naive T cells circulate through
the secondary lymphoid organs, whereas Ag-experienced T cells can
circulate through nonlymphoid tissues, allowing the latter population
to respond rapidly at the site of Ag entry. Naive and Ag-experienced T
cells also differ with respect to lymphokine production. Naive cells
produce primarily IL-2, which is thought to facilitate the clonal
expansion of initially rare T cells that express the appropriate TCRs,
whereas Ag-experienced cells synthesize less IL-2 and produce effector
lymphokines such as IL-4 and IFN-
, which control the microbicidal
activities of other cells (3).
The means by which these functional changes are maintained in Ag-experienced T cells are poorly understood. In vivo activation of viral Ag-specific CD8 T cells produces cells with the function and cell surface phenotype of Ag-experienced cells that persist for long time periods in the absence of detectable Ag (4, 5, 6). This suggests that immunological memory is maintained because of an irreversible differentiation step in Ag-experienced T cells. On the other hand, some work with CD4 T cells suggests that the functional (7) and phenotypic (8) changes displayed by Ag-experienced cells at early times after exposure to Ag are only maintained as long as Ag persists in the host. Thus, the changes ascribed to Ag-experienced CD4 T cells may be reversible and may not even be intrinsic properties of the T cells themselves; rather, they may be imposed by extrinsic factors present in the environment.
We addressed the roles of intrinsic changes and environmental factors in the behavior of Ag-experienced cells using an adoptive transfer model in which CD4 T cells of known specificity from TCR transgenic mice were tracked in immune or naive recipients. Our results suggest that the clonal expansion potential of Ag-experienced CD4 T cells is influenced by extrinsic factors present in the immune environment.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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DO11.10 (9) TCR transgenic mice were bred in a specific pathogen-free facility under National Institutes of Health guidelines and screened for the TCR transgenes as previously described (10). These mice have been bred onto the BALB/c background for >15 generations and are histocompatible with normal BALB/c mice. BALB/c mice were purchased from the National Cancer Institute (Frederick, MD) and housed in a conventional facility. DO11.10 SCID donors were obtained by crossing DO11.10 BALB/c mice for two generations with SCID BALB/c mice (purchased from The Jackson Laboratory, Bar Harbor, ME) and selecting offspring that expressed the DO11.10 TCR but lacked B cells in the peripheral blood. B cell-deficient (µMT) mice (11), provided by Dr. Steven L. Reiner (University of Chicago, Chicago, IL), were bred to BALB/c mice for five generations.
Adoptive transfer and immunization
The behavior of naive T cells was studied by adoptive transfer (via i.v. injection) of lymph node and spleen cells from naive wild-type or SCID DO11.10 donors, containing 2.5 x 106 CD4+, KJ1-26+ cells, into unirradiated normal BALB/c mice (10). In cases where B cell-deficient mice were used as recipients, lymph node and spleen cells from the DO11.10 donors were exhaustively depleted of B cells using an anti-B220 mAb (RA3-3A1/6.1, American Type Culture Collection, Manassas, VA) and rabbit complement (Accurate Chemical and Scientific, Westbury, CT). The elimination of B cells from the cell preparation was confirmed before transfer by staining with FITC-labeled goat anti-mouse Ig (Caltag, South San Francisco, CA). Recipients were immunized 1 to several days after transfer by injection of 100 µg of chicken OVA (Sigma, St. Louis, MO) or 300 µg of OVA peptide 323–339 emulsified in CFA or IFA into two s.c. sites on the back or by i.v. injection of 300 µg of OVA peptide 323–339 plus 150 µg of LPS (serotype Escherichia coli 026:B6; Difco, Detroit, MI). In some experiments bromodeoxyuridine was offered to mice in the drinking water (0.8 mg/ml) ad libitum, beginning on the day of immunization. In other experiments recipients of naive DO11.10 T cells were passively immunized by i.p. injection every day for 4 days with 1.2 mg of purified IgG from OVA/CFA-primed or naive mice, ending on the day before immunization with OVA/IFA. The titers of anti-OVA IgG1 and IgG2a Abs in the passively immunized animals were similar to those of DO11.10 recipients immunized 3 wk previously with OVA/CFA. In some cases animals received a second set of s.c. injections of 100 µg of OVA or 300 µg of OVA peptide 323–339 in CFA or IFA at several different sites on the back, 9–50 days after the first injection.
