bluuebunny/crossref_metadata_2025 huggingface dataset:
Naïve and memory CD4+ T cells and T cell activation markers in HIV-1 infected children on HAART
Authors: Resino S , Navarro J , Bellón J M , Gurbindo D , León J A , Muñoz-Fernández M A
Published: March 2003

The objective of this study was to investigate the relationship between peripheral blood CD4+ T cell subsets and routine viro-immunological markers in vertically HIV-1-infected children undergoing highly active antiretroviral therapy (HAART). CD4+ and CD8+ T cell subsets were examined by three-colour flow cytometry. Plasma viraemia was quantified by a standardized molecular assay. A negative correlation between the %CD4+ T cells and both viral load and the %CD8+ T cells was observed. A strong positive correlation between the %CD4 T cells and naïve, CD38+ and non-activated CD4+ T cell subsets was found, whereas the %CD4 T cells correlated negatively with the numbers of memory, activated and memory-activated CD4+ T cell subsets. Elevated percentages of CD8 T cells were associated with increased memory and CD4+ CD62L-T cell subsets, whereas the naïve and CD4+ HLA-DRCD38+ subsets negatively correlated with the CD8%. Co-expression of CD62L on memory CD4+ cells and high expression of HLA-DR (but not of CD38) were associated with high viral load. No association between viral load and naïve CD4+ T cells was observed. Specific CD4+ T cell subsets may be more informative than routine surrogate markers in defining the evolution of HIV infection and immune reconstitution in children.
Autoantibodies to human cytosol: a marker of sporadic porphyria cutanea tarda
Authors: Ma Y , Fracanzani A L , Sampietro M , Mattioli M , Cheeseman P , Williams R , Mieli-Vergani G , Vergani D , Fargion S
Published: March 2003

The enzymes potentially involved in the pathogenesis of sporadic porphyria cutanea tarda (PCT) reside in liver cytosoles and microsomes. PCT is frequently associated with hepatitis C virus (HCV) infection, which is in turn associated with autoimmune manifestations. To investigate whether autoimmune reactions, possibly triggered by HCV, are involved in the pathogenesis of PCT, we measured by immunoblot autoantibodies to human cytosolic and microsomal liver fractions in 82 patients with PCT (77% with HCV infection), 105 with other liver disorders and 40 healthy subjects. Anti-liver cytosolic antibodies were more frequent in PCT patients (38/82, 46%) than in pathological controls (P < 0·05–P < 0·001) or in healthy subjects (3/40, 8%, P < 0·001). Among PCT patients, anticytosolic antibodies were more frequent in HCV positive (36/63, 57%) than in HCV negative (2/19, 11%, P < 0·05) cases. Reactivity to a 40-kDa cytosolic polypeptide was present in 20 PCT patients (19 HCV positive), being more frequent than in all pathological controls (P < 0·01–P < 0·0001). Histological activity index (P = 0·04) and antibodies to HCV (P = 0·027) – but not HCV RNA – were associated independently with anticytosolic antibodies as assessed by multivariate analysis. In contrast, frequency of antiliver microsomal antibodies was similar in PCT patients (24/82, 29%) and pathological controls (8–26%), being higher in the autoimmune hepatitis control group (23/23, 100%, P < 0·0001). In conclusion, anticytosolic antibodies, particularly to a 40-kDa polypeptide, are frequent in PCT and associated with HCV infection and severity of liver damage.
Variations in serum IL-7 and 90K/Mac-2 binding protein (Mac−2 BP) levels analysed in cohorts of HIV-1 patients and correlated with clinical changes following antiretroviral therapy
Authors: Darcissac E C A , Vidal V , De La Tribonniere X , Mouton Y , Bahr G M
Published: March 2003

Serum levels of interleukin-7 (IL-7), a non-redundant cytokine that plays a crucial role in lymphopoiesis, are known to be elevated in HIV-1-infected subjects. To examine further the association between levels of IL-7, CD4+ cell counts and viraemia, we analysed these parameters in a large cohort of HIV-1 patients along with serum levels of 90K, a marker of disease severity but with no established involvement in lymphopoiesis. While IL-7 levels were only found to correlate with CD4+ cell counts, 90K levels presented strong correlations with both CD4+ cell numbers and with plasma viral loads (VLs). These correlations were maintained in patients naive to treatment with antiretrovirals (n = 38) but were abolished when the analysis was restricted to the group receiving highly active antiretroviral therapy (HAART, n = 82). Moreover, although 90K levels were significantly reduced in patients on HAART, IL-7 levels continued to be elevated despite successful treatment. The influence of HAART on the variations in these serum parameters was further assessed in a longitudinal study on 32 subjects. The HAART-induced decrease in VLs and increase in CD4+ counts were found to correlate with a reduced serum level of 90K and IL-7, respectively. Nevertheless, following a median period of 33 months of immunological and virological successful HAART, serum levels of IL-7 continued to be significantly elevated compared with those detected in healthy controls. These findings suggest that immunotherapy with IL-7, aimed to replenish T-cell stock in HAART-treated subjects, may have a limited impact on the process of immune reconstitution.
Anti-CD38 autoimmunity in patients with chronic autoimmune thyroiditis or Graves' disease
Authors: Antonelli A , Fallahi P , Nesti C , Pupilli C , Marchetti P , Takasawa S , Okamoto H , Ferrannini E
Published: March 2003

