| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Original Articles |
Department of Medicine, University of Sheffield, Clinical Sciences Center, Northern General Hospital (R.A.M., R.S.M., A.P.W.), Sheffield, United Kingdom S5 7AU; and the Department of Immunology, Imperial College School of Medicine, Hammersmith Hospital (F.M.-B., G.L., R.L.), London, United Kingdom W12 0NN
Address all correspondence and requests for reprints to: Prof. A. P. Weetman, Department of Medicine, University of Sheffield Clinical Sciences Center, Northern General Hospital, Sheffield, United Kingdom S5 7AU
Abstract
Thyroid follicular cells (TFC) are a common target of autoimmune
attack, but the role they play in inciting and maintaining this attack
is unclear. TFC express cytokines, adhesion molecules, and class I and
II major histocompatibility complex molecules, but without additional
signals that costimulate T cells, they may down-regulate, rather than
stimulate, T cell function. In this report, we have investigated
whether TFC can express the CD40 molecule, which plays a crucial role
in the reciprocal two-way communication between T and B cells. We have
shown by immunohistochemistry and flow cytometry that CD40 is expressed
by TFC in vivo and in vitro in both
autoimmune and nonautoimmune glands. CD40 expression was up-regulated
by interleukin-1
and interferon-
, but not by TSH. Although there
was no significant effect of CD40 ligation on cAMP synthesis or
[3H]thymidine incorporation, there was a significant
increase in interleukin-6 release by TFC. Thus, although TFC do not
express members of the B7 family of T cell costimulators, they do
express CD40, indicating the possibility of mutually stimulatory T
cell-TFC interaction. This has important implications, both for TFC
synthesis of immunological mediators and for the biasing of T cell
behavior toward a T helper 2-type phenotype.
THE CD40 antigen and its ligand (CD40L; also called CD154 and gp39) are one of several receptor-ligand pairs crucial for immune function and, in particular, humoral immune responses (reviewed in Refs. 1–4). CD40 is expressed by a number of cell types, principally cells of the immune system, but also thymic epithelial cells, proximal tubule epithelial cells, endothelial cells, keratinocytes, fibroblasts, and synoviocytes (2, 3, 5, 6, 7, 8, 9, 10). CD40 expressed by nonimmune cell types can interact functionally with T cell-expressed CD40L (2, 3, 4, 6, 9). CD40L is expressed principally on activated CD4+ T cells and is up-regulated in response to T cell receptor signaling (2, 3, 4). CD40L/CD40 signaling mediates T cell maturation (11, 12), enhances cytokine production (13), and induces expression of other costimulatory molecules on antigen-presenting cells (4, 14, 15, 16).
The role that thyroid follicular cells (TFC) have in inducing and maintaining thyroid autoimmunity is currently contentious (reviewed in Refs. 17 and 18). TFC express class I and class II human leukocyte antigens (HLA), which led to the hypothesis that this class II expression might induce autoimmunity (19). Alternatively, it may down-regulate the autoimmune response, because TFC may not provide necessary costimulatory signals (18). TFC express intercellular adhesion molecule-1 (ICAM-1; CD54) and leukocyte function-associated antigen-3 (CD58; reviewed in Ref.17), which can both play a role in T cell activation (1). However, they do not express B7 (B7–1, CD80) (20, 21, 22, 23) or B7–2 (CD86) (21, 22) (Metcalfe, R. A., R. S. McIntosh, and A. P. Weetman, manuscript in preparation), which play a more important role in T cell costimulation (24). An immunohistological survey of tissue CD40 expression identified positive staining in human thyroid, but without identifying the cells responsible (25), and CD40 has also been reported on thyroid endothelial cells (6). Recently, CD40 expression has been reported on TFC juxtaposed to lymphoid infiltrates (23). We have, therefore, investigated more closely CD40 expression on TFC, including the effects of TSH, cytokines, and antithyroid drugs on CD40 expression and the effect of CD40 ligation on TFC function.
