bluuebunny/crossref_metadata_2025 huggingface dataset:
T cell responses to vaccines in infants: defective IFNγproduction after oral polio vaccination
Authors: VEKEMANS J , OTA M O C , WANG E C Y , KIDD M , BORYSIEWICZ L K , WHITTLE H , MCADAM K P W J , MORGAN G , MARCHANT A
Published: March 2003

The immaturity of the neonatal immune system is associated with an increased susceptibility to infections. Studies in mice indicate that neonatal immune responses are biased towards the T helper 2 type, but little is known about helper T cell responses in human newborns. In this study, the oral polio vaccine was used as a model of early immunization to investigate the capacity of young infants to develop cellular immune responses. We show that neonatal immunization with oral polio vaccine induces the production of high titres of neutralizing antibodies but reduced proliferative and IFNγ responses to polio antigens compared to immune adults. These data suggest that specific strategies will be required to immunize newborns against pathogens controlled by Th1 type immune responses.
Lack of muco-cutaneous signs of toxic shock syndrome when T cells are absent:S. aureusshock in immunodeficient adults with multiple myeloma
Authors: KAMEL N S , BANKS M C , DOSIK A , URSEA D , YARILINA A A , POSNETT D N
Published: March 2003

Staphylococcal toxic shock syndrome (TSS) is an acute life threatening disease. The diagnosis can be made clinically based on diagnostic criteria. The clinical manifestations are caused in large part by the release of high levels of T-cell-derived cytokines as a result of potent toxins, also called superantigens (SAg), produced by Staphylococcus aureus, but it is not clear which clinical symptoms/signs are strictly T-cell dependent. Here, we report on three adults with multiple myeloma (MM) presenting with S. aureus sepsis/shock, and two patients with typical TSS. The MM patients had compromised humoral immunity because of depression of normal immunoglobulin (Ig) levels at the expense of the M protein. In addition, their T cells were absent due to high dose chemotherapy initiated for bone marrow transplantation. The MM cases lacked mucosal hyperemia, erythroderma and desquamation, but were otherwise indistinguishable from the TSS cases. All patients grew S. aureus and in each case, SAg genes were detected by PCR. In several cases, the plasma contained biological SAg activity resulting in Vβ specific proliferation of indicator T cells in vitro. The same specific activity was observed with the supernatant fluids of S. aureus broth cultures from the respective bacterial isolates. This confirms the presence of bio-active toxins in the plasma but did not lead to full blown TSS when T cells were lacking. Thus, S. aureus sepsis/shock can be clinically distinguished from typical TSS, and we suggest that muco-cutaneous manifestations of TSS are the most telling signs of massive T-cell-dependent cytokine release.
Rotavirus infections and development of diabetes-associated autoantibodies during the first 2 years of life
Authors: BLOMQVIST M , JUHELA S , ERKKILÄ S , KORHONEN S , SIMELL T , KUPILA A , VAARALA O , SIMELL O , KNIP M , ILONEN J
Published: March 2003

Rotavirus, the most common cause of childhood gastroenteritis, has been implicated as one of the viral triggers of diabetes-associated autoimmunity. To study the possible association between rotavirus infections and the development of diabetes-associated autoantibodies, we measured the prevalence of rotavirus antibodies in serum samples collected at 3–6-month intervals up to the age of 2 years from 177 children selected from consecutive newborns because they carried HLA-DQB1 alleles associated with increased risk for type 1 diabetes. Twenty-nine of the children developed at least two of four diabetes-associated autoantibodies (ICA, IAA, GADA or IA-2A) during the first 2 years of life (the cases), whereas 148 children remained autoantibody-negative matched with the cases for date of birth, gender, living region and HLA-DQB1 alleles. The temporal association between the development of the first-appearing diabetes-associated autoantibody and rotavirus infections was studied by analysing whether the cases had a diagnostic increase in rotavirus antibody titre more often during the 6-month period that preceded seroconversion to autoantibody positivity than the controls. By the age of 12 months one of the 13 case children (7%), who had a serum sample drawn at that age and who had developed at least one type of diabetes-associated autoantibodies, had experienced a rotavirus infection, while 12 of the 61 (20%) autoantibody-negative control children had had a rotavirus infection. By 18 months, four of the 22 autoantibody-positive cases (18%) and 18 of the 89 controls (20%) had rotavirus antibodies, and by the age of 24 months the respective numbers were five of the 27 cases (19%) and 32 of the 113 (28%) controls. A rotavirus infection occurred during the 6 months preceding the sample which was positive for an autoantibody in four of the 25 periods (16%) for which both necessary samples were available, while the controls had a rotavirus infection during 55 of the 370-such periods (15%). Accordingly, our data suggest that rotavirus infections are unlikely triggers of beta-cell autoimmunity in young children with genetic susceptibility to type 1 diabetes.
Modification of bacteraemia by specific antibodies and relation with mortality in a pneumococcal mouse sepsis model
Authors: YUSTE J , JADO I , GIMÉNEZ M J , AGUILAR L , MOLERO F , FENOLL A , CASAL J
Published: March 2003

