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IMMUNOBIOLOGY
From the Fondazione Andrea Cesalpino, Dipartimento di
Medicina Interna, Università di Roma La Sapienza, Rome, Italy;
Dipartimento di Pediatria and Istituto di Medicina Molecolare Angelo
Nocivelli, Università di Brescia, Italy; and Istituto
Pasteur-Cenci Bolognetti, Rome, Italy.
Conflicting results obtained from animal studies suggest that
B cells play a role in maintaining long-term T-cell memory and in
skewing T-cell response toward a T-helper 2 (TH2)
phenotype. X-linked agammaglobulinemia (XLA) is a genetic human disease
characterized by the lack of circulating B cells due to the mutation of
Bruton tyrosine kinase. This disease thus represents a unique
model for studying the role of B lymphocytes in regulating T-cell
functions in humans. To this aim, we analyzed hepatitis B envelope
antigen (HBenvAg)-specific T-cell memory in a series of
XLA patients vaccinated against hepatitis B virus (HBV). We found
HBenvAg-specific T lymphocytes producing interferon- X-linked agammaglobulinemia (XLA) is an
X-chromosome-linked recessive genetic disease characterized by a lack
of mature B cells, which results in primary
immunodeficiency.1,2 Affected subjects show a defective
humoral immune response that renders them susceptible to recurrent
bacterial infections.3 Early intravenous
immunoglobulin replacement therapy is effective in preventing
severe acute bacterial infections.4 The T-cell counts and
function are generally normal in these patients,5 and this explains why these patients cope normally with viral infections during
infancy. However, in some patients severe echovirus-dependent encephalitis and meningitis may develop.6
The molecular basis of this disorder is mutations in Bruton tyrosine
kinase (btk) gene, whose malfunction leads to an arrest of
B-cell differentiation.7-10 This tyrosine kinase is
expressed in myeloid cells as well as in B-lineage cells, but the
effects of mutations in Btk appear to have no clinically significant
effects on myeloid cell function.11
It has been suggested that B lymphocytes not only are involved in
humoral immune response, but may also have a role in influencing different facets of T-cell immunity. First, B cells have been shown to
function as efficient antigen-presenting cells (APCs), owing to their
peculiar capacity to uptake soluble antigens (Ags) at very low
concentration through their surface immunoglobulins.12,13 Second, it has been hypothesized that B lymphocytes are involved in
maintaining long-term T-cell memory.14-16 Third, the role
played by B cells in influencing the cytokine pattern of T-helper
(TH) cells has been analyzed in a variety of studies. As is
well known, T lymphocytes recognize Ags by engaging the T-cell receptor
(TCR) with peptide-major histocompatibility complexes (MHCs) displayed on the surface of APCs.17 Following TCR triggering,
CD4+ T lymphocytes polarize toward TH1 or
TH2 cells that produce different sets of cytokines, such as
interferon- However, the majority of data on the B-cell role in influencing T-cell
responses derives from animal models because of the evident
difficulties in addressing these complex issues directly in humans.
Therefore, XLA may represent a unique human model of B-cell deficiency
for studying the role of B lymphocytes in influencing Ag-specific
responses of memory-resting and memory-effector T cells. To
this aim, we evaluated here the hepatitis B envelope Ag
(HBenvAg)-specific T-cell response in a series of XLA patients vaccinated against hepatitis B virus (HBV) up to at least 24 months after completion of the immunization protocol, comparing them with
healthy age-matched immunized volunteers.