A serial adoptive transfer protocol was used for study of Ag-experienced DO11.10 T cells in a naive environment. Naive DO11.10 T cells (5–7.5 x 106/mouse) were injected i.v. into naive BALB/c recipients, which were then immunized with 100 µg of chicken OVA in CFA as described above. After 9–12 days, draining lymph node cells (cervical, brachial, axillary, and inguinal) were recovered, and the CD4 T cells were enriched by depletion of CD8 T cells with anti-CD8 mAb (clone 83.12.5, American Type Culture Collection) and rabbit complement (Accurate Chemical and Scientific) and of B cells and adherent cells using a Cellect T cell column (Biotex, Edmonton, Canada). The enriched CD4 T cells, which contained 3–5% Ag-experienced DO11.10 T cells, were then transferred to naive BALB/c recipients such that each recipient received 1.25–2.5 x 106 DO11.10 T cells. Recipient mice were then injected with OVA in CFA as described above.
In some cases lymph node cells from naive DO11.10 mice were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE;3 Molecular Probes, Eugene, OR) before transfer, using a modification of a previously described technique (12). Briefly, cells were washed once in HBSS (Biofluids, Rockville, MD), adjusted to a final concentration of 5 x 107 cells/ml in HBSS, and incubated with 5 µM CFSE for 10 min in a 37°C water bath. An equal volume of cold HBSS containing 10% FCS was added to stop the labeling, and the cells were washed before adoptive transfer. The number of cell divisions that the CFSE-labeled DO11.10 T cells experienced after Ag activation in vivo was determined as previously described (13), based on the assumption that the CFSE signal is reduced by half at each cell division.
Detection of TCR transgenic T cells
DO11.10 T cells were identified in adoptive recipients by two-color flow cytometry as previously described (10). The analysis was performed on lymph nodes cells or enriched CD4 T cells that were prepared from lymph nodes as described above. Before addition of labeled mAb, cells were incubated on ice for 10 min with anti-FcR mAb in 10% rat and mouse serum to block FcRs. When unlabeled DO11.10 T cells were transferred, cells were stained with PE-labeled anti-CD4 mAb (PharMingen, San Diego, CA), biotinylated KJ1-26 mAb (which recognizes the DO11.10 TCR) (14), and FITC-labeled streptavidin (SA; Caltag). When CFSE-labeled DO11.10 T cells were transferred, cells were stained with CyChrome-labeled anti-CD4 mAb (PharMingen), biotinylated KJ1-26 mAb, and PE-labeled SA (Caltag). Thirty-thousand events were collected on a FACScan flow cytometer (Becton Dickinson, Mountain View, CA) and analyzed using CellQuest software. DO11.10 T cells were identified as CD4+, KJ1-26+ events. The number of DO11.10 T cells present in the draining lymph nodes was determined by multiplying the total number of viable cells recovered by the percentage of CD4+, KJ1-26+ events obtained by flow cytometry.
The size of the DO11.10 T cells was estimated from the forward light scatter values of 1000–5000 CD4+, KJ1-26+ events.
The cell surface phenotype of DO11.10 T cells was determined by staining lymph node cells with allophycocyanin-labeled anti-CD4 mAb, biotin-labeled KJ1-26, peridinin chlorophyl protein (PerCP)-labeled SA (Becton Dickinson, San Jose, CA), and PE-labeled anti-CD45RB or anti-LFA-1 mAbs; with digoxygenin-labeled KJ1-26, sheep anti-digoxygenin Ab, PE-labeled donkey anti-sheep Ab, CyChrome-labeled anti-CD4 mAb, biotin-labeled anti-CD45RB mAb, and allophycocyanin-labeled SA; or with biotin-labeled KJ1-26, CyChrome-labeled anti-CD4 mAb, PerCP-labeled SA, and FITC-labeled anti-CD45RB mAb (all mAb except KJ1-26 were from PharMingen). Five hundred to 1000 CD4+, KJ1-26+ events were collected using a FACScaliber flow cytometer. The fluorescence intensity related to CD45RB or LFA-1 expression was measured in the appropriate channel.
