| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
*
Division of Pulmonary and Critical Care Medicine, Oregon Health Sciences, University/Portland Veterans Affairs Medical Center, Portland, OR 97201; and
Corixa Corporation, Seattle, WA 98104
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
enzyme-linked
immunospot assay was used to determine the frequency of Mtb-reactive
CD8+ T cells directly from PBMC. The effector cell
frequency among five healthy purified protein derivative-positive
subjects was 1/7,600 ± 4,300 compared with 1/16,000 ± 7,000
in six purified protein derivative-negative controls. To determine the
frequencies of classically, CD1-, and nonclassically restricted cells,
a limiting dilution analysis was performed. In one purified protein
derivative-positive subject, 192 clones were generated using
Mtb-infected dendritic cells (DC). Clones were assessed for reactivity
against control autologous DC, Mtb-infected autologous DC, and
HLA-mismatched CD1+ targets (DC), as well as HLA-mismatched
CD1- targets (macrophages). Of the 96 Mtb-reactive
CD8+ T cell clones, four (4%) were classically restricted
and 92 (96%) were nonclassically restricted. CD1-restricted cells were
not detected. Of the classically restricted cells, two were HLA-B44
restricted and one was HLA-B14 restricted. These results suggest that
while classically restricted CD8+ lymphocytes can be
detected, they comprise a relatively small component of the overall
CD8+ T cell response to Mtb. Further definition of the
nonclassical response may aid development of an effective vaccine
against tuberculosis. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
In tuberculosis, there is abundant evidence to support an important role for CD4+ T cell-mediated immunity (2, 3). However, several lines of evidence support a role for CD8+ CTL as well. MHC class I-deficient mice, and thus CD8+ T cell-deficient mice, in which the gene for ß2-microglobulin has been disrupted, are more susceptible to Mtb infection than their wild-type littermates (4), as are mice deficient in TAP (5). Silva et al. found that CD8+ CTL clones generated to the Mtb heat shock protein (hsp65) could confer partial immunity to Mtb infection in mice (6). Immunization of mice with plasmids expressing Mtb Ags such as hsp65 (7), Ag 85a (8), or the 38-kDa (9) Ag have resulted in protection from subsequent challenge with Mtb and have been associated with the generation of Ag-specific CD8+ CTL. In addition, Stenger et al. have demonstrated that human CD1b-restricted CD8+ CTL are able to inhibit the growth of Mtb in vitro (10). Finally, CD8+ T cells have been shown to localize preferentially to the mouse lung following infection with Mtb (11, 12).
In the host response to tuberculosis infection, the role of
CD8+ T cells may be in the recognition and
destruction of heavily infected macrophages or in the elimination of
infected MHC class II-negative cells such as endothelial cells and
fibroblasts. CD8+ CTL may play a protective role
through several mechanisms. First, these cells produce potent
anti-bacterial cytokines such as IFN- and TNF-
in response to
antigenic stimulation. The importance of these cytokines has been
illustrated by the susceptibility of mice to Mtb challenge in which the
genes for IFN- (13) and TNF-R have been disrupted
(14). Additionally, CD8+ CTL may
play a unique role in host defense to Mtb either by virtue of
preferential presentation of HLA-I-associated Ags in heavily infected
cells or through the enhanced release of granular constituents. Because
it has been suggested that apoptosis selectively inhibits mycobacterial
growth (15, 16), CTL may play an important role by
inducing apoptosis in infected cells. However, mice deficient in the
expression of perforin, granzyme, or CD95 (Fas) are still able to
contain infection with Mtb (17, 18). These data have been
used to argue that in the mouse model, CD8+ CTL
may play a role in host defense to Mtb through the secretion of
macrophage-activating cytokines such as IFN- and TNF-.
Alternatively, other components of the cytotoxic granule such as
granulysin may play a direct role in the inhibition of Mtb growth
(10, 19, 20).