Vaccination against HBV is highly effective in preventing both acute
and chronic HBV infection. Protection is believed to be mediated by the
presence of both protective HBenvAg-specific antibodies (Abs) and by
the induction and expansion of HBenvAg-specific memory CD4+
T lymphocytes.27 In particular, we decided to study XLA
patients immunized against HBV, as this vaccine has been proved to
reduce serum HBV DNA in chronically infected patients in the absence of
neutralizing Abs,28 indicating that specific
CD4+ T-cell-mediated response plays a pivotal role in
directly suppressing HBV replication.29,30
Patients
Purification of PBMC subpopulations
Proliferative response All proliferation assays were set up by seeding 2 × 105 PBMCs per well in 96-well round-bottomed plates, as previously described.32 Cultures were incubated for 5 days at 37°C in 5% CO2, in the presence of different concentrations of recombinant (r) HBenvAg (a d subtype) (10, 5, 1, and 0 µg/mL) containing the 15-52 sequence of pre-S1 domain, the 133-145 sequence of pre-S2 domain, and the entire S domain (SmithKline Beecham). Cells were then pulsed with 1 µCi (.037 MBq) tritiated thymidine (Amersham, Buckinghamshire, United Kingdom) per well for 18 hours. Plates were harvested, and tritiated thymidine incorporation was assessed by means of a liquid scintillation counter (MicroBeta Plus) (Wallac, Turku Finland). All assays were performed in triplicate.Enzyme-linked immunospot assay HBenvAg-specific IFN- - and IL-4-secreting T cells were
detected by enzyme-linked immunospot (ELISPOT) assay, as previously described.31,33 This assay allows the frequencies of
cytokine-secreting cells to be precisely estimated by directly counting
the percentage of cytokine-specific spots formed upon Ag stimulation.
Briefly, 96-well nitrocellulose-backed plates (MAHA S4510) (Millipore, Bedford, MA) were coated with 5 µg/mL mouse antihuman IFN- or IL-4
mAb (PharMingen). Plates were washed with phosphate-buffered saline
(PBS) supplemented with 0.25% Tween 20 (Sigma, St Louis, MO)
(PBS/0.25% Tween 20) and blocked with PBS/10% human serum. PBMCs were
deposited in the wells in duplicate at 1 × 105 cells per
well in the presence of different concentrations of HBenvAg (10, 5, 1, and 0 µg/mL). After 48 hours' incubation at 37°C, plates were
washed with PBS/0.25% Tween 20, and then 2 µg/mL biotinylated mouse
antihuman IFN- or IL-4 mAb (PharMingen) was added to the well. After
2 hours' incubation at RT, plates were washed 5 times; 50 µL
streptavidin-horseradish peroxidase conjugate (PharMingen) (dilution
1:500) was added to the wells; and plates were incubated for a further
90 minutes at RT. Colorimetric reaction was obtained by means of
P-nitroblue tetrazolium/ 5-bromo-4-chloro-3-indolylphosphatase as a
substrate. Spots were quantified by means of an AID ELISPOT Reader
(AID, Strassberg, Germany), a computer-based system allowing the
semiautomatic interpretation of ELISPOT microtiter plates. Once background or artifacts had been eliminated, the counting software
quantitated the number of spots in the well, and the data were exported
for analysis. Potential cell ability to produce IFN- or IL-4 was
verified by stimulating cells with mitogen phytohemoagglutinin (PHA)
(Wellcome, Beckenham, United Kingdom).
Intracellular cytokine staining by flow cytometry PBMCs from 6 XLA patients and 2 healthy controls were either cultured in the presence or absence of HBenvAg 10 µg/mL for 18 hours at 37°C or treated with phorbolmyristate acetate/ionomycin (Sigma-Aldrich, Milan, Italy) (20 ng/mL/1 µg/mL) for 6 hours. At 2 hours after starting cultures, 10 µg/mL brefeldin-A (Sigma-Aldrich) was added. Triple staining was then performed to detect IFN-- and
IL-4-producing CD4+ T cells as follows: cells were
incubated with human -globulins for 15 minutes at RT and then
stained with CyChrome-conjugated anti-CD4 mAb (PharMingen) at 4°C for
30 minutes. Cells were washed twice with fluorescence-activated cell
sorter (FACS) buffer (PBS 1 ×, 2% fetal calf serum, 0.01% sodium
azide), fixed and permeabilized with Cytofix/Cytoperm solution
(Cytofix/Cytoperm Kit) (PharMingen) at 4°C for 20 minutes, followed
by 2 washes with Perm Wash Buffer (PharMingen). Fluorescein
isothiocyanate-conjugated antihuman IFN-- and
phycoerythrin-conjugated antihuman IL-4 mAbs (PharMingen) were added,
and cells were incubated for 30 minutes at 4°C. Cells were then
washed twice with FACS buffer, acquired with a FACScan flow
cytometer (Becton Dickinson, San Jose, CA), and anlyzed by means of
CellsQuest software (BD, Palo Alto, CA). Negative controls were
obtained by staining cells with an irrelevant
isotype-matched mAb.