Autoantibodies directed against human CD38 (an enzyme catalysing the interconversion of NAD+ and cyclic ADP-ribose) have been demonstrated recently in patients with type 2 diabetes. We tested 220 consecutive Caucasian patients with autoimmune chronic thyroiditis, 104 patients with Graves' disease, 220 subjects from the general population (control I) and 78 healthy control subjects not affected by thyroid autoimmune disorders (control II) for the presence of anti-CD38 autoimmunity. Using Western blot analysis and optical densitometry, a specific band corresponding to human recombinant CD38 was identified in the serum of several subjects. By defining anti-CD38 positivity as a standardized optical reading > 3 s.d. higher than the mean value of control I, 10·4% of patients with thyroiditis and 7·7% of Graves' patients were anti-CD38 positive (P = 0·0009 versus 1·8% of control I). Similarly, 13·1% of patients with thyroiditis and 10·5% of Graves' patients had a standardized optical reading > 3 s.d. higher than the mean value of the subjects not affected by thyroid autoimmune disorders (P = 0·002 versus 1·2% of control II). Anti-CD38 autoimmunity did not differ between euthyroid, hyperthyroid or hypothyroid patients or between patients with or without thyroid hypoechogenicity. Anti-CD38 autoantibodies were associated with higher levels of circulating antithyroid-peroxidase antibodies (P = 0·03) and they were more frequent in Graves' patients with ophthalmopathy (P < 0·05). Anti-CD38 autoantibodies are a new autoimmune marker in chronic autoimmune thyroiditis and Graves' disease. The specific role of CD38 and its autoantibodies in the modulation of thyroid cell function or growth remains to be investigated.
Increased expression of pro-inflammatory cytokines and metalloproteinase-1 by TGF-β1 in synovial fibroblasts from rheumatoid arthritis and normal individuals
Authors: CHEON H , YU S -J , YOO D H , CHAE I J , SONG G G , SOHN J
Published: March 2003

Transforming growth factor (TGF)-β1 is expressed abundantly in the rheumatoid synovium. In this study, the inflammatory effect of TGF-β1 in rheumatoid arthritis (RA) was investigated using cultured fibroblast-like synoviocytes (FLS) from RA and osteoarthritis (OA) patients, as well as non-arthritic individuals. mRNA expressions of IL-1β, tumour necrosis factor (TNF)-α, IL-8, macrophage inflammatory protein (MIP)-1α and metalloproteinase (MMP)-1 were increased in RA and OA FLS by TGF-β1 treatment, but not in non-arthritic FLS. Enhanced protein expression of IL-1β, IL-8 and MMP-1 was also observed in RA FLS. Moreover, TGF-β1 showed a synergistic effect in increasing protein expression of IL-1β and matrix metalloproteinase (MMP)-1 with TNFα and IL-1β, respectively. Biological activity of IL-1 determined by mouse thymocyte proliferation assay was also enhanced by 50% in response to TGF-β1 in the culture supernatant of RA FLS. DNA binding activities of nuclear factor (NF)-κB and activator protein (AP)-1 were shown to increase by TGF-β1 as well. These results suggest that TGF-β1 contributes for the progression of inflammation and joint destruction in RA, and this effect is specific for the arthritic synovial fibroblasts.
Down-regulated donor-specific T-cell reactivity during successful tapering of immunosuppression after kidney transplantation
Authors: VAN BESOUW N M , VAN DER MAST B J , DE KUIPER P , GREGOOR P J H SMAK , VAESSEN LENARD M B , IJZERMANS J N M , VAN GELDER T , WEIMAR W
Published: March 2003

Stable cadaveric renal transplant patients were routinely converted from cyclosporin A (CsA) to either azathioprine (AZA) or mycophenolate mofetil (MMF) 1 year after transplantation to reduce the side effects of long-term immunosuppressive therapy. Thereafter, the AZA and MMF dose was gradually tapered to 50% at 2 years after transplantation. We questioned whether a reduction of immunosuppressive treatment results in a rise of donor-specific T-cell reactivity. Before transplantation (no immunosuppression), 1 year (high dose immunosuppression) and 2 years (low dose immunosuppression) after transplantation, the T-cell reactivity of peripheral blood mononuclear cells (PBMC) against donor and third-party spleen cells was tested in mixed lymphocyte cultures (MLC) and against tetanus toxoid (TET) to test the general immune response. We also measured the frequency of donor and third-party reactive helper (HTLpf) and cytotoxic (CTLpf) T-lymphocyte precursors in a limiting dilution assay. Donor-specific responses, calculated by relative responses (RR = donor/third-party reactivity), were determined. Comparing responses after transplantation during high dose immunosuppression with responses before transplantation (no immmunosuppression), the donor-specific MLC-RR (P = 0·04), HTLp-RR (P = 0·04) and CTLp-RR (P = 0·09) decreased, while the TET-reactivity did not change. Comparing the responses during low dose with high dose immunosuppression, no donor- specific differences were found in the MLC-RR, HTLp-RR and CTLp-RR, although TET-reactivity increased considerably (P = 0·0005). We observed a reduction in donor-specific T-cell reactivity in stable patients after renal transplantation during in vivo high dose immunosuppression. Tapering of the immunosuppressive load had no rebound effect on the donor-specific reactivity, while it allowed recovery of the response to nominal antigens.
Granulocyte macrophage colony stimulating factor is elevated in alveolar macrophages from sheep naturally infected with maedi-visna virus and stimulates maedi-visna virus replication in macrophagesin vitro
Authors: ZHANG Z , HARKISS G D , HOPKINS J , WOODALL C J
Published: March 2003