At present, the role of MHC class I-restricted
CD8+ CTL in human immunity to tuberculosis
remains largely unexplored. We have previously demonstrated the
existence of human Mtb-reactive CD8+ CTL to Mtb.
These cells are present preferentially in persons infected with Mtb,
are cytolytic, and are capable of IFN- production in response to
Mtb-infected targets. While partially inhibited by anti-MHC class I
Ab, they are not restricted to the MHC class I A, B, or C alleles. We
demonstrated that these cells recognize a protein Ag that is generated
in the proteasome, but that does not require transport through the
Golgi endoplasmic reticulum. Our data suggested the possible use of
nonpolymorphic MHC class Ib Ag-presenting structures other than the
group 1 CD1 Ags (21). Interestingly, Canaday et al.
(22) have described Mtb-reactive
CD8+ T cells that while inhibited by
anti-class I Ab were not inhibited by brefeldin A. Whether or not
these cells were classically or nonclassically restricted was not
investigated (22).
Monocyte-derived dendritic cells (DC) pulsed with mycobacterial chloroform/methanol extract (23, 24) have been used to elicit Mtb-reactive CD1-restriced CD8+ T cells. Similarly, HLA-B44-, B52-, and A*0201-predicted binding peptides for ESAT-6 (25) or an HLA-A*0201-predicted binding peptide for the 19-kDa Ag (26) have been used to elicit CD8+ T cells that are reactive with Mtb-infected DC in an HLA-Ia-restricted manner. One limitation of these studies is that they have relied upon T cells stimulated in vitro with synthetic Ag. Hence, the relative contribution of these restriction specificities in the human host response to mycobacterially infected cells is unknown.
In this report, an IFN- enzyme-linked immunospot
(ELISPOT)-based limiting dilution analysis (LDA) is described and
is used to determine the distribution of both classically and
nonclassically restricted Mtb-reactive CD8+ T
cells in two healthy purified protein derivative (PPD)-positive
subjects. This method will allow for the detailed analysis of
Mtb-specific T cell responses in a wide variety of subjects. Healthy,
Mtb-infected individuals have developed a successful host response to
ongoing mycobacterial infection. It is hoped that a more detailed
understanding of this response in comparison to those with active
disease may elucidate the immunopathogenesis of tuberculosis and hence
allow for more rational vaccine design.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Subjects were recruited from employees at Harborview Medical Center, The Fred Hutchinson Cancer Research Center, Corixa Corporation, and Oregon Health Sciences University. PPD responses were determined by the employee health service at the respective institutions. Protocols for venipuncture and apheresis were Institutional Review Board approved. HLA typing was performed on PBMC by the Puget Sound Blood Center.
Monoclonal Abs and reagents
Culture medium consisted of RPMI 1640 supplemented with 10% FBS (BioWhittaker, Walkersville, MD), 50 µg/ml gentamicin sulfate (BioWhittaker), 5 x 10-5 M 2 ME (Sigma, St. Louis, MO), and 2 mM glutamine (Life Technologies, Grand Island, NY). For the elicitation of Mtb-reactive T cell clones, RPMI 1640 was supplemented with 10% human serum (HS). Mtb (H37Rv) was obtained from American Type Culture Collection (Manassas, VA) and grown in modified Middlebrook 7H9 media (Difco, Detroit, MI). After the preparation of glycerol stocks, aliquots were frozen and subsequently titered on Middlebrook 7H10 plates (Becton Dickinson Microbiology Systems, Cockeysville, MD).
