bluuebunny/crossref_metadata_2025 huggingface dataset:
Inflammatory cytokines in small intestinal mucosa of patients with potential coeliac disease
Authors: WESTERHOLM-ORMIO M , GARIOCH J , KETOLA I , SAVILAHTI E
Published: March 2003

T helper cell type 1 (Th1) response to gluten has been implicated in the pathogenesis of coeliac disease (CD). To characterize immunological activation and mild inflammations leading to overt CD in potential coeliac patients, jejunal biopsies were obtained from family members of patients with CD or dermatitis herpetiformis (DH). Nine family members and one latent CD, eight CD patients and eight normal controls furnished jejunal biopsy specimens. Immunohistochemical staining of sections for interleukin-1α (IL-1α), IL-2, IL-4, interferon-γ (IFN-γ), tumour necrosis factor α (TNF-α), CD3, γδ-T cell receptor (γδ-TCR), and αβ-TCR was carried out with monoclonal antibodies. Further, expression of IL-4 and IFN-γ messenger RNA was detected by radioactive in situ hybridization in these same samples. In lamina propria, CD patients and potential CD patients had higher densities of IL-2 (P = 0·028, P = 0·043), IL-4 (P = 0·021, P = 0·034) and IFN-γ positive cells (P = 0·000, P = 0·009) than did controls. Moreover, CD patients showed a higher density of TNF-α positive cells (P = 0·012, P = 0·001) than the other two groups, and expression of IFN-γ mRNA (P = 0·035) was higher in them than in the other two study groups. Additionally, higher densities of TNF-α and IFN-γ positive cells occurred in potential CD patients with high γδ-TCR+ intraepithelial lymphocytes (IELs). Our findings support the hypothesis that lamina propria T cells and macrophages, through their secretion of cytokines, play a central role in the pathogenesis of coeliac disease. The inflammatory cytokines found in potential CD specimens strongly suggest that these inflammatory markers can be identified long before visible villous changes have occurred.
Effect of CD14 promoter polymorphism andH. pyloriinfection and its clinical outcomes on circulating CD14
Authors: KARHUKORPI J , YAN Y , NIEMELÄ S , VALTONEN J , KOISTINEN P , JOENSUU T , SAIKKU P , KARTTUNEN R
Published: March 2003

CD14 is a pattern recognition receptor on the membranes of monocytes and macrophages for several microbial products, of which lipopolysaccharide (LPS) is the best known. A shed form of CD14 is present in serum. As the CD14 gene promoter polymorphism –159C/T and some bacterial infections may affect the sCD14 levels, we compared the impact of both the CD14 promoter polymorphism and Helicobacter pylori infection on serum sCD14 levels in 201 dyspeptic patients (group 1) who had undergone gastroscopy, and 127 staff members (group 2) with no endoscopy. sCD14 was measured from the sera by a commercial enzyme immunoassay (EIA), and CD14 genotyping was carried out with PCR. Helicobacter pylori infection was detected by serology and/or culture or PCR. sCD14 levels were elevated in the subjects carrying the T allele (CT or TT genotype) in both groups when compared with subjects with the CC genotype. Overall, H. pylori-positive subjects tended to have higher sCD14 levels compared with H. pylori-negative subjects. In group 1 consisting of dyspeptic patients, those with gastric ulcer, gastric erosion or duodenal ulcer had significantly elevated levels of sCD14 compared with the patients with normal endoscopic findings or macroscopic gastritis. The recent use of NSAIDs was also associated with enhanced sCD14. Thus, we were able to show several factors, one genetic and the other environmental (H. pylori infection and mucosal lesion), to have an impact on sCD14.
Macrophage Inflammatory Protein 3α–CC chemokine receptor 6 interactions play an important role in CD4+T-cell accumulation in periodontal diseased tissu
Authors: HOSOKAWA Y , NAKANISHI T , YAMAGUCHI D , TAKAHASHI K , YUMOTO H , OZAKI K , MATSUO T
Published: March 2003

The regulatory role of chemokines and chemokine receptors on specific lymphocyte recruitment into periodontal diseased tissue is poorly characterized. We observed that lymphocytes infiltrating inflamed gingival tissue expressed marked levels of CCR6. In periodontal diseased tissue, the expression of MIP-3α mRNA was detected by RT-PCR and further, MIP-3α was distributed in the basal layer of gingival epithelial cells, microvascular endothelial cells and the areas of inflammatory cells as shown by immunohistochemistry. Moreover, CCR6-expressing cells infiltrated into periodontal diseased tissue, and the proportion of CCR6-positive CD4+ T cells was significantly elevated in periodontal diseased tissue compared with peripheral blood in the same patients. Furthermore, gingival lymphocytes isolated from patients showed migration toward MIP-3α in an in vitro chemotaxis assay in which migration was abrogated by specific antibody to CCR6. Thus, these findings suggested that CCR6 and the corresponding chemokine, MIP-3α may have an important regulatory role in specific lymphocyte migration into inflamed periodontal tissue.
Generation of a recombinant Fab antibody reactive with the Alzheimer's disease-related Aβ peptide
Authors: TAMMER A H , COIA G , CAPPAI R , FULLER S , MASTERS C L , HUDSON P , UNDERWOOD J R
Published: March 2003