Infection by maedi-visna virus, a lentivirus of sheep, leads to chronic inflammatory reactions of various tissues. In this report we have analysed the role of specific cytokines in the disease process. A significant increase in expression of interleukin-6, interleukin-10, granulocyte macrophage-colony stimulating factor (GM-CSF) and transforming growth factor-β1 mRNA was observed in alveolar macrophages isolated from the lungs of naturally infected animals when compared with lungs of seronegative controls. Levels of GM-CSF mRNA expression in alveolar macrophages correlated with the presence of lung lesions, but there was no correlation of interleukin-10, interleukin-6, tumour necrosis factor-α and transforming growth factor-β1 mRNA levels in alveolar macrophages from animals with pulmonary lesions. In vitro investigation showed that GM-CSF in the range 0·1–10ng/ml induced a significant increase in viral p25 production after 7 days in acutely infected blood monocyte-derived macrophages. The production of p25 peaked between 7 and 14 days exposure to 10ng/ml of GM-CSF. Quantitative polymerase chain reaction showed that the level of viral DNA in monocyte-derived macrophages was dose-dependent following GM-CSF treatment in the range 0·1–100ng/ml after 7 days. Viral mRNA expression was also enhanced. These findings indicate a role for GM-CSF in the pathogenesis of lymphoid interstitial pneumonia in infected animals.
Up-regulation of MMP-8 and MMP-9 activity in the BALB/c mouse spinal cord correlates with the severity of experimental autoimmune encephalomyelitis
Authors: NYGÅRDAS P T , HINKKANEN A E
Published: March 2003

Induction of EAE can be inhibited or repressed by administration of soluble metalloproteinase inhibitors. We studied the matrix metalloproteinase (MMP) and their tissue inhibitor (TIMP) expression pattern in experimental autoimmune encephalomyelitis (EAE) of the resistant Th2 prone BALB/c mouse, where the disease can be induced with ultrasound-emulsified antigen/adjuvant (son-ag), but not with conventional technique (syr-ag). We found highly elevated expression of MMP-8 (neutrophil collagenase) mRNA and protein in diseased son-ag challenged mice, colocalizing to neutrophil infiltrates found in brain and extensively in the spinal cord submeningeal space. MMP-8 expression has not been found previously in sensitive mouse strains. The infiltrates stained positive also for MMP-9 protein, and brain homogenates from corresponding mice showed MMP-9 activity during overt disease (days 12–16 post-immunization). TIMP-1 gene expression could be detected in CNS samples from diseased son-ag challenged mice but not in syr-ag or control mice, and the TIMP-1 protein colocalized with GFAP-staining. In contrast, in syr-ag mice both TIMP-2 and TIMP-3 gene expression in the spinal cords was elevated. The results show that sonication, but not extrusion, creates an adjuvant formula potent in activating the matrix metalloproteinase cascade similar to sensitive mouse strains, strongly implicating their role in EAE induction in this Th2 prone strain. The study provides the basis for establishment of MMP-specific therapy in this model.
Simultaneous analysis of T cell clonality and cytokine production in rheumatoid arthritis using three-colour flow cytometry
Authors: BAKAKOS P , PICKARD C , WONG W M , AYRE K R , MADDEN J , FREW A J , HODGES E , CAWLEY M I D , SMITH J L
Published: March 2003

In this study we examined the cytokine production by T cells and TCRVβ subsets in peripheral blood (PB) and synovial fluid (SF) from six RA patients and PB from 10 normal subjects, using three-colour flow cytometry. In two RA subjects we assessed T cell clonality by RT PCR using TCRBV family-specific primers and analysed the CDR3 (complementarity determining region 3) length by GeneScan analysis. A high percentage of IFN-γ- and IL-2- producing cells was observed among the PB T cells in both the RA patients and normal controls and among the SF T cells in RA patients. In contrast, the percentage of T cells producing IL-4 and IL-5 was small among PB T cells in both RA patients and normal controls and among SF T cells in RA patients. There was no significant difference in the production of IFN-γ, IL-2 and IL-5 between the two compartments (PB and SF); however, there were significantly more IL-4-producing cells in SF. Molecular analysis revealed clonal expansions of four TCRBV families in SF of two of the RA patients studied: TCRBV6·7, TCRBV13·1 and TCRBV22 in one and TCRBV6·7, TCRBV21·3 and TCRBV22 in the second. These expansions demonstrated cytokine expression profiles that differed from total CD3+ cells, implying that T cell subsets bearing various TCR-Vβ families may have the potential to modulate the immune response in RA patients.
Conserved CDR 3 region of T cell receptor BV gene in lymphocytes from bronchoalveolar lavage fluid of patients with idiopathic pulmonary fibrosis
Authors: SHIMIZUDANI N , MURATA H , KEINO H , KOJO S , NAKAMURA H , MORISHIMA Y , SAKAMOTO T , OHTSUKA M , SEKISAWA K , SUMIDA M , SUMIDA T , MATSUOKA T
Published: March 2003

Idiopathic pulmonary fibrosis (IPF) is an inflammatory lung disease characterized by the accumulation of inflammatory cells and deposition of collagen, resulting in lung remodelling. High numbers of T cells are present in bronchoalveolar lavage fluid (BALF) of IPF patients, although the characteristics of these cells are yet to be determined. To elucidate the pathogenic mechanisms of IPF, we analysed the T cell receptor (TCR) of BALF lymphocytes in three patients with IPF and three healthy subjects as control. TCR repertoire of BALF lymphocytes and T cell clonality were examined by family PCR and Southern blot analysis, and single-strand conformation polymorphism (SSCP), respectively. We observed that the TCR repertoire in the lung was heterogeneous, both in the control subjects and three patients with IPF. SSCP analysis demonstrated an increase in the number of accumulated T cell clones in BALF of two of the three patients, but not in the healthy subject. Furthermore, junctional sequence analysis showed the presence of conserved amino acid motifs (ETGRSG, LAxG, QGQ, GxQP, GRxG, VAR, PGT, GTI, GGT, TGR, LxLxQ, SGQ) in the TCR-CDR 3 region of BAL lymphocytes in patients with IPF, whereas only two amino acid motifs (VTTG, GGE) were found in the control. Our findings suggest that T cells in BALF of patients with IPF expand oligoclonally in the lung, suggesting antigen stimulation of these cells.
Preventive effect of sulphated colominic acid on P-selectin-dependent infiltration of macrophages in experimentally induced crescentic glomerulonephritis
Authors: OGAWA D , SHIKATA K , MATSUDA M , OKADA S , WADA J , YAMAGUCHI S , SUZUKI Y , MIYASAKA M , TOJO S , MAKINO H
Published: March 2003