Intracellular IL-2 was detected by flow cytometry as initially described by Openshaw et al. (15) with the modifications reported by Khoruts et al. (16). Naive recipients of DO11.10 T cells or recipients of DO11.10 T cells that were immunized with OVA plus LPS 17 days previously were injected i.v. with 300 µg of OVA peptide to stimulate IL-2 production in vivo. Spleen cells were stained with CyChrome-labeled anti-CD4 mAb and biotinylated KJ1-26, followed by FITC-labeled SA. The cells were then washed in PBS and fixed for 20 min at room temperature in PBS containing 2% formaldehyde, permeabilized with two washes in staining buffer containing 0.5% saponin (Sigma), and then incubated for 30 min at room temperature with PE-labeled anti-IL-2 mAb or a similarly labeled control mAb of the same isotype (PharMingen). The cells were then washed twice with saponin buffer and twice with PBS. The PE channel fluorescence of at least 1000 CD4+, KJ1-26+ events was measured on a FACScan flow cytometer.
Bromodeoxyuridine was detected in DO11.10 cells by flow cytometry as described by Pape et al. (17).
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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OVA peptide I-Ad-specific CD4 T cells from the DO11.10 TCR transgenic mouse line were transferred into normal recipients, which were then immunized with OVA to produce a uniform population of Ag-experienced DO11.10 T cells. Although most CD4 T cells from the normal DO11.10 strain express the DO11.10 TCR and a naive phenotype (CD45RBhigh), these mice contain a small population of CD4 T cells that express the DO11.10 TCR and a memory/activated phenotype (CD45RBlow). These T cells express an endogenous TCR and presumably became Ag-experienced cells via recognition of an environmental Ag (18). To eliminate any contribution of these cells, DO11.10 BALB/c SCID donors were used for the initial experiments because they contain only naive CD4 T cells that express the DO11.10 TCR.
Lymph node and spleen cells from DO11.10 BALB/c SCID donors were
labeled with the cytoplasmic fluorochrome CFSE before transfer, so that
their cell division history could be monitored in the recipient.
Recipient mice were immunized via s.c. injection of OVA in CFA. Within
1 day of transfer, KJ1-26+ DO11.10 T cells were
identified in unimmunized recipients at low, but consistent, levels in
the secondary lymphoid organs of recipient mice (Fig. 1
A). These T cells uniformly
expressed the high level of CD45RB characteristic of naive T cells
(Fig. 1B), lacked the CD45RBlow
population of Ag-experienced CD4 T cells found in normal mice (Fig. 1B), and retained large amounts of CFSE, indicating that
they had not entered the cell cycle (Fig. 1D). In contrast,
most of the DO11.10 T cells that were present in the draining lymph
nodes 3 days after injection of OVA/CFA were blasts (Fig. 1G) and contained low amounts of CFSE (Fig. 1, D
and F) as would be expected if these cells had divided many
times. This possibility was supported by the finding that the number of
DO11.10 T cells present in the lymph nodes that drained the OVA/CFA
injection site increased dramatically, such that 20-fold more cells
were present on day 5 after Ag injection than were present before
immunization (Fig. 1E). The number of DO11.10 T cells in the
draining lymph nodes then fell dramatically by day 7 and slowly
thereafter to a level on day 25 that was about twice the starting
level. The majority of DO11.10 T cells that remained in the lymph nodes
21–25 days after immunization were no longer blasts (Fig. 1G), showed evidence of at least four cell divisions (Fig. 1F), and expressed the lower levels of CD45RB (Fig. 1C) characteristic of Ag-experienced T cells. Identical
results were obtained when normal DO11.10 mice were used as donors
(data not shown). Thus, the small population of pre-existing
Ag-experienced cells present in normal DO11.10 donors had no detectable
effect on the clonal expansion and contraction of naive DO11.10 T cells
or on the production of OVA-experienced DO11.10 T cells during the
primary response to OVA. For this reason, normal DO11.10 donors were
used for subsequent experiments. Similar results were obtained when
OVA/LPS or OVA/IFA were used as the s.c. immunogens instead of OVA/CFA
(10, 19).