The relationship between mortality and the bacteraemic profile was investigated in a pneumococcal (serotype 6B) sepsis BALB/c mouse model where animals received protection by specific hyperimmune serum. A single intraperitoneal dose of hyperimmune serum obtained from mice immunized with the heat-inactivated strain was administered (non-diluted or diluted to 1/4 or to 1/16) to 5-mice study groups 1 h prior to intraperitoneal inoculation with the infective inoculum (3·57 × 108 cfu/ml). Blood cultures were performed daily over 15 days, with 8 μl of blood being collected from the tail vein; the samples were resuspended in Todd-Hewitt broth containing 10% trisodium citrate and plated onto blood agar for colony counting. Animals included in the control group received placebo (PBS). Mortality was 100% in control animals within the first 48 h. Hyperimmune serum decreased and delayed mortality in a dose-related trend, producing 100%, 80%, 60% and 40% survival rates at 72, 96, 144 and 360 h, with non-diluted serum. Bacteraemic profiles with maximum colony counts ≥5 × 107 cfu/ml in blood during the follow-up period were related to ≥65% probability of death, regardless of the serum dilution administered.
Human proteinase 3 can inhibits LPS-mediated TNF-alpha production through CD14 degradation: lack of influence of antineutrophil cytoplasmic antibodies
Authors: YARD B A , WILLE A-I , HAAK M , V AN D ER WOUDE F J
Published: March 2003

The present study was conducted to investigate if proteinase-3 (PR3) is able to influence lipopolysaccharide (LPS) responses of monocytes via degradation of CD14 and if antineutrophil cytoplasmic antibodies (ANCA) may modify this process. Recombinant (r) CD14 and CD14 expressed on monocytes were investigated for PR3 mediated degradation by SDS-PAGE and FACS analysis, respectively. TNF-α production in whole blood was used to determine functional consequences of CD14 degradation. PR3 degraded rCD14 in a dose- and time-dependent fashion. Major degradation products were found with apparent molecular weight of 45, 25 and 10 kDa. Treatment of PR3 with PMSF completely abolished CD14 degradation. ANCA IgG did not inhibit CD14 degradation. In whole blood, addition of PR3 resulted in diminished CD14 expression on monocytes. In contrast, CD14 was increased in a subpopulation of cells that expressed major histocompatibility (MHC) class II and PR3, but lacked expression of CD64 and CD16. LPS mediated TNF-α production in whole blood was significantly inhibited when preincubated with PR3. This study demonstrates that PR3 can degrade rCD14 and that PR3 differentially affects CD14 expression in subsets of monocytes. ANCA IgG does not play a significant role herein.
CD161+ T (NT) cells exist predominantly in human intestinal epithelium as well as in liver
Authors: IIAI T , WATANABE H , SUDA T , OKAMOTO H , ABO T , HATAKEYAMA K
Published: March 2003

