Previous Article | Next Article 
Journal of Virology, May 2001, p. 4907-4911, Vol. 75, No. 10
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.10.4907-4911.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Persistence of Human Immunodeficiency Virus Type
1-Specific Cytotoxic T-Lymphocyte Clones in a Subject with Rapid
Disease Progression
Sabina A.
Islam,1
Christine M.
Hay,2
Kelly E.
Hartman,1
Suqin
He,1
Amy K.
Shea,1
Alicja K.
Trocha,1
Mark J.
Dynan,1
Neha
Reshamwala,1
Susan P.
Buchbinder,3
Nesli O.
Basgoz,1 and
Spyros A.
Kalams1,*
Partners AIDS Research Center & ID Division,
Massachusetts General Hospital, Harvard Medical School, Boston,
Massachusetts 021141; New York
Presbyterian Hospital, College of Physicians and Surgeons, Columbia
University, New York, New York 100322; and
AIDS Office, Department of Public Health, San Francisco,
California 941403
Received 12 July 2000/Accepted 20 February 2001
 |
ABSTRACT |
We longitudinally measured T-cell receptor transcript frequencies
of human immunodeficiency virus type 1 (HIV-1) specific cytotoxic T
lymphocytes (CTL) in an individual with rapidly progressive disease and
high levels of viremia. CTL clones elicited during acute HIV-1
infection were present at the time of death, despite absent functional
CTL responses, arguing against clonal deletion as a mechanism for the
decline of CTL responses observed during HIV-1 infection.
| |
TEXT |
Virus-specific cytotoxic T
lymphocytes (CTL) appear in acute human immunodeficiency virus type 1 (HIV-1) infection coincident with control of the initial intense burst
of viral replication (4, 13). Evidence that these CTL
expansions are oligoclonal is supported by studies demonstrating major
oligoclonal or monoclonal expansions of CD8+ T cells having
a predominant V
usage (17). In HIV-1 infection, some of
these V expansions have been shown to contain HIV-specific CTL
(17), but the long-term fate of these cells has not been determined. Despite the appearance of these early robust immune responses, in most individuals there is a decline in CTL function as
the disease progresses (6, 12). Clonal exhaustion,
resulting in physical deletion of CTL clones from chronic exposure to
high levels of antigen, has been proposed as a mechanism for the
decline in CTL activity seen during HIV-1 disease progression
(15, 18).
Moskophidis et al. (15) have shown that the presence of
rapidly replicating, noncytopathic lymphocytic choriomeningitis virus
throughout the lymphoid system in acute murine infection can initiate
large expansions of antiviral CTL followed, after a short period of
anergy, by clonal deletion. Pantaleo et al. (18), by using
CDR3-specific PCR to demonstrate a rapid disappearance of two expanded
clonotypes, suggested that clonal exhaustion occurred during acute
HIV-1 infection in two humans. Whether the decline in CTL function seen
in late-stage HIV-1 disease is because CTL clones, elicited during
early stages of disease, undergo clonal exhaustion in the face of a
chronically elevated antigen burden remains to be seen.
We have quantitated the magnitude and duration of individual
HIV-1-specific CTL expansions based on the molecular analysis of CTL
clones obtained by limiting dilution assays. Using oligonucleotide probes complementary to the unique CDR3 region of CTL clones, we
evaluated the T-cell receptor (TCR) transcript frequencies in
peripheral blood in the chronic stages of disease in an intermediate progressor and during acute- and late-stage disease in an individual with rapidly progressive infection.
Longitudinal tracking of TCR transcripts in PBMC of a chronic
HIV-1-infected subject.
To determine the fate of individual CTL
clonal frequencies during chronic stable infection we first
longitudinally tracked CTL clones in subject 115, an individual with
chronic stable disease and persistent functional CTL responses. Subject
115 was known to have been HIV-1 positive since 1987 and had been
started on antiretroviral therapy (zidovudine, lamivudine, and
indinavir) in July 1996. CTL clones were isolated and maintained by
limiting dilution cloning performed on either fresh or thawed
cryopreserved peripheral blood mononuclear cells (PBMC)
(21). Table 1 summarizes the
HLA restriction, epitope recognition, and TCR V gene usage of four
CTL clones isolated from this individual. The method used for
sequencing the TCR of CTL clones has been previously described in
detail (11).