Generation of peripheral blood DCs and macrophages
Monocyte-derived DCs were prepared according to the method of Romani et al. (27). Briefly, PBMC were isolated from heparinized blood by centrifugation over Ficoll-Hypaque (Sigma) and washed three times with culture medium. Alternatively, PBMC were obtained via leukapharesis. Cells were resuspended in AIM-V medium (BioWhittaker) and allowed to adhere to a T-75 (Costar, Cambridge, MA) flask at 37°C for 1 h in the presence of 10 ng/ml of GM-CSF (Immunex, Seattle, WA). After gentle rocking, nonadherent cells were removed, and 30 ml of AIM-V containing 10 ng/ml of IL-4 (Immunex) and 30 ng/ml of GM-CSF (Immunex) was added. After 18 h, the media was removed and centrifuged, and the cell-conditioned media was placed on the adherent cells. After 5–7 days, cells were harvested with cell-dissociation media (Sigma). To generate macrophages, PBMC were adhered to a T-75 flask as described above and cultured in the absence of cytokine. When prepared in AIM-V, peripheral blood-derived DC were CD1a positive and CD14 negative, whereas macrophages were CD14 positive and CD1a negative.
Flow cytometry
Cells to be analyzed for cell-surface marker expression were first incubated at 4°C in a blocking solution of PBS containing 2% normal rabbit serum (Sigma), 2% normal goat serum (Sigma), and 2% human serum to prevent nonspecific binding of mouse Ig. Cells were washed in FACS buffer (PBS containing 0.5% FBS and 0.02% sodium azide) and incubated with either FITC-conjugated anti-CD1a, anti-CD1b, CD4, anti-CD8, anti-CD14, anti-CD56 Abs (5 µg/ml), or an FITC-IgG1 control (Becton Dickinson Immunocytometry Systems, San Jose, CA; 5 µg/ml) for 30 min at 4°C in a total volume of 50 µl. Cells were then washed, flow cytometry was performed using a FACSCalibur (Becton Dickinson), and data were collected on 104 viable cells.
IFN- ELISPOT assay
Mtb-specific effectors were detected from purified
CD8+ T cells by ELISPOT, as described with
minor modifications (28). Briefly, 96-well
nitrocellulose-backed plates (MAHA S4510; Millipore, Bedford, MA) were
coated as recommended by the manufacturer with 10 µg/ml capture mouse
anti-IFN- mAb (1-D1K; Mabtech, Nacka, Sweden) overnight at room
temperature. Plates were then washed six times with PBS/0.05% Tween 20
(Sigma), blocked with RPMI 1640/10% HS for 1 h at room
temperature. Irradiated autologous Mtb-infected DC (2 x
104) were used as APC. Autologous purified
CD8+ T cells were then added, and the plate was
incubated overnight at 37°C. After washing with PBS/0.05% Tween 20,
100 µl of 1 µg/ml biotinylated secondary anti-IFN- mAb
(7B6-1; Mabtech) was added. After 2 h of incubation at room
temperature, plates were washed six times, 100 µl avidin/biotinylated
enzyme (HRP) complex (Vectastain ABC Elite Kit; Vector Laboratories,
Burlingame, CA) was added to wells, and the plates were incubated for a
further 2 h. Then, plates were washed six times, and 100 µl
3-amino-9-ethylcarbazole substrate (Vectastain AEC substrate kit,
Vector Laboratories) was added. After 4–7 min, the colorimetric
reaction was stopped by washing with distilled water, and plates were
air dried. Spots were quantitated using a Zeiss Axioplan 2 microscope
with 3200 K incident illumination equipped with a Epiplan Neofluar
5x/0.15 objective, Sony DXC 950 CCD camera, Märzhäuser
scanning stage, MCP4 control unit, Pentium PC computer, and KS ELISPOT
software (Carl Zeiss Vision, Hallbergmoos, Germany).
Rapid generation of Mtb-reactive CD8+ T cell clones
A total of 1 x 106 monocyte-derived DCs were cultured overnight in the presence of Mtb (H37Rv; multiplicity of infection (MOI), 5) in low-adherence 16-mm wells (no. 3473; Costar). After 18 h, the cells were harvested and resuspended in RPMI 1640/10% HS. A total of 2 x 104 Mtb-infected DC were seeded into each cloning well.