Leucocytes infiltrate into renal tissue and are involved in the pathogenesis of crescentic glomerulonephritis. The initial event in the process of leucocyte infiltration is characterized by selectin-mediated leucocyte rolling on endothelial surface. Role of selectins in pathogenesis of glomerulonephritis has still been controversial. Sulphated glycolipids and sulphated polysaccharides interfere with the binding of P- and L-selectin with carbohydrate ligands on endothelial cells or on leucocytes. Here we evaluated the role of selectins and the preventive effects of sulphated colominic acid (SCA), a synthetic sulphated polysaccharide, on experimental crescentic glomerulonephritis in Wistar-Kyoto (WKY) rats. Crescentic glomerulonephritis was induced by injection of nephrotoxic serum (NTS) in WKY rats. Rats subsequently received intraperitoneal injection of saline, neutralizing or non-neutralizing monoclonal antibody (mAb) to rat P-selectin and L-selectin, SCA (5 or 10mg/kg/day) or nonsulphated colominic acid (CA) (10mg/kg/day) for 2 weeks. Localization of P-, E-selectin, ligands for L-selectin and intraglomerular leucocytes was examined by immunohistochemistry. Gene expression of platelet-derived growth factor (PDGF) B chain in glomeruli was quantified using real-time RT-PCR. P-selectin was highly expressed on glomerular endothelial cells after injection of NTS, whereas E-selectin and L-selectin ligands were not detected. Anti-P-selectin mAb, but not anti-L-selectin mAb, significantly reduced glomerular infiltration of macrophages, crescent formation, and proteinuria. SCA also reduced proteinuria, macrophage infiltration, and crescent formation in a dose-dependent manner. Furthermore, SCA suppressed gene expression of PDGF B chain in glomeruli. Our results indicate that P-selectin partially mediate glomerular infiltration of macrophage in experimental crescentic glomerulonephritis. Moreover, SCA may inhibit intraglomerular infiltration of macrophages by interfering with P-selectin-dependent adhesion pathway, and progression of experimental crescentic glomerulonephritis.
Antibody responses to the repetitivePlasmodium falciparumantigen Pf332 in humans naturally primed to the parasite
Authors: AHLBORG N , HADDAD D , SIDDIQUE A B , ROUSSILHON C , ROGIER C , TRAPE J F , TROYE-BLOMBERG M , BERZINS K
Published: March 2003

Antibodies to the degenerate repeats of EB200, a part of the Plasmodium falciparum antigen Pf332, are protective in monkeys. To analyse the prevalence, magnitude and specificity of antibodies to EB200 in malaria-exposed humans, the IgG antibody reactivity with recombinant EB200 protein as well as with crude malaria antigen was determined in Senegalese donors (n = 100; 4–87 years). Antibody reactivity with EB200 was low or absent in children below 15 years but was prevalent and significantly higher in older donors. In comparison, all individuals displayed reactivity with a crude malaria antigen preparation, which also increased with age. The reactivity with the crude malaria antigen was correlated to the reactivity with EB200, suggesting that the low levels of IgG to EB200 found in some adult donors reflected a limited degree of recent exposure to parasites rather than a selective non-responsiveness to Pf332. Comparison of serological and clinical data showed that high levels of antibodies to crude malaria antigen and to EB200 were predictive of fewer future clinical attacks of malaria. A reactivity pattern very similar to that found in Senegalese donors was observed in Liberian adults where 80% of the sera showed reactivity with EB200 and all peptides were recognized by between 60 and 100% of the donors. This strong reactivity with EB200-derived overlapping peptides suggests that the epitopes in EB200, to a large extent, are linear. In the light of previous data on the parasite neutralizing capacity of antibodies to Pf332, the present results emphasize the potential interest of Pf332-derived sequences for inclusion in a subunit vaccine against P. falciparum malaria.
Differential killing of pre-B acute lymphoblastic leukaemia cells by activated NK cells and the NK-92 ci cell line
Authors: REID G S D , BHARYA S , KLINGEMANN H-G , SCHULTZ K R
Published: March 2003

The use of NK cells in adoptive therapy for malignant disease is an area of great potential. Currently the only NK cell line in clinical trials is NK-92, an activated NK cell line with a broad range of cytotoxicity against malignant cells. The activity of NK-92 against pre-B acute lymphoblastic leukaemias, however, is highly variable. In this study we compare the cytotoxic mechanisms and signalling pathways utilized by NK-92 ci and IL-2 activated NK cells to mediate killing of pre-B acute lymphoblastic leukaemia cell lines. Deficiencies in TNF family mediated apoptosis, phosphoinositide-3 kinase dependent and phosphoinositide-3 kinase independent killing limit the efficiency of NK-92 ci against pre-B acute lymphoblastic leukaemia cells. Importantly, treatment of the poorly killed leukaemia cells with TNF-α augmented both phosphoinositide-dependent and -independent cytolysis.
Loss of cytomegalovirus-specific immunological memory in a patient with thymoma
Authors: HUISSOON A P , DAVIES G , COX R A , SLOPER C M L , THOMSON B J , ROBINS R A
Published: March 2003