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The clonal expansion of Ag-experienced DO11.10 T cells during a
secondary response was tested by challenging recipient mice with OVA at
various times after primary immunization. In the experiment shown in
Fig. 2A, recipients were
immunized with OVA peptide/CFA, and as expected, the initially naive
DO11.10 T cells increased 30-fold in the draining lymph nodes on day 5
and then fell to a lower level on day 12 that was still significantly
greater than the starting level. When these recipients were injected
with OVA again on day 12, the now Ag-experienced DO11.10 T cells
increased in the draining lymph nodes 3 days later to a maximal level
that was only 3-fold higher than the number present on the day of
secondary challenge. Large numbers of DO11.10 T cells were not present
in the thoracic duct lymph (assessed by cannulation of the thoracic
duct as described in Ref. 20), at the s.c. challenge site,
or in the spleen at any time after secondary challenge (R. Merica,
unpublished observation), indicating that rapid migration out of the
lymph nodes was not responsible for the poor accumulation of
Ag-experienced DO11.10 T cells. On the contrary, poor proliferation was
involved because DO11.10 T cells in mice that received primary
immunization 2 wk earlier, incorporated less bromodeoxyuridine into DNA
after secondary challenge than naive DO11.10 T cells did after primary
exposure to OVA (Fig. 2B). Therefore, although the
Ag-experienced DO11.10 T cells accumulated maximally in the lymph nodes
more rapidly after Ag challenge than naive DO11.10 T cells, the
magnitude of their proliferation was much smaller.
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. The fold increase in DO11.10 T cells
present in the draining lymph nodes after OVA injection is shown to
correct for the different numbers of DO11.10 T cells present at the
time of primary (for naive cells) or secondary (for Ag-experienced
cells) immunization. Again, naive DO11.10 T cells underwent a 20-fold
expansion in the draining lymph nodes that peaked 5 days after Ag
injection. In contrast, DO11.10 T cells that experienced primary
immunization 2 wk before secondary challenge increased only 5-fold,
with maximal accumulation occurring on day 3. Ag-experienced DO11.10
cells accumulated to a greater extent after secondary challenge if 10
wk passed between the time of the first and second injections of OVA,
although again the peak of clonal expansion was lower than that
achieved by naive cells and occurred on day 3 instead of day 5 (Fig. 3A). Notably, DO11.10 T cells that experienced Ag 2 wk
earlier possessed the memory/activated phenotype
(CD45RBlow, LFA-1high),
whereas the DO11.10 T cells that experienced Ag 10 wk earlier could not
be distinguished from naive T cells based on surface phenotype (Fig. 3B). Therefore, the improvement in clonal expansion
potential by Ag-experienced DO11.10 T cells between 2 and 10 wk
correlated with reversion to a naive cell surface phenotype.
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A). In addition,
Ag-experienced DO11.10 T cells produced IL-2 more rapidly in response
to in vivo Ag challenge than did naive DO11.10 T cells. As shown in
Fig. 4B, naive DO11.10 T cells produced IL-2 in the lymph
nodes within 3 h of i.v. injection of OVA peptide, with maximal
production occurring at 6 h. Approximately 25% of the DO11.10 T
cells contained intracellular IL-2 at the peak of production. Naive
DO11.10 T cells stopped producing IL-2 by 12 h after OVA peptide
injection. In contrast, DO11.10 T cells in mice immunized 17 days
earlier achieved peak production of IL-2 by 1.5 h after i.v.
challenge with OVA peptide, and production declined rapidly thereafter.
Therefore, although Ag-experienced DO11.10 T cells displayed a reduced
capacity for clonal expansion, these cells were very sensitive to Ag
stimulation and were rapid lymphokine producers in vivo.