A recombinant Fab antibody, designated 1E8–4b, which reacts with the Alzheimer's disease (AD)-related Aβ peptides, Aβ[1–40], Aβ[1–42] and Aβ[1–43] has been developed. The 1E8–4b Fab was constructed by cloning the VHCH1 and VLCL domains from the parent hybridoma 1E8 antibody, reported previously to recognize these Aβ peptides. Briefly, a C-terminal Flag tag sequence was incorporated into this construct, which was ligated into the vector pHFA2 and expressed in Escherichia coli. Following purification on an M2 anti-Flag affinity column, the 1E8–4b recombinant Fab antibody was shown to bind plaques within sections of brain tissue from CERAD-defined AD patients by immunohistochemistry. ELISA, epitope mapping and immunoblotting confirmed the recognition of the Aβ1–40/42/43] peptides by the 1E8–4b Fab. The 1E8–4b Fab did not recognize APP695 or APP770 which contain the Aβ sequence. The Aβ specificity of the recombinant 1E8–4b Fab antibody was identical to the parent 1E8 monoclonal antibody.
Fractalkine expression on human renal tubular epithelial cells: potential role in mononuclear cell adhesion
Authors: CHAKRAVORTY S J , COCKWELL P , GIRDLESTONE J , BROOKS C J , SAVAGE C O S
Published: March 2003

Fractalkine (CX3CL1) is a transmembrane molecule with a CX3C chemokine domain attached to an extracellular mucin stalk which can induce both adhesion and migration of leucocytes. Mononuclear cell infiltration at renal tubular sites and associated tubular epithelial cell damage are key events during acute renal inflammation following renal allograft transplantation. Using northern and Western blot analysis, we have demonstrated the expression of fractalkine message and protein by renal tubular epithelial cells in vitro. The expression was up-regulated by TNF-α, a key proinflammatory cytokine in acute rejection. Investigation of surface expression of fractalkine on cultured proximal tubular epithelial cells revealed only a subpopulation of positively staining cells. Immunohistochemistry revealed that only a proportion of tubules in renal allograft biopsies showed induction of fractalkine expression. Studies using a static model of adhesion demonstrated CX3CR1/fractalkine interactions accounted for 26% of monocytic THP-1 cell and 17% of peripheral blood natural killer cell adhesion to tubular epithelial cells, suggesting that fractalkine may have a functional role in leucocyte adhesion and retention, at selected tubular sites in acute renal inflammation. Thus, fractalkine blockade strategies could reduce mononuclear cell mediated tubular damage and improve graft survival following kidney transplantation.
Respiratory syncytial virus induces prostaglandin E2, IL-10 and IL-11 generation in antigen presenting cells
Authors: BARTZ H , BÜNING-PFAUE F , TÜRKEL Ö , SCHAUER U
Published: March 2003

Bronchiolitis caused by respiratory syncytial virus (RSV) infection is a major cause of hospitalization in children under 1 years of age. The disease characteristically does not induce protective immunity. However, a mononuclear peribronchiolar and perivascular infiltrate during RSV infection is suggestive of an immune-mediated pathogenesis. Macrophages and dendritic cells (DCs) play an essential role in the initiation and maintenance of immune response to pathogens. To analyse interactions of RSV and immune cells, human cord blood derived macrophages and dendritic cells were infected with RSV. Both cells were found to be infected with RSV resulting in the activation of macrophages and maturation of dendritic cells as reflected by enhanced expression of several surface antigens. In the next set of experiments, generation of mediators was compared between cells infected with RSV, parainfluenza (PIV3) and influenza virus as well as ultracentrifuged virus free supernatant. Whereas the supernatant did not induce release of mediators, all three live virus infections induced IL-6 production from macrophages and DC. Influenza virus infection induced predominantly IL-12 p75 generation in DC. In contrast, RSV induced strong IL-11 and prostaglandin E2 release from both macrophages and DCs. In addition, RSV but not influenza and parainfluenza virus induced a strong IL-10 generation particularly from macrophages. Since IL-10, IL-11 and PGE2 are known to act immunosuppressive rather than proinflammatory, these mediators might be responsible for the delayed protective RSV specific immune response.
Treatment of oral pemphigoid with intravenous immunoglobulin as monotherapy. Long-term follow-up: influence of treatment on antibody titres to human α6 integrin
Authors: SAMI N , BHOL K C , AHMED A R
Published: March 2003

Oral pemphigoid (OP) is a chronic autoimmune disease, involving the oral cavity, characterized by a homogenous linear deposition of immunoglobulins, complement, or both along the basement membrane zone (BMZ) and a subepithelial blister formation. The α6/β4 heterodimer is an integrin family of adhesion receptors, which mediates basal cell to matrix interactions. Recent evidence suggests a pathophysiologic role for antibodies against human α6 integrin in blister formation in OP, in organ culture studies. Fifty percent of OP patients have been reported to experience disease progression to involve other mucosal tissues, including the eye and larynx. To prevent this extension of disease, systemic therapy with systemic corticosteroids, dapsone, and immunosuppressive agents has been recommended. The use of intravenous immunoglobulin (IVIg) in the treatment of pemphigoid has been recently described. In this study, we present the use of IVIg, in a group of seven patients, with severe OP, in whom systemic conventional treatment was contraindicated. To determine the influence of treatment on antibodies to human α6 integrin in OP, seven patients with OP treated with IVIg therapy and a comparable control group of seven patients with OP, treated with conventional therapy, were evaluated at monthly intervals, for a 12 consecutive month treatment period. An effective clinical response was observed in all seven patients treated with IVIg therapy, after a mean treatment period of 4·5 months. IVIg therapy induced a prolonged and sustained clinical remission in all seven patients after a mean treatment period of 26·9 months. A statistically significant difference was observed in the quality of life pre- and post-IVIg therapy (P < 0·001). Both the study and the control groups had a very similar initial serological response to treatment. A statistically significant reduction in the antibody titres was observed after four months of treatment, in both groups (P = 0·015). Thereafter, patients treated with IVIg therapy had a faster rate of decline in the antibody titres, and the difference in the rate of decline between the study and control groups became statistically significant after six months of treatment (P = 0·03). The use of IVIg therapy resulted in reduction of antiα6 antibody titres and in inducing and maintaining both a sustained, clinical and serological remission.
Reduced virus specific T helper cell induction by autologous dendritic cells in patients with chronic hepatitis B – restoration by exogenous interleukin-12
Authors: LÖHR H F , PINGEL S , BÖCHER W O , BERNHARD H , HERZOG-HAUFF S , ROSE-JOHN S , GALLE P R
Published: March 2003