Cytomegalovirus (CMV) retinitis is a re-activation infection associated with severely impaired T cell-mediated immunity. We describe a patient with long-standing Crohn's disease and thymoma who developed severe CMV retinitis. While thymoma can be associated with impaired humoral immunity and a quantitative CD4+ T helper cell deficiency, these were not evident in our patient. However, more detailed investigation of anti-CMV responses showed absence of specific T cell responses to CMV antigen. Normal CMV seropositive controls have detectable proliferation and interferon-γ production by T cells in response to stimulation with CMV antigen, but this was absent in this patient both during the acute infection and in convalescence. Other measures of T cell function were normal. Since CMV retinitis is due to reactivation of latent CMV infection, it appears that selective loss of CMV-specific immunity had occurred, perhaps secondary to a thymoma. The causes of thymoma-associated immune impairment are not understood, but this case demonstrates that selective defects can occur in the absence of global T cell impairment. Opportunistic infections should therefore be suspected in patients with thymoma even in the absence of quantitative immune deficiencies.
Long-term serological evaluation of patients with cystic echinococcosis treated with benzimidazole carbamates
Authors: RIGANÒ R , IOPPOLO S , ORTONA E , MARGUTTI P , PROFUMO E , ALI M D , VICO B D I , TEGGI A , SIRACUSANO A
Published: March 2003

Seeking better immunological markers indicating the long-term outcome of cystic echinococcosis (CE) after chemotherapy we studied 23 patients receiving albendazole, clinically followed for 8 years, and grouped ultrasonographically according to therapeutic outcome. Antibody responses against a partially purified fraction of hydatid fluid (HFF) and antigen B (AgB) were evaluated by indirect haemagglutination (IHA), ELISA and immunoblotting (IB). Although IHA titres varied over the course of treatment, differences in mean antibody titres to HFF between groups were significant only at 4 years (P = 0·031). IgG isotype expression remained unchanged during follow-up whereas IgE expression decreased in patients with cured or stable disease. AgB disclosed higher IgG4 expression (P < 10–4; P = 0·025) and lower IgG1 expression than HFF (P < 10–4; P = 0·022). IHA antibody titres were higher in patients with progressive than in those with cured or stable disease, even in those with the same cyst type. ELISA isotype profiles differed between groups, particularly for type CE 3, 4 and 5 cysts: higher serum IgG1 and IgG3, lower IgG4 and IgE in patients with cured or stable disease. Although combined serological testing provides scarce information on the long-term outcome of CE after chemotherapy it may be useful for reviewing in a retrospective study the outcome of a cyst and for assessing the host-parasite relationship.
The Th2-response in mercuric chloride-induced autoimmunity requires continuing costimulation via CD28
Authors: MACPHEE I A M , TURNER D R , YAGITA H , OLIVEIRA D B G
Published: March 2003

Mercuric chloride (HgCl2)-induced autoimmunity in Brown Norway (BN) rats is a highly polarized polyclonal Th2-driven autoimmune response with increased IgE production, lymphoproliferation, vasculitis and proteinuria. The increase in serum IgE concentration is clearly measurable by day 4 after the first HgCl2 injection and peaks between days 15 and 20. Treatment with CD80 and CD86 antibodies prior to administration of HgCl2 completely suppresses the autoimmune process. To determine whether interruption of CD28 signalling after initial stimulation of the Th2-response would be suppressive, antibody treatment was delayed. BN rats were given 5 doses of HgCl2 subcutaneously on alternate days. CD80 and CD86 antibodies, or an isotype control, were given daily for 3 days and then on alternate days until day 12 commencing either on the day of the first HgCl2 injection (day 0) or on days 4 or 8. Treatment from day 0 reduced serum IgE concentrations to below baseline (median 9·34μg/ml on day 0 versus 4·6μg/ml, on day 5, P = 0·03) suggesting that ongoing costimulation via CD28 is required to maintain basal serum IgE production. Delaying treatment until day 4 or day 8 after the first HgCl2 injection resulted in significant inhibition of IgE secretion, lymphoproliferation, and vasculitis, although less markedly than when treatment was commenced on day 0.

These data indicate that CD28-mediated costimulation is not only required for the initiation of the Th2-response but is required for maintenance of a maximal response, making this an attractive therapeutic target for antibody-mediated autoimmune diseases.
Changes in thymus volume in adult HIV-infected patients under HAART: correlation with the T-cell repopulation
Authors: RUBIO A , MARTÍNEZ-MOYA M , LEAL M , FRANCO J M , RUIZ-MATEOS E , MERCHANTE E , SÁNCHEZ-QUIJANO A , LISSEN E
Published: March 2003

An important thymus role has been suggested in T-cell repopulation after HAART in adult HIV-1 infected patients. Thymus volume increase after treatment has been described in HIV-1 infected children but not in adult patients. The objective of this work was to evaluate the effect of HAART on the thymic volume of adult HIV-1 infected patients and its relation with the T-cell repopulation. Twenty-one adult patients following 24 weeks under HAART were included in the study. All patients underwent a thoracic computed tomography (CT) evaluation for the measurement of thymic volumes at weeks 0, 12 and 24.

Baseline thymus volume showed a significant correlation with the patient's age. Thymic volume significantly increased after 24 weeks of HAART. Besides, a significant correlation between changes in the thymus volume and changes in both total and naïve CD4+ cell counts was found. Only patients with increases ≥100 CD4+ cell counts after treatment significantly increased the thymic volume.

