bluuebunny/crossref_metadata_2025 huggingface dataset:
Improved outcome of chronicPseudomonas aeruginosalung infection is associated with induction of a Th1-dominated cytokine response
Authors: MOSER C , JENSEN P O , KOBAYASHI O , HOUGEN H P , SONG Z , RYGAARD J , KHARAZMI A , HOBY N
Published: March 2003

Repeated challenge with antigen is involved in the pathogenesis of a variety of pulmonary diseases. Patients with cystic fibrosis (CF) experience recurrent pulmonary colonization with Pseudomonas aeruginosa before establishment of chronic lung infection. To mimic recurrent lung infections in CF patients, the lungs of susceptible BALB/c mice were re-infected with P. aeruginosa 14 days after the initial infection. Singly-infected BALB/c mice, as well as non-infected mice, were used as controls. Decreased mortality and milder lung inflammation in re-infected BALB/c mice, as well as a tendency for improved clearance of bacteria, was observed when compared with singly-infected mice. The improved outcome in re-infected mice correlated with changes in CD4 cell numbers. Surface expression of LFA-1 on pulmonary CD4 cells was increased in re-infected compared with singly-infected mice. Moreover, resistance to re-infection was paralleled by a shift towards a Th1-dominated response and increased IL-12 production. No significant increase in serum IgG was observed in the re-infected mice. In conclusion, these results indicate a protective role for a Th1-dominated response, independent of antibody production, in chronic P. aeruginosa lung infection in CF.
A type 2 response in lipopolysaccharide (LPS)-stimulated whole blood cell cultures from periodontitis patients
Authors: FOKKEMA S J , LOOS B G , SLEGTE C , VELDEN U VAN DER
Published: March 2003

It is acknowledged that periodontitis results from the interaction of the host immune response with bacteria accumulating on the tooth surfaces. Although bacteria are essential, they are insufficient to cause the disease. Despite this knowledge it remains unclear why certain individuals are more susceptible to periodontitis than others. Therefore the present study investigated whether differences exist in the actual immune response between periodontitis patients and controls after stimulation of peripheral blood cells. Whole blood cell cultures (WBCC) were stimulated with LPS from Escherichia coli during 18 h and the release of prostaglandin E2 (PGE2), IL-1β, IL-6, IL-8, IL-10, IL-12p40, IL-12p70 and tumour necrosis factor-alpha (TNF-α) was measured. The levels of PGE2 were two-fold higher in the WBCC from periodontitis patients than from controls. In contrast, the levels of IL-12p70 in WBCC from patients were two-fold lower. Furthermore, WBCC from patients secreted lower levels of IL-1β and higher levels of IL-8 when compared with WBCC from controls. No differences were observed with respect to IL-6, IL-10, IL-12p40 and TNF-α production. It is known from the literature that LPS-stimulated WBCC reflect specifically the behaviour of the monocytes and that monocytes are peripheral precursors of antigen-presenting cells (APC). Therefore it is concluded that the monocytes in the present WBCC from periodontitis patients are responsible for the higher levels of PGE2 and lower levels of IL-12p70. Since it is has been shown that APC-derived IL-12p70 induces type (Th1) cells that promote cellular immunity, while APC-derived PGE2 induces type 2-helper (Th2) cells that promote humoral immunity, it is postulated that APC from periodontitis patients may have a bias in directing Th2 responses and thereby promoting the humoral immunity in periodontitis.
Transfection of single-stranded hepatitis A virus RNA activates MHC class I pathway
Authors: SUZUKI K , YANAGI M , MORI-AOKI A , MORIYAMA E , ISHII K J , KOHN L D
Published: March 2003

Although infection of single-stranded RNA viruses can enhance expression of major histocompatibility complex (MHC) class I genes, the mechanism underlying this process remains unclear. Recent studies have indicated that exposure of non-immune cells to double-stranded deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) of viral origin can directly increase the expression of MHC class I and related molecules without immune cell interaction. In this report, we show that transfection of single-stranded hepatitis A virus RNA into cultured hepatocytes results in the induction of genes for MHC class I, LMP2 and transporter for antigen processing (TAP1), in addition to the generation of viral proteins. We suggest that this stimulatory effect is due to the double-stranded RNA formed during replication of single-stranded viral RNA, and involves both double-stranded, RNA-dependent protein kinase PKR and the secretion of IFNβ.
The arthritogenic adjuvant squalene does not accumulate in joints, but gives rise to pathogenic cells in both draining and non-draining lymph nodes
Authors: HOLM B C , SVELANDER L , BUCHT A , LORENTZEN J C
Published: March 2003

A single intradermal injection of the adjuvant-oil squalene induces T cell-mediated arthritis in DA rats. The chain of events leading from non-specific provocation of the immune system to arthritis, with clinical similarities to rheumatoid arthritis, is largely undetermined. Here, we combined in vivo tracking of tritium-labelled squalene with lymph node (LN) cell transfer experiments to determine where critical activation events may take place. The majority of squalene remained at the injection site (79%). The amounts recovered in peripheral joints (<1%) were equal to that recovered in other organs that can be targets in autoimmune diseases. This argues that arthritis does not develop as a consequence of adjuvant accumulation in joints. In contrast, substantial amounts of squalene were recovered in hyperplastic LN draining the injection site (1–13%). The adjuvant was deposited to a larger extent in cells than in extracellular matrix. The draining LN cells could transfer arthritis to naïve irradiated DA rats following in vitro stimulation with conA. Interestingly, non-draining LN were also hyperplastic and harboured arthritogenic cells, although they contained low amounts of squalene (<1%). Consequently, the amount of arthritogenic adjuvant in a particular LN is not closely linked to the development of pathogenic cells. The distribution pattern of squalene was similar in MHC-identical but arthritis-resistant PVG.1AV1 and LEW.1AV1 rats, and it was unaffected by T cell depletion with a monoclonal antibody (R73). Thus, T cells and non-MHC genes do not regulate dissemination of squalene, but rather determine arthritis development at the level of adjuvant response.
Penetration of engineered antibody fragments into the eye
Authors: THIEL M A , COSTER D J , STANDFIELD S D , BRERETON H M , MAVRANGELOS C , ZOLA H , TAYLOR S , YUSIM A , WILLIAMS K A
Published: March 2003