View this table:
[in this window]
[in a new window]
|
TABLE 1.
Summary of HLA restriction, epitope recognition, and TCR
gene usage of CTL clones obtained from subject 115
|
|
For each longitudinal time point of interest, total RNA was isolated
from 5 × 10
6 cryopreserved PBMC. Two types of cDNA
libraries were constructed
from the RNA. The first library,
representing all rearranged TCR
transcripts obtained from the
subjects' cDNA, was generated by
anchored PCR (
14). The
second cDNA library of clone-specific
V
transcripts was generated by
using the respective V
primer
and a TCR
-chain constant region
primer (
5) for PCR amplification.
The V
-specific
primers used have been described previously (
11).
Colonies were picked from each library and transferred to nylon
membranes (Colony/Plaque Screen [137 mm], NEF-978Y; NEN Life
Science
Products, Boston, Mass.). Denatured membranes were hybridized
with
V
- or CDR3-specific biotin-labeled oligonucleotide probes.
Hybridization conditions were stringently optimized, and membranes
were
developed according to the manufacturer's specifications
(Tropix,
Bedford, Mass.). The CDR3-specific probes had 100% specificity
and
greater than 99% sensitivity on test membranes gridded with
positive
and negative controls. Controls for the entire procedure
were run in
parallel, and positive and negative colonies were
sequenced for
confirmation.
The library containing total PBMC TCR transcripts by anchored PCR was
probed with an oligonucleotide specific for the V
region
of interest
to obtain the frequency of clone-specific V
-chain
transcripts among
all TCR
-chain transcripts. The library containing
only TCR
transcripts of this particular V
was then probed with
an
oligonucleotide probe specific to the CDR3 region of the relevant
CTL
clone to obtain the frequency of clone-specific CDR3 sequences
among
the V
amplified transcripts. The final clone-specific frequency
among all TCR transcripts was obtained by multiplying these two
frequencies. Five hundred to 1,500 colonies of PBMC or of the
V
of
interest were
probed.
Four individual CTL clones from subject 115, an individual with chronic
stable HIV-1 infection, had frequencies ranging from
130 to 40,000 transcripts/million PBMC over 6 years of longitudinal
follow-up
extending from years 6 to 11 of the disease course (Fig.
1). These translate into clonal
frequencies ranging from 0.012%
of the total T-cell population for
clone 115 p175b at 9 years
postinfection to 3.78% of the total T-cell
population for clone
115 M21 at 6 years postinfection. Frequencies of
individual clonal
responses varied by up to 100-fold, but all responses
persisted
during the time of longitudinal follow-up in the presence of
ongoing
detectable viremia and during almost 2 years of follow-up on
highly
active antiretroviral therapy (HAART) during which viral load
remained below the limits of detection by commercial RNA PCR (<50
copies/ml). The measured transcript frequencies were detected
at
frequencies 2 to 5 logs higher than the limit of detection
for the
assay (Fig.
1).
View larger version (17K):
[in this window]
[in a new window]
|
FIG. 1.
Longitudinal CTL TCR clone transcript frequencies in a
chronically infected subject. Longitudinal CTL transcript frequencies
of CTL clones M21, E15, D87, and p175b from chronically infected
subject 115. CTL clones M21 and E15 recognize the B14 restricted Env
epitope 588K (ERYLKDQQL). Clone D87 recognizes the B14
restricted Env variant epitope 588Q (ERYLQDQQL). Clone p175b
recognizes the A2 restricted p17 Gag epitope SL9 (SLYNTVATL).
The black arrow indicates the time of institution of
antiretroviral therapy in subject 115. For each time point the
longitudinal limit of detection of the assay is shown for the least
prevalent clonotype. The limit of detection at a particular time point
for a clonotype is defined by the following formula: (1/total PBMC TCR
-chain transcripts) × (1 CTL clone CDR3 transcript/all V
transcripts). The diversity region-specific probes are identified in
footnote a of Table 1. CDR3 region-specific biotin-labeled
probes designed for each individual clonotype were 27 to 30 nucleotides
in length and spanned the D region with some overlap of the adjacent
V and J regions.
|
|
To validate the reproducibility of our V
and CDR3 probing techniques
we also did a series of spiking experiments. The HIV-specific
clone 115 M21 was mixed with either PBMC from an HIV-1-seronegative
subject or
with an unrelated CTL clone at frequencies ranging
from 0.1 to 10%.