Insufficient stimulatory capacities of autologous dendritic cells (DC) may contribute in part to impaired T cell stimulation and therefore viral persistence in patients with chronic hepatitis B virus (HBV) infection. In order to characterize the antigen presenting functions of DC from chronic HBV carriers and controls antigen specific T cell responses were analysed. CD34+ peripheral blood progenitor cells were differentiated to immature DC in the presence of GM-CSF, IL-6/IL-6R fusion protein and stem cell factor. Proliferative CD4+ T cell responses and specific cytokine release were analysed in co-cultures of DC pulsed with HBV surface and core antigens or tetanus toxoid and autologous CD4+ T cells. Cultured under identical conditions DC from chronic HBV carriers, individuals with acute resolved hepatitis B and healthy controls expressed similar phenotypical markers but chronic HBV carriers showed less frequent and weaker HBV antigen specific proliferative T helper cell responses and secreted less interferon-γ while responses to the tetanus toxoid control antigen was not affected. Preincubation with recombinant IL-12 enhanced the HBV specific immune reactivities in chronic HBV patients and controls. In conclusion, the weak antiviral immune responses observed in chronic hepatitis B may result in part from insufficient T cell stimulating capacities of DC. Immunostimulation by IL-12 restored the HBV antigen specific T cell responses and could have some therapeutical benefit to overcome viral persistence.
Exposure to Mycobacterium avium primes the immune system of calves for vaccination with Mycobacterium bovis BCG
Authors: HOWARD C J , KWONG L S , VILLARREAL-RAMOS B , SOPP P , HOPE J C
Published: March 2003

The objective of the investigation was to provide data on how a prior exposure of cattle to Mycobacterium avium, used here as a model of exposure to an environmental mycobacterium, affected the cellular immune response that follows vaccination with Mycobacterium bovis BCG. The assessment of cellular immune responses included lymphocyte proliferation assays, the delayed hypersensitivity skin test and IFN-γ synthesis in whole blood cultures. One group of calves was inoculated subcutaneously with M. avium followed 12 weeks later by M. bovis-BCG. The other group was vaccinated subcutaneously with BCG alone. Calves previously exposed to M. avium responded more rapidly, as assessed in the in vitro assays, to purified protein derivative (PPD) from M. avium (PPD-A) or M. bovis (PPD-B) than did calves inoculated with BCG only, indicating that the exposure to M. avium had primed the immune response in these calves. Following inoculation of BCG the intensity of the in vitro responses and the delayed hypersensitivity skin test to PPD-A was higher for the M. avium-primed animals while the responses to PPD-B were similar in the M. avium-primed and BCG-only groups. The results are consistent with a model in which prior exposure to environmental mycobacteria does not necessarily inhibit the immune response to the vaccine strain, BCG. They suggest that M. avium infection primes the immune system of calves and that the detection of an immune response specific for M. bovis BCG is masked by reactivity to antigens also present in M. avium.
Autoantibodies against small nucleolar ribonucleoprotein complexes and their clinical associations
Authors: VAN EENENNAAM H , VOGELZANGS J H P , BISSCHOPS L , TE BOOME L C J , SEELIG H P , RENZ M , DE ROOIJ D-J , BROUWER R , PLUK H , PRUIJN G J M , VAN VENROOIJ W J , VAN DEN HOOGEN F H J
Published: March 2003

Sera from patients suffering from systemic autoimmune diseases such as systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) have been shown to contain reactivities to nuclear components. Autoantibodies specifically targeting nucleolar antigens are found most frequently in patients suffering from SSc or SSc overlap syndromes. We determined the prevalence and clinical significance of autoantibodies directed to nucleolar RNA-protein complexes, the so-called small nucleolar ribonucleoprotein complexes (snoRNPs). A total of 172 patient sera with antinucleolar antibodies were analysed by immunoprecipitation. From 100 of these patients clinical information was obtained by chart review. Autoantibodies directed to snoRNPs were detected not only in patients suffering from SSc and primary Raynaud's phenomenon (RP), but also in patients suffering from SLE, rheumatoid arthritis (RA) and myositis (PM/DM). Antibodies against box C/D small snoRNPs can be subdivided in antifibrillarin positive and antifibrillarin negative reactivity. Antifibrillarin-positive patient sera were associated with a poor prognosis in comparison with antifibrillarin negative (reactivity with U3 or U8 snoRNP only) patient sera. Anti-Th/To autoantibodies were associated with SSc, primary RP and SLE and were found predominantly in patients suffering from decreased co-diffusion and oesophagus motility and xerophthalmia. For the first time autoantibodies that recognize box H/ACA snoRNPs are described, identifying this class of snoRNPs as a novel autoantigenic activity. Taken together, our data show that antinucleolar patient sera directed to small nucleolar ribonucleoprotein complexes are found frequently in other diseases than SSc and that categorization of diagnoses and clinical manifestations based on autoantibody profiles seems particularly informative in patient sera recognizing box C/D snoRNPs.
Murine glomerular mesangial cell uptake of apoptotic cells is inefficient and involves serum-mediated but complement-independent mechanisms
Authors: CORTES-HERNANDEZ J , FOSSATI-JIMACK L , CARUGATI A , POTTER P K , WALPORT M J , COOK H T , BOTTO M
Published: March 2003