It has been reported that human CD161 (NKR-P1A)+ T cells are counterparts of murine natural T (NT) cells and predominantly accumulate in the liver. However, NT cells in the human intestine have not been well analysed. The aim of this study was to assess the existence of NT cells in human intestinal epithelium and determine their phenotypical characterization. Intra-epithelial lymphocytes (IEL) were isolated from surgical specimens (jejunum, ileum and colon). The surface phenotype of IEL was analysed using a FACScan and compared with that of mononuclear cells (MNC) from other organs. CD161+ T cells were abundant in human intestinal epithelium as well as the liver. The majority of CD161+ T cells in IEL were CD8+ cells. About 50% of CD161+ T cells in hepatic lymphocytes (HL) expressed CD56, whereas only 14% of CD161+ T cells in IEL expressed CD56. The jejunum showed the greatest abundance of CD161+ T cells among the intestinal regions investigated. These results suggest that CD161+ T (NT) cells predominantly exist in human intestinal epithelium and may play an important role in local immunity.
β7 Integrin expression is not required for the localization of T cells to the intestine and colitis pathogenesis
Authors: SYDORA B C , WAGNER N , LÖHLER J , YAKOUB G , KRONENBERG M , MÜLLER W , ARANDA R
Published: March 2003

β7 Integrins have been shown to have an important role in the localization of T cells to the intestine. Utilizing two different experimental mouse models of inflammatory bowel disease (IBD), this study was undertaken to determine if β7 integrin expression is critical for T cell localization to the intestine and colitis pathogenesis. Transfer of CD4+ CD45RBhigh cells into immunodeficient mice results in colitis. To examine the role of β7 integrins, donor cells were obtained from β7 integrin gene-deficient animals and disease induction was examined following transfer into severe combined immunodeficiency (SCID) mice. Additionally, β7 integrin gene-deficient animals were crossed to IL-2-deficient mice and the onset of spontaneous colitis that normally occurs in IL-2-deficient animals was examined. No differences in the onset or severity of spontaneous colitis was noted in animals that were deficient in both β7 integrin and IL-2. In contrast, the onset of colitis in recipients of T cells from β7 integrin-deficient donors was delayed significantly. In mice receiving β7 integrin negative cells, the initial lack of colitis appeared to correlate with fewer numbers of CD3+β7 integrin –/– donor lymphocytes present in the host colon. The eventual development of disease, however, was associated with increased numbers of donor β7 integrin –/– lymphocytes. These results show that β7 integrin expression is not absolutely required for T cell localization to the intestine and colitis pathogenesis.
Antinuclear autoantibodies (ANA) in Gulf War-related illness and chronic fatigue syndrome (CFS) patients
Authors: SKOWERA A , STEWART E , DAVIS E T , CLEARE A J , UNWIN C , HULL L , ISMAIL K , HOSSAIN G , WESSELY S C , PEAKMAN M
Published: March 2003

It is established that veterans of the 1991 Gulf War have an increased frequency of experiencing multiple symptoms. The underlying mechanism of these ailments is unclear, although they do not correspond to any clearly defined syndrome. The most common symptoms overlap with those of chronic fatigue syndrome (CFS). CFS was recently associated with a novel subtype of antinuclear autoantibody (ANA) that reacts with nuclear envelope (NE) antigens. NE autoantibodies are not known to be linked with any distinct clinical condition, but have been observed in patients with unusual mixed chronic autoimmune disorders and connective tissue diseases. In this study we examined whether NE ANAs are a feature of patients with CFS and symptomatic Gulf War veterans (sGWV). We studied the prevalence of ANA in 130 sGWV, 90 well Gulf War veterans (wGWV), 128 symptomatic Bosnia and Era veterans (sBEV), 100 CFS patients, and 111 healthy control subjects matching for age and sex. We found no significant difference in the prevalence of ANAs between any of the groups. None of the patients/or veterans we studied had ANA of the NE type. Our results show that multisymptom illness due to CFS or related to Gulf War service is not associated with antinuclear autoimmunity.
Nitric oxide: a regulator of mast cell activation and mast cell-mediated inflammation
Authors: COLEMAN J W
Published: March 2003

Nitric oxide (NO) plays diverse roles in physiological and pathological processes. During immune and inflammatory responses, for example in asthma, NO is generated at relatively high and sustained levels by the inducible form of nitric oxide synthase (NOS-2). NOS-2 derived NO regulates the function, growth, death and survival of many immune and inflammatory cell types. In the case of mast cells, NO suppresses antigen-induced degranulation, mediator release, and cytokine expression. The action of NO on mast cells is time dependent, requiring several hours, and noncGMP mediated, most probably involving chemical modification of proteins. NO inhibits a number of mast cell-dependent inflammatory processes in vivo, including histamine mediated vasodilatation, vasopermeation and leucocyte-endothelial cell attachment. In human asthma and animal models of lung inflammation the role of NO is harder to define. However, although there are conflicting data, the balance of evidence favours a predominantly protective role for NO. Mimicking or targeting NO dependent pathways may prove to be a valuable therapeutic approach to mast cell mediated diseases.
X-chromosome inactivation analysis in a female carrier of FOXP3 mutation
Authors: TOMMASINI A , FERRARI S , MORATTO D , BADOLATO R , BONIOTTO M , PIRULLI D , NOTARANGELO L D , ANDOLINA M
Published: March 2003