Antibodies are powerful immunotherapeutic agents but their use for treating ocular disorders is limited by their poor penetration into the eye. We hypothesized that antibody fragments of relatively small size might penetrate the cornea more readily. Monovalent single chain variable region (scFv) antibody fragments and divalent miniantibodies were engineered from existing monoclonal antibodies, expressed in a bacterial expression system, and purified by metal ion affinity chromatography. Corneoscleral preparations from normal pig and cat eyes were mounted in a corneal perfusion chamber. Intact antibodies and antibody fragments were applied topically to the anterior corneal surface over 12-h periods, and samples were collected from the artificial anterior chamber. Similar experiments were performed with whole enucleated pig and human eyes. Penetration of antibodies and fragments was quantified by high-sensitivity flow cytometry on appropriate target cells. Both monovalent scFv and divalent miniantibody fragments (but not whole immunoglobulin molecules) passed through de-epithelialized and intact corneas after topical administration, and could be detected by antigen binding. Addition of 0·5% sodium caprate facilitated penetration through intact corneas. Topically-applied scFv was found to penetrate into the anterior chamber fluid of rabbit eyes in vivo. The engineered fragments were stable and resistant to ocular proteases. Monovalent and divalent antibody constructs of molecular weight 28 kD and 67 kD, respectively, can penetrate through intact corneas into the anterior chamber, with retention of appropriate antigen-binding activity. Such constructs may form novel therapeutic agents for topical ophthalmic use.
Analysis of mitochondrial antigens reveals inner membrane succinate dehydrogenase flavoprotein subunit as autoantigen to antibodies in anti-M7 sera
Authors: CICEK G , SCHILTZ E , HESS D , STAIGER J , BRANDSCH R
Published: March 2003

The role of mitochondrial proteins as antigens to antibodies of anti-M7 sera was analysed by flavin fluorescence, one- and two-dimensional Western blots and blue native gel electrophoresis. Flavin fluorescence of succinate dehydrogenase (SucDH, complex II of the respiratory chain) of rat liver inner mitochondrial membranes correlated with the immunoreactivity of a representative anti-M7 myocarditis serum. Antigens of isolated bovine heart mitochondria reacting with antibodies of myocarditis serum on two-dimensional Western blots were identified by MALDI-TOF and NanoESI mass spectrometry as myosin heavy chain β and as dihydrolipoamide dehydrogenase of the mitochondrial 2-oxoacid dehydrogenase complexes. The SucDH-flavoprotein was not resolved as a discrete protein spot on two-dimensional polyacrylamide gels. However, separation of the rat liver inner mitochondrial membrane complexes by blue native gel electrophoresis followed by Western blotting, and Western blots of purified Escherichia coli SucDH complex revealed that anti-M7 sera contained antibodies directed against the SucDH-flavoprotein subunit.
Characterization of autoantibodies from patients with Goodpasture's disease using a resonant mirror biosenso
Authors: DOUGAN T , LEVY J B , SALAMA A , GEORGE A J T , PUSEY C D
Published: March 2003

Goodpasture's disease is characterized by the binding of IgG autoantibodies to the glomerular basement membrane, leading to glomerular inflammation. The autoantigen has been identified as the noncollagenous domain of the α3 chain of type IV collagen (α3(IV)NC1). We have used the IAsys resonant mirror biosensor to analyse the extent and affinity of binding of anti-GBM antibodies from sera of patients to purified α3(IV) NC1. α3(IV) NC1 monomers were immobilized to a carboxylate cuvette, with the simultaneous use of a control well. The binding of serum from patients with Goodpasture's disease (n = 12), normal controls (n = 14) and disease controls with vasculitis (n = 14) was analysed. Antibody binding was detected in sera from all patients with Goodpasture's disease but not from controls. IAsys measurements of binding correlated with antibody levels assessed by the standardized ELISA used for clinical assays. Both ELISA and biosensor measurements showed declining antibody levels in serial serum samples from treated patients; however, the biosensor detected antibody recrudescence when ELISA remained negative. Autoantibodies from patients’ serum had average affinity constants (Kd) of 6·5 × 10–11m to 52·07 × 10–10m, as determined by an inhibition assay, indicating high affinity. Sips analysis showed that the antibody response was relatively homogeneous (values of 0·46–1). Biosensor techniques can therefore be used to detect and characterize anti-GBM antibodies in serum from patients, with high sensitivity and without need for antibody purification. This technique may be useful in diagnosis and monitoring of patients with Goodpasture's disease, and may be applicable to other autoantibody mediated diseases.
Lupus-specific kidney deposits of HSP90 are associated with altered IgG idiotypic interactions of anti-HSP90 autoantibodies
Authors: KENDEROV A , MINKOVA V , MIHAILOVA D , GILTIAY N , KYURKCHIEV S , KEHAYOV I , KAZATCHKINE M , KAVERI S , PASHOV A
Published: March 2003