Statistical analysis Differences between groups were analyzed by a 2-tailed, 2-sample Student t test or by contingency tables ( 2)
with 2 degrees of confidence. A value of P < .05 was
considered to be statistically significant.
PBMC proliferative response to HBenvAg PBMCs isolated from patients or healthy controls either before or after vaccination against HBV were stimulated in vitro with HBenvAg. The results presented here were obtained with the use of HBenvAg at 10 µg/mL, a concentration that proved to be optimal in our experiments. At basal time, no patient or control subject had a significant proliferative response to HBenvAg, corresponding to a stimulation index (SI) below 3. SI was defined as the ratio of cpm from HBenvAg-stimulated PBMCs to unstimulated PBMCs. At 1 month after the end of the vaccination cycle, all individuals from both study populations had a significantly positive SI. Mean HBenvAg-specific SIs ± SEM for PBMCs obtained from XLA patients were as follows: 1.2 ± 0.29; 12 ± 2.73; 11.50 ± 3.05; 21.64 ± 3.14, assessed before and 1, 12, and 24 months after immunization, respectively. These results were not significantly different from controls (1.09 ± 0.15; 18.68 ± 7.98; 16.05 ± 5.50; 24.62 ± 9.32). Individual results for both XLA and control groups are shown in Figure 1.
Frequencies of HBenvAg-specific IFN- - or IL-4-producing
HBenvAg-specific T-cell frequencies in the peripheral blood of either
XLA patients or healthy subjects were carried out by ELISPOT assay. Spots were analyzed after 48 hours of culture in the presence or
absence of HBenvAg. Potential cell capability to produce IFN- or
IL-4 was verified by stimulating cells with mitogen PHA. Cells from all
the subjects tested produced similar amounts of cytokines after this
nonspecific stimulus (not shown). Before vaccination, no XLA patient or
control subject showed any significant specific response, arbitrarily
defined as the presence of at least 5 spots in the stimulated wells
after subtraction of spots in the nonstimulated ones.
Conversely, in response to HBenvAg, all the XLA patients, as well as
all the healthy subjects, produced significant numbers of both IFN-
and IL-4 spots (Figure 2). Frequencies of
cytokine-producing cells were expressed as number of positive T cells
per 1 × 106 total PBMCs and varied individually. Mean
HBenvAg-specific cytokine-producing cells per 1 × 106
PBMCs in XLA patients ± SEM, calculated after subtraction of spots in unstimulated wells from spots in HBenvAg-stimulated ones were
as follows: for IFN-, 3.66 ± 0.66; 356.44 ± 123.07;
231 ± 82.37; 233 ± 64.12; and for IL-4, 3.77 ± 0.72;
138 ± 65.47; 139 ± 46.27; 100.66 ± 19.16, as assessed before
and 1, 12, and 24 months after anti-HBV immunization, respectively.
None of these data differed significantly from those obtained in
control subjects (for IFN-, 2.5 ± 1.11; 143 ± 40.16;
229.5 ± 107.19; 200.83 ± 91.82; for IL-4, 7 ± 2;
51.83 ± 15.92; 124 ± 40.51; 83.16 ± 23.47).
Intracellular cytokine staining In order to substantiate that memory-effector (ME) T cells with TH1, TH2, or TH0 phenotype are long-lived, intracellular cytokine analysis by flow cytometry was determined in both XLA patients and healthy subjects at 24 months after the last vaccination boost. Specific intracytoplasmic IFN- and IL-4
productions were differently distributed in the peripheral T cells of
the 2 study populations, upon 18 hours' incubation with HBenvAg in
vitro (Figure 3; Table 2). This supports the hypothesis that
memory T cells with prompt TH1, TH2, or
TH0 function are long-lived in humans and that their maintenance does not require the presence of B cells. Moreover, these data confirm those obtained with ELISPOT assay, showing the absence of a selective polarization toward the TH1 cell
phenotype in XLA patients.