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It was possible that a factor(s) present in the immune environment
was responsible for the reduced clonal expansion potential of
Ag-experienced DO11.10 T cells at early times after primary
immunization. If this supposition was correct, then Ag-experienced T
cells would have been expected to recover the capacity for robust
clonal expansion after transfer into naive recipients. This was tested
using a serial adoptive transfer method. Naive DO11.10 T cells were
transferred into naive recipients, which were immunized with OVA/CFA.
Several weeks later, purified CD4 T cells (containing
106 Ag-experienced DO11.10 T cells) from these
mice were transferred into a new set of naive recipients that had never
been exposed to OVA. One week after cell transfer, the recipient mice
were challenged with OVA. As shown in Fig. 5A, the Ag-experienced DO11.10
cells that resided in a naive environment for 1 wk displayed the robust
clonal expansion pattern of naive DO11.10 T cells. The recovery of
clonal expansion potential in a naive environment was accompanied by
reversion of the Ag-experienced DO11.10 cells to a naive cell surface
phenotype (Fig. 5B). However, the Ag-experienced DO11.10
cells that resided for 1 wk in naive hosts did not behave like naive T
cells with respect to IL-2 production. Like Ag-experienced DO11.10 T
cells that remained in immune hosts, the Ag-experienced DO11.10 T cells
that resided in naive hosts produced IL-2 more quickly than naive cells
during the first 1.5 h after in vivo challenge with OVA peptide
(Fig. 5C). In addition, a larger fraction of the
Ag-experienced DO11.10 T cells that resided in naive hosts produced
IL-2 than did Ag-experienced DO11.10 T cells that remained in the
immune environment. Therefore, Ag-experienced CD4 T cells that resided
in a naive environment for 1 wk reverted to the surface phenotype and
robust clonal expansion behavior of naive T cells, but retained the
capacity to produce IL-2 rapidly.
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C) like Ag-experienced cells
that remained in immune hosts (Fig. 6B), not less than one
cell division like naive cells (Fig. 6A). Thus, although the
DO11.10 T cells from immunized mice that were parked in naive hosts
possessed a naive surface phenotype and clonal expansion potential,
these cells clearly experienced Ag in the past based on their cell
division history.
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Ag-specific Ab could have been the factor in immune hosts that
limited the clonal expansion potential of Ag-experienced T cells by
rapidly clearing the OVA after challenge injection (21).
However, DO11.10 T cells that were immunized with OVA in normal or B
cell-deficient hosts underwent limited clonal expansion compared with
naive DO11.10 cells, 3 (data not shown) or 5 days after secondary
challenge with OVA (Fig. 7A). Thus, Ag-specific Abs cannot
account for the factor in immune mice that limits the clonal expansion
potential of Ag-experienced T cells. This conclusion was supported by
passive immunization experiments. Naive DO11.10 T cells were
transferred into recipient mice that had been injected with
anti-OVA Abs and were then immunized with OVA/CFA. As shown in Fig. 7B, the clonal expansion of
naive DO11.10 T cells was not inhibited in recipients that had been
passively immunized with anti-OVA Abs. On the contrary, the
OVA-specific Abs increased the tempo of the expansion of the DO11.10 T
cells, such that a maximal level was reached on day 3 instead of
day 5.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The poor accumulation of Ag-experienced T cells in the draining lymph nodes at early times after immunization could not be explained by rapid migration of these cells from the lymph nodes to the spleen or site of secondary immunization. Ag clearance by Ag-specific Ab was also ruled out as a cause of the reduced response, because Ag-experienced CD4 T cells accumulated poorly during a secondary response in B cell-deficient, and thus Ab-deficient, recipients. Because the Ag-experienced CD4 T cells displayed limited DNA synthesis in the lymph nodes in response to secondary immunization, it is more likely that poor proliferation was responsible for their reduced accumulation. This was not easily explained by poor production of T cell growth factors, because Ag-experienced CD4 T cells produced IL-2 quickly and in nearly the same quantity as naive T cells. It is therefore more likely that the blunted clonal expansion is related to a block in proliferation in response to T cell growth factors. Th1 clones that are exposed to high concentrations of Ag produce IL-2 but do not proliferate despite expression of the IL-2R (27). Therefore, it is conceivable that Ag-experienced CD4 T cells are chronically exposed to residual Ag derived from the primary immunization, and that this situation promotes rapid T cell growth factor production, but prevents the T cells from proliferating maximally in response to the growth factor. The finding that Ag-experienced T cells undergo robust clonal expansion when challenged in naive hosts or in immune hosts long after primary immunization supports the possibility that exposure to residual Ag is the factor that limits the proliferation of Ag-experienced CD4 T cells.