Previous studies have shown that autoantibodies to heat shock protein 90 (HSP90) are elevated in a significant proportion of patients with systemic lupus erythematosus (SLE) who are more likely to have renal disease and a low C3 level. Using samples from 24 patients, we searched for glomerular deposits of HSP90 in renal biopsy specimens from seven patients with lupus nephritis and 17 cases of glomerulonephritis from patients without SLE. Positive glomerular immunofluorescent staining for HSP90 was observed in six of seven cases of SLE and positive tubular staining in two of seven SLE patients. The staining for HSP90 was granular in nature and was located in subepithelial, subendothelial and mesangial areas. None of the non-SLE renal biopsies revealed positive staining for HSP90 deposition. Further we showed the presence of anti-HSP90 IgG autoantibodies in IgG from sera of patients with SLE as well as in normal human IgG (IVIg). In normal IgG this autoreactivity could be adsorbed almost completely on F(ab′)2 fragments from the same IgG preparation, coupled to Sepharose and could be inhibited by the effluent obtained after subjecting normal IgG to HSP90 affinity column. These findings indicate that anti-HSP90 natural autoantibodies are blocked by idiotypic interactions within the IgG repertoire. Unlike natural autoantibodies, anti-HSP90 IgG from SLE patients’ sera were only moderately adsorbed on F(ab′)2 fragments of normal IgG. These results demonstrate that immunopathogenesis of lupus nephritis is associated with HSP90 (as an autoantigen) and that the pathology is associated with altered idiotypic regulation of the anti-HSP90 IgG autoantibodies.
GM-CSF and IL-12 production by malignant plasma cells promotes cell-mediated immune responses against monoclonal Ig determinants in a light chain myeloma model
Authors: GALEA H R , COGNÉ M
Published: March 2003

Immune responses towards malignant plasma cells have clearly been demonstrated in the course of monoclonal B cell dyscrasias and shown to be mostly specific for idiotypic determinants of the monoclonal immunoglobulin (Ig). These responses are specifically efficient against lymphoma cells expressing a membrane form of the monoclonal Ig. In myeloma, such immune responses are often weak and a number of strategies are currently assayed in order to boost the cell-mediated responses against the secreted monoclonal Ig. The use of cytokines promoting Th1 responses could be helpful for the induction of anti-tumour immunity and the control of residual disease in patients treated with myeloablative therapy, and such strategies need to be evaluated.

In a light chain myeloma model where the monoclonal Ig can only be secreted, we tried to induce protective immune responses through immunization of animals with transfected malignant plasma cells. An expression plasmid encoding GM-CSF and IL-12 proved to be highly efficient for the induction of both cytotoxic and proliferative responses after immunization of animals with transfected and irradiated tumour cells. Anti-tumour immunization according to this protocol was successful in protecting 93·4% of the animals against a subsequent tumour challenge.
Role of reactive oxygen species in neutrophil apoptosis following ingestion of heat-killedStaphylococcus aureus
Authors: YAMAMOTO A , TANIUCHI S , TSUJI S , HASUI M , KOBAYASHI Y , KOBAYASHI Y
Published: March 2003

Neutrophils, short-lived leucocytes that die by apoptosis, play an important role in the first stage of defense against bacterial infections. It has been reported that phagocytosis of intact bacteria or Candida albicans can accelerate neutrophil apoptosis. However, the mechanism of phagocytosis-mediated neutrophil apoptosis is not well characterized. In this study, we evaluated whether ingestion of heat-killed Staphylococcus aureus (S. aureus) enhances neutrophil apoptosis and whether this type of apoptosis is mediated by oxidative stress by using antioxidants and polymorphonuclear leucocytes (PMNs) from patients with chronic granulomatous disease (CGD). Co-culture of PMNs with varying doses of S. aureus resulted in accelerated PMN death in a dose- and time-dependent manner. Increased PMN apoptosis was observed by both Annexin V and PI staining. Similar results were observed in PMNs of CGD patients. Dimethyl sulphoxide (DMSO, an OH• scavenger) did not significantly inhibit either S. aureus-ingested PMN apoptosis or spontaneous PMN apoptosis. On the other hand glutathione (GSH, an H2O2 scavenger) significantly inhibited both types of apoptosis.

Our findings suggest that oxygen-independent pathways may mainly operate in the process of phagocytosis-induced apoptosis.
T-cell reactivity to glutamic acid decarboxylase in stiff-man syndrome and cerebellar ataxia associated with polyendocrine autoimmunity
Authors: COSTA M , SAIZ A , CASAMITJANA R , CASTAÑER FERNÁNDEZ M , SANMARTÍ A , GRAUS F , JARAQUEMADA D
Published: March 2003

Antibodies to glutamic acid decarboxilase (GAD-Abs) are present in the serum of 60–80% of newly diagnosed type 1 diabetes (DM1) patients and patients with autoimmune polyendocrine syndrome (APS) associated with DM1. Higher titre of GAD-Abs are also present in the serum of 60% of patients with stiff-man syndrome (SMS) and all reported patients with cerebellar ataxia associated with polyendocrine autoimmunity (CAPA). Several studies suggest that GAD-Abs may play a critical role in the pathogenesis of SMS and CAPA but little is known about T-cell responsiveness to GAD-65 in these neurological diseases. To analyse cell-mediated responses to GAD, we studied the peripheral blood lymphocyte proliferation and cytokine responses to recombinant human GAD-65 in 5 patients with SMS, 6 with CAPA, 9 with DM1, 8 with APS and 15 control subjects. GAD-65-specific cellular proliferation was significantly higher in SMS than in CAPA, DM1, APS or controls. In contrast, only T cells from CAPA patients showed a significantly high production of interferon-γ after GAD stimulation, compared to all other patients and controls. No differences were found for IL-4 production. These results suggest that, despite similar humoral autoreactivity, cellular responses to GAD are different between SMS and CAPA, with a greater inflammatory response in CAPA, and this difference may be relevant to the pathogenesis of these diseases.
Intramolecular epitope spreading among anti-caspase-8 autoantibodies in patients with silicosis, systemic sclerosis and systemic lupus erythematosus, as well as in healthy individuals
Authors: UEKI A , ISOZAKI Y , TOMOKUNI A , HATAYAMA T , UEKI H , KUSAKA M , SHIWA M , ARIKUNI H , TAKESHITA T , MORIMOTO K
Published: March 2003

Dysregulation of apoptosis through the Fas-Fas ligand pathway is relevant in autoimmune disease onset. We recently reported elevated serum levels of sFas in patients with silicosis, systemic sclerosis (SSC) and systemic lupus erythematosus (SLE), and proposed a block of apoptosis in the pathogenesis. The disturbance of apoptosis in lymphocytes including autoreactive clones could induce autoantibody production. Since autoantibodies directed against unknown antigens are present in the sera of these patients, the sera samples were examined for the presence of autoantibodies directed to caspase-8.