ELISPOT analysis in CD27+ and CD27 As indicated in Figure 4, both IFN-
Owing to mutations of Btk genes, subjects affected by XLA lack circulating B cells and represent an excellent and unique model to study the role of these lymphocytes in modulating T-cell memory in humans. Indeed, although the current opinion is that B cells are unable to activate naive T lymphocytes or are even tolerogenic,36-38 several animal studies suggest that they might be required to maintain long-term T-cell memory.14-16 There is very little and contrasting data on T-cell memory in XLA patients. In one study, an impaired delayed cutaneous hypersensitivity reaction in XLA patients was reported.39 However, normal T-cell proliferation and cytokine production in response to either mitogens or tetanus toxoid (TT) in XLA patients up to 6 months after TT immunization boost have been subsequently shown.40 The conditions required to maintain T-cell memory are still the subject of intense debate and investigation. Various evidence suggests that neither B cells nor Ag persistence is required for maintaining either CD4+ or CD8+ T-cell memory, as these are both long-lived cell populations.14,41-43 On the other hand, different reports have proposed that memory CD4+ T cells require continual stimulation by APCs expressing MHC/Ag complexes to survive.14,44 According to this view, follicular DCs (FDCs) retain antigen on their surface in the form of immune complexes (ICs) for a very long time, functioning as an Ag reservoir for continual stimulation of memory CD4+ T cells.14,44 Hence, the presence of B cells producing specific Abs would be strictly necessary not only for producing the Abs required for IC formation, but also for providing an environment in which Ags may be trapped and retained.44 In order to define the role of B cells in influencing memory T-cell immunity in humans, we analyzed the HBenvAg-specific CD4+ T-cell responses in terms of proliferation, frequency, and cytokine production. This study was performed on 9 XLA patients immunized against HBV who were compared with age-matched controls at different times after completion of the vaccination protocol. We found that HBenvAg-specific proliferative responses did not differ from those of controls, up to at least 24 months after vaccination. Proliferative assay quantifies the expansion of circulating-specific T cells, as assessed 6 days after incubation with HBenvAg and thus evaluates mainly the presence of memory T cells requiring Ag restimulation in order to perform their effector functions. An important factor is that memory T cells from XLA patients showed either a TH1 or a TH2 phenotype, as detected by both ELISPOT and intracytoplasmic cytokine analyses, with frequencies that did not differ from those of healthy controls up to at least 24 months after the last vaccination boost. All together, these data suggest that in contrast with most, but not all, animal models,22,23,26,36 neither T-cell memory nor the balance between TH1- and TH2-cell responses are impaired by severe B-cell deficiency in humans. Furthermore, these findings might be only apparently in disagreement with a recent study that indicates a dominant TH1 polarization in TT-specific T-cell clones derived from the same type of patient.45 Indeed, our experimental procedure, monitoring antigen-specific T-cell responses ex vivo for a 24-month period after vaccination, may mainly detect early memory T cells with both TH1 and TH2 phenotype. Conversely, the Del Prete protocol, which studies the responses of long-term T-cell lines isolated more than 10 years after the first vaccination boost, may identify late memory T cells with strong TH1 polarization.45 This hypothesis may be reinforced by the possibility that the frequent bacterial infections that affect XLA patients during childhood may select a preferential TH1 polarization in the long run.46 Nevertheless, our data clearly indicate that XLA patients are capable of mounting TH1- and/or TH2-cell responses, and this has important implications for setting up protective vaccination protocols against both intracellular and extracellular pathogens. It could be argued that since XLA patients receive periodic administration of intravenous immunoglobulins, these might include specific Abs that form ICs with the immunizing HBenvAg and thus represent an important factor in maintaining the FDC-dependent memory.14 However, this possibility seems unlikely, since it has been shown that a lack of B cells cannot support the development of FDCs47 and that lymph node and splenic architecture are disrupted in both XLA patients and transgenic mice expressing nonfunctional mutated forms of Btk.48,49 We also studied the role of B cells in influencing both ME and MR
T-cell immunity in humans, by performing a series of experiments using
CD27-enriched and CD27-depleted T-cell
populations.31,34,50 CD27 is a Traf-linked tumor necrosis
factor receptor family member receptor,51 whose presence
on the cell membrane has been strictly related to naive or MR T cells
requiring Ag-driven proliferation for differentiating in
effectors.21,31,34,50,52,53 MR T cells, also defined as
"central memory," are trapped in the secondary lymphoid organs by
the lymph node homing receptors cc chemokine receptor-7 (CCR7)
and L-selectin and require antigen-driven restimulation in order to
differentiate into ME cells.21 In contrast, the CD27 In conclusion, in this report we demonstrate that XLA patients immunized against HBV show a robust antigen-specific T-cell response in vitro, which is retained for at least 24 months after vaccination. Analysis of HBenvAg-specific cytokine production performed at the single-cell level by ELISPOT or by flow cytometry revealed that both MR and ME CD4+ T cells with TH1, TH2, or TH0 phenotype are long-lived in these patients, as well as in healthy controls. All these data lead us to conclude that severe B-cell deficiency in humans does not impair the generation of an effective Ag-specific T-cell memory. The detection of HBenvAg-specific MR and ME T cells with either TH1 or TH2 phenotype may provide important building blocks for setting up highly protective, T-cell-based vaccination protocols in XLA patients and for devising innovative strategies for the manipulation of antimicrobial responses in these subjects.