Immune dysregulation, polyendocrinopathy and enteropathy with X-linked inheritance (IPEX) is a serious disease arising from mutations in FOXP3. This gene codifies for a transcription factor whose dysfunction results in hyperactivation of T cells. It is not clear, however, why an intermediate phenotype is not seen in heterozygous females, who are completely healthy. In order to address this question, we investigated X-chromosome inactivation in peripheral blood lymphocytes from a heterozygous female with a child affected by IPEX. No preferential inactivation was shown in freshly sorted CD4+, CD8+, CD19+ cells or in IL-2 cultured CD4 and CD8 T cells, indicating that peripheral blood lymphocytes in these women are randomly selected. Moreover, only one single FOXP3 transcript was expressed by CD4 T cell clones analysed by RT-PCR, confirming that this gene is subject to X- inactivation. We hypothesize that hyper-activation of T cell in carriers of FOXP3 mutations is regulated by the presence of normal regulatory T cells.
Inflammatory cytokine (interleukin 6 and tumour necrosis factor alpha) release in a human whole blood system in response toStreptococcus pneumoniaeserotype 14 and its capsular polysaccharide
Authors: JAGGER M P , HUO Z , RICHES P G
Published: March 2003

Gram-positive bacteria, which lack lipopolysaccharide (LPS), produce a septic-shock-like condition, accompanied by release of pro-inflammatory cytokines. Various components of the bacteria may be responsible for this. We stimulated a whole blood system with heat-inactivated Streptococcus pneumoniae serotype 14 (S14) bacteria, with pneumococcal S14 capsular polysaccharide (PPS S14) and with PPS S14 coated on to latex beads, to compare interleukin 6 (IL-6) and tumour necrosis factor alpha (TNFα) production over a six hour period, to ascertain the contribution of PPS to the inflammatory response. This was compared with the response to LPS. After sonication of the bacteria, their PPS content was estimated by an enzyme-linked immunoabsorbent assay, to compare this with the concentration of free PPS needed to generate cytokine release. The whole bacteria elicited a much larger cytokine response than the equivalent amount of PPS alone, whereas the PPS-coated beads gave minimal response. The different cytokine responses to PPS and LPS suggest that there are differences in the receptors and/or signalling pathways for Gram-negative and Gram-positive bacteria. We conclude that the estimated amount of PPS in the bacteria is not enough to account for the large cytokine response we observed. Since PPS could not be shown to contribute significantly to cytokine induction, specific antibodies to PPS would not play any significant role in combating cytokine release associated with pneumococcal infection and possible septic shock. This needs to be considered in production of future vaccines.
Differential effects of prophylactic, concurrent and therapeutic lactoferrin treatment on LPS-induced inflammatory responses in mice
Authors: KRUZEL M L , HARARI Y , MAILMAN D , ACTOR J K , ZIMECKI M
Published: March 2003