Using Western blotting, autoantibodies against caspase-8 were detected in healthy individuals and in over 60% of patients. Using epitope mapping employing 12 amino acid polypeptides with SPOTs system, a minimum of 4 epitopes and a maximum of 13 were found, which implied that epitope spreading was in progress. It is noteworthy that two important catalytic cystein residues were included within the epitopes; firstly the active site cystein Cys287, and secondly Cys360 located in the unique pentapeptide motif QACQG.

Using recombinant human caspase-8 linked protein chip array, autoantibodies were identified and molecular weight determined. The antibodies were mainly IgG; 80% were subclass IgG1λ; 20% were IgG4κ. Despite the ratio of human light chain κ:λ = 2:1, the predominance of IgG1λ is noticeable.

Anti-caspase-8 autoantibodies are detectable in healthy individuals and in patients suffering silicosis, SSc or SLE. A few epitopes were detected in healthy individuals compared to those suffering autoimmune diseases, indicating the intramolecular epitope spreading. Relationship of autoantibodies and the clinical background of the patients requires clarification.
Inducible nitric oxide synthase is expressed in synovial fluid granulocytes
Authors: CEDERGREN J , FORSLUND T , SUNDQVIST T , SKOGH T
Published: March 2003

The objective of the study was to evaluate the NO-producing potential of synovial fluid (SF) cells. SF from 15 patients with arthritis was compared with blood from the same individuals and with blood from 10 healthy controls. Cellular expression of inducible nitric oxide synthase (iNOS) was analysed by flow cytometry. High-performance liquid chromatography was used to measure l-arginine and l-citrulline. Nitrite and nitrate were measured colourimetrically utilizing the Griess’ reaction. Compared to whole blood granulocytes in patients with chronic arthritis, a prominent iNOS expression was observed in SF granulocytes (P < 0·001). A slight, but statistically significant, increase in iNOS expression was also recorded in lymphocytes and monocytes from SF. l-arginine was elevated in SF compared to serum (257 ± 78 versus 176 ± 65 µmol/l, P = 0·008), whereas a slight increase in l-citrulline (33 ± 11 versus 26 ± 9 µmol/l), did not reach statistical significance. Great variations but no significant differences were observed comparing serum and SF levels of nitrite and nitrate, respectively, although the sum of nitrite and nitrate tended to be elevated in SF (19·2 ± 20·7 versus 8·6 ± 6·5 µmol/l, P = 0·054). Synovial fluid leucocytes, in particular granulocytes, express iNOS and may thus contribute to intra-articular NO production in arthritis.
Down-regulation of multiple low dose streptozotocin-induced diabetes by mycophenolate mofetil
Authors: MAKSIMOVIC-IVANIC D , TRAJKOVIC V , MILJKOVIC D J , STOJKOVIC M MOSTARICA , STOSIC-GRUJICIC S
Published: March 2003

The new immunosuppressive agent mycophenolate mofetil (MMF) has been shown recently to exert a protective effects in certain animal models of autoimmunity, including diabetes in diabetes-prone bio-breeding (BB) rats. In the present study, the immunomodulatory potential of MMF was investigated in autoimmune diabetes induced by multiple low doses of streptozotocin (MLD-STZ) in genetically susceptible DA rats 20 mg STZ/kg body weight (b.w.) for 5 days] and CBA/H mice (40 mg STZ/kg b.w. for 5 days). In both species, short time treatment of animals with MMF (25 mg/kg) during the early development of the disease, as well as continuous MMF treatment, prevented the appearance of hyperglycaemia and inflammatory infiltrates in the pancreatic tissue. Moreover, clinical manifestations of diabetes were suppressed by application of the drug after the onset of clinical symptoms. Treatment with guanosine (1 mg/kg) in parallel with MMF completely reversed MMF activity in vivo, indicating that inhibition of inosine monophosphate dehydrogenase (IMPDH) was responsible for the observed suppressive effects. MMF-mediated protection from diabetes correlated with reduced ex vivo spontaneous spleen mononuclear cell (MNC) proliferation and defective adhesive cell interactions. MMF-treated animals also had lower local production of IFN-γ, as well as IL-12 and nitric oxide (NO) production by peripheral tissues (spleen and peritoneal cells), compared to that in control diabetic groups, while IL-10 level was elevated. Together, these data demonstrate that MMF interferes with autoimmune process in streptozotocin-induced diabetes at multiple levels, including lymphocyte proliferation and adhesion, as well as pro/anti-inflammatory cytokine balance.
Definition of the HLA-A2 restricted peptides recognized by human CD8+ effector T cells by flow-assisted sorting of the CD8+ CD45RA+ CD28– T cell subpopulation
Authors: HÖHN H , JÜLCH M , PILCH H , KORTSIK C , TULLY G , NEUKIRCH C , FREITAG K , MAEURER M
Published: March 2003

In response to antigenic stimulation, naive MHC-class I restricted and antigen-specific CD8+ CD45RA+ CD28+ T cells undergo clonal expansion, differentiate into CD8+ CD45RO+ memory T cells and convert to CD8+ CD45RA+ CD28− T cells displaying potent immune effector functions upon re-encounter with the nominal antigen. We show that the effector CD8+ CD45RA+ CD28– T cell subset is expanded in peripheral blood lymphocytes (PBL) from patients with human papilloma virus (HPV)+ cervical lesions as well as in PBL from patients with pulmonary tuberculosis. Flow-cytometric cell sorted CD8+ CD45RA+ CD28– and CD8+ CD45RA+ CD28– T cells were tested for recognition of HLA-A2 restricted peptides derived either from the human papillomavirus (HPV)16-E7 gene product, or from M. tuberculosis antigens. Mostly CD8+ CD45+ CD28– T cells define antigen/peptide-specific and MHC-restricted responses. These data were confirmed in PBL from patients with tuberculosis using HLA-A2 tetramer-complexes loaded with a peptide from the M. tuberculosis Ag85b antigen by flow cytometry. The sorting of this T cell subset enables to determine the fine specificity of CD8+ effector T cells without the need for in vitro manipulation.
Lymphocyte and mast cell counts are increased in the nasal mucosa in symptomatic natural colds
Authors: ALHO O-P , KARTTUNEN T J , KARTTUNEN R , TUOKKO H , KOSKELA M , UHARI M
Published: March 2003