An increased number of apoptotic bodies have been detected in glomeruli of non-nephritic kidneys of C1q-deficient mice. In these mice an in vivo impaired uptake of apoptotic cells by peritoneal macrophages was also demonstrated. Here we investigated whether C1q plays a role in the in vitro clearance of apoptotic cells by glomerular mesangial cells. Phagocytosis was assessed using a novel flow cytometric assay that was validated by immunofluorescence studies. The uptake of apoptotic cells by mesangial cells, measured as percentage of mesangial cells ingesting apoptotic cells, was ∼25%, 10% and 10% for a T cell lymphoma line (RMA), thymocytes and neutrophils, respectively. The uptake reached a plateau phase after 3 h, was specific for apoptotic cells and was mediated by serum but not by complement components C1q or C3. The phagocytosis of apoptotic cells was significantly inhibited by Arg-Gly-Asp-Ser (RGDS), a peptide capable of blocking the interaction of thrombospondin with CD36 or the vitronectin receptor. Pretreatment of the mesangial cells with dexamethasone (200 nm) but not with LPS increased the uptake markedly. These findings indicate that murine mesangial cells are capable of taking up syngeneic apoptotic cells, although much less efficiently than professional phagocytic cells. They also show that serum proteins other than complement components mediate the removal of apoptotic cells by murine mesangial cells in vitro.
Butyrate modulates intestinal epithelial cell-mediated neutrophil migration
Authors: BÖCKER U , NEBE T , HERWECK F , HOLT L , PANJA A , JOBIN C , ROSSOL S , SARTOR R B , SINGER M V
Published: March 2003

Butyrate, a short-chain fatty acid released by colonic bacteria and administered therapeutically in inflammatory bowel diseases, exerts immunomodulatory properties. The aim of the study was to determine the functional consequences of butyrate exposure on the proinflammatory responsiveness of human intestinal epithelial cells (IEC). IL-8 promoter activity in IEC pretreated with butyrate then exposed to proinflammatory stimuli was assayed by transfection of luciferase constructs. IL-8 secretion was determined by ELISA and neutrophil migration by flow cytometry. Receptor mRNA was assessed by reverse transcriptase–polymerase chain reaction (RT-PCR). Butyrate modulated proinflammatory IL-8 secretion differentially in Caco-2 and HT-29 cells on the transcriptional level. Pointing to the potentially underlying mechanism of increased IL-1β-stimulated IL-8 secretion in HT-29 cells, butyrate up-regulated IL-1RI mRNA but not IL-1RII. Butyrate pretreatment of IEC lines stimulated by IL-1β modulated neutrophil migration significantly: reduction towards Caco-2 and enhancement towards HT-29/p cells. Pharmacological inhibition of protein tyrosine phosphatases or treatment with mesalamine or sulphasalazine diminished IL-1β-stimulated IL-8 secretion by butyrate-exposed HT-29 cells substantially. Immunomodulatory effects of butyrate on IEC are functionally relevant for neutrophil migration. Pharmacological inhibition of enhanced IL-1β-mediated IL-8 secretion in a subpopulation of IEC may improve the clinical efficacy of butyrate.
Decreases in plasma TNF-αlevel and IFN-γmRNA level in peripheral blood mononuclear cells (PBMC) and an increase in IL-2 mRNA level in PBMC are associated with effective highly active antiretroviral therapy in HIV-infected patients
Authors: BRAZILLE P , DEREUDDRE-BOSQUET N , LEPORT C , CLAYETTE P , BOYER O , VILDÉ J-L , DORMONT D , BENVENISTE O
Published: March 2003

In this study, we investigated the cytokine profiles of 14 treatment-naive HIV-infected patients on the initiation of highly active antiretroviral therapy (HAART). At baseline, plasma levels of TNF-α and its mRNA in peripheral blood mononuclear cells (PBMC) were highest in the most severely immunocompromised patients (<200 CD4+ cells/mm3). After 12 months of HAART, the virus was undetectable in the plasma of all patients (<200 copies/ml), and median CD4 T cell counts had increased (+164 cells/mm3). We also observed a gradual decrease in the number of proviral DNA copies in PBMC and in immune activation, with lower levels of IFN-γ mRNA in PBMC associated with weaker activation of CD8+ T cells and lower levels of plasma TNF-α. IL-2 mRNA levels in PBMC were found to increase in parallel. The decrease in TNF-α and IFN-γ levels and the increase in IL-2 production appear to be correlated with the efficacy of HAART in naive immunocompromised HIV-infected individuals.
Increased circulating levels of proteinase 3 in patients with anti-neutrophilic cytoplasmic autoantibodies-associated systemic vasculitis in remission
Authors: OHLSSON S , WIESLANDER J , SEGELMARK M
Published: March 2003