These data show the first evidence of an early change in thymic volume of adult HIV-1 infected patients under HAART. This increase was related to the rise of both total and naïve CD4+ cell counts suggesting a functional role of thymic volume increase.
Elevated levels of soluble CD163 in sera and fluids from rheumatoid arthritis patients and inhibition of the shedding of CD163 by TIMP-3
Authors: MATSUSHITA N , KASHIWAGI M , WAIT R , NAGAYOSHI R , NAKAMURA M , MATSUDA T , HOGGER P , GUYRE P M , NAGASE H , MATSUYAMA T
Published: March 2003

The aim of the present study was to evaluate levels of soluble CD 163 in sera and fluids from rheumatoid arthritis (RA) patients and elucidate the mechanism that regulates the shedding of CD163. Levels of soluble CD163 in sera and fluids from RA patients were examined by a sandwich enzyme immunoassay and Western blotting. To determine the effects of tissue inhibitors of metalloproteinase (TIMPs) on the shedding of CD163 from monocytes/macrophages, levels of soluble CD163 in cultures of monocytes/macrophages and the expression of CD163 on monocytes/macrophages in the presence or absence of TIMPs were examined by a sandwich enzyme immunoassay and flow cytometry, respectively. The clinical marker that was most associated with serum levels of soluble CD163 was levels of CRP. TIMP-3, but not TIMP-1 or TIMP-2, inhibited the shedding of CD163 from monocytes/macrophages. It was shown that serum levels of soluble CD163 are a sensitive and reliable marker to monitor activated macrophages in synovitis from RA patients and the results imply that the responsible proteinase for the shedding of CD163 is not a member of the matrix metalloproteinases, but is likely to be a member of ADAMs.
A potassium ion channel is involved in cytokine production by activated human macrophages
Authors: QIU M R , CAMPBELL T J , BREIT S N
Published: March 2003

Macrophages play an important role in immune and inflammatory responses, largely through secretion of bioactive molecule such as cytokines. While calcium is known to be an important regulator of this process, less is known about the role of other ions and the ion channels that regulate them. We have previously implicated an outwardly rectifying potassium channel (Kor) in this process and for this reason we have investigated the role of potassium (K+) and K+ channels in the regulation of tumour necrosis factor-α (TNF-α)and interleukin (IL)-8 production by activated human culture-derived macrophages. The effect of blockade of Kor is to inhibit phorbol myristate acetate (PMA)-induced cytokine production by translational or post-translational mechanisms, an effect that is duplicated by increasing extracellular K+. By contrast, the effects of K+ on LPS-stimulated cells are far more complex and are probably mediated through the change of osmolality and occur largely at the mRNA level. This data directly implicates K+, and its regulation through Kor, in early events following PMA stimulation of these cells.
Transient elevation of interleukin-16 levels at the initial stage of meningitis in children
Authors: SUZUKI T , ISHIGURO A , SHIMBO T
Published: March 2003

IL-16 is an immunomodulatory cytokine that is characterized by chemotactic activity and stimulation of proinflammatory cytokine expression in monocytic cells. We studied IL-16 using ELISA in children with meningitis. When meningeal symptoms existed, IL-16 levels were high in the cerebrospinal fluid (CSF) of both bacterial (939 ± 877 ng/l, n = 20) and aseptic (341 ± 371 ng/l, n = 23) meningitis. The values in the CSF were significantly higher than those in non-meningitis controls (29 ± 8 ng/l, n = 22, P < 0·0001). After meningeal symptoms disappeared, IL-16 levels in bacterial (191 ± 149 ng/l, n = 10, P = 0·0042) and aseptic (159 ± 188 ng/l, n = 13, P = 0·0118) meningitis were lower than those during the symptomatic stage. IL-16 levels were the highest before day 5 of the illness and then gradually fell. Significant correlations were found between IL-16 levels and both G-CSF levels (r = 0·783, n = 11, p = 0·0029) and IL-6 levels (r = 0·818, n = 12, P = 0·0005) in the CSF of bacterial and aseptic meningitis. IL-16 levels in all CSF samples from non-meningitis controls were lower than those in serum. In contrast, IL-16 levels in the CSF in six of 16 samples from bacterial meningitis and two of 18 samples from aseptic meningitis were higher than those in serum. Serum levels of IL-16 did not fluctuate throughout the course of meningitis. These data indicate that IL-16 levels rise transiently in CSF at the initial stage of meningitis. We speculate that IL-16 may promote inflammatory responses during meningitis in concert with other proinflammatory cytokines.
Elevated serum prolactin concentrations in phenylketonuric patients on a ‘loose diet’
Authors: Schulpis  , Papakonstantinou  , Michelakakis  , Theodoridis  , Papandreou  , Constantopoulos
Published: March 2003

OBJECTIVE

High levels of phenylalanine (Phe) have been shown to reduce dopamine (DA) and noradrenaline (NA) production. We have therefore evaluated whether increased serum Phe concentrations lead to increased prolactin (PRL) secretion in phenylketonuria.

PATIENTS

Forty‐two patients with classical phenylketonuria were divided into two groups. Group A (n = 19) adhered very strictly to their special diet (Phe = 0.09 ± 0.039 mmol/l) and Group B (n = 23) were on a ‘loose diet’ (Phe = 0.77 ± 0.45 mmol/l). Thirty healthy children of comparable age served as controls.

MEASUREMENTS

PRL was measured by RIA, and Phe and tyrosine (Tyr) with an amino‐acid autoanalyser. Adrenaline (A), noradrenaline (NA), and dopamine (DA) were measured by HPLC.

RESULTS

Serum PRL in group B patients (1595 ± 57 mU/l) was significantly raised compared to Group A patients (243 ± 57 mU/l) and controls (201 ± 55 mU/l) (P < 0.001). Serum DA, A and NA in group B were lower than in group A patients and controls. Serum Phe in group B patients was significantly correlated to serum PRL (r = 0.59) and DA (r = − 0.41). Irregular menses were observed in 1/10 women from group A but in 10/13 in group B.

CONCLUSION

High serum phenylalanine concentrations in phenylketonuric patients not strictly adhering to their diets are correlated with high serum PRL and low serum dopamine concentrations, and a high prevalence of menstrual irregularities.
Overnight urinary free cortisol determination: A screening test for the diagnosis of Cushing's syndrome
Authors: Corcuff Jean‐Benoît , Tabarin Antoine , Rashedi Michel , Duclos Martine , Roger Patrick , Ducassou Dominique
Published: March 2003

OBJECTIVE

The collection of urine over 24 h to measure free cortisol (UFC) is used to diagnose Cushing's syndrome. However, a complete collection of urine is not easy to achieve and the sampling is frequently inaccurate, so a 24 h collection for the determination of UFC excretion is used as a confirmatory rather than a screening test for Cushing's syndrome. Our objective was to evaluate a more convenient urine collection for screening patients suspected of Cushing's syndrome.