Knowledge of the virus-induced immune response is important in understanding the pathophysiology of respiratory virus infections. Data on the cellular immune response is still limited and based mainly on experimental studies. Natural colds may differ in their pathophysiology from experimentally induced ones. To evaluate the inflammatory cell responses in the upper respiratory tract during natural colds we counted the number of lymphocytes, mast cells and macrophages in the nasal mucosa. Nasal biopsies were taken from 22 adult volunteers during the acute (2–4 days of symptoms) and convalescent phases (day 21) of the cold, and the numbers of cells were counted with immunohistochemical methods. Viral aetiology was identified in 14 (64%) subjects by using viral isolation, antigen detection and rhino-polymerase chain reaction assays. The number of T lymphocytes was increased in the nasal epithelium and that of T and B lymphocytes and mast cells in the subepithelial layer in the acute phase compared to the convalescent phase. Intraepithelial T lymphocyte counts were significantly higher in the subjects who had a proven viral infection or a finding of pathogenic bacteria in the nasopharynx compared to the subjects without such findings (P = 0·005 and P = 0·04, respectively). Contrary to the earlier experimental studies, we found that viruses cause accumulation of T and B lymphocytes and mast cells during the first days of a symptomatic naturally acquired respiratory infection.
γ/δT cell subsets in patients with activeMycobacterium tuberculosisinfection and tuberculin anergy
Authors: SZEREDAY L , BALIKO Z , SZEKERES-BARTHO J
Published: March 2003

Earlier data suggest that γ/δ T cells may play an important role in the immune response to Mycobacterium tuberculosis. The aim of this study was to determine the percentage of different γ/δ subsets in peripheral blood of active tuberculosis patients with a positive or negative tuberculin reaction. Thirty-eight patients infected with M. tuberculosis and 22 healthy controls were included in the study. Venous blood was taken before starting antimycobacterial treatment. Lymphocytes were reacted with monoclonal antibodies specific for different γ/δ V chains (Vδ1, Vδ2, Vγ9 and Vγ4). The results were analysed in the context of tuberculin reactivity and X-ray findings. Our results revealed a selective loss of Vγ9/Vδ2 T cells in the peripheral blood of tuberculin-negative patients with active tuberculosis compared to healthy controls, while the ratio of Vγ9/Vδ2 T cells in the peripheral blood of patients with a positive skin test did not differ from that of healthy controls. These findings demonstrate a relationship between the loss of the major M. tuberculosis-reactive subset of γδ T cells and the absence of tuberculin reactivity. The data are consistent with the hypothesis that γδ T cells play a role in the protective immune response to M. tuberculosis infection.
Interleukin 10 induced augmentation of delayed-type hypersensitivity (DTH) enhances Mycobacterium bovis bacillus Calmette-Guérin (BCG) mediated antitumour activity
Authors: NADLER R , LUO Y , ZHAO W , RITCHEY J K , AUSTIN J C , COHEN M B , O'DONNELL M A , RATLIFF T L
Published: March 2003

Intravesical BCG therapy is effective in the treatment of superficial bladder cancer. Both clinical and experimental results suggest a role for cytokines and delayed-type hypersensitivity (DTH) in BCG-induced antitumour immunity. We characterized the modulatory effects of BCG on bladder cytokine expression and determined the relationship between DTH and BCG antitumour activity. The bladders of mice were instilled with BCG through a catheter. Bladder tissue RNA and urine were collected for evaluation of cytokine expression using reverse transcriptase-polymerase chain reaction (RT-PCR) and/or ELISA. IFN-γ and TNF-α, the two major cytokines associated with DTH, were efficiently induced by BCG. IL10, an important down-regulator of DTH, was also induced by BCG. Constitutive levels of IL4 and IL5 were observed, but neither IL4 nor IL5 were modulated by BCG. Similar results were observed in the kinetic analysis of urinary cytokines in patients after intravesical BCG therapy. Production of Th1 (T helper type 1) cytokines (IFN-γ, IL2 and IL12) preceded that of the Th2 (T helper type 2) cytokine IL10. A tendency toward higher ratios of IFN-γ versus IL10 for BCG responders also was observed. In animal studies the absence of IL10 abrogated either by antibody inhibition or the use of genetically modified, IL10 deficient (IL10–/–) mice resulted in enhanced DTH responses. Under conditions of enhanced DTH, a significant enhancement in antitumour activity was observed. These data demonstrate that DTH and its associated mononuclear infiltration and cytokine production are important to the antitumour activity of intravesical BCG therapy, and suggest that effects to diminish IL10 production may have therapeutic value.
Role of thyroid‐stimulating blocking antibody in patients who developed hypothyroidism within one year after 131I treatment for Graves' disease
Authors: Yoshida Katsumi , Aizawa Yoshihiko , Kaise Nobuko , Fukazawa Hiroshi , Kiso Yoshinori , Sayama Noriyo , Hori Hironobu , Nakazato Naoki , Tani Jun‐ichi , Abe Keishi
Published: March 2003

OBJECTIVE

We recently reported that thyroid‐stimulating blocking antibody (TSBAb) may not contribute to the development of hypothyroidism more than six years after 131I treatment. In the present study, we attempted to determine whether hypothyroidism that develops within a shorter period of time following 131I therapy is associated with TSBAb.

DESIGN

Retrospective study.