Mice injected with endotoxin develop endotoxaemia and endotoxin-induced death, accompanied by the oxidative burst and overproduction of inflammatory mediators. Lactoferrin, an iron binding protein, provides a natural feedback mechanism to control the development of such metabolic imbalance and protects against deleterious effects of endotoxin. We investigated the effects of intraperitoneal administration of human lactoferrin on lipopolysaccharide (LPS)-induced release of tumour necrosis factor alpha (TNF-α), interleukin 6 (IL-6), interleukin 10 (IL-10) and nitric oxide (NO) in vivo. Lactoferrin was administered as a prophylactic, concurrent or therapeutic event relative to endotoxic shock by intravenous injection of LPS. Inflammatory mediators were measured in serum at 2, 6 and 18 h post-shock induction. Administration of lactoferrin 1 h before LPS resulted in a rather uniform inhibition of all mediators; TNF by 82%, IL-6 by 43%, IL-10 by 47% at 2 h following LPS injection,and reduction in NO (80%) at 6 h post-shock. Prophylactic administration of lactoferrin at 18 h prior to LPS injection resulted in similar decreases in TNF-α (95%) and in NO (62%), but no statistical reduction in IL-6 or IL-10. Similarly, when lactoferrin was administered as a therapeutic post-induction of endotoxic shock, significant reductions were apparent in TNF-α and NO in serum, but no significant effect was seen on IL-6 and IL-10. These results suggest that the mechanism of action for lactoferrin contains a component for differential regulation of cellular immune responses during in vivo models of sepsis.
Distinct antigenic features of linear epitopes at the N-terminus and C-terminus of 65 kDa glutamic acid decarboxylase (GAD65): implications for autoantigen modification during pathogenesis
Authors: AL-BUKHARI T A M A , RADFORD P M , BOURAS G , DAVENPORT C , TRIGWELL S M , BOTTAZZO G-F , LAI M , SCHWARTZ H L , TIGHE P J , TODD I
Published: March 2003

Autoantibodies to 65 kDa glutamic acid decarboxylase (GAD65) are produced in many patients with autoimmune polyendocrine syndrome type II (APS-II) or stiff-man syndrome (SMS) and are heterogeneous in their epitope specificities, recognizing both conformational and linear determinants. Major linear epitopes of GAD, which are recognized by autoantibodies in a minority of these patients, occur in the N-terminal and C-terminal regions. We have investigated antibody recognition of the N- and C-termini of GAD65 in relation to their structural features as an approach to understanding what modifications to the native GAD structure may occur that facilitate the generation of antibodies specific to linear epitopes in these regions during the autoimmune pathogenesis. A monoclonal antibody specific to the N-terminus of GAD65 bound both native and denatured GAD in ELISA, whereas monoclonal and polyclonal antibodies specific to the C-terminus of GAD bound only denatured GAD. These antibodies were epitope mapped using random peptide phage-display libraries and the epitopes related to a previously proposed structural model of GAD65. This has led us to propose that the α-helical secondary structure of the C-terminus of GAD65 must be denatured to generate linear epitopes. In contrast, the N-terminus is both surface exposed and linear in the native structure, but may be masked by membrane interactions, which must be broken to facilitate recognition by B cells.
Increased expression of mucosal addressin cell adhesion molecule-1 (MAdCAM-1) and lymphocyte recruitment in murine gastritis induced byHelicobacter pylori
Authors: HATANAKA K , HOKARI R , MATSUZAKI K , KATO S , KAWAGUCHI A , NAGAO S , SUZUKI H , MIYAZAKI K , SEKIZUKA E , NAGATA H , ISHII H , MIURA S
Published: March 2003

Although  T  cell  involvement  in  Helicobactor  pylori-induced  gastritis  is  known,  mechanism  about  T cell recruitment is not understood. In this study we examined how mucosal addressin cell adhesion ­molecule-1 (MAdCAM-1) is involved in lymphocyte recruitment in murine chronic gastritis induced by H. pylori. C57 BL/6 mice were infected with Sydney strain (SS1). Six months after infection, the stomach was removed. The expression of adhesion molecules, MAdCAM-1, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), and the cell surface antigens CD4, CD8, CD45R/B220 or β7-integrin were determined by immunohistochemistry. A significant increase in CD4 lymphocytes was observed in the body portion of stomach in SS1-infected mice and most of these CD4 cells express β7-integrin, a known counter ligand for MAdCAM-1 molecule. Strong MAdCAM-1 expression was observed adjacent to these cells in the lamina propria as well as in the submucosa of SS1-infected stomach. Quantitative analysis showed that the area of MAdCAM-1 expression well correlated with the infiltration of β7-integrin positive lymphocytes. On the other hand, expression of ICAM-1 or VCAM-1 in the lamina propria was few even in the SS1-infected stomach. Increased expression of MAdCAM-1 was well correlated to the location of lymphocytes, which express CD4 and β7-integrin. These results suggest the possibility that MAdCAM-1 may be largely involved in the lymphocyte recruitment in the gastritis mucosa with H. pylori.
Nasal administration of CTB-insulin induces active tolerance against autoimmune diabetes in non-obese diabetic (NOD) mice
Authors: ASPORD C , THIVOLET C
Published: March 2003