Subjects and Methods
Patient details
Samples of thyroid tissue were obtained from patients undergoing subtotal thyroidectomy with Graves’ disease (GD; n = 14), Hashimoto’s thyroiditis (HT; n = 2), and multinodular goiter (MNG; n = 12). All patients were women except for two male patients each with GD and MNG. In all cases, diagnosis was confirmed by thyroid histology. All but one patient with GD had received antithyroid drugs before surgery, and the two patients with HT were taking T4 at the time of surgery.
Cytokines and other modulators
Cytokines and modulators used in the study were bovine TSH
(preparation 53/11, National Institute for Biological Standards and
Control, Hertfordshire, UK), interleukin-1 (IL-1; provided by
Hoffmann-La Roche, Nutley, NJ), tumor necrosis factor- (TNF;
Calbiochem, Nottingham, UK), interferon- (IFN; Boehringer
Mannheim, East Sussex, UK), methimazole, and propylthiouracil (both
from Sigma Chemical Co., Dorset, UK).
Preparation and culture of human thyroid cells and cell lines
TFC were prepared from thyroidectomy specimens by collagenase/dispase digestion as described previously (26), and semiintact follicles were cultured overnight in 75-cm2 tissue culture flasks (Costar Corp., Cambridge, MA) in RPMI 1640 medium (Life Technologies, Paisley, UK) supplemented with 10% FCS (Labtech International, East Sussex, UK), 2 mmol/L L-glutamine, 100 U/mL penicillin, 100 U/mL streptomycin, 1.25 µg/mL amphotericin, and 50 µg/mL neomycin (all from Life Technologies). The adherent TFC were washed and detached from the flasks using trypsin-ethylenediamine tetraacetate (trypsin-EDTA; Life Technologies), washed in medium, and counted. TFC were then plated out immediately for assay or stored in liquid nitrogen. The human thyroid cell line HTori3 was cultured as previously described (27).
Immunohistochemistry
Samples of tissue were snap-frozen in liquid nitrogen for sectioning. Frozen sections were washed in phosphate-buffered saline (PBS) and quenched using 3% hydrogen peroxide for 5 min at room temperature. After washing, the sections were incubated with isotype-matched negative control, anti-CD40 (clone EA-5), or anti-HLA-ABC (positive control) monoclonal antibodies (mAb; all from Serotec, Oxford, UK) diluted 1:50 in PBS for 1 h at room temperature. The derivation and reactivity of the EA-5 anti-CD40 mAb have been previously described; in particular, this antibody stimulates CD40 signaling (28, 29). After washing, sections were developed using a LSAB2 peroxidase kit (Dako, High Wycombe, UK), counterstained using Harris’s hematoxylin, washed, dehydrated, cleared with xylene, and mounted using DPX mountant (Raymond A Lamb, London, UK). TFC staining was divided into three categories: strong, positive, and weak, with strong being staining equivalent to the positive control mAb, weak showing more staining than the negative control mAb but insufficient to be considered unequivocally positive, and positive being intermediate staining. The observer was blinded as to the identity of, and the mAb used in, each section.
Flow cytometry
After incubation with modulators, TFC were detached from the flasks using a HEPES-EDTA buffer (10 mmol/L HEPES, EDTA 2 mg/mL in PBS), washed in PBS, and resuspended in PBS-1% BSA. One hundred microliters of this cell suspension were stained with 10 µL of either anti-CD40:R-phycoerythrin (RPE; clone B-B20), anti-HLA-ABC:fluorescein isothiocyanate (FITC), anti-HLA-DP+DR+DQ:RPE (all from Serotec), or anti-ICAM1:FITC (R&D Systems, Abingdon, UK) mAb. B-B20 mAb was used for flow cytometric studies, as it was available directly conjugated. Thyroid peroxidase (TPO) was detected using previously described anti-TPO Fab fragments (30) and murine anti-Fab:FITC (Sigma). The staining was controlled using isotype-matched FITC- and RPE-conjugated negative control mAb (Serotec). After incubation for 30 min on ice, the cells were washed twice with PBS and analyzed using a FACScan running CellQuest acquisition and analysis software (both from Becton Dickinson, Oxford, UK). Gating for TFC was carried out using forward and side scatter parameters. Results were analyzed as the percentage of cells staining positive compared to those staining for the negative control mAb, and, as an indication of the intensity of staining, as the peak channel number.