The frequencies obtained for this clone with
this method were linear
over this range (
R = 0.95 and
P = 0.003).
When seronegative PBMC were probed with two CDR3 region
probes
from two separate HIV-1-specific CTL clones, no positive
colonies
were detected, despite detectable colonies for the
corresponding
V
.
Comparison of CTL TCR transcript frequencies to persistent
functional CTL responses in chronic HIV-1 infection.
In order to
address the relationship between transcript frequencies and functional
CTL assays, gamma interferon (IFN-
) enzyme-linked immunospot
(ELISPOT) assays and peptide-stimulated limiting dilution precursor
frequency assays were performed on the four clones from subject 115.
Precursor frequencies of HIV-1 epitope-specific CTL were estimated by
performing limiting dilution on freshly isolated PBMC
followed by in
vitro stimulation with peptide-pulsed autologous
PBMC and irradiated
PBMC from an HIV-1 seronegative donor, as
previously described
(
10,
11). Cells secreting IFN-
in an
antigen-specific
manner were detected using a standard ELISPOT
assay, as previously
described (
2,
3,
16,
20).
The frequencies of spot-forming cells (SFC) obtained by the IFN-
ELISPOT assay and peptide-stimulated CTL precursors (CTLp)
were
comparable and, in general, lower than the corresponding
CTL TCR
transcript frequencies measured at each time point for
the four clones
(Fig.
2). The functional responses and
the transcript
frequencies persist in the absence of therapy and also
after nearly
2 years of continuous viral suppression (Fig.
2). These studies
indicate that clonal
CTL responses, once generated, persist over
time. This occurs even when
HAART results in a decline in viral
load to below the limits of
detection and suggests that a new
steady state of memory CTL is
achieved, as has been described
for the chronic phase of other
persistent viral infections (
1,
16,
22). These comparative
studies provide a reference range
for the TCR transcript frequencies
when CTL functional responses
are present in an individual.
View larger version (37K):
[in this window]
[in a new window]
|
FIG. 2.
Comparative analysis of transcript frequency, SFC per
million and CTLp per million PBMC, in subject 115 for CTL clones M21,
E15, D87, and p175b. Frequencies of CTL transcripts per million PBMC
are longitudinally compared to CTL responses measures by IFN-
ELISPOT assay as SFC per million and compared to CTLp
frequencies measured by peptide-stimulated precursor frequency assay
for CTL clone M21 (a) and CTL clone E15 (b), clones which recognize the
B14 restricted Env epitope ERYLKDQQL, CTL clone D87 (c),
which recognizes the variant epitope ERYLQDQQL, and CTL
clone p175b (d), which recognizes the A2 restricted Gag epitope
SLYNTVATL. The black arrow indicates the time of institution
of antiretroviral therapy.
|
|
View larger version (41K):
[in this window]
[in a new window]
|
FIG. 3.
Longitudinal transcript frequencies of CTL clonotypes of
the rapid progressor. Longitudinal transcript frequencies of clonotypes
CDR3 B1, CDR3 B2, and CDR3 B3 in the PBMC of the rapid progressor at 3 months
( )
(February 1993, the time of seroconversion) and 45 months
( )
(August 1996, the time of death) after acute retrovirus infection.
|
|
Evaluation of the fate of clonal CTL responses over the course of
rapidly progressive infection.
The above data do not address the
fate of clonal responses generated in acute infection, nor do they
address the fate of such responses over the entire course of infection
when functional CTL responses are eventually lost. To address these
issues, we evaluated an individual (subject 053i) with rapidly
progressive HIV-1 disease who had been identified with acute HIV-1
infection, developed AIDS 13 months after seroconversion, and died
within 4 years of infection (8).
Previous studies had shown that this individual had both strong in vivo
activated CTL responses and in vitro stimulated memory
CTL responses
narrowly directed against a dominant B7 restricted
epitope in Env, gp41
IPRRIRQGL, and a subdominant B7 restricted
epitope in Pol,
SPAIFQSSM (
8). These functional responses were
lost with disease progression. No sequence variation occurred
within
the targeted epitopes (
8). Clones specific for the Env
epitope were generated 3, 8, and 39 months after presentation
with
acute HIV-1 infection. TCR cDNA libraries generated from
PBMC obtained
at two time points, during seroconversion at 3 months
(February 1993)
and just prior to death at 45 months (August 1996),
were screened with
CDR3-and V
-specific probes to quantitate the
frequency of these
clones in the
circulation.