In systemic small vessel vasculitides, patients form autoantibodies against neutrophil granular proteins, anti-neutrophilic cytoplasmic autoantibodies (ANCA). Some correlation is seen between ANCA titre and disease activity, but whether this is cause or effect is still unknown. It has been reported that levels of proteinase 3 (PR3), one of the main ANCA antigens, are increased in patients with active disease. An increased level of circulating antigen could mean a predisposition to autoimmunity. In order to explore this we measured PR3 levels in patients with stable disease. In addition we measured neutrophil gelatinase-associated lipocalin (NGAL) as a specific marker of neutrophil degranulation, cystatin C as a marker of renal function as well as C-reactive protein (CRP), IL-6 and sTNFr1 as markers of inflammation. PR3, NGAL, IL-6 and sTNFr1 were measured in plasma by the ELISA technique. In the PR3 ELISA, we used anti-PR3 monoclonal antibodies as capture-antibodies and affinity-purified rabbit-anti-PR3 antibodies for detection. PR3-ANCA, myeloperoxidase (MPO)-ANCA, CRP and cystatin C were measured by routine methods. PR3 was significantly raised (P < 0·0001) in vasculitis patients (median 560 µg/l, range 110–3940, n = 59) compared with healthy blood donors (350 µg/l, 110–580, n = 30) as well as disease controls (360, 110–580, n = 46). No correlation was seen with disease activity, inflammation or renal function. The raised NGAL levels correlated strongly with decreased renal function (r = 0·8, P < 0·001). After correcting for this, slightly increased levels (110, 42–340, n = 59) were observed compared with healthy blood donors (81, 38–130, n = 25), but not compared with the disease controls (120, 57–260, n = 48). In the disease controls, there was a significant correlation between NGAL and proteinase 3 (r = 0·3, p < 0·05), but this was not the case in the vasculitis patients. Whether patients had PR3-ANCA or MPO-ANCA was of no significance. In our measurements, we found significantly raised levels of PR3 in plasma from patients with small vessel vasculitis, regardless of ANCA specificity. This was not due to decreased renal function, ongoing inflammation or neutrophil activation. Plausible mechanisms for this include defects in the reticuloendothelial system, genetic factors and selective neutrophil degranulation or leakage.
Differential cytokine expression of human retinal pigment epithelial cells in response to stimulation by C5a
Authors: FUKUOKA Y , STRAINIC M , MEDOF M E
Published: March 2003

Human retinal pigment epithelial (RPE) cells form part of the blood–retina barrier where they potentially can regulate leucocyte function. RPE cells are known to secrete several cytokines in response to stimulation by other cytokines. Anaphylatoxin C5a, a potent inflammatory mediator produced during complement activation, binds to G-protein coupled C5a receptors (C5aR) on monocytes/macrophages and releases various cytokines from the cells. We previously reported that the human RPE cell line ARPE-19 possesses C5aR and expresses IL-8 mRNA in response to C5a stimulation. In this study, we used a primary human RPE cell line (RPE43) and found that C5a induces increased expression of IL-1β, IL-6, MCP-1 and GM-CSF mRNAs as well as IL-8 mRNA. ARPE-19 cells showed similar increases in the same cytokines. Interestingly, the kinetics of expression of the various cytokines differed. These results provide further evidence that C5a stimulation of RPE cells may play a role in regulating leucocyte function during ocular inflammation in which there is complement activation.
Erythrocyte insulin‐like growth factor‐I binding in younger and older males
Authors: Moromisato David Y. , Roberts Jr Charles , Brasel Jo Anne , Mohan Subburaman , Cowles Elizabeth , King Stephen M. , Cooper Dan M.
Published: March 2003

OBJECTIVE

Insulin‐like growth factor‐I (IGF‐I) levels are lower in older compared with younger subjects. We tested the hypothesis that the reduction in circulating IGF‐I would be accompanied by upregulation in tissue IGF‐I binding in at least some tissues. We tested erythrocyte IGF‐I binding since blood is an accessible tissue in humans, and there is growing evidence to suggest that erythrocyte IGF‐I binding is influenced by circulating IGF‐I.

DESIGN AND PATIENTS

We compared 9 healthy older males (61–68 years old) with 9 healthy younger males (15–19 years old).

MEASUREMENTS

Standard techniques were used to assay circulating IGF‐I and IGF binding proteins 1–5 (IGFBPs 1–5). Erythrocyte IGF‐I binding was first measured by studies in which native [125I]‐IGF‐I was displaced with unlabelled native IGF‐I. In order to determine a possible role for IGF binding proteins (IGFBP), native [125I]‐IGF‐I was displaced with des‐(1‐3)IGF‐I, which binds with IGF receptors but not IGFBPs.

RESULTS

As expected, circulating IGF‐I was significantly lower in the older compared with the younger subjects. In addition, IGFBP‐3 and 5 were significantly lower, and IGFBP‐4 higher, in older compared with younger subjects. When native [125I]‐IGF‐I was displaced with unlabelled native IGF‐I, the number of IGF‐I binding sites per erythrocyte was higher in the older subjects (43 ± 5 vs. 18 ± 2, older vs. younger, respectively; P < 0.05). In contrast, when native [125I]‐IGF‐I was displaced with des‐(1‐3), IGF‐I binding capacity was not different between the two age groups.

CONCLUSIONS

Erythrocyte IGF binding was increased in older compared with younger subjects. Surprisingly, the mechanism of the increase may not be a simple up regulation of IGF‐I receptors in response to reduced circulating IGF‐I, but possibly by an increase in the levels of as yet unidentified erythrocyte membrane‐associated IGF binding proteins.
Normalization of serum thyrotrophin by means of radioiodine treatment in subclinical hyperthyroidism: effect on bone loss in postmenopausal women
Authors: Faber  , Jensen  , Petersen  , Nygaard  , Hegedüs  , Siersbæk‐Nielsen
Published: March 2003

BACKGROUND

Patients with subclinical hyperthyroidism (reduced serum TSH and normal free T4 and T3 concentrations) have slightly increased bone turnover and might have reduced bone mass, especially among postmenopausal women (due to concomitantly reduced oestrogen production), as also seen during suppressive L‐T4 treatment.

OBJECTIVE

We have evaluated whether normalization of serum TSH using radioiodine treatment (RAI) in postmenopausal women with a nodular goitre and subclinical hyperthyroidism, protects against bone loss?