Nasal administration of beta cell-derived auto-antigens has been reported to suppress the development of autoimmune diabetes. We investigated the tolerogenic effects of insulin conjugated to the B subunit of cholera toxin (CTB). Nasal administration of 1 µg of CTB-insulin significantly delayed the incidence of diabetes in comparison to CTB treated mice. However, administration of 4 or 8 µg of the conjugate had no protective effect. Protection induced by CTB-insulin was transferred to naive recipients by splenic CD4+ T cells. This result favours an active cellular mechanism of regulation, which was lost using higher (4–8 µg) or lower (0·1–0·5 µg) amounts of the conjugate. When co-administered with diabetogenic  T  cells,  splenic  T  cells  from  CTB-insulin-treated  mice  reduced  the  lymphocytic  infiltration  of the islets. Reverse transcription-polymerase chain reaction analysis of recipients’ pancreatic glands revealed an increase of TGF-β and IL-10 transcripts after donor mice tolerization, while levels of IFN-γ and IL-4 RNAs were unchanged. We observed a significant increase of T cell proliferation after unspecific stimulation in the spleen and pancreatic lymph nodes 24 h after CTB-insulin administration in ­comparison to control treatment. Higher amounts of IL-4 and IFN-γ   were noticed in pancreatic lymph nodes of tolerized mice upon in vitro stimulation. Antigen-specific unresponsiveness after immunization and upon subsequent in vitro exposure to homologous antigen was obtained in nasally treated animals. Our results underlined the importance of nasal mucosa as an inducing site of tolerance and provided evidence for similar mechanisms of action to what has been described for the oral route, which favoured a CTB-insulin specific effect.
Up-regulation of the dendritic cell marker CD83 on polymorphonuclear neutrophils (PMN): divergent expression in acute bacterial infections and chronic inflammatory disease
Authors: IKING-KONERT C , WAGNER C , DENEFLEH B , HUG F , SCHNEIDER M , ANDRASSY K , HÄNSCH G M
Published: March 2003

Upon cultivation with interferon-γ (IFN-γ) and granulocyte/macrophage-colony stimulating factor (GM-CSF) polymorphonuclear neutrophils (PMN) acquire characteristics of dendritic cells, including expression of major histocompatibility complex (MHC) class II antigens, of the co-stimulatory antigens CD80, CD86 and of CD83, the latter considered to be specific for dendritic cells. Dendritic-like PMN were also able to present to T cells antigens in a MHC class II-restricted manner. To assess whether dendritic-like PMN are also generated in vivo, cells of patients with acute bacterial infections and of patients with chronic inflammatory diseases (primary vasculitis) were tested. During acute infection up to 80% of PMN acquired CD83, but remained negative for MHC class II, CD80 or CD86. PMN of patients with primary vasculitis expressed MHC class II antigens, CD80 and CD86, but not CD83, indicating that up-regulation of MHC class II and of CD83 are not necessarily linked to each other. Indeed, parallel studies with PMN of healthy donors showed that while IFN-γ and granulocyte/macrophage colony stimulating factor (GM-CSF) induced both, MHC class II and CD83, tumour necrosis factor (TNF)-α selectively induced de novo synthesis of CD83. The function of CD83 on PMN is still elusive. A participation in the MHC class II-restricted antigen presentation could be ruled out, consistent with the segregation of MHC class II and CD83 expression. Regardless, however, of its function, CD83 expression could serve as a marker to differentiate between acute and chronic inflammation.
Induced sputum-retrieved matrix metalloproteinase 9 and tissue metalloproteinase inhibitor 1 in granulomatous diseases
Authors: FIREMAN E , KRAIEM Z , SADE O , GREIF J , FIREMAN Z
Published: March 2003