At 3 months after presentation, seven IPRRIRQGL-specific CTL
clones isolated were analyzed for TCR gene usage (Table
2).
The majority population of CTL clones
isolated from this time
utilized V
6 and J
2.7 in their
-chains and had the TCR
CDR3
sequence WAASS, a clonotype
designated CDR3 B1. The minority population
had
-chain sequences
matching the IPRRIRQGL-specific CTL clone
isolated at 8 months with the CDR3 sequence ERSPPGD, a clonotype
designated CDR3 B2. The CDR3 sequence for the clone isolated at
39 months is PTAAG, a clonotype designated CDR3 B3.
View this table:
[in this window]
[in a new window]
|
TABLE 2.
TCR -chain sequences of B7 restricted Env (gp41/843-
851)-specific CTL clones isolated longitudinally from rapid
progressor subject
053ia
|
|
These clonotypes exhibited varying degrees of expansion over the course
of the illness (Table
3). With regard to
the first
clonotype, V
6 constituted 10.99% of all TCR transcripts
at seroconversion
and the CDR3 B1 clonotype was 32.26% of all V
6 transcripts.
The overall clonotype frequency was 3.54% (1 of 28) of
all TCR
transcripts, indicating an expansion of a single effector
clonotype
specific for the dominant CTL response during acute HIV-1
infection.
At 45 months, the time of death, when no in vivo activated
CTL
activity was detected, this clonotype was still present at a fairly
high frequency of 0.18% (1 of 547) of PBMC transcripts. The second
clonotype, CDR3 B2, isolated 8 months after presentation, constituted
87.2% of all V
16 transcripts at seroconversion and represented
4.29% (1 of 23) of all TCR transcripts in PBMC. At the time of
death,
the CDR3 B2 clonotype was 2.6% of V
16 transcripts, yielding
a
persistent but lower clonotype frequency of 0.0063% (1 of 15,873)
of
all TCR transcripts. The third clonotype, CDR3 B3, constituted
1.5% of
V
14 and 0.12% (1 of 833) of all TCR transcripts at the
time of
seroconversion. At the time of death this clonotype made
up 2.11% of
V
14, with a clonotype frequency of 0.02% (1 of 5,618)
of T cells.
Figure
3 graphically summarizes the longitudinal CTL
transcript
frequencies of clonotypes CDR3 B1, CDR3 B2, and CDR3
B3 in subject 053i
at seroconversion and at the time of death.
View this table:
[in this window]
[in a new window]
|
TABLE 3.
Summary of the number of positive colonies obtained out
of total number of colonies screened after hybridization of PBMC and
V-specific cDNA libraries with biotin-labeled V- and
CDR3-specific probes used for calculation of logitudinal TCR
frequences in the rapid progressor at seroconversion and at the
time of death
|
|
Our results indicate that once clonal CTL responses are established,
they persist in vivo, both in settings of persistent
high-level viremia
and when HAART lowers viremia to undetectable
levels. These studies
also extend an earlier study of rapidly
progressive infection
(
8), demonstrating that identical clones
generated in
acute infection persisted over the course of infection
and were still
present, albeit at lower levels, at the time of
death despite the loss
of these epitope-specific CTL functional
responses over time. Since no
sequence variation occurred within
the targeted epitope
(
8), and yet the CTL response declined,
the data suggest
both an in vivo defect in the ability to expand
to a cognate stimulus
and weak immune selection pressure mediated
through some CTL
responses.
We have demonstrated the cytolytic activity of the V
expansions
identified in acute HIV-1 infection by isolating functional
CTL clones
from these expansions in infected persons. In the CTL
clones we studied
at seroconversion, we found that the frequencies
of the expanded CDR3
B1 and CDR3 B2 clonotypes were 3.54 and 4.29%
of PBMC and that the
frequencies of the respective expanded V
6 and V
16 were 10.99 and 4.88% of PBMC. Our data demonstrate
that the composite frequency
of three CTL clones specific for
the same B7 restricted epitope was
7.95% of PBMC at the time of
acute HIV-1 infection. These data
indicate that early CTL expansions
can be large but still may be
insufficient to maintain viral
control.