CD8+ T cells were purified from PBMC first by negative selection using CD4+ Ab-coated magnetic beads and then positive selection using CD8 Ab-coated magnetic beads per the manufacturer’s instructions (Miltenyi Biotec, Auburn, CA). Flow cytometric analysis confirmed that the cells were >98% CD8 positive. Ag-specific T cells were seeded by limiting dilution at 25 cells per well in the presence of 1 x 105 irradiated (3500 rad using a 137Cs source) autologous PBMC and rIL-2 (10 ng/ml). Cell culture media consisted of 200 µl of RPMI 1640 supplemented with 10% HS. Wells were assessed for growth between 10–14 days. Wells exhibiting growth were selected and transferred to a new plate for further analysis.
Evaluation of clonal responses to Mtb-infected APCs
DC and macrophages were prepared in AIM-V, harvested, and
incubated in the presence of Mtb (MOI, 50). After 18 h, cells were
harvested, washed, and seeded at 2–4 x 104
cells per well in RPMI 1640/10% HS in ELISPOT plates coated with
IFN- mAb. Aliquots of each clone (25 µl) were then added and the
ELISPOT assay completed after 18 h incubation at 37°C. Assays
were performed in the presence of IL-2 (1 ng/ml).
Expansion of T cell clones
To expand the CD8+ T cell clones, a rapid expansion protocol using anti-CD3 mAb stimulation was used (29). T cell clones were cultured in the presence of irradiated allogeneic PBMC (25 x 106), irradiated allogeneic lymphoblastoid cell line (5 x 106) and anti-CD3 mAb (30 ng/ml; Chiron, Emeryville, CA) in RPMI 1640 media with 10% HS in a T-25 upright flask in a total volume of 30 ml. The cultures were supplemented with IL-2 (1 ng/ml) on days +1, +4, +7, and +10 of culture. The cell cultures were washed on day +4 to remove remaining soluble anti-CD3 mAb.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Previous work by others and ourselves has suggested that
Mtb-specific CD8+ T cells are found
preferentially in persons infected with Mtb (21, 25, 26, 30, 31). These studies have relied on the generation of lines or on
peptide-specific responses and thus may be limited either by their
sensitivity or in their ability to reflect a diverse immune response.
To determine the frequency of Mtb-specific CD8+
effector cells, monocyte-derived DC were generated from a panel of five
healthy Mtb-infected individuals and six PPD-negative controls.
Monocyte-derived DC were infected with Mtb and used as APC in an
IFN- ELISPOT assay using autologous magnetic bead-purified
CD8+ T lymphocytes as effectors. Results from two
experiments are presented in Fig. 1
, and
the derived effector cell frequencies are presented in Table I. All five of the Mtb-infected
individuals had strong Mtb-specific CD8+ T cells
responses, with effector frequencies ranging from 1:4,000 to 1:16,000
(1:7,600 ± 4,300). Interestingly, all six of the PPD-negative
individuals also had demonstrable Mtb-reactive
CD8+ T cells (range, 1:2,900 to 1:29,000; mean,
1:16,000 ± 11,000; p > 0.6, Student’s
t test). These responses were all at least 2 SDs above the
uninfected DC control and were within the reported sensitivity of the
IFN- ELISPOT-based LDA (28). Thus, IFN- ELISPOT can
be used to determine effector cell frequencies in both Mtb-infected and
naive individuals and would suggest that an LDA-based approach can be
used to further characterize the CD8+ T cell
response in both subject groups.