The increased potential for clonal expansion by Ag-experienced CD4 T
cells long after primary immunization or after transfer into naive
recipients coincided with reversion to a naive cell surface phenotype.
Reversion of Ag-experienced CD4 T cells to a naive surface phenotype
has been observed in other studies (28, 29, 30). Garcia et al.
(31) recently reported that Ag-experienced CD4 T cells
retained the capacity to produce IFN- long after APC capable of
presenting residual Ag disappeared. This result together with our
finding that Ag-experienced DO11.10 T cells retained the capacity for
rapid IL-2 production after transfer into the naive environment
demonstrates that not all functions of Ag-experienced CD4 T cells
revert to the naive state in the naive environment.
The Ag-experienced CD4 T cells that regained the capacity for robust clonal expansion 10 wk after primary immunization still displayed a peak response on day 3 instead of day 5 like truly naive T cells. This response pattern was similar to that displayed by naive T cells that were exposed to Ag in passively immunized recipients. Thus, 10 wk after immunization, Ag-experienced T cells may be inherently capable of the slower clonal expansion rate of naive T cells, but may respond more quickly than naive T cells because Ag-specific Abs bind the challenge Ag and efficiently direct it to APC. Ab-mediated facilitation of Ag presentation may also contribute to the capacity of Ag-experienced cells to respond to lower Ag doses than naive T cells. Ag-Ab complexes have been reported to enhance Ag presentation by binding to Fc or complement receptors (21, 32, 33, 34) on APC.
Our results are consistent with the following model of Ag-experienced T cell function. Naive CD4 T cells are stimulated by peptide-MHC-bearing APC in the lymphoid tissue to proliferate and help B cells produce Abs. Many of the activated T cells then die, leaving behind a population of Ag-experienced T cells. These cells are chronically or periodically stimulated by APC bearing small numbers of peptide-MHC molecules derived from the immunogen. This chronic signaling causes the Ag-experienced T cells to express the memory/activated phenotype and exist in a state characterized by rapid lymphokine production potential but poor proliferative capacity. If the host is challenged with Ag during this period, the Ag is rapidly bound by Ab and targeted to complement or Fc receptor-bearing APC that activate the Ag-experienced T cells to produce IL-2 rapidly. The combined effects of Ab-facilitated Ag presentation and rapid lymphokine production lead to rapid clonal expansion of the Ag-experienced T cells. However, the poor capacity of these cells to proliferate in response to growth factors limits the magnitude of the expansion. As residual peptide-MHC molecules derived from the primary immunization disappear, the Ag-experienced T cells revert to the naive surface phenotype and slower response status of naive T cells. If the host is challenged with Ag during this period, Ag-specific Ab, which remains elevated in immune hosts for life, will direct the Ag to APC efficiently and enhance the activation of the Ag-experienced T cells that retain rapid lymphokine production potential but whose proliferation is not longer suppressed by residual Ag. If this scenario is correct, then, as proposed by Bell and colleagues (35), CD4 T cell-mediated protection after the primary immunogen has disappeared may be due to the concerted action of Ag-specific Ab and an expanded population of Ag-experienced T cells that share some functional properties with naive cells.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Marc K. Jenkins, Department of Microbiology and Center for Immunology, University of Minnesota, Box 334 Mayo, 420 Delaware Street S.E., Minneapolis, MN 55455.
3 Abbreviations used in this paper: CFSE, carboxyfluorescein diacetate succinimidyl ester; PerCP, peridinin chlorophyl protein; SA, streptavidin: CD62L, CD62 ligand.
Received for publication November 16, 1999. Accepted for publication February 22, 2000.