These studies also provide strong evidence that CTL clonal deletion is
not the predictable result of persistent high-level
viremia in
late-stage HIV-1 infection. In murine lymphocytic choriomeningitis
virus infection, despite the induction of vigorous CTL responses
during
initial antigen exposure, exposure to high levels of antigen
can lead
to clonal exhaustion as a result of excessive stimulation
provided by
the persistence of elevated viral antigen (
15).
Additionally, Gallimore et al. and Zajac et al. have shown that
in some
situations where there is excessive antigen, antigen-specific
cells
detected by tetramers may have a diminished capacity to
produce IFN-
and lytic activity in vitro (
7,
22). Our data
support the
possibility that clonal deletion of CTL may not predictably
occur in
late-stage HIV-1 disease, pointing rather to an inability
of CTL clones
to expand and retain functional activity for the
cognate epitope. At
least one explanation for this would be a
lack of adequate T helper
cell function, since there is an association
between CTL and T helper
cell magnitude (
9,
19). Since the
method described here
discriminates among clones based on the
CDR3 region, it can
unambiguously define the magnitude of clonal
responses and should thus
be helpful in defining the relationship
between clonal CTL responses
and disease
progression.
| |
ACKNOWLEDGMENTS |
We thank Bruce D. Walker for critical review of the manuscript. We
thank Debbie J. Ruhl for excellent technical support. We thank M. Gately and Hoffman-LaRoche for the generous gift of interleukin-2.
This research was supported by NIH grants AI-39966 and AI-28568
(S.A.K.). S. A. Islam was supported by a Howard Hughes Medical Institute Postdoctoral Research Fellowship.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: MGH-East, CNY
5212, 149 13th St., Charlestown, MA 02129. Phone: (617) 726-8166. Fax: (617) 726-4691. E-mail:
Kalams{at}helix.mgh.harvard.edu.
| |
REFERENCES |
| 1.
|
Ahmed, R., and D. Gray.
1996.
Immunological memory and protective immunity: understanding their relation.
Science
272:54-60[Abstract].
|
| 2.
|
Altfeld, M.,
E. S. Rosenberg,
R. Shankarappa,
J. S. Mukherjee,
F. M. Hecht,
R. L. Eldridge,
M. M. Addo,
S. H. Poon,
M. N. Phillips,
G. K. Robbins,
P. E. Sax,
S. Boswell,
J. O. Kahn,
C. Brander,
P. J. Goulder,
J. A. Levy,
J. I. Mullins, and B. D. Walker.
2001.
Cellular immune responses and viral diversity in individuals treated during acute and early HIV-1 infection.
J. Exp. Med.
193:169-180[Abstract/Free Full Text].
|
| 3.
|
Altfeld, M. A.,
B. Livingston,
N. Reshamwala,
P. T. Nguyen,
M. M. Addo,
A. Shea,
M. Newman,
J. Fikes,
J. Sidney,
P. Wentworth,
R. Chesnut,
R. L. Eldridge,
E. S. Rosenberg,
G. K. Robbins,
C. Brander,
P. E. Sax,
S. Boswell,
T. Flynn,
S. Buchbinder,
P. J. R. Goulder,
B. D. Walker,
A. Sette, and S. A. Kalams.
2001.
Identification of novel HLA-A2-restricted human immunodeficiency virus type 1-specific cytotoxic T-lymphocyte epitopes predicted by the HLA-A2 supertype peptide-binding motif.
J. Virol.
75:1301-1311[Abstract/Free Full Text].
|
| 4.
|
Borrow, P.,
H. Lewicki,
B. H. Hahn,
G. M. Shaw, and M. B. A. Oldstone.
1994.
Virus-specific CD8+ cytotoxic T-lymphocyte activity associated with control of viremia in primary human immunodeficiency virus type 1 infection.
J. Virol.
68:6103-6110[Abstract/Free Full Text].
|
| 5.
|
Brander, C.,
P. J. Goulder,
K. Luzuriaga,
O. O. Yang,
K. E. Hartman,
N. G. Jones,
B. D. Walker, and S. A. Kalams.
1999.