cAMP assay
TFC were plated out into 24-well tissue culture plates at 2 x 104 cells/well in RPMI 1640 medium supplemented as described above for 3 days. Two alternative methods of labeling cAMP were used. For preloading, the cells were incubated for 2 h at 37 C with medium containing 37 kilobecquerels (kBq) [3H]adenine (DuPont, Hertfordshire, UK), washed with PBS, and incubated in medium containing modulators for an additional 1–3 days. For postloading, TFC were incubated with medium containing modulators for 1–3 days and then incubated for 2–5 h with 37 kBq [3H]adenine. For both methods, the medium was removed, and 100 µL cold (-20 C) ethanol were added to each well. The medium and cell extract were stored at -20 C or assayed immediately for cAMP content (31). For cAMP assay, 900 µL nucleotide carrier mixture containing 1 mmol/L adenine, ADP, and ATP (all from Sigma), and 1 mmol/L AMP and cAMP (both from Boehringer Mannheim) were added to each sample. Samples were then applied to a neutral alumina (Sigma) column prewashed with 10 mL 5 mmol/L hydrochloric acid, washed through with 10 mL 5 mmol/L hydrochloric acid, and allowed to drain completely. Bound cAMP was eluted with 6 mL 0.1 mol/L ammonium acetate and counted using Ultima Gold XR scintillation fluid (Canberra Packard, Berkshire, UK).
[3H]Thymidine incorporation
TFC were plated out into 24-well tissue culture plates at 2 x 104 cells/well in RPMI 1640 medium supplemented as described above for 3 days. The medium was replaced with fresh medium plus isotype-matched control mAb, anti-CD40 mAb (clone EA-5), or TSH. The cells were cultured for an additional 3 days with the addition of 18.5 kBq [3H]thymidine (Amersham International, Aylesbury, UK) for the final 18 h. The medium was then removed, and the cells were washed twice with ice-cold PBS and twice with ice-cold 10% trichloroacetic acid. The cells were then solubilized with 1 mol/L sodium hydroxide at 37 C and counted using Ultima Gold XR scintillation fluid.
IL-6 ELISA
IL-6 was detected using a sandwich ELISA as previously described (32). Briefly, ELISA plates were coated with antihuman IL-6 mAb (2.3 µg/mL; 50 µL/well; clone 8, Eurogenetics UK, London, UK), blocked, and washed, and samples and standards (recombinant human IL-6, Boehringer Mannheim) were added. Bound IL-6 was detected with a biotinylated polyclonal anti-human IL-6 (diluted 1:500; Eurogenetics), followed by streptavidin-conjugated alkaline phosphatase (1:1000; Amersham) and phosphatase substrate (Sigma 104 substrate). The sensitivity of the assay was 30 U/mL (300 pg/mL) IL-6.
Statistics
Statistical analysis was carried out using Student’s paired t tests.
Results
Immunohistochemistry
Immunohistochemistry using the EA-5 anti-CD40 and control mAb was
carried out on samples from 12 patients with GD and 12 patients with
MNG. Representative sections are shown in Fig. 1
. Strong staining of TFC was observed in
sections from 2 of the 12 GD patients, 1 of whom was untreated with
antithyroid drugs between the times of diagnosis and surgery, and from
2 of the 12 MNG patients. Positive staining of TFC was observed in
sections from 3 of 12 GD patients and from 7 of 12 MNG patients, and
weak staining in sections from 7 of 12 GD and 3 of 12 MNG patients.