DESIGN

We have studied the possibility of using night‐time (from 2000 h to 0800 h) UFC excretion as a screening test for Cushing's syndrome (‘overnight UFC’).

PATIENTS

Thirty patients with proven Cushing's syndrome were studied before treatment (21 cases of Cushing's disease, two cases of ectopic ACTH secretion, seven of adrenal adenoma). The results were compared to those from 150 control obese subjects.

MEASUREMENTS

Overnight UFC, and creatininuria (UC), were evaluated at least once in the patients and in all subjects. The 24 h‐UFC and UC were determined at least once in the patients and in 56 control subjects.

RESULTS

The overnight UFC/UC ratio clearly separated all but one patient of the two groups: 194 ± 386 vs 5.7 ± 3.4 nmol/μmol (P < 0.0005) (ranges 16.2–2024 vs 0.6–17.4, Cushing's syndrome vs controls, respectively). The only patient with Cushing's syndrome who had an overnight UFC/UC ratio that overlapped with that of controls suffered from renal failure. The 24 h UFC/UC ratio of the patients differed from the ratio of controls: 184 ± 365 vs 8.4 ± 4.1 nmol/μmol (P < 0.0005) (ranges 14–1639 vs 1.5–21.2, respectively). Four patients with Cushing's syndrome had 24 h UFC/UC ratios lower than the upper control ratio.

CONCLUSION

The overnight urinary sampling is a simple procedure compared to the 24 h urine collection performed in out‐patients. For a sensitivity set at 100%, the specificities were 97% for the overnight UFC/UC ratio and 87% for the 24 h‐UFC/UC ratio. This therefore appears to provide a good screening method for Cushing's syndrome.
Serum collagen crosslinks as markers of bone turn‐over during GH replacement therapy in growth hormone deficient adults
Authors: Rodríguez‐Arnao  , James  , Jabbar  , Trainer  , Perrett  , Besser  , Ross
Published: March 2003

OBJECTIVES

Bone metabolism is an important target for GH replacement therapy. However, in adults, treatment periods exceeding 12 months are required for a positive effect of GH on bone mineral density. Thus, to detect an early effect of GH on bone, markers of bone turn‐over are important. Pyridinoline (PYR) and deoxypyridinoline (DPYR) are well‐defined sensitive markers of bone resorption, but to date only urinary assays have been available. We report the use of a novel assay to measure changes in serum PYR and DPYR in GH deficient (GHD) adults during GH replacement therapy.

STUDY DESIGN

The study consisted of a 6‐month randomized, double‐blind, placebo‐controlled study of the administration of GH (Genotropin®) (0.25 IU/Kg/week (0.125 IU/kg/week for the first four weeks)) followed by a 6‐month open phase of GH therapy.

PATIENTS

Thirty‐five GHD adults (17 women; mean age 39.8 years; range 21.1–59.9) on conventional hormone replacement therapy as required, were studied.

MEASUREMENTS

Bone formation was analysed using serum bone alkaline phosphatase (BAP) and serum osteocalcin (OC). Bone resorption was analysed using serum pyridinoline (PYR) and serum deoxypyridinoline (DPYR). Bone mineral density (BMD) was determined by dual energy X‐ray absorptiometry (DEXA).

RESULTS

After 6 months placebo treatment there were no significant changes in any of the bone markers analysed, nor in BMD. In the active arm of the study there was a significant increase in serum OC, BAP, PYR and DPYR (P = 0.03, P = 0.004, P = 0.003 and P = 0.01, respectively), remaining significantly elevated over their baseline levels for the subsequent 6 months of treatment (P = 0.04, P = 0.009, P = 0.003 and P = 0.04, respectively). No changes were observed in BMD in any of the groups after 6 months GH treatment. In the active arm of the study, after 12 months GH treatment there was a significant increase in BMD at both the lumbar spine and femoral neck (P = 0.01 for both sites).

CONCLUSIONS

In summary, the present study confirms that administration of GH treatment to GHD adult patients significantly activates bone remodelling, with the effect of GH both in bone formation and bone resorption markers being maximal after 6 months of treatment. The serum assay for PYR and DPYR has a number of practical and theoretical advantages over the urine assay and gave similar results to those previously reported for the urine assay.
Aldosterone production despite absence or defectiveness of the CYP21 genes in two patients with salt‐losing congenital adrenal hyperplasia caused by steroid 21‐hydroxylase deficiency
Authors: Koppens Paul F. J. , Hoogenboezem Theo , Drop Stenvert L. S. , De Muinck Keizer‐Schrama Sabine M. P. F. , Degenhart Herman J.
Published: March 2003

Aldosterone and cortisol were found in plasma samples from two patients with salt‐losing congenital adrenal hyperplasia caused by steroid 21‐hydroxylase deficiency. One patient had a CYP21 gene deletion on one chromosome and a mutation causing erroneous mRNA splicing on the other. The other patient had a CYP21 gene deletion on one chromosome and a large scale conversion of CYP21 to CYP21P on the other. All CYP21P‐like genes in these patients were defective, since they carried a deleterious 8 bp deletion in the third exon. After HPLC purification of the patients' plasma samples, cortisol was no longer detectable in the radioimmunoassay, but aldosterone levels were still within or slightly above the normal reference range. Aldosterone dropped to very low levels after steroid replacement therapy had taken effect. In at least one of these patients, the genetic defect rules out normal functioning of the adrenocortical steroid 21‐hydroxylase, which implies involvement of an alternative enzyme system.
Can intra‐operative GH measurement in acromegalic subjects predict completeness of surgery?
Authors: Van Den Berg Gerrit , Van Dulken Hans , Frölich Marijke , Meinders A. Edo , Roelfsema Ferdinand
Published: March 2003

OBJECTIVE

Results of trans‐sphenoidal pituitary surgery, in terms of long‐term cure, vary considerably between centres. Additional techniques, which can assist the neurosurgeon in deciding whether surgery is complete or not, might therefore be important. One such potential tool is the intra‐operative measurement of GH and calculating the plasma half‐life from the plasma samples obtained after the presumed complete resection of the adenoma.