Persistent HIV-1-specific CTL clonal expansion despite high viral burden post in utero HIV-1 infection.
J. Immunol.
162:4796-4800[Abstract/Free Full Text].
|
| 6.
|
Carmichael, A.,
X. Jin,
P. Sissons, and L. Borysiewicz.
1993.
Quantitative analysis of the human immunodeficiency virus type 1 (HIV-1)-specific cytotoxic T lymphocyte (CTL) response at different stages of HIV-1 infection: differential CTL responses to HIV-1 and Epstein-Barr virus in late disease.
J. Exp. Med.
177:249-256[Abstract/Free Full Text].
|
| 7.
|
Gallimore, A.,
A. Glithero,
A. Godkin,
A. C. Tissot,
A. Pluckthun,
T. Elliott,
H. Hengartner, and R. Zinkernagel.
1998.
Induction and exhaustion of lymphocytic choriomeningitis virus-specific cytotoxic T lymphocytes visualized using soluble tetrameric major histocompatibility complex class I-peptide complexes.
J. Exp. Med.
187:1383-1393[Abstract/Free Full Text].
|
| 8.
|
Hay, C. M.,
D. J. Ruhl,
N. O. Basgoz,
C. C. Wilson,
J. M. Billingsley,
M. P. DePasquale,
R. T. D'Aquila,
S. M. Wolinsky,
J. M. Crawford,
D. C. Montefiori, and B. D. Walker.
1999.
Lack of viral escape and defective in vivo activation of human immunodeficiency virus type 1-specific cytotoxic T lymphocytes in rapidly progressive infection.
J. Virol.
73:5509-5519[Abstract/Free Full Text].
|
| 9.
|
Kalams, S. A.,
S. P. Buchbinder,
E. S. Rosenberg,
J. M. Billingsley,
D. S. Colbert,
N. G. Jones,
A. K. Shea,
A. K. Trocha, and B. D. Walker.
1999.
Association between virus-specific cytotoxic T-lymphocyte and helper responses in human immunodeficiency virus type 1 infection.
J. Virol.
73:6715-6720[Abstract/Free Full Text].
|
| 10.
|
Kalams, S. A.,
R. P. Johnson,
M. J. Dynan,
K. E. Hartman,
T. Harrer,
E. Harrer,
A. K. Trocha,
W. A. Blattner,
S. P. Buchbinder, and B. D. Walker.
1996.
T cell receptor usage and fine specificity of human immunodeficiency virus 1-specific cytotoxic T lymphocyte clones: analysis of quasispecies recognition reveals a dominant response directed against a minor in vivo variant.
J. Exp. Med.
183:1669-1679[Abstract/Free Full Text].
|
| 11.
|
Kalams, S. A.,
R. P. Johnson,
A. K. Trocha,
M. J. Dynan,
H. S. Ngo,
R. T. D'Aquila,
J. T. Kurnick, and B. D. Walker.
1994.
Longitudinal analysis of T cell receptor (TCR) gene usage by human immunodeficiency virus 1 envelope-specific cytotoxic T lymphocyte clones reveals a limited TCR repertoire.
J. Exp. Med.
179:1261-1271[Abstract/Free Full Text].
|
| 12.
|
Klein, M. R.,
C. A. van Baalen,
A. M. Holwerda,
S. R. Kerkhof Garde,
R. J. Bende,
I. P. Keet,
J. K. Eeftinck-Schattenkerk,
A. D. Osterhaus,
H. Schuitemaker, and F. Miedema.
1995.
Kinetics of Gag-specific cytotoxic T lymphocyte responses during the clinical course of HIV-1 infection: a longitudinal analysis of rapid progressors and long-term asymptomatics.
J. Exp. Med.
181:1365-1372[Abstract/Free Full Text].
|
| 13.
|
Koup, R. A.,
J. T. Safrit,
Y. Cao,
C. A. Andrews,
G. McLeod,
W. Borkowsky,
C. Farthing, and D. D. Ho.
1994.
Temporal association of cellular immune responses with the initial control of viremia in primary human immunodeficiency virus type 1 syndrome.
J. Virol.
68:4650-4655[Abstract/Free Full Text].
|
| 14.
|
Loh, E. Y.,
J. F. Elliot,
S. Cwirla,
L. L. Lanier, and M. M. Davis.
1989.