Matrix metalloproteinases (MMPs) capable of degrading various components of connective tissue matrices, and tissue inhibitor metalloproteinases (TIMPs) are considered important in lung parenchymal remodeling and repair processes in pulmonary diseases. Induced sputum (IS) is a reliable noninvasive method to investigate pathogenesis, pathophysiology and treatment of lung disease. This study was designed to determine whether IS-MMP9/TIMP1 levels demonstrate lung parenchymal remodeling in sarcoidosis (SA) and Crohn's disease (CRD) patients. Sputum was induced and processed conventionally in 13 SA patients, 18 CRD patients and 9 controls. Two-hundred cells were counted on Giemsa-stained cytopreps, and T lymphocytes subsets (CD4 = T helper and CD8 = T suppressor cytotoxic cells) were analysed by FACS using monoclonal antibodies.MMP-9 and TIMP-1 were measured using commercial ELISA kits. MMP-9 concentrations, but not those of TIMP-1, were significantly greater in the sputum supernatant in SA and CRD patients compared to controls (P = 0·018 and P= 0·0019, respectively). The molar ratio, MMP-9/TIMP-1, was significantly higher in SA and CRD patients compared to controls (P = 0·008 and P= 0·024, respectively). Gelatinase species having a molecular weight similar to that of MMP-9 were demonstrated by zymographic analysis. MMP-9 levels were highly correlated with the CD4/CD8 ratio and DLCO capacity in SA but less in CRD patients. MMP-9 levels in IS provide a sensitive marker for pulmonary damage.
Interferon-gamma levels in nasopharyngeal secretions of infants with respiratory syncytial virus and other respiratory viral infections
Authors: JOSHI P , SHAW A , KAKAKIOS A , ISAACS D
Published: March 2003

Respiratory syncytial virus (RSV) infection, one of the most common causes of hospitalization of children in developed countries, has been implicated as a cause of asthma. We aimed to characterize the cytokine profile in nasopharyngeal aspirates (NPAs) taken from infants during upper respiratory tract infection to investigate whether RSV induced a unique immune response as compared with other viruses. Additionally, we sought to determine whether this profile was influenced by the infants’ atopic status. A prospective birth cohort of babies at high risk of atopy was recruited. Ratios of a T-helper 1 (Th1) cytokine, interferon gamma (IFN-γ) and a T-helper 2 (Th2)-like cytokine, interleukin-10 (IL-10), in NPAs were determined during episodes of respiratory tract infections in the first year. The viral aetiology of the respiratory tract infections was determined using polymerase chain reaction (PCR), culture and immunofluorescence. Atopic status was ascertained at 1 year of age using skin prick tests. Participants were recruited antenatally and subsequently followed in the community. Sixty babies with one or both parents atopic were enrolled into the study. IFN-γ : IL-10 ratios in NPAs during upper respiratory tract infections and their correlation with viral aetiology and atopic status were the main outcome measures. The mean IFN-γ : IL-10 ratio was significantly lower (due to lower IFN-γ) during RSV infections than during infections with other viruses (P = 0·035). The cytokine ratio, however, did not differ between infants with or without wheeze during URTIs (P = 0·44), or between infants who were atopic or non-atopic (P = 0·49). This study suggests that RSV is associated with lower IFN-γ production in young babies, regardless of their atopic status, compared to upper respiratory tract infections where either another virus is detected or where no viral identification is made.
Influence ofMycobacterium tuberculosison differential activation of helper T-cells
Authors: TALREJA J , BHATNAGAR A , JINDAL S K , GANGULY N K
Published: March 2003

Host defence against tuberculosis infection involves T-lymphocyte mediated cellular immune responses. In this study we assessed T-cell activation by studying the early signal transduction events and production of cytokines by human CD4+ T-cells. The study constituted of five groups of subjects: (a) untreated acid fast bacilli (AFB)+ve TB patients who have not started anti-tuberculosis therapy (ATT) [New]; (b) patients who have taken ATT for two months [2T]; (c) patients who have taken ATT for six months [6T]; (d) mantoux positive healthy controls [T+ve]; (e) mantoux negative healthy controls [T−ve]. We found that mantoux positive healthy controls produced significantly higher levels of IP3, intracellular Ca2+ and presented increased PKC activity when CD4+ T-cells were stimulated with M. tuberculosis H37Rv cell lysate as compared to mantoux negative controls. Furthermore, decreased expression of CD54 (ICAM-1) and reduced [Ca2+]i were seen in TB patients as compared to T+ve healthy controls. TB patients showed significantly lower levels of IL-2 and IFNγ and higher levels of IL-4 as compared to normal healthy controls, suggesting a diminished Th1 response. Thus, the reciprocal changes in cytokines, reduced [Ca2+]i levels, and CD54 expression in patients imply phenotype shifting of Th precursors to Th2 type in TB patients.
Analysis of gene expression in peripheral blood eosinophils from patients with atopic dermatitis andin vitrocytokine-stimulated blood eosinophils
Authors: OGAWA K , HASHIDA R , MIYAGAWA M , KAGAYA S , SUGITA Y , MATSUMOTO K , KATSUNUMA T , AKASAWA A , TSUJIMOTO G , SAITO H
Published: March 2003