|
|
To determine the frequencies of classically, CD1-, and
nonclassically restricted cells, a modified LDA was performed. To avoid
bias that might be introduced by repeated in vitro stimulation of T
cells lines, we devised a strategy that would allow for the evaluation
of clonal T cell responses after a single in vitro stimulation with
Mtb-infected DC and would allow for subsequent expansion and
characterization of these clones. In brief, magnetic bead-purified
CD8+ T cells were seeded at 25 cells/well into
96-well cloning plates containing Mtb-infected DC as APC (MOI, 5),
irradiated autologous PBMC feeders, and IL-2. After 12 days, 192 wells
exhibiting growth were transferred to a second 96-well plate to
facilitate subsequent analysis. IFN- ELISPOT was chosen for the
analysis because of its ability to detect small numbers of T cells (in
our hands, <20 cells using both CD4 and CD8 T cell clones; data not
shown). T cell clones were classified as either classically or
nonclassically restricted based upon recognition of Mtb-infected
autologous or HLA-A, B-mismatched targets (Fig. 2). To distinguish group 1 CD1-restricted
from nongroup 1 CD1-restricted responses, peripheral blood-derived DC
and 5-day adherent macrophages were generated from the HLA-A,
B-mismatched donor in AIM-V serum-free media. AIM-V was used to ensure
expression of group 1 CD1 Ags. Peripheral blood-derived DC were
confirmed by flow cytometry to be CD1a and CD1b positive, whereas
macrophages were CD1a and CD1b negative (data not shown). One-eighth of
each clone was assessed for its ability to recognize HLA-matched and
-mismatched Mtb-infected targets (Fig. 3). Of 192 clones tested, 70 did not
produce IFN-. Six produced IFN- in response to uninfected DC and
were termed nonspecific. Twenty-four were HLA restricted, in that
IFN- was produced only in response to autologous Mtb-infected DCs.
The remaining 92 were nonclassically, nongroup 1 CD1 restricted in that
IFN- was produced in response to all Mtb targets, including the
group 1 CD1-negative macrophages; representing 81% of the Mtb-reactive
clones. As an example, the clone tested in position B2 (Fig. 3)
responds only to D160 Mtb-infected APC and thus would be termed HLA
restricted, whereas the clone tested in A4 responds to all Mtb-infected
APC and is thus nonclassically restricted.
|
|
ß TCR and were
negative for NK markers CD16 and CD56. All expanded clones were
retested for specificity against HLA-matched and -mismatched DC and
macrophages. None of the nonclassically restricted cells exhibited
reactivity against uninfected mismatched DC.
To define a restricting MHC class I allele for classically restricted
Mtb-reactive CD8+ T clones, a panel of DCs was
generated that matched the CD8+ T cells at a
single HLA-A or B locus. As shown in Fig. 4, clone 1-6F is HLA-B14 restricted
because it is reactive with D44-infected DC who match D160 at HLA-B14.
Clones 1-1B and 1-4F are HLA-B44 restricted because they are reactive
with D131 and D184 DC, both of which share the HLA-B44 allele with
D160. Interestingly, one of the clones is neither HLA-A nor HLA-B
restricted.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
ELISPOT-based analysis to
estimate the frequency of Mtb-reactive CD8+ T
cells from peripheral blood and to define the relative frequencies of
both classically and nonclassically restricted Mtb-reactive
CD8+ T cells. All of the Mtb-infected donors who
were tested have strong CD8+ T cell responses to
Mtb-infected DCs, suggesting these responses may represent a recall
response to Mtb. Interestingly, the PPD-
individuals had demonstrable Mtb-reactive CD8+ T
cells. It is possible that these responses represent exposure to
atypical mycobacteria or that they are qualitatively different from
those found in Mtb-infected donors. Although the Mtb-specific effector
cell frequencies were not statistically different between the
PPD-positive and -negative groups, larger subject groups will be
assessed to determine whether or not CD8+ T cell
responses in those who are Mtb infected are distinct from naive
individuals. The prevalence of Mtb-reactive CD8+
lymphocytes in PPD- individuals may have been
previously underestimated by others and ourselves due to the fact that
ELISPOT-based estimates are more sensitive than those derived from the
generation of T cell lines.