Polymerase chain reaction with single-sided specificity: analysis of T cell receptor d chain.
Science
243:217-220[Abstract/Free Full Text].
|
| 15.
|
Moskophidis, D.,
F. Lechner,
H. Pircher, and R. M. Zinkernagel.
1993.
Virus persistence in acutely infected immunocompetent mice by exhaustion of antiviral cytotoxic effector T cells.
Nature
362:758-761[CrossRef][Medline]. (Erratum, 364:262.)
|
| 16.
|
Murali-Krishna, K.,
J. D. Altman,
M. Suresh,
D. J. Sourdive,
A. J. Zajac,
J. D. Miller,
J. Slansky, and R. Ahmed.
1998.
Counting antigen-specific CD8 T cells: a reevaluation of bystander activation during viral infection.
Immunity.
8:177-187[CrossRef][Medline].
|
| 17.
|
Pantaleo, G.,
J. F. Demarest,
H. Soudeyns,
C. Graziosi,
F. Denis,
J. W. Adelsberger,
P. Borrow,
M. S. Saag,
G. M. Shaw,
R. P. Sekaly, et al.
1994.
Major expansion of CD8+ T cells with a predominant V beta usage during the primary immune response to HIV.
Nature
370:463-467[CrossRef][Medline].
|
| 18.
|
Pantaleo, G.,
H. Soudeyns,
J. F. Demarest,
M. Vaccarezza,
C. Graziosi,
S. Paolucci,
M. Daucher,
O. J. Cohen,
F. Denis,
W. E. Biddison,
R. P. Sekaly, and A. S. Fauci.
1997.
Evidence for rapid disappearance of initially expanded HIV-specific CD8+ T cell clones during primary HIV infection.
Proc. Natl. Acad. Sci. USA
94:9848-9853[Abstract/Free Full Text].
|
| 19.
|
Rosenberg, E. S.,
J. M. Billingsley,
A. M. Caliendo,
S. L. Boswell,
P. E. Sax,
S. A. Kalams, and B. D. Walker.
1997.
Vigorous HIV-1-specific CD4+ T cell responses associated with control of viremia.
Science
278:1447-1450[Abstract/Free Full Text].
|
| 20.
|
Taguchi, T.,
J. R. McGhee,
R. L. Coffman,
K. W. Beagley,
J. H. Eldridge,
K. Takatsu, and H. Kiyono.
1990.
Detection of individual mouse splenic T cells producing IFN-V and IL-5 using the enzyme-linked immunospot (ELISPOT) assay.
J. Immunol. Methods
128:65-73[CrossRef][Medline].
|
| 21.
|
Walker, B. D.,
S. Chakrabarti,
B. Moss,
T. J. Paradis,
T. Flynn,
A. G. Durno,
R. S. Blumberg,
J. C. Kaplan,
M. S. Hirsch, and R. T. Schooley.
1987.
HIV-specific cytotoxic T lymphocytes in seropositive individuals.
Nature
328:345-348[CrossRef][Medline].
|
| 22.
|
Zajac, A. J.,
J. N. Blattman,
K. Murali-Krishna,
D. J. Sourdive,
M. Suresh,
J. D. Altman, and R. Ahmed.
1998.
Viral immune evasion due to persistence of activated T cells without effector function.
J. Exp. Med.
188:2205-2213[Abstract/Free Full Text].
|
Journal of Virology, May 2001, p. 4907-4911, Vol. 75, No. 10
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.10.4907-4911.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Ramalingam, R. K., Meyer-Olson, D., Shoukry, N. H., Bowen, D. G., Walker, C. M., Kalams, S. A.
(2008). Kinetic Analysis by Real-Time PCR of Hepatitis C Virus (HCV)-Specific T Cells in Peripheral Blood and Liver after Challenge with HCV. J. Virol.
82: 10487-10492
[Abstract]
[Full Text]
-
Derre, L., Bruyninx, M., Baumgaertner, P., Devevre, E., Corthesy, P., Touvrey, C., Mahnke, Y. D., Pircher, H., Voelter, V., Romero, P., Speiser, D. E., Rufer, N.