DESIGN

Prospective, non‐randomized study, out‐patients, 2 years follow‐up.

PATIENTS

Postmenopausal women with a nodular goitre, biochemically subclinical hyperthyroidism (TSH < 0.2 mU/l, and signs of a growing goitre or compression symptoms. Sixteen were treated with RAI (median dose 555 MBq) (+RAI), whereas 12 were followed without treatment (−RAI).

MEASUREMENTS

Serum TSH (third generation technology), free T4 and T3 indices, and bone mass (BMD) as measured by Dual Photon Absorptiometry (4 in each group) (only spine) or Dual X‐ray Absorptiometry (DEXA) (both spine and hip), were measured yearly for up to 2 years.

RESULTS

The two groups did not differ regarding age, thyroid hormone parameters, and absolute levels of BMD at spine and hip. RAI resulted in normalization of TSH in all 16 women, and FT4I as well as FT3I decreased to 78% after one year (P < 0.01). These parameters did not change in the untreated group, thus serum TSH remained reduced.

BMD at the spine tended to increase (n.s.) after RAI to (median) 101.9% after one year, and 101.5% after 2 years. In contrast the −RAI group experienced a continued fall in BMD to 97.3% after one year, and 95.5% after 2 years, both reduced as compared to the +RAI group (P < 0.02). BMD of the hip also increased after RAI, to 102.3% after one year, and 101.7% after 2 years. In contrast BMD in the −RAI group decreased to 94.8% after one year, and 98.0% after 2 years, both lower than in the +RAI group (P < 0.01).

CONCLUSIONS

Subclinical hyperthyroidism due to a nodular goitre in postmenopausal women resulted in a continued loss of bone mass of about 2% per year. Radioiodine treatment resulting in normalization of serum TSH prevented this continued bone loss for at least 2 years. Our study supports earlier intervention in such patients.
Effect of long‐term treatment with GH on bone metabolism, bone mineral density and bone elasticity in GH‐deficient adults
Authors: Kann P. , Piepkorn B. , Schehler B. , Andreas J. , Lotz J. , Prellwitz W. , Beyer J.
Published: March 2003

OBJECTIVE

Adults with GH deficiency (GHD) commonly have subnormal bone mineral density (BMD), and have been reported to have an increased risk of fractures. It has been suggested that GH replacement therapy may have beneficial effects on bone in such patients. The aim of this study was to investigate the effects of long‐term GH replacement therapy on bone metabolism, BMD and bone elasticity in adults with GHD.

DESIGN

At the start of the study, 20 adults with GHD were randomized to receive either GH, 0.25 IU/kg/week (the ‘GH group’) or placebo (the ‘placebo group’). After 6 months, patients in the placebo group were switched to GH therapy, and all patients received GH for a further 42 months.

PATIENTS

Of the 20 patients included in the study, 11 were male and nine were female. Mean age at the start of the study was 42.5 ± 10.1 years. All patients had been GH‐deficient for at least 2 years before the start of the study.

MEASUREMENTS

Rates of bone resorption and formation were assessed by measuring serum levels of type I collagen carboxyterminal cross‐linked telopeptide (ICTP) and carboxyterminal propeptide of type I procollagen (PICP), respectively. BMD was measured at the lumbar spine by dual‐photon absorptiometry (DPA) and at the non‐dominant forearm by single‐photon absorptiometry (SPA). Bone elasticity was assessed by measuring apparent phalangeal ultrasound transmission velocity (APU).

RESULTS

The main results in the GH group were as follows. The rate of bone resorption increased significantly during the first 6 months of treatment and remained significantly elevated above its baseline level thereafter. The rate of bone formation also rose during the first 6 months of treatment and remained elevated thereafter, but was significantly higher than at baseline only after 24 months of treatment. At both sites measured, BMD was subnormal at baseline, decreased during the first 6 months of treatment, and increased progressively for the rest of the study, eventually rising well above its baseline level. Bone elasticity decreased during the first 6 months of treatment, but had returned to its baseline level after 24 months.

CONCLUSIONS

Our results support previous findings that BMD is subnormal in adults with GHD, that GH replacement therapy can stimulate bone turnover in such adults and that, in the long term, such stimulation results in a significant increase in BMD. In addition they show, for the first time, that BMD may continue to rise even after GH replacement therapy has been administered for 4 years, and indicate that bone elasticity is not adversely affected by long‐term GH therapy.
Effects of four years' treatment with biosynthetic human growth hormone (GH) on glucose homeostasis, insulin secretion and lipid metabolism in GH‐deficient adults
Authors: Al‐Shoumer Kamal A. S. , Gray Robert , Anyaoku Victor , Hughes Carol , Beshyah Salem , Richmond William , Johnston Desmond G.
Published: March 2003

OBJECTIVE

To study the effects of long‐term growth hormone (GH) treatment on lipid metabolism and carbohydrate tolerance in GH‐deficient adults.

DESIGN

Open trial of GH treatment for 4 years. GH dose was (median, range) 0.025 (0.010–0.050) IU/kg daily.

PATIENTS

Thirteen GH‐deficient hypopituitary adults (seven men, six women), aged (median, range) 47 (24–65) years were followed for 4 years.

MEASUREMENTS

Fasting lipids, lipoproteins, apolipoproteins, glucose and insulin concen_trations were measured at yearly intervals during GH therapy. A 75‐g oral glucose tolerance test (OGTT) was also performed yearly, during which circulating glucose and insulin were measured at 30‐minute intervals for 3 h.