Immunohistochemistry could not be carried out on sections from patients
with HT because of lack of available tissue.
|
Initial studies were carried out to evaluate the best procedures
for detection of CD40 on TFC by flow cytometry. Three methods were
compared for removing adherent cells from flasks, trypsin-EDTA, versene
(Life Technologies), and HEPES-EDTA, indicating that HEPES-EDTA gave
stronger and more consistent signals. Comparison of TFC analyzed
directly from liquid nitrogen stocks and after a 3-day period of
culture indicated that the latter gave more consistent results,
possibly as a result of loss of cell surface antigens after proteolytic
enzyme digestion (33, 34). Dual staining of TFC cultures with anti-TPO
Fab fragments indicated that 60–80% of CD40+ events were
also TPO+ (Fig. 2).
|
). Representative flow
cytometry profiles are shown in Fig. 3. A
large number of TFC (>40%) stained with the B-B20 mAb in six of seven
GD cultures, two of two HT cultures, and two of two MNG cultures (Table 1). The peak channel number indicated that the staining was relatively
weak (Table 1).
|
|
CD40 expression was also confirmed in the immortalized human thyroid
cell line HTori3. HTori3 cells stained strongly positive for CD40 (Fig. 4), with 91.2% of the cells present
expressing CD40 and a peak channel number (of 43) comparable to that in
primary cultured TFC (Table 1
).
|
TFC were treated with TSH or the cytokines IL-1 and IFN to
assess potential mechanisms responsible for the in vivo
control of CD40 expression. Modulators were added to 3-day cultures
either at a single concentration (Table 1) or at a range of
concentrations. The percentage of TFC staining and peak channel number
were not significantly affected by 1 mU/mL TSH, but were significantly
increased by 100 U/mL IL-1 and 100 U/mL IFN (Table 1). Over a
range of concentrations, TSH (0.1–10 mU/mL; n = 4; Fig. 5), IL-1 (1–100 U/mL; n = 4;
Fig. 5), and TNF (0.1–10 ng/mL; n = 2) had no significant
effect on the number of TFC expressing CD40. IFN (n = 4) caused
significantly increased CD40 expression at 10 U/mL (P =
0.024) and 100 U/mL (P = 0.005), but not at 1 U/mL.
Peak channel number was significantly increased by 10 U/mL IL-1
(P = 0.010), 10 and 100 U/mL IFN (P
= 0.035 and 0.047, respectively), and significantly decreased by 10
mU/mL TSH (P = 0.047), but was otherwise not
significantly affected.
|
(Fig. 4). Staining with a positive control mAb, specific for
HLA-DR, was negligible using untreated HTori3, but was strongly
positive with IFN-treated HTori3 (Fig. 4). Effects of antithyroid drugs on CD40 expression
Both the direct effects of methimazole and propylthiouracil on
CD40 expression in unstimulated 3-day TFC cultures and the effects of
the drugs on IL-1- or IFN-stimulated up-regulation of CD40
expression were investigated. Neither drug caused a significant
difference in CD40 expression, either as a percentage of the TFC
stained (untreated: mean, 71.7%; SD, 14.5%;
10-4 mol/L methimazole: mean, 68.8%; SD,
14.4%; 10-3 mol/L methimazole: mean, 71.4%;
SD, 13.3%; 10-4 mol/L propylthiouracil: mean,
68.8%; SD, 15.7%; 10-3 mol/L
propylthiouracil: mean, 65.1%; SD, 15.5%; n = 4 for
all cultures) or as peak channel number (data not shown). TFC were also
cultured with the antithyroid drugs for 4–5 days, with the final 3–4
days of culture in the presence of 10 U/mL IL-1 or IFN. Neither
drug showed any significant effect on CD40 expression (n = 3; data
not shown).
Effect of CD40 stimulation on cAMP synthesis and [3H]thymidine incorporation
To study the cAMP response in TFC after CD40 ligation, TFC were
cultured for 1–3 days untreated or treted with TSH or IFN, with a
range of concentrations (0–1 µg/mL) of the EA-5 anti-CD40 mAb. After
the addition of [3H]adenine to label cAMP, both
intracellular and extracellular cAMP were measured (postlabeled; n
= 3). Alternatively, cells were labeled with [3H]adenine
and cultured for 1–3 days with modulators before analyzing
intracellular and extracellular cAMP (prelabeled; n = 6).