Submitted July 19, 2001; accepted November 1, 2001.
Supported by Ministero della Sanità-Istituto Superiore di Sanità (Progetti AIDS 1998-2000 and Epatite Virale 1997-1999) (V.B.); Ministero dell'Università e della Ricerca Scientifica e Tecnologica (MURST), 40% 1997-1999 (V.B.); Progetto Associazione Italiana Sclerosi Multipla (AISM), 1997-1999 (V.B.); Progetto Finalizzato CNR Biotecnologie, European Community contract BMH4-CT98-3703 (V.B.); and MURST, 40% 1999 (A.P.).
M.P. and D.A. contributed equally to this work.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Vincenzo Barnaba, Fondazione Andrea Cesalpino, Dipartimento di Medicina Interna, Policlinico Umberto I, V le del Policlinico 155, 00161, Rome, Italy; e-mail: vincenzo.barnaba{at}uniroma1.it.
1. Kinnon C, Hinshelwood S, Levinsky RJ, Lovering RC. X-linked agammaglobulinemia: gene cloning and future prospects. Immunol Today. 1993;14:554-558[CrossRef][Medline] [Order article via Infotrieve]. 2. Conley ME. B cells in patients with X-linked agammaglobulinemia. J Immunol. 1985;134:3070-3074[Abstract]. 3. Ochs HD, Smith CI. X-linked agammaglobulinemia: a clinical and molecular analysis. Medicine (Baltimore). 1996;75:287-299[CrossRef][Medline] [Order article via Infotrieve]. 4. Quartier P, Debre M, De Blic J, et al. Early and prolonged intravenous immunoglobulin replacement therapy in childhood agammaglobulinemia: a retrospective survey of 31 patients. J Pediatr. 1999;134:589-596[CrossRef][Medline] [Order article via Infotrieve]. 5. Waldman TA. Immunodeficiency diseases: primary and acquired. In: Samter M,Talmage DW,Frank MM,Austen KF,Claman HN, eds. Immunological Diseases. Boston, MA: Little, Brown; 1988:411-465. 6. Misbah SA, Spickett GP, Ryba PC, et al. Chronic enteroviral meningoencephalitis in agammaglobulinemia: case report and literature review. J Clin Immunol. 1992;12:266-270[CrossRef][Medline] [Order article via Infotrieve].
7.
de Weers M, Mensink RG, Kraakman ME, Schuurman RK, Hendriks RW.
Mutation analysis of the Bruton's tyrosine kinase gene in X-linked agammaglobulinemia: identification of a mutation which affects the same codon as is altered in immunodeficient xid mice.
Hum Mol Genet.
1994;3:161-166 8. Conley ME, Parolini O, Rohrer J, Campana D. X-linked agammaglobulinemia: new approaches to old questions based on the identification of the defective gene. Immunol Rev. 1994;138:5-21[CrossRef][Medline] [Order article via Infotrieve]. 9. Lovering RC, Sweatman A, Genet SA, et al. Identification of deletions in the btk gene allows unambiguous assessment of carrier status in families with X-linked agammaglobulinaemia. Hum Genet. 1994;94:77-79[CrossRef][Medline] [Order article via Infotrieve]. 10. Khan WN, Alt FW, Gerstein RM, et al. Defective B cell development and function in Btk-deficient mice. Immunity. 1995;3:283-299[CrossRef][Medline] [Order article via Infotrieve].