PATIENTS

Sera were obtained from 8 patients who developed hypothyroidism within 6 months after 131I therapy (Group 1), 8 patients who became euthyroid one year after 131I therapy (Group 2), and 7 patients who developed transient hypothyroidism (Group 3).

MEASUREMENTS

Thyroid stimulating antibody (TSAb) activity was measured as the amount of cyclic adenosine monophosphate (cAMP) produced by cultured FRTL‐5 cells, and TSBAb activity as the inhibition of cAMP produced in response to 100 mU/l bovine TSH.

RESULTS

At about 3 months after 131I treatment, TSAb activity increased significantly in Groups 2 and 3, but did not change in Group 1. In contrast, TSBAb activity in Group 1 increased significantly and was positive in 6 patients at that time. At 12–18 months after 131I treatment, TSBAb activity tended to decrease and remained positive in 3 patients but became negative in 3 patients. It did not change in the patients in Groups 2 and 3. The patients in Group 1 were treated with levothyroxine, 75–125 μg/day. Levothyroxine was discontinued in the 3 patients whose TSBAb activity disappeared. Two of them remained euthyroid, and 1 became hypothyroid.

CONCLUSION

Results indicate that the hypothyroidism that develops within a short time after 131I treatment may be caused by TSBAb activity. Thyroid function may be recovered when TSBAb activity disappears.
Gonadal status is an important determinant of bone density in acromegaly
Authors: Lesse  , Fraser  , Farquharson  , Hipkin  , Vora
Published: March 2003

OBJECTIVE

To measure bone mineral density (BMD) in patients with acromegaly and to look for clinical features which may explain previously reported discrepant results.

DESIGN

Prospective case controlled observational study.

PATIENTS

18 patients with acromegaly (seven women, 11 men; mean age 53.2±3.5 years). Eight patients had active disease and 10 had controlled acromegaly. Growth hormone and insulin‐like growth factor‐1 (IGF‐1) concentrations were measured to assess disease activity.

MEASUREMENTS

BMD was measured at four sites on the lumbar spine, three at the femoral neck and at Ward's triangle using a Hologic dexa scanner. Results were compared to a locally determined control population (n=1800). In eight patients with active acromegaly, urinary free pyridinoline and deoxypyridinoline, and serum osteocalcin, propeptide of type 1 procollagen, vitamin D, 1000 h parathyroid hormone, bone specific alkaline phosphatase, calcium and phosphate concentrations were measured before and after 6 months treatment with octreotide.

RESULTS

The mean BMD for all acromegalic patients was not significantly different from the control population except at the femoral neck where it was increased (P=0.05). At all sites, the BMD of patients who had been hypogonadal (n=12/18) was significantly lower (P<0.05–0.01) than that of patients who had been eugonadal (n=6/18). BMD for hypogonadal patients was lower than the control population at Ward's triangle (P=0.03). Eugonadal acromegalic patients had BMD greater than non‐ acromegalic controls at all sites. Patients with controlled acromegaly had a higher BMD than non‐acromegalic controls, but there were no differences in BMD between patients with active and controlled acromegaly. Serum IGF‐1 concentrations decreased from 64.5±5.1 nmol/l to 37.5±6.9 nmol/l (P=0.02) after 6 months treatment with octreotide, but there was no change in any of the biochemical markers of bone turnover.

CONCLUSIONS

Eugonadal acromegalic patients have increased lumbar spine and femoral neck BMD compared to hypogonodal acromegalic patients and the general population, but it is reduced if patients have been hypogonadal.
Lean body mass as a determinant of thyroid size
Authors: Wesche Maria F. T. , Wiersinga Wilmar M. , Smits Nico J.
Published: March 2003

OBJECTIVE

Males have a larger thyroid gland than females, and this has been related to the difference in body weight. In view of the different body composition of men and women, we hypothesized that lean body mass is a better determinant of thyroid volume than body weight.

DESIGN

A cross‐sectional study in an area not deficient in iodine.

SUBJECTS

44 non‐obese healthy adults (group I, 21 men, 23 women with equal distribution of sexes in age groups between 21 and 70 years) and 20 adults with marked obesity (group II, 8 men, 12 women, BMI > 30 kg/m2) were studied. None used medication and all had normal thyroid function tests.

MEASUREMENTS

Thyroid volume was measured by ultrasonography, and lean body mass with a body impedance analyser.

RESULTS

The thyroid volume in men was larger than in women in both groups; it was also larger in the obese than in the non‐obese subjects. In the non‐obese subjects, thyroid volume was related both to body weight (r = 0.42, P < 0.005) and to lean body mass (r = 0.55, P = 0.0001). In the obese subjects, thyroid volume was no longer related to body weight (r = 0.23, NS) but was still correlated with lean body mass (r = 0.54, P = 0.01). Taking both groups together, the correlation between thyroid volume and lean body mass (r = 0.64, P < 0.001) was stronger than between thyroid volume and body weight (r = 0.50, P < 0.001). Thyroid volume was also related to body length (group I, r = 0.42, P < 0.005; group II, r = 0.54, P = 0.01), but to body‐surface area only in the non‐obese subjects (group I, r = 0.45, P < 0.01; group II, r = 0.38, NS). The larger thyroid size in the obese was associated with slightly but significantly higher TSH and lower free T4 serum concen_trations as compared to the non‐obese subjects.

CONCLUSIONS

In healthy adults, lean body mass rather than body weight explains the differences in thyroid volume between males and females and between obese and non‐obese subjects. Lean body mass appears to be a major determinant of thyroid size.
The low dose ACTH test in chronic fatigue syndrome and in health
Authors: Scott Lucinda V. , Medbak Sami , Dinan Timothy G.
Published: March 2003

OBJECTIVE

A number of dynamic tests of the hypothalamic–pituitary–adrenal axis provide evidence for a mild cen_tral adrenal insufficiency in chronic fatigue syndrome (CFS). The 1 μg adrenocorticotropin (ACTH) test has been proposed to be more sensitive than the standard 250 μg ACTH test in the detection of subtle pituitary–adrenal hypofunctioning. We aimed to establish whether the 1 μg ACTH test would support such a dysregulation in CFS, and also, given the relative novelty of this test in clinical practice and the uncertainty with regard to appropriate cut‐off values for normality, to compare our healthy volunteer data with those of previous studies.