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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K. Attanavanich and J. F. Kearney Marginal Zone, but Not Follicular B Cells, Are Potent Activators of Naive CD4 T Cells J. Immunol., January 15, 2004; 172(2): 803 - 811. [Abstract] [Full Text] [PDF]
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K. A. Pape, V. Kouskoff, D. Nemazee, H. L. Tang, J. G. Cyster, L. E. Tze, K. L. Hippen, T. W. Behrens, and M. K. Jenkins Visualization of the Genesis and Fate of Isotype-switched B Cells during a Primary Immune Response J. Exp. Med., June 16, 2003; 197(12): 1677 - 1687. [Abstract] [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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C. T. Moses, K. M. Thorstenson, S. C. Jameson, and A. Khoruts Competition for self ligands restrains homeostatic proliferation of naive CD4 T cells PNAS, February 4, 2003; 100(3): 1185 - 1190. [Abstract] [Full Text] [PDF]
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E. Ingulli, D. R. Ulman, M. M. Lucido, and M. K. Jenkins In Situ Analysis Reveals Physical Interactions Between CD11b+ Dendritic Cells and Antigen-Specific CD4 T Cells After Subcutaneous Injection of Antigen J. Immunol., September 1, 2002; 169(5): 2247 - 2252. [Abstract] [Full Text] [PDF]
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K. M. Smith, F. McAskill, and P. Garside Orally Tolerized T Cells Are Only Able to Enter B Cell Follicles Following Challenge with Antigen in Adjuvant, but They Remain Unable to Provide B Cell Help J. Immunol., May 1, 2002; 168(9): 4318 - 4325. [Abstract] [Full Text] [PDF]
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 M. Sester, U. Sester, B. Gartner, B. Kubuschok, M. Girndt, A. Meyerhans, and H. Kohler Sustained High Frequencies of Specific CD4 T Cells Restricted to a Single Persistent Virus J. Virol., March 19, 2002; 76(8): 3748 - 3755. [Abstract] [Full Text] [PDF]
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K. E. Foulds, L. A. Zenewicz, D. J. Shedlock, J. Jiang, A. E. Troy, and H. Shen Cutting Edge: CD4 and CD8 T Cells Are Intrinsically Different in Their Proliferative Responses J. Immunol., February 15, 2002; 168(4): 1528 - 1532. [Abstract] [Full Text] [PDF]
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N. N. Iwakoshi, T. G. Markees, N. Turgeon, T. Thornley, A. Cuthbert, J. Leif, N. E. Phillips, J. P. Mordes, D. L. Greiner, and A. A. Rossini4 Skin Allograft Maintenance in a New Synchimeric Model System of Tolerance J. Immunol., December 1, 2001; 167(11): 6623 - 6630. [Abstract] [Full Text] [PDF]
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K. M. Thorstenson and A. Khoruts Generation of Anergic and Potentially Immunoregulatory CD25+CD4 T Cells In Vivo After Induction of Peripheral Tolerance with Intravenous or Oral Antigen J. Immunol., July 1, 2001; 167(1): 188 - 195. [Abstract] [Full Text] [PDF]
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H. N. Shi, H. Y. Liu, and C. Nagler-Anderson Enteric Infection Acts as an Adjuvant for the Response to a Model Food Antigen J. Immunol., December 1, 2000; 165(11): 6174 - 6182. [Abstract] [Full Text] [PDF]
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T. Zell, A. Khoruts, E. Ingulli, J. L. Bonnevier, D. L. Mueller, and M. K. Jenkins Single-cell analysis of signal transduction in CD4 T cells stimulated by antigen in vivo PNAS, September 11, 2001; 98(19): 10805 - 10810. [Abstract] [Full Text] [PDF]
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) or low levels of LFA-1
(
) in each group at the time of OVA/IFA injection.
) or CFA alone (
). A, The
mean fold expansion values over a prechallenge baseline ± SEM
(n = 3–16 for each group and at each time point)
of DO11.10 T cells from each group, present in the draining lymph nodes
at the indicated times after s.c. challenge with OVA/CFA.
B, The percentages of DO11.10 T cells expressing high
levels of CD45RB (