(2007). In Vivo Persistence of Codominant Human CD8+ T Cell Clonotypes Is Not Limited by Replicative Senescence or Functional Alteration. J. Immunol.
179: 2368-2379
[Abstract]
[Full Text]
-
Frater, A. J., Brown, H., Oxenius, A., Gunthard, H. F., Hirschel, B., Robinson, N., Leslie, A. J., Payne, R., Crawford, H., Prendergast, A., Brander, C., Kiepiela, P., Walker, B. D., Goulder, P. J. R., McLean, A., Phillips, R. E., and the Swiss HIV-Cohort Study,
(2007). Effective T-Cell Responses Select Human Immunodeficiency Virus Mutants and Slow Disease Progression. J. Virol.
81: 6742-6751
[Abstract]
[Full Text]
-
Meyer-Olson, D., Brady, K. W., Bartman, M. T., O'Sullivan, K. M., Simons, B. C., Conrad, J. A., Duncan, C. B., Lorey, S., Siddique, A., Draenert, R., Addo, M., Altfeld, M., Rosenberg, E., Allen, T. M., Walker, B. D., Kalams, S. A.
(2006). Fluctuations of functionally distinct CD8+ T-cell clonotypes demonstrate flexibility of the HIV-specific TCR repertoire. Blood
107: 2373-2383
[Abstract]
[Full Text]
-
Emu, B., Sinclair, E., Favre, D., Moretto, W. J., Hsue, P., Hoh, R., Martin, J. N., Nixon, D. F., McCune, J. M., Deeks, S. G.
(2005). Phenotypic, Functional, and Kinetic Parameters Associated with Apparent T-Cell Control of Human Immunodeficiency Virus Replication in Individuals with and without Antiretroviral Treatment. J. Virol.
79: 14169-14178
[Abstract]
[Full Text]
-
Meyer-Olson, D., Shoukry, N. H., Brady, K. W., Kim, H., Olson, D. P., Hartman, K., Shintani, A. K., Walker, C. M., Kalams, S. A.
(2004). Limited T Cell Receptor Diversity of HCV-specific T Cell Responses Is Associated with CTL Escape. JEM
200: 307-319
[Abstract]
[Full Text]
-
Wherry, E. J., Ahmed, R.
(2004). Memory CD8 T-Cell Differentiation during Viral Infection. J. Virol.
78: 5535-5545
[Full Text]
-
Karlsson, A. C., Deeks, S. G., Barbour, J. D., Heiken, B. D., Younger, S. R., Hoh, R., Lane, M., Sallberg, M., Ortiz, G. M., Demarest, J. F., Liegler, T., Grant, R. M., Martin, J. N., Nixon, D. F.
(2003). Dual Pressure from Antiretroviral Therapy and Cell-Mediated Immune Response on the Human Immunodeficiency Virus Type 1 Protease Gene. J. Virol.
77: 6743-6752
[Abstract]
[Full Text]
-
Wherry, E. J., Blattman, J. N., Murali-Krishna, K., van der Most, R., Ahmed, R.
(2003). Viral Persistence Alters CD8 T-Cell Immunodominance and Tissue Distribution and Results in Distinct Stages of Functional Impairment. J. Virol.
77: 4911-4927
[Abstract]
[Full Text]
-
Meyer-Olson, D., Brady, K. W., Blackard, J. T., Allen, T. M., Islam, S., Shoukry, N. H., Hartman, K., Walker, C. M., Kalams, S. A.
(2003). Analysis of the TCR {beta} Variable Gene Repertoire in Chimpanzees: Identification of Functional Homologs to Human Pseudogenes. J. Immunol.
170: 4161-4169
[Abstract]
[Full Text]
-
Hodges, E, Krishna, M T, Pickard, C, Smith, J L
(2003). Diagnostic role of tests for T cell receptor (TCR) genes. J. Clin. Pathol.
56: 1-11
[Abstract]
[Full Text]
-
McKay, P. F., Schmitz, J. E., Barouch, D. H., Kuroda, M. J., Lifton, M. A., Nickerson, C. E., Gorgone, D. A., Letvin, N. L.
(2002). Vaccine Protection Against Functional CTL Abnormalities in Simian Human Immunodeficiency Virus-Infected Rhesus Monkeys. J. Immunol.
168: 332-337
[Abstract]
[Full Text]
Top
Abstract
Text