PATIENTS AND DESIGN

Twenty subjects with CFS, diagnosed according to Cen_tres for Disease Control and Prevention criteria, were compared with 20 healthy volunteer subjects. All participants underwent a 1 μg ACTH test beginning at 1400 h. Plasma samples for cortisol estimation were drawn at 0, +30 and +40 min.

RESULTS

Baseline cortisol values did not differ between CFS patients and healthy subjects. The δ cortisol (maximum increment from baseline) value was significantly lower in the CFS than the volunteer group (P < 0.05). Comparison of the + 30 min cortisol values revealed no significant differences. Using an incremental cortisol of > 250 nmol/l as an arbitrary cut‐off point, two (10%) of the healthy subjects and nine (45%) of the CFS subjects failed the test on this basis (χ2 = 4.3, df = 38, P < 0.05).

CONCLUSIONS

This study provides further evidence for a subtle pituitary–adrenal insuffiency in subjects with chronic fatigue syndrome compared to healthy volunteers. Disparities between our healthy volunteer data and those of other groups using the 1 μg ACTH test suggest that the test may not be as reliable as previously indicated.
Elevated serum granulocyte colony‐stimulating factor levels in patients with Graves' disease
Authors: Iitaka Makoto , Noh Jaeduk Yoshimura , Kitahama Shinji , Fukasawa Nobuhiko , Miura Shiro , Kawakami Yoshito , Kawasaki Satomi , Yamanaka Kayo , Ishii Jun , Katayama Shigehiro , Ito Kunihiko
Published: March 2003

OBJECTIVE

Agranulocytosis is a serious side effect of anti‐thyroid drugs (ATD). Granulocyte colony‐stimulating factor (G‐CSF) is one of the cytokines that increase granulocyte number. The aim of this study was to examine the sequential variation of serum G‐CSF levels in patients with Graves' disease before and after ATD therapy.

PATIENTS

Sixty‐three patients with Graves' disease were studied before, during and after treatment with methimazole (MMI). Serum samples from 71 healthy subjects were used as controls.

DESIGN AND MEASUREMENTS

Serum levels of G‐CSF were measured by a novel chemiluminescent enzyme immunoassay, which was sensitive enough to determine G‐CSF levels in healthy subjects. Blood granulocyte counts, serum, thyroid hormone and TSH levels, and titres of thyroid autoantibodies were also measured.

RESULTS

Serum G‐CSF levels in Graves' patients before and 2 weeks after MMI were significantly higher than in healthy subjects. There was a significant correlation between serum G‐CSF levels and granulocyte counts in untreated patients with Graves' disease. Untreated patients with granulocyte counts less than 2 × 109/l had significantly lower serum G‐CSF levels as compared with other untreated patients. Serum G‐CSF levels gradually decreased thereafter. No correlation was observed between serum G‐CSF levels and serum thyroid hormone levels or titres of thyroid autoantibodies. After ATD treatment, no correlation was found between serum G‐CSF levels and granulocytes counts. There was no significant correlation between the change of serum G‐CSF levels and that of granulocyte counts before and after MMI treatment. Graves' patients with mild agranulocytosis had normal or elevated serum G‐CSF levels.

CONCLUSIONS

Significantly elevated serum G‐CSF levels were observed in patients with Graves' hyperthyroidism. During ATD therapy, deficiency of G‐CSF was not identified as a cause of agranulocytosis in this study.
Determinants of serum insulin‐like growth factor I in growth hormone deficient adults as compared to healthy subjects
Authors: Jørgensen Jens O. L. , Vahl Nina , Hansen Tony B. , Skjærbæk Christian , Fisker Sanne , Ørskov Hans , Hagen Claus , Christiansen Jens. S.
Published: March 2003

OBJECTIVE

Growth hormone status is an important determinant of serum IGF‐I but it is well known that hypopituitary adults with pronounced GH‐deficiency (GHDA) may exhibit normal IGF‐I levels. To elucidate possible causes of this apparent paradox we compared the significance of putative IGF‐I predictors in GHDA and normal subjects.

DESIGN

A cross‐sectional study.

SUBJECTS

Twenty‐seven GHDA (9 females, 18 males, mean ± SE age 44 ± 1 years) and 27 healthy control subjects (9 females, 18 males, mean ± SE age 43 ± 2 years).

RESULTS

Serum IGF‐I and IGFBP‐3 were significantly lower in GHDAs, but a considerable overlap existed (IGF‐I (μg/l) 87 ± 12 (GHDA) vs 177 ± 10 (Control) (P < 0.001)). In both Controls and GHDA, IGF‐I was higher in males than females (Control: 196 ± 12 vs 138 ± (P = 0.004); GHDA: 97 ± 16 vs 56 ± 11 (P = 0.05)). In GHDA, males on testosterone substitution had the highest IGF‐I concentrations. The molar IGF‐I:IGFBP‐3 ratio was significantly lower in GHDAs (0.18 ± 0.01 vs 0.23 ± 0.02 (P = 0.002)). IGFBP‐1 (μg/l) was significantly elevated in GHDAs (6.28 ± 1.11 vs 3.07 ± 0.32 (P < 0.001)) despite comparable fasting insulin levels. Percentage total body fat (TBF, DEXA, waist/hip ratio, and intra‐abdominal fat (CT) were all elevated in GHDAs. IGF‐I correlated positively with lean body mass (DEXA) and negatively with TBF and IGFBP‐1 in both groups. IGF‐I correlated negatively with age in CON but not in GHDAs, whereas IGF‐I correlated positively with IGFBP‐3 only in GHDAs. Multiple regression analysis revealed that age and IGFBP‐1 were the only significant predictors of IGF‐I in CON, whereas IGFBP‐3 and, to a lesser extent TBF, were the only independent predictors of IGF‐I in GHDAs. Neither peak stimulated GH, nor physical fitness contributed in any equations in the two groups.