RESULTS

Fasting total and low density lipoprotein (LDL) cholesterol concen_trations decreased on GH therapy, but no change was observed in fasting triglyceride or high density lipoprotein (HDL) concen_trations. Compared to pretreatment values, total and LDL cholesterol levels were significantly lower at 1 year (mean ± SEM) (6.39 ± 0.46 vs. 5.71 ± 0.38 mmol/l, P < 0.05; 4.46 ± 0.36 vs. 3.24 ± 0.20 mmol/l, P < 0.01, respectively) and the reductions were maintained for the 4 years. Apolipoproteins A‐1 and B did not differ significantly from the pretreatment levels. Fasting plasma glucose increased significantly at the first year (4.9 ± 0.1 vs. 5.3 ± 0.1 mmol/l, P < 0.05) but it returned to the pretreatment value in the following years. Fasting plasma insulin increased significantly at 1 year (4.3 (1.0–13.6) vs. 11.9 (1.2–26.9) mU/l, P < 0.05) and showed a progressive downward trend but remained significantly raised throughout the subsequent years. The 3‐h area under the glucose curve (AUC) during the OGTT tended to be increased at the first year (P = 0.07) and it returned to the pretreatment level in the following years. The AUC of plasma insulin was significantly raised at 1 year (P = 0.024) and it returned to the pretreatment level in the following years.

CONCLUSIONS

Four years of GH therapy in GH‐deficient adults resulted in a sustained improvement in total and LDL cholesterol concen_trations. Mild fasting hyperinsulinaemia persisted, although an initial deterioration in glucose tolerance, associated with post‐glucose hyperinsulinaemia, was not sustained.
Serum concentrations of insulin, insulin‐like growth factor(IGF)‐I, IGF binding protein (IGFBP)‐1 and ‐3 and growth hormone binding protein in obese children: fasting IGFBP‐1 is suppressed in normoinsulinaemic obese children
Authors: Saitoh Hisako , Kamoda Tomohiro , Nakahara Satoko , Hirano Takeki , Nakamura Norimasa
Published: March 2003

OBJECTIVE

Simple obesity is characterized by normal or accelerated growth in the presence of reduced serum levels of GH, whereas its detailed mechanism remains unknown. We, therefore, evaluated interrelationships among serum levels of insulin, IFG‐I, IGF binding protein (IGFBP)‐1 and ‐3 and growth hormone binding protein (GHBP) in prepubertal obese children.

SUBJECTS

Prepubertal 20 obese children and 20 age‐matched control children were included in the study.

RESULTS

Serum levels of insulin, IGF‐I and IGFBP‐3 in obese children did not differ from those in controls. The serum level of IGFBP‐1 was significantly lower in obese children (22.1 ± 18.4 μg/l, P<0.001) than in control children (76.0 ± 62.9 μg/l). No relationship was found between the serum levels of insulin and IGF‐I, IGFBP‐1, or IGFBP‐3 in obese subjects. The serum level of GHBP in obese children was significantly elevated as compared with that in controls and was positively correlated with body mass index (BMI). No relationship was found between the serum levels of GHBP and IGF‐I in obese subjects.

CONCLUSIONS

The present study showed for the first time that the fasting IGFBP‐1 level was suppressed in prepubertal obese children with fasting normoinsulinaemia. We speculate that the hyperinsulinaemia which cannot be detected in the fasting state may have suppressed hepatic production of IGFBP‐1. Alternatively, the reduced IGFBP‐1 is likely to be a compensatory response to impaired insulin sensitivity. Thus, the IGFBP‐1 level may be a useful predictor for the early identification in the development of insulin resistance in prepubertal obese children.
Usefulness of surface phenotype study of intrathyroidal lymphocytes obtained by fine needle aspiration cytology in autoimmune thyroid disease and malignant lymphoma of the thyroid
Authors: Fujikawa Megumi , Okamura Ken , Sato Kaori , Mizokami Tetsuya , Tanabe Shoko , Ikenoue Hiroshi , Okamura Seiichi , Ohta Masayuki , Fujishima Masatoshi
Published: March 2003

OBJECTIVES

The surface phenotypes of intrathyroidal lymphocytes have been studied in various thyroid diseases. In most of the previous reports, intrathyroidal lymphocytes were obtained by surgical operation. We evaluated the usefulness of surface phenotype study of intrathyroidal lymphocytes obtained by fine needle aspiration in the diagnosis of Graves' disease, chronic thyroiditis, and malignant lymphoma of the thyroid.

PATIENTS AND DESIGN

Eighty‐seven untreated patients including 24 with Graves' disease, 59 with chronic thyroiditis, and 4 with malignant lymphoma of the thyroid, and 2 treated patients with malignant lymphoma of the thyroid were studied. Surface phenotypes of the peripheral lymphocytes and the intrathyroidal lymphocytes obtained by fine needle aspiration were analyzed using a FACScan and the monoclonal antibodies: anti‐Leu5b/CD2, Leu4/CD3, Leu3a/CD4, Leu2a/CD8, and Leu12/CD19. Percentages of cells positive for each monoclonal antibody were calculated. In one case with malignant lymphoma, monoclonal antibodies to surface‐immunoglobulin markers were also studied.

RESULTS

In peripheral lymphocytes, the percentage of positive cells in each phenotype was almost normal in each disease. In intrathyroidal lymphocytes, the percentage of CD19 positive cells was increased, and the percentage of CD2 and CD3 positive cells was reduced compared to those of peripheral lymphocytes in each disease. The percentage of intrathyroidal CD19 positive cells was remarkably high in malignant lymphoma, constituting more than 70% of cells. In Graves' disease, a relative decrease in the percentage of intrathyroidal CD4 positive cells and an increase in CD8 positive cells compared to peripheral lymphocytes were observed. In 2 treated patients with malignant lymphoma of the thyroid, the intrathyroidal CD19 positive cells almost disappeared in a patient shortly after successful treatment, and reappeared to constitute 76% of cells in a patient in relapse. In one case with malignant lymphoma in which monoclonal antibodies to surface‐immunoglobulin markers were studied, the clonality of the affected cells could be demonstrated; more than 70% of the cells were positive for κ light‐chain and μ and δ heavy‐chain. This finding was proved by subsequent immunohistochemical study based on open biopsy.