There was no clear relationship between the dose of EA-5 in the culture
medium and the amount of cAMP generated (Table 2). When the two labeling methods were
analyzed together, there was a significant shift in intracellular cAMP
levels only with the highest concentration (1 µg/mL) of EA-5 in both
the presence and absence of IFN (P = 0.045 and
0.029, respectively); no significant change was observed in the
corresponding extracellular cAMP levels. When analyzed separately,
there were no significant changes in levels of cAMP in response to any
of the modulators.
|
were added to parallel sets of cultures (n = 2). The
EA-5 mAb showed no statistically significant effect in either the
presence or absence of the other modulators (data not shown). CD40-induced IL-6 expression
IL-6 release from TFC is stimulated by a number of modulators
(reviewed in Ref.35). Analysis was, therefore, performed of the effect
of TFC CD40 ligation on the release of IL-6 by TFC. EA-5 mAb (0, 0.01,
0.1, and 1 µg/mL) was added to 3-day TFC cultures with or without the
addition of 1 mU/mL TSH, 100 U/mL IL-1, or 100 U/mL IFN (Table 3). IL-6 release was significantly
increased with the highest concentration of EA-5 (but not by 1 µg/mL
isotype-matched control mAb) in the absence of other modulators and in
the presence of TSH and IFN, but not in the presence of IL-1,
which itself caused a large increase in IL-6 synthesis (Table 3).
|
We have shown that TFC express CD40 and that its expression is
increased by IL-1 and IFN, but that TSH and antithyroid drugs had
little or no effect on expression. In addition, TFC CD40 signals
functionally, significantly increasing IL-6 release, but not inducing
the expression of HLA class I, class II, or ICAM-1 (data not shown).
Four pieces of information suggest that CD40 expression is on TFC and
not contaminating lymphoid cells. Firstly, although primary TFC
cultures are known to be contaminated with small numbers of lymphoid
cells, this typically amounts to less than 1% in our preparations, and
we consistently observed CD40 expression on a far greater number of
cells than this. Secondly, lymphoid cells and TFC have different
forward scatter/side scatter profiles in flow cytometry, and the
CD40+ population was from the region typical of TFC.
Thirdly, we observed clear positive staining of TFC in thyroid
sections. Fourthly, dual staining with antihuman TPO and anti-CD40
showed CD40 staining to be predominantly on TPO+ cells. We
also observed strong staining of HTori3 cells, which, being
immortalized, cannot be contaminated with lymphocytes. However, being
an immortalized cell line, the pattern of expression may not reflect
that shown by primary TFC. In addition, significant contamination of
cultures with fibroblasts, known to express CD40, was not revealed
during routine microscopic examination or by absence of staining with
anti-TPO Fab.
We have detected CD40 expression on TFC by both immunohistochemistry and flow cytometry. CD40 expression was detectable by immunohistochemical analysis in all thyroid sections analyzed, whether from patients with clinical autoimmune disease or MNG. There was no discernible correlation between clinical disease state and CD40 expression in vitro as detected by flow cytometry, although the lowest percentage of unstimulated expression was seen on TFC from GD patients.
We have shown that TFC up-regulate CD40 expression after culture with
IL-1 and IFN, but that TNF and TSH have little effect on
levels of expression. Both IL-1 and IFN are found in thyroid
tissue from patients with autoimmune thyroid disease, although there
are also data suggesting their absence in some patients (reviewed in
Ref.35). CD40 expression on other nonimmune cell types is typically
increased by IL-1, TNF, and IFN, although responses to
different cytokines are heterogeneous (2, 5, 8, 9, 10). All nonimmune cell
types tested respond by up-regulating CD40 expression in response to
IFN. CD40 expression was also increased in two cultures by treatment
of TFC with PMA (data not shown). PMA has been previously reported to
enhance the potency of TFC in presenting antigen to T cells (36).