CONCLUSIONS

1) IGF‐I levels are regulated by several variables in addition to GH status 2) age per se is an independent negative determinant in healthy subjects but not in GHDA 3) It is probable that some cases of paradoxically high IGF‐I levels in GHDA are secondary to inappropriately elevated IGFBP‐3 levels. 4) in mid‐adulthood males have higher IGF‐I levels than females and it is likely that testosterone directly stimulates IGF‐I. The influence of gender and sex steroids must therefore be accounted for when comparing IGF‐I levels between hypopituitary and healthy subjects.
Effect of corticotrophin‐releasing hormone on arginine vasopressin and atrial natriuretic factor in patients with Cushing's disease
Authors: Colao Annamaria , Pivonello Rosario , Ferone Diego , Faggiano Antongiulio , Facciolli Giuseppina , Di Somma Carolina , Boudouresque Françoise , Oliver Charles , Lombardi Gaetano
Published: March 2003

OBJECTIVE

To evaluate the effect of CRH administration on plasma AVP and ANF concentration in patients with Cushing's disease and healthy subjects.

SUBJECTS

Fifteen patients with Cushing's disease and 15 sex‐ and age‐matched healthy subjects entered the study.

STUDY DESIGN

All subjects were randomly given i.v. 100 μg hCRH and placebo (NaCl 0.9%) on two non consecutive days. Blood samples for plasma AVP and ANF assay were taken before and every 15 min for 2 hr after CRH or placebo. Five out of the 15 patients with Cushing's disease underwent a repeat CRH test after surgical cure of the disease.

RESULTS

At baseline evaluation, plasma ANF concentrations were significantly higher in patients with Cushing's disease compared to healthy subjects (15.0 ± 0.8 vs 10.8 ± 0.6 pmol/l, P < 0.001) whereas plasma AVP concentrations were similar between the two groups of subjects (8.0 ± 0.4 vs 6.8 ± 0.6 pmol/l). CRH administration induced a significant increase in plasma ANF concentrations both in patients with Cushing's disease (24.0 ± 1.6 pmol/l, P < 0.05) and healthy subjects (29.4 ± 1.5 pmol/l, P < 0.05). Conversely, a significant increase in plasma AVP concentrations was found only in patients with Cushing's disease (14.7 ± 1.4 vs 8.0 ± 0.4 pmol/l, P < 0.05). In addition, patients with Cushing's disease had an increase in ANF levels after CRH significantly lower than that observed in healthy subjects (peak/basal ratio: 1.7 ± 0.1 vs 3.1 ± 0.4, P < 0.01). In 5 patients re‐tested after disease cure, CRH administration significantly increased plasma ANF levels (26.8 ± 1.1 vs 10.7 ± 0.4 pmol/l, P < 0.05) but it did not modify plasma AVP levels (8.4 ± 0.4 vs 7.3 ± 0.5 pmol/l). ANF response to CRH in patients cured from Cushing's disease was similar to that recorded in healthy subjects (peak/basal ratio: 2.6 ± 0.2 vs 3.1 ± 0.4).

CONCLUSION

CRH stimulates ANF secretion both in patients with Cushing's disease and healthy subjects. Plasma ANF response to CRH is blunted in patients with Cushing's disease compared to patients cured from Cushing's disease and healthy subjects, probably because of hypercortisolism. By contrast, CRH stimulates AVP secretion only in patients with Cushing's disease, but not in patients cured from Cushing's disease and healthy subjects. This phenomenon could be related to the activity of the disease.
Effect of isoprenaline on plasma leptin and lipolysis in humans
Authors: Pinkney Jonathan H. , Coppack Simon W. , Mohamed‐Ali Vidya
Published: March 2003

OBJECTIVE

The sympathetic nervous system may play a central role in the regulation of both lipolysis and leptin production. Therefore, we investigated the effect of intravenous infusions of the β‐adrenergic agonist isoprenaline on plasma concentrations of leptin and nonesterified fatty acids.

DESIGN AND PATIENTS

Eight lean, healthy human volunteers, (4M:4F; median (interquartile range) age 36.5 (30.8‐40.0) years; BMI 22.9 (20.1‐29.2)kg.m−2; % body fat 24.5 (17.9–26.3)), were studied following an overnight fast. Intravenous infusion of isoprenaline was carried out for 3 h, followed by a 1 hour recovery phase. The isoprenaline infusion rates (0.5–3.5 μg.min−1) were titrated individually for each subject in order to achieve similar biological sympathetic responses based on heart rate (target heart rates were > 100 min−1 but < twice resting heart rate).

MEASUREMENTS

Plasma leptin was determined using an in‐house radioimmunoassay, nonesterified fatty acids estimated with an enzymatic colourimetric assay and insulin concentrations were assayed using a specific, two‐site immunoenzymometric assay.

RESULTS

Fasting preinfusion plasma leptin concentrations (6.3 (3.0–12.8)μg/l) correlated with percentage body fat measured by bioimpedance (r = 0.95; P < 0.001). Plasma leptin concentrations were rapidly suppressed by isoprenaline, with maximal suppression (20.5 (15.0–25.0)% of preinfusion levels (Wilcoxon rank sum test; P < 0.05)), observed after 2 h. In the recovery period, plasma leptin concentrations rapidly returned to preinfusion levels (postinfusion vs maximally suppressed leptin concentrations P < 0.05; vs preinfusion leptin concentrations P = NS). Plasma nonesterified fatty acids and insulin concentrations showed opposite changes to those observed with leptin.

CONCLUSION

Plasma leptin concentrations are rapidly and reversibly suppressed by the infusion of isoprenaline in humans in vivo.