CONCLUSIONS

Surface phenotype study of intrathyroid lymphocytes obtained by fine needle aspiration has limited utility in the evaluation or diagnosis of Graves' disease and chronic thyroiditis. However, this simple rapid method is very helpful in the diagnosis and follow up of malignant lymphoma of the thyroid.
Determination of a common genetic variant of luteinizing hormone using DNA hybridization and immunoassays
Authors: Nilsson Christel , Jiang Min , Pettersson Kim , Iitiä Antti , Mäkelä Minna , Simonsen Henrik , Easteal Simon , Herrera René J. , Huhtaniemi Ilpo
Published: March 2003

OBJECTIVE

An immunologically anomalous form of LH, due to two point mutations in codons 8 and 15 of the LHβ gene, has previously been described. LH status, i.e. the discrimination between wild‐type (WT) and variant (V) LH, is usually determined by immunoassays, which can be unreliable at low serum concentrations of LH. A DNA hybridization assay was therefore developed to score the LH genotype in all subjects, independent of their serum LH concentrations. To evaluate the performance of the hybridization method, and to expand our observations of the worldwide occurrence of the V‐LH, we determined its frequency in additional populations. To confirm the connection between the anomalous immunoreactivity and the V‐LHβ gene, we also sequenced the LHβ subunit gene of a homozygous person.

DESIGN

According to the ratio of two immunoassays, one detecting only WT‐LH and the other detecting equally WT and V‐LH, individuals can be classified as homozygotes for the V‐LHβ allele, heterozygotes or WT. DNA samples from persons with known LH status, according to the immunoassays, were used for the development and evaluation of a new allele‐specific DNA hybridization assay. This assay, and PCR and restriction fragment length polymorphism analysis, were used to determine the frequency of the V‐LHβ allele in DNA samples obtained from eight populations.

PATIENTS

Ambulatory adult men and women, apparently healthy and with no endocrine disorders.

RESULTS

The LH genotyping by immunoassays and by the new hybridization method gave identical results with all samples analysed (n = 25). The V‐LHβ subunit was observed to always have the two point mutations, and to be identical with the ones previously reported. The V‐LHβ carrier frequency in the DNA samples collected from various populations varied between 0 and 53.5%.

CONCLUSIONS

The immunoassay technique and the hybridization assay can be used as alternatives to determine the LH status. A great variation in carrier frequency of the V‐LHβ allele is observed in different populations.
Increased incidence of neoplasia in patients with pituitary adenomas
Authors: Popovic V. , Damjanovic S. , Micic D. , Nesovic M. , Djurovic M. , Petakov M. , Obradovic S. , Zoric S. , Simic M. , Penezic Z. , Marinkovic J. , The Pituitary Study Group
Published: March 2003

OBJECTIVE

The goal of our study was to determine the rate of neoplasms in patients with other pituitary adenomas (non‐functioning and prolactinomas) in comparison with acromegaly which is known to favour the development of neoplasia.

DESIGN AND PATIENTS

We reviewed clinical records for 220 patients with acromegaly, 151 patients with non‐functioning pituitary adenoma (NF) and 98 patients with prolactinomas. Incidence rates of cancer for patients with pituitary tumours were calculated per person‐years of follow‐up study. These rates were then compared with sex and age adjusted incidence rates reported by National Tumour Registry. An internal control group of 163 subjects with a non‐neoplastic condition, i.e. Graves' disease followed chronically in the same clinic was also studied. The ratios observed to expected were expressed as standardized incidence rates (SIR). The only significant difference between the acromegalic and other pituitary tumours patients was in hypopituitarism, present in 18.2% (acromegaly) 47% (NF) and 18.6% (prolactinomas).

RESULTS

Twenty‐three malignant tumours were registered in 19 acromegalics (1 Hodgkin disease, 1 myelogenous leukaemia, 1 lymphocytic leukaemia , 3 papillary thyroid carcinomas, 1 ovarian carcinoma, 2 colorectal carcinoma, 1 renal cell carcinoma, 4 cervical carcinoma, 2 skin cancers, 2 pancreatic carcinoma, 4 breast carcinoma, 1 bladder carcinoma). Three acromegalics harboured two malignancies. Patients with acromegaly had a 3.39‐fold increased rate of malignant tumours compared with the general population and a 3.21‐fold increased rate compared with our internal control group. Eleven malignant tumours were found in patients with NF‐pituitary adenomas and 2 in prolactinoma patients (1 lymphoma, 1 multiple myeloma, 1 colonic cancer, 1 renal cell cancer, 1 stomach cancer, 2 lung cancers, 1 cervix carcinoma, 1 breast cancer, 1 testicular carcinoma and 3 melanoma). Patients with NF pituitary adenomas had a 3.91‐fold increased rate of malignant tumours compared with the general population and 4.07‐fold increase compared with the internal control group. Patients harbouring prolactinomas did not have an increased incidence rate of malignancy compared with the general population or our internal controls. Female patients with acromegaly and male patients with NF‐pituitary adenoma had higher incidences of neoplasia.

CONCLUSION

We have demonstrated that the overall incidence of malignant tumours in patients with non‐functioning pituitary adenomas and acromegaly is significantly higher than expected for general population and for our internal control group.