However, it is unlikely that CD40, already present without PMA
stimulation, is a candidate for the factor causing PMA-enhanced
presentation.
The weakest staining for CD40 by both immunohistochemical and flow cytometric analysis came from GD patients. This may be related in part to the common use of antithyroid drug treatment of patients with GD in preparation for surgery, although we could not detect any significant direct effect of antithyroid drugs on CD40 expression. Indirect effects may, nevertheless, occur in vivo, with the reduced lymphocytic infiltrate resulting in reduced exposure of TFC to cytokines, leading to reduced CD40 expression.
CD40 signaling is induced by binding to the trimeric CD40L complex,
inducing multimerization of the CD40 antigen; this is mimicked by
several of the mAb reactive to CD40 (3). Although the most important
effects of CD40 are probably modulated by tyrosine kinases and
transcription factor activation, CD40 ligation can also lead to
activation of protein kinase A and increased intracellular cAMP (2, 3, 4).
We observed both increases and decreases in the accumulation of cAMP in
TFC in response to CD40 ligation, with no statistically significant
change other than for increased intracellular cAMP levels with the
highest concentration of anti-CD40 mAb in unstimulated and
IFN-stimulated cultures. In addition, CD40 ligation did not lead to
any significant change in the [3H]thymidine incorporation
by TFC. CD40 stimulation of synoviocytes and fibroblasts leads to
increased proliferation (2, 7, 8), whereas in keratinocytes, CD40
signaling induces differentiation (10).
We have shown that the highest levels of the EA-5 anti-CD40 mAb induced
an increased level of IL-6 synthesis, except in the presence of
IL-1, which itself stimulates IL-6 synthesis. Although this level of
CD40 stimulation may be greater than that present in vivo,
IL-6 synthesis by TFC is up-regulated by a number of modulators (35),
and in vivo these presumably synergize in the control of
IL-6 synthesis. CD40-induced IL-6 synthesis has been reported in
several immune and nonimmune cell types (2, 4, 9, 37). In addition to
IL-6, TFC have been implicated in the production of a number of other
cytokines: for example, IL-1, IL-8, IL-12, and TNF (35).
Secretion of IL-8, TNF, and other cytokines has been reported in
CD40-stimulated keratinocytes, dendritic cells, and monocytes (4, 10),
and CD40-induced TFC cytokine production may, therefore, extend beyond
IL-6.
There is substantial evidence for the involvement of the CD40/CD40L signaling pathway in both the development and the perpetuation of autoimmune disease (reviewed in Ref.38). In an animal model of GD, using severe combined immunodeficient (SCID) mice with GD thyroid explants, anti-CD40L-treated mice had fewer lymphocytes resident in the gland and significantly reduced levels of thyroid-reactive antibodies and TFC ICAM-1 expression (39). Similarly, in a thyroglobulin/adjuvant-induced model of thyroiditis, treatment with anti-CD40L greatly reduced the severity of the thyroiditis and the induction of antithyroglobulin IgG (40). Although these models are probably detecting the effects of reduced CD40/CD40L signaling between T and B cells, they nevertheless indicate the potential importance of this pathway in initiating and perpetuating thyroid autoimmune responses.
There is a strong correlation between IFN synthesis and class II
expression in the thyroid (17, 26, 35), and our results suggest that
these in vivo IFN-stimulated TFC should also express
enhanced amounts of CD40, as confirmed recently (23). However, in
postulating a role for CD40 in thyroid autoimmunity, account must be
taken of the high level of CD40 detected in our MNG samples, which is
thought to represent a disease of predominantly nonautoimmune pathology
(17). Although CD40 was detected in all of our MNG samples, albeit
weakly in many, it was not described in the majority of control samples
in another report, in which a different CD40 mAb was used, and
detection was by indirect immunofluorescence rather than
immunohistochemistry (23). As with the expression of HLA class II by
TFC (18, 19), CD40 expression may play a role in the perpetuation or
regulation of an existing autoimmune response, but appears unlikely to
be the cause of autoimmunity. Nevertheless, the CD40/CD40L signaling
pathway is directly involved in the development and perpetuation of
autoimmune responses, and functional CD40 expression on TFC may,
therefore, contribute to thyroid autoimmunity. In particular, ligation
of CD40 on TFC may result in increased synthesis of proinflammatory
cytokines such as IL-6, whereas stimulation of CD40L on intrathyroidal
T cells could result in the stimulation of T cell maturation and
alteration of cytokine synthesis patterns (22).