11.
Futatani T, Miyawaki T, Tsukada S, et al.
Deficient expression of Bruton's tyrosine kinase in monocytes from X-linked agammaglobulinemia as evaluated by a flow cytometric analysis and its clinical application to carrier detection.
Blood.
1998;91:595-602 12. Lanzavecchia A. Antigen-specific interaction between T and B cells. Nature. 1985;314:537-539[CrossRef][Medline] [Order article via Infotrieve]. 13. Barnaba V, Franco A, Alberti A, Benvenuto R, Balsano F. Selective killing of hepatitis B envelope antigen-specific B cells by class I-restricted, exogenous antigen-specific T lymphocytes. Nature. 1990;345:258-260[CrossRef][Medline] [Order article via Infotrieve]. 14. Ahmed R, Gray D. Immunological memory and protective immunity: understanding their relation. Science. 1996;272:54-60[Abstract]. 15. Dutton RW, Bradley LM, Swain SL. T cell memory. Annu Rev Immunol. 1998;16:201-223[CrossRef][Medline] [Order article via Infotrieve]. 16. Zinkernagel RM. On immunological memory. Annu Rev Immunol. 1996;14:333-367[CrossRef][Medline] [Order article via Infotrieve]. 17. Germain RN. MHC-dependent antigen processing and peptide presentation: providing ligands for T lymphocyte activation. Cell. 1994;76:287-299[CrossRef][Medline] [Order article via Infotrieve]. 18. Mosmann TR, Coffman RL. TH1 and TH2 cells: different patterns of lymphokine secretion lead to different functional properties. Annu Rev Immunol. 1989;7:145-173[CrossRef][Medline] [Order article via Infotrieve]. 19. Paul WE, Seder RA. Lymphocyte responses and cytokines. Cell. 1994;76:241-251[CrossRef][Medline] [Order article via Infotrieve]. 20. Romagnani S. Lymphokine production by human T cells in disease states. Annu Rev Immunol. 1994;12:227-257[CrossRef][Medline] [Order article via Infotrieve]. 21. Sallusto F, Mackay CR, Lanzavecchia A. The role of chemokine receptors in primary, effector, and memory immune responses. Annu Rev Immunol. 2000;18:593-620[CrossRef][Medline] [Order article via Infotrieve]. 22. Macaulay AE, DeKruyff RH, Goodnow CC, Umetsu DT. Antigen-specific B cells preferentially induce CD4+ T cells to produce IL-4. J Immunol. 1997;158:4171-4179[Abstract].
23.
Macaulay AE, DeKruyff RH, Umetsu DT.
Antigen-primed T cells from B cell-deficient JHD mice fail to provide B cell help.
J Immunol.
1998;160:1694-1700
24.
Moulin V, Andris F, Thielemans K, Maliszewski C, Urbain J, Moser M.
B lymphocytes regulate dendritic cell (DC) function in vivo: increased interleukin 12 production by DCs from B cell-deficient mice results in T helper cell type 1 deviation.
J Exp Med.
2000;192:475-482
25.
Langhorne J, Cross C, Seixas E, Li C, von der Weid T.
A role for B cells in the development of T cell helper function in a malaria infection in mice.
Proc Natl Acad Sci U S A.
1998;95:1730-1734
26.
Mukhopadhyay S, Sahoo PK, George A, Bal V, Rath S, Ravindran B.
Delayed clearance of filarial infection and enhanced Th1 immunity due to modulation of macrophage APC functions in xid mice.
J Immunol.
1999;163:875-883 27. Barnaba V, Franco A, Paroli M, et al. Selective expansion of cytotoxic T lymphocytes with a CD4+CD56+ surface phenotype and a T helper type 1 profile of cytokine secretion in the liver of patients chronically infected with hepatitis B virus. J Immunol. 1994;152:3074-3087[Abstract]. 28. Couillin I, Pol S, Mancini M, et al. Specific vaccine therapy in chronic hepatitis B: induction of T cell proliferative responses specific for envelope antigens. J Infect Dis. 1999;180:15-26[CrossRef][Medline] [Order article via Infotrieve].
29.
Guidotti LG, Guilhot S, Chisari FV.
Interleukin-2 and alpha/beta interferon down-regulate hepatitis B virus gene expression in vivo by tumor necrosis factor-dependent and -independent pathways.