Mtb-infected DC were used in a modified LDA to generate a panel of T
cell clones from peripheral blood from one of two
PPD+ donors. Using the IFN- ELISPOT, each
clone was assessed for its ability to recognize HLA-matched and
-mismatched targets and thus to infer its restriction specificity. In
these individuals, the majority of the Mtb-reactive
CD8+ T cells were nonclassically and nongroup
1-CD1 restricted, suggesting that these cells may represent the
immunodominant CD8+ T cell response to Mtb.
Therefore, if these results can be extended to additional Mtb-infected
healthy individuals, then further characterization of these cells may
facilitate the rational design of an effective vaccine.
For the classically restricted clones, and a subset of the nonclassically restricted clones from one donor, these results were confirmed by expansion and further characterization of these Mtb-specific T cell clones. While in the minority, Mtb-infected DC also stimulated HLA-Ia-restricted T cells. This represents the first report of the elicitation and cloning of MHC class Ia-restricted cells directly from Mtb-infected APC. One advantage of this approach is that we have allowed the immune system to select the Ag specificity and restricting allele, thus strengthening the argument that these responses reflect recall responses to Mtb. Interestingly, two clones were HLA-B44 restricted and one was HLA-B14 restricted, whereas none were restricted by HLA-A alleles. It is possible that the one remaining clone is HLA-C restricted. Whether or not HLA-B and possibly C restriction plays a unique role in the host response to Mtb will be the subject of future investigations, although HLA-A*0201-specific responses to Ags ESAT-6 and the 19-kDa Ag have been previously reported (25, 26).
Group 1 CD1-restricted T cell clones were not identified. In previous
reports, CD1-restricted T cells have been identified from both
Mtb-infected and PPD- individuals
(32). Whether or not CD1-restricted responses represent a
recall response to Mtb remains an important and unresolved question.
Thus, it is possible that the individuals analyzed are deficient in
group 1 CD1-restricted responses or that the effector frequency was
below the limit of detection of this analysis. Alternatively, it is
possible that the in vitro culture conditions were not favorable to the
generation of group 1 CD1-restricted responses. Stenger et al. have
recently reported that Mtb infection can decrease group 1 CD1
expression (33). Thus, there may have been inadequate CD1
expression on our Mtb-infected DC to stimulate CD1-restricted T cells.
Finally, clones were analyzed initially on the basis of IFN-
production. Thus, clones that grew in response to Mtb, but did not
produce IFN-, would not have been detected.
The approach described in this report is well suited to the analysis of populations of Ag- or pathogen-stimulated T cells from peripheral blood with minimal in vitro manipulation. In this report, we were able to detect and characterize Mtb-specific HLA-Ia-restricted T cells that comprised a relatively small component of the overall CD8+ T cell response to Mtb. Conventional bulk culture-based techniques are not well suited to such subpopulation analysis. With regard to Mtb, it is hypothesized that individuals who are healthy Mtb infected (PPD+), Mtb infected with active tuberculosis (PPD-), and those infected with bacillus Calmette-Guérin may have distinct CD8+ T cell responses. Thus, this analysis will be extended to these subject groups in future studies. Alternatively, the approach is well suited to any situation where T cells can be distinguished on the basis of selective responses to APC. Examples include restriction specificity as described herein or antigenic specificity using protein or peptide pulsed APC.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. David Lewinsohn, R&D 11, Portland Veterans Affairs Medical Center, 3710 U.S. Veterans Road, Portland, OR 97207.
3 Abbreviations used in this paper: Mtb, Mycobacterium tuberculosis; DC, dendritic cell; ELISPOT, enzyme-linked immunospot; LDA, limiting dilution analysis; PPD, purified protein derivative; HS, human serum; MOI, multiplicity of infection.