Acknowledgments
The authors thank Mr. B. J. Harrison and Mr. G. Jacob for
provision of surgically removed tissue, Mrs. S. K. Justice, Mrs.
R. Davies, Mrs. L. Fleming and Miss C. Findlay for excellent technical
assistance, and Hoffmann-La Roche for providing IL-1.
Footnotes
1 This work was supported by the Wellcome Trust. 
2 Current address: Division of Molecular and Cellular Immunology,
Department of Clinical Laboratory Sciences, Floor A, West Block,
Queen’s Medical Center, Nottingham, United Kingdom NG7 2UH.
Received September 8, 1997.
Revised December 5, 1997.
Accepted December 31, 1997.
References
anti-thyroid peroxidase antibodies from different
tissues in Hashimoto’s thyroiditis. J Clin Endocrinol Metab. 82:3818–3825.B
mobilisation and IL-6 production. J Immunol. 55:4588–4595.
This article has been cited by other articles:
 |
|
 |
|
  E. C. Ridgway, Y. Tomer, and S. M. McLachlan Update in Thyroidology J. Clin. Endocrinol. Metab., October 1, 2007; 92(10): 3755 - 3761. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. G. Gianoukakis, R. S. Douglas, C. S. King, W. W. Cruikshank, and T. J. Smith Immunoglobulin G from Patients with Graves' Disease Induces Interleukin-16 and RANTES Expression in Cultured Human Thyrocytes: A Putative Mechanism for T-Cell Infiltration of the Thyroid in Autoimmune Disease Endocrinology, April 1, 2006; 147(4): 1941 - 1949. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. M. Jacobson, E. Concepcion, T. Oashi, and Y. Tomer A Graves' Disease-Associated Kozak Sequence Single-Nucleotide Polymorphism Enhances the Efficiency of CD40 Gene Translation: A Case for Translational Pathophysiology Endocrinology, June 1, 2005; 146(6): 2684 - 2691. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Kimura, M. Kimura, S.-C. Tzou, Y.-C. Chen, K. Suzuki, N. R. Rose, and P. Caturegli Expression of Class II Major Histocompatibility Complex Molecules on Thyrocytes Does Not Cause Spontaneous Thyroiditis but Mildly Increases Its Severity after Immunization Endocrinology, March 1, 2005; 146(3): 1154 - 1162. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-S. Li, N. Kanamoto, Y. Hataya, K. Moriyama, H. Hiratani, K. Nakao, and T. Akamizu Transgenic Mice Producing Major Histocompatibility Complex Class II Molecules on Thyroid Cells Do Not Develop Apparent Autoimmune Thyroid Diseases Endocrinology, May 1, 2004; 145(5): 2524 - 2530. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. G. Gianoukakis, L. J. Martino, N. Horst, W. W. Cruikshank, and T. J. Smith Cytokine-Induced Lymphocyte Chemoattraction from Cultured Human Thyrocytes: Evidence for Interleukin-16 and Regulated upon Activation, Normal T Cell Expressed, and Secreted Expression Endocrinology, July 1, 2003; 144(7): 2856 - 2864. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Bourcier, Magda De Saint-Jean, F. Brignole, A. Goguel, and C. Baudouin Expression of CD40 and CD40 Ligand in the Human Conjunctival Epithelium Invest. Ophthalmol. Vis. Sci., January 1, 2000; 41(1): 120 - 126. [Abstract] [Full Text]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Endocrinology | Endocrine Reviews | J. Clin. End. & Metab. |
| Molecular Endocrinology | Recent Prog. Horm. Res. | All Endocrine Journals |