J Virol.
1994;68:1265-1270 30. Franco A, Guidotti LG, Hobbs MV, Pasquetto V, Chisari FV. Pathogenetic effector function of CD4-positive T helper 1 cells in hepatitis B virus transgenic mice. J Immunol. 1997;159:2001-2008[Abstract].
31.
Scognamiglio P, Accapezzato D, Casciaro MA, et al.
Presence of effector CD8+ T cells in hepatitis C virus-exposed healthy seronegative donors.
J Immunol.
1999;162:6681-6689 32. Barnaba V, Franco A, Alberti A, Balsano C, Benvenuto R, Balsano F. Recognition of hepatitis B virus envelope proteins by liver-infiltrating T lymphocytes in chronic HBV infection. J Immunol. 1989;143:2650-2655[Abstract]. 33. Prezzi C, Casciaro MA, Francavilla V, et al. Virus-specific CD8(+) T cells with type 1 or type 2 cytokine profile are related to different disease activity in chronic hepatitis C virus infection. Eur J Immunol. 2001;31:894-906[CrossRef][Medline] [Order article via Infotrieve].
34.
Hamann D, Baars PA, Rep MH, et al.
Phenotypic and functional separation of memory and effector human CD8+ T cells.
J Exp Med.
1997;186:1407-1418 35. De Jong R, Brouwer M, Hooibrink B, Van der Pouw-Kraan T, Miedema F, Van Lier RA. The CD27-subset of peripheral blood memory CD4+ lymphocytes contains functionally differentiated T lymphocytes that develop by persistent antigenic stimulation in vivo. Eur J Immunol. 1992;22:993-999[Medline] [Order article via Infotrieve].
36.
Epstein MM, Di Rosa F, Jankovic D, Sher A, Matzinger P.
Successful T cell priming in B cell-deficient mice.
J Exp Med.
1995;182:915-922
37.
Fuchs EJ, Matzinger P.
B cells turn off virgin but not memory T cells.
Science.
1992;258:1156-1159
38.
Bennett SR, Carbone FR, Toy T, Miller JF, Heath WR.
B cells directly tolerize CD8(+) T cells.
J Exp Med.
1998;188:1977-1983 39. Crockard AD, Boyd NA, McNeill TA, McCluskey DR. CD4 lymphocyte subset abnormalities associated with impaired delayed cutaneous hypersensitivity reactions in patients with X-linked agammaglobulinaemia. Clin Exp Immunol. 1992;88:29-34[Medline] [Order article via Infotrieve]. 40. Plebani A, Fischer MB, Meini A, Duse M, Thon V, Eibl MM. T cell activity and cytokine production in X-linked agammaglobulinemia: implications for vaccination strategies. Int Arch Allergy Immunol. 1997;114:90-93[Medline] [Order article via Infotrieve].
41.
Murali-Krishna K, Lau LL, Sambhara S, Lemonnier F, Altman J, Ahmed R.
Persistence of memory CD8 T cells in MHC class I-deficient mice.
Science.
1999;286:1377-1381
42.
Swain SL, Hu H, Huston G.
Class II-independent generation of CD4 memory T cells from effectors.
Science.
1999;286:1381-1383
43.
Di Rosa F, Matzinger P.
Long-lasting CD8 T cell memory in the absence of CD4 T cells or B cells.
J Exp Med.
1996;183:2153-2163
44.
van Essen D, Dullforce P, Brocker T, Gray D.
Cellular interactions involved in Th cell memory.
J Immunol.
2000;165:3640-3646 45. Amedei A, Romagnani C, Benagiano M, et al. Preferential Th1 profile of T helper responses in X-linked (Bruton's) agammaglobulinemia. Eur J Immunol. 2001;31:1927-1934[CrossRef][Medline] [Order article via Infotrieve].
46.
Shirakawa T, Enomoto T, Shimazu S, Hopkin JM.
The inverse association between tuberculin responses and atopic disorder.
Science.