Received for publication January 14, 2000. Accepted for publication May 2, 2000.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
interferon-secreting CD8+, as well as CD4+, T cells in lungs following aerosol infection with Mycobacterium tuberculosis. Infect. Immun. 67:3242. in resistance to Mycobacterium tuberculosis infection. J. Exp. Med. 178:2249. is required in the protective immune response against Mycobacterium tuberculosis in mice. Immunity 2:561.[Medline]
ß TCR+ cells by Mycobacterium tuberculosis via an alternate class I MHC antigen-processing pathway. J. Immunol. 162:372.ß T cell pool. J. Immunol. 162:366.-secreting CD8+ T lymphocytes specific for Mycobacterium tuberculosis. Proc. Natl. Acad. Sci. USA 95:270.This article has been cited by other articles:
|
|
 |
|
  A. Tvinnereim and B. Wizel CD8+ T Cell Protective Immunity against Chlamydia pneumoniae Includes an H2-M3-Restricted Response That Is Largely CD4+ T Cell-Independent J. Immunol., September 15, 2007; 179(6): 3947 - 3957. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Das, J. Das, P. Eynott, Y. Zhang, A. L.M. Bothwell, L. V. Kaer, and Y. Shi Pivotal roles of CD8+ T cells restricted by MHC class I-like molecules in autoimmune diseases J. Exp. Med., November 27, 2006; 203(12): 2603 - 2611. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. W. Wieland, M. E. Kerver, S. Florquin, M. A. Nolte, J. Borst, R. van Lier, M. H. J. van Oers, and T. van der Poll CD27 contributes to the early systemic immune response to Mycobacterium tuberculosis infection but does not affect outcome Int. Immunol., November 1, 2006; 18(11): 1531 - 1539. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Ewer, K. A. Millington, J. J. Deeks, L. Alvarez, G. Bryant, and A. Lalvani Dynamic Antigen-specific T-Cell Responses after Point-Source Exposure to Mycobacterium tuberculosis Am. J. Respir. Crit. Care Med., October 1, 2006; 174(7): 831 - 839. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Carranza, E. Juarez, M. Torres, J. J. Ellner, E. Sada, and S. K. Schwander Mycobacterium tuberculosis Growth Control by Lung Macrophages and CD8 Cells from Patient Contacts Am. J. Respir. Crit. Care Med., January 15, 2006; 173(2): 238 - 245. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S Stenger Immunological control of tuberculosis: role of tumour necrosis factor and more Ann Rheum Dis, November 1, 2005; 64(suppl_4): iv24 - iv28. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Buettner, C. Meinken, M. Bastian, R. Bhat, E. Stossel, G. Faller, G. Cianciolo, J. Ficker, M. Wagner, M. Rollinghoff, et al. Inverse Correlation of Maturity and Antibacterial Activity in Human Dendritic Cells J. Immunol., April 1, 2005; 174(7): 4203 - 4209. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. B. Kamath, J. Woodworth, X. Xiong, C. Taylor, Y. Weng, and S. M. Behar Cytolytic CD8+ T Cells Recognizing CFP10 Are Recruited to the Lung after Mycobacterium tuberculosis Infection J. Exp. Med., December 6, 2004; 200(11): 1479 - 1489. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. L. Gervassi, K. H. Grabstein, P. Probst, B. Hess, M. R. Alderson, and S. P. Fling Human CD8+ T Cells Recognize the 60-kDa Cysteine-Rich Outer Membrane Protein from Chlamydia trachomatis J. Immunol., December 1, 2004; 173(11): 6905 - 6913. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Shams, P. Klucar, S. E. Weis, A. Lalvani, P. K. Moonan, H. Safi, B. Wizel, K. Ewer, G. T. Nepom, D. M. Lewinsohn, et al. Characterization of a Mycobacterium tuberculosis Peptide That Is Recognized by Human CD4+ and CD8+ T Cells in the Context of Multiple HLA Alleles J. Immunol., August 1, 2004; 173(3): 1966 - 1977. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. A. Lewinsohn, A. S. Heinzel, J. M. Gardner, L. Zhu, M. R. Alderson, and D. M. Lewinsohn Mycobacterium tuberculosis-specific CD8+ T Cells Preferentially Recognize Heavily Infected Cells Am. J. Respir. Crit. Care Med., December 1, 2003; 168(11): 1346 - 1352. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. L. Gervassi, P. Probst, W. E. Stamm, J. Marrazzo, K. H. Grabstein, and M. R. Alderson Functional Characterization of Class Ia- and Non-Class Ia-Restricted Chlamydia-Reactive CD8+ T Cell Responses in Humans J. Immunol., October 15, 2003; 171(8): 4278 - 4286. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. A. R. Tobian, N. S. Potter, L. Ramachandra, R. K. Pai, M. Convery, W. H. Boom, and C. V. Harding Alternate Class I MHC Antigen Processing Is Inhibited by Toll-Like Receptor Signaling Pathogen-Associated Molecular Patterns: Mycobacterium tuberculosis 19-kDa Lipoprotein, CpG DNA, and Lipopolysaccharide J. Immunol., August 1, 2003; 171(3): 1413 - 1422. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Kawashima, Y. Norose, Y. Watanabe, Y. Enomoto, H. Narazaki, E. Watari, S. Tanaka, H. Takahashi, I. Yano, M. B. Brenner, et al. Cutting Edge: Major CD8 T Cell Response to Live Bacillus Calmette-Guerin Is Mediated by CD1 Molecules J. Immunol., June 1, 2003; 170(11): 5345 - 5348. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. B. Urdahl, D. Liggitt, and M. J. Bevan CD8+ T Cells Accumulate in the Lungs of Mycobacterium tuberculosis-Infected Kb-/-Db-/- Mice, But Provide Minimal Protection J. Immunol., February 15, 2003; 170(4): 1987 - 1994. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. S. Heinzel, J. E. Grotzke, R. A. Lines, D. A. Lewinsohn, A. L. McNabb, D. N. Streblow, V. M. Braud, H. J. Grieser, J. T. Belisle, and D. M. Lewinsohn HLA-E-dependent Presentation of Mtb-derived Antigen to Human CD8+ T Cells J. Exp. Med., December 2, 2002; 196(11): 1473 - 1481. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Lazarevic and J. Flynn CD8+ T Cells in Tuberculosis Am. J. Respir. Crit. Care Med., October 15, 2002; 166(8): 1116 - 1121. [Full Text] [PDF]
|
|
|
|
|
|
D. A. Lewinsohn, R. A. Lines, and D. M. Lewinsohn Human Dendritic Cells Presenting Adenovirally Expressed Antigen Elicit Mycobacterium tuberculosis-Specific CD8+ T Cells Am. J. Respir. Crit. Care Med., September 15, 2002; 166(6): 843 - 848. [Abstract] [Full Text]
|
|
|
|
 |
|
 K. Hashimoto, Y. Maeda, H. Kimura, K. Suzuki, A. Masuda, M. Matsuoka, and M. Makino Mycobacterium leprae Infection in Monocyte-Derived Dendritic Cells and Its Influence on Antigen-Presenting Function Infect. Immun., September 1, 2002; 70(9): 5167 - 5176. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. V. Serbina, V. Lazarevic, and J. L. Flynn CD4+ T Cells Are Required for the Development of Cytotoxic CD8+ T Cells During Mycobacterium tuberculosis Infection J. Immunol., December 15, 2001; 167(12): 6991 - 7000. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. M. Lewinsohn, L. Zhu, V. J. Madison, D. C. Dillon, S. P. Fling, S. G. Reed, K. H. Grabstein, and M. R. Alderson Classically Restricted Human CD8+ T Lymphocytes Derived from Mycobacterium tuberculosis-Infected Cells: Definition of Antigenic Specificity J. Immunol., January 1, 2001; 166(1): 439 - 446. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