METHODS

GH half‐life was calculated from 5–10 min plasma samples after adenomectomy in 20 patients. GH was measured with a sensitive and rapid IFMA, and the results could be reported within 30 min, but were not used in this study for per‐operative decisions. Cure was defined by a glucose suppressed plasma GH concentration below 1 mU/l (0.38 μg/l) during follow‐up studies and a normal plasma IGFI concentration.

RESULTS

In 13 cured patients the plasma half‐life was 22.2 ± 1.9 min (range 14–40.6). In three non‐cured patients the plasma half‐life could not be calculated, and in four other patients the plasma half‐life was 35.8 ± 5.9 min (range 25.8–51 min). By applying 25 min as the upper normal limit for the GH plasma half‐life, the sensitivity was 77%, specificity 100%, and positive predictive value 100%.

CONCLUSION

Per‐operative plasma GH monitoring is a potentially useful tool for determining the completeness of trans‐sphenoidal surgery in acromegaly.
Functional evidence for two types of parathyroid adenoma
Authors: D'Amour P. , Weisnagel J. , Brossard J. H. , Ste‐Marie L. G. , Rousseau L. , Lepage R.
Published: March 2003

OBJECTIVE

The carboxyterminal parathyroid hormone (C‐PTH)/intact (I‐) PTH ratio is influenced by serum calcium concentrations in man, increasing to a maximum value in hypercalcaemia and decreasing to a minimum value in hypocalcaemia. We decided to use this ratio to screen for parathyroid tumour with a normal sensitivity to calcium, symptomatic mainly through a mass effect.

DESIGN AND SUBJECTS

Nineteen patients with hypercalcaemia and elevated or inappropriate PTH, were studied in the basal state and during CaCl2 and Na2EDTA infusion and compared with 26 normal individuals. They all had one parathyroid adenoma removed surgically, and two remained hypercalcaemic.

RESULTS

In the basal state, the patients were hypercalcaemic (ionized calcium 1.44 ± 0.12 vs. 1.23 ± 0.03 mmol/l, P < 0.001) and had elevated PTH levels (I‐PTH: 10.8 ± 8.0 vs. 2.3 ± 0.6 pmol/l, P < 0.001; C‐PTH: 31.6 ± 38.9 vs. 5.25 ± 1.11 pmol/l, P < 0.001) when compared with normals. Their mean C‐PTH/I‐PTH ratio was similar to normals (2.7 ± 1.3 vs. 2.4 ± 0.6, NS) but, when individual values were considered, three patients had elevated values at 4.9, 5.3 and 5.8 (normal = 1.2–3.6). The regression line between basal C‐ and I‐PTH revealed a significantly higher slope in these patients (P  < 0.0001). The 16 patients with a normal basal C‐PTH/I‐PTH ratio had, as a group, an increased set point of I‐ or C‐PTH stimulation by calcium and increased values of stimulated and non‐suppressible I‐ and C‐PTH, but these abnormalities were not all present in the smaller tumours (≤200 mg). Only three tumours in that group were larger than 1000 mg. Serum calcium concentration was related to the increased set point and non‐suppressible fraction of I‐PTH in these patients (r2 = 0.797). The three patients with a high basal C‐PTH/I‐PTH ratio had large tumours (2346, 4364 and 17 300 mg) and were more difficult to study, requiring a larger decrease in calcium concentration to achieve maximal stimulation. In the basal state, they were already expressing a non‐suppressible level of I‐ or C‐PTH and already had a maximal C‐PTH/I‐PTH ratio. Our data further suggest a normal set point of I‐ and C‐PTH stimulation in the two patients who achieved sufficient hypocalcaemia and a normal set point of C‐PTH/I‐PTH ratio modulation in these three patients. Their hypercalcaemia was essentially related to the non‐suppressible fraction of PTH. Furthermore, larger tumours were less active than smaller ones and produced less stimulated I‐PTH/100 mg of tissue.

CONCLUSIONS

These data indicate two types of parathyroid tumours when calcium sensitivity is considered: (1) a majority of small tumours with abnormal sensitivity to calcium, symptomatic through an abnormal set point and an increased non‐suppressible fraction and (2) a smaller number of larger tumours, with normal sensitivity to calcium and an increased non‐suppressible fraction, of PTH.
Riedel's thyroiditis presenting with hypothyroidism and hypoparathyroidism: dramatic response to glucocorticoid and thyroxine therapy
Authors: Lo Joan C. , Loh Keh‐Chuan , Rubin Alan L. , Cha Imok , Greenspan Francis S.
Published: March 2003

Riedel's thyroiditis is a rare fibro‐inflammatory process originating in the thyroid gland with progressive extension and invasion of surrounding tissues. Patients frequently present with a stony hard thyroid mass suggestive of anaplastic carcinoma. We report a striking case of Riedel's thyroiditis associated with hypothyroidism, hypoparathyroidism and bilateral vocal cord paralysis. A dramatic response to high dose prednisone and levothyroxine therapy was seen, with recovery of parathyroid and vocal cord function. Our case suggests that early initiation of combination therapy may be important, particularly in the presence of severe disease.