1997;275:77-79 47. Kapasi ZF, Burton GF, Shultz LD, Tew JG, Szakal AK. Cellular requirements for functional reconstitution of follicular dendritic cells in SCID mice. Adv Exp Med Biol. 1993;329:383-386[Medline] [Order article via Infotrieve]. 48. Dingjan GM, Maas A, Nawijn MC, et al. Severe B cell deficiency and disrupted splenic architecture in transgenic mice expressing the E41K mutated form of Bruton's tyrosine kinase. EMBO J. 1998;17:5309-5320[CrossRef][Medline] [Order article via Infotrieve]. 49. Conley ME, Cooper MD. Genetic basis of abnormal B cell development. Curr Opin Immunol. 1998;10:399-406[CrossRef][Medline] [Order article via Infotrieve]. 50. Hendriks J, Gravestein LA, Tesselaar K, van Lier RA, Schumacher TN, Borst J. CD27 is required for generation and long-term maintenance of T cell immunity. Nat Immunol. 2000;1:433-440[CrossRef][Medline] [Order article via Infotrieve]. 51. Gravestein LA, Borst J. Tumor necrosis factor receptor family members in the immune system. Semin Immunol. 1998;10:423-434[CrossRef][Medline] [Order article via Infotrieve]. 52. Sallusto F, Lenig D, Forster R, Lipp M, Lanzavecchia A. Two subsets of memory T lymphocytes with distinct homing potentials and effector functions. Nature. 1999;401:708-712[CrossRef][Medline] [Order article via Infotrieve]. 53. Lanzavecchia A, Sallusto F. From synapses to immunological memory: the role of sustained T cell stimulation. Curr Opin Immunol. 2000;12:92-98[CrossRef][Medline] [Order article via Infotrieve].
54.
Bachmann MF, Kundig TM, Hengartner H, Zinkernagel RM.
Protection against immunopathological consequences of a viral infection by activated but not resting cytotoxic T cells: T cell memory without "memory T cells"?
Proc Natl Acad Sci U S A.
1997;94:640-645 55. Reinhardt RL, Khoruts A, Merica R, Zell T, Jenkins MK. Visualizing the generation of memory CD4 T cells in the whole body. Nature. 2001;410:101-105[CrossRef][Medline] [Order article via Infotrieve].
56.
Masopust D, Vezys V, Marzo AL, Lefrancois L.
Preferential localization of effector memory cells in nonlymphoid tissue.
Science.
2001;291:2413-2417
© 2002 by The American Society of Hematology.
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  B. Morales-Aza, S. J. Glennie, T. P. Garcez, V. Davenport, S. L. Johnston, N. A. Williams, and R. S. Heyderman Impaired maintenance of naturally acquired T-cell memory to the meningococcus in patients with B-cell immunodeficiency Blood, April 30, 2009; 113(18): 4206 - 4212. [Abstract] [Full Text] [PDF]
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 M. R. Ehrenstein and C. Mauri If the treatment works, do we need to know why?: the promise of immunotherapy for experimental medicine J. Exp. Med., October 1, 2007; 204(10): 2249 - 2252. [Abstract] [Full Text] [PDF]
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 C. Barbey, E. Pradervand, N. Barbier, and F. Spertini Ex Vivo Monitoring of Antigen-Specific CD4+ T Cells after Recall Immunization with Tetanus Toxoid Clin. Vaccine Immunol., September 1, 2007; 14(9): 1108 - 1116. [Abstract] [Full Text] [PDF]
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 S. S. Neelapu and L. W. Kwak Vaccine Therapy for B-Cell Lymphomas: Next-Generation Strategies Hematology, January 1, 2007; 2007(1): 243 - 249. [Abstract] [Full Text] [PDF]
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 J. Bayry, S. Lacroix-Desmazes, V. Donkova-Petrini, C. Carbonneil, N. Misra, Y. Lepelletier, S. Delignat, S. Varambally, E. Oksenhendler, Y. Levy, et al. Natural antibodies sustain differentiation and maturation of human dendritic cells PNAS, September 28, 2004; 101(39): 14210 - 14215. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
T cells, which promptly produced cytokines in
response to antigen (Ag), and memory-resting CD27+ T cells,
which required Ag restimulation to perform their functions, were
maintained in both XLA patients and controls for up to 24 months after
the last vaccination boost. These data strongly suggest that B
cells are not an absolute requirement for the generation of effective
T-cell memory in humans, nor do they seem to influence TH1/TH2 balance.
(Blood. 2002;99:2131-2137)