Investigation of differentially expressed genes in eosinophils of patients with allergic diseases such as atopic dermatitis (AD) will provide important information for elucidating possible mechanisms of pathology. To identify novel genes that are expressed in AD, we compared gene expression in samples of peripheral blood eosinophils from AD patients and healthy volunteers. RNA was extracted from peripheral blood eosinophils. The expression of various genes, such as those for cytokine receptors, eosinophil activation marker, platelet activating factor (PAF) receptor, eosinophil-specific granular proteins and apoptosis-related genes, was confirmed using real-time reverse transcription–polymerase chain reaction (RT-PCR). Peripheral blood eosinophils of healthy volunteers were also isolated and stimulated for introduction of various cytokines. RNA was extracted and gene expression was monitored. Several genes, such as those for cytokine receptors (granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor α and β chain and interleukin (IL)-3 receptor α chain), CD44 and PAF receptor were expressed at significantly higher levels in AD patients than in healthy volunteers. In addition, the anti-apoptotic genes, bcl-2 and bcl-xL, were expressed at increased levels in AD patients. No single gene expression correlated with clinical markers, such as eosinophil count or IgE levels. Expression of GM-CSF receptor β chain and IL-3 receptor α chain in isolated blood eosinophils of healthy volunteers was stimulated by IL-5, IL-4, interferon (IFN)-γ and GM-CSF. Expression of bcl-2 and bcl-xL was also increased after stimulation with IL-5, IL-4 or IFN-γ. The in vitro enhancement of cytokine-stimulated gene expression correlated well with the enhancement observed in clinical samples of eosinophils, suggesting that cytokines may affect gene expression in vivo in eosinophils of patients with AD.
LiveLeishmaniapromastigotes can directly activate primary human natural killer cells to produce interferon-gamma
Authors: NYLÉN S , MAASHO K , SÖDERSTRÖM K , ILG T , AKUFFO H
Published: March 2003

Natural killer (NK) cells have been implicated in the natural protection and healing of leishmaniasis by their ability to secrete the macrophage activating cytokine interferon (IFN)γ. Previous studies have demonstrated that early production of interleukin (IL)-12 triggers IFNγ secretion by NK cells. Here we report that live Leishmania promastigotes (the form that is injected by the vector) can directly induce human peripheral blood NK cells from healthy donors to IFNγ secretion in the absence of IL-12 and professional antigen presenting cells. Killing of promastigotes abolishes this property. This novel mechanism of activation of the innate immune response may be relevant for establishment of infection and thus also the design of vaccines against leishmaniasis.
Prolactinomas express human heparin‐binding secretory transforming gene (hst) protein product: marker of tumour invasiveness
Authors: Shimon Ilan , Hinton David R. , Weiss Martin H. , Melmed Shlomo
Published: March 2003

BACKGROUND AND OBJECTIVE

We have shown previously that heparin‐binding secretory transforming gene (hst) overexpression in rat pituitary cells mediates lactotroph tumour growth and stimulates PRL transcription, and that transforming sequences of the gene, which encode fibroblast growth factor‐4 (FGF‐4), are expressed in human prolactinomas. To further determine the role of hst in human PRL‐secreting adenoma pathogenesis we studied the presence of hst protein in these tumours and other types of human pituitary adenoma.

PATIENTS AND DESIGN

Pituitary adenoma tissue samples were obtained at surgery from 14 patients with PRL‐secreting adenomas, 5 patients with GH‐secreting tumours, 3 with ACTH‐secreting, and 13 patients with nonfunctioning tumours. Two normal pituitary tissue specimens were also studied. Clinical data, including tumour invasiveness assessed by preoperative MRI studies, were available. For hst protein immunolocalization, tumour frozen sections were immunostained with antihuman FGF‐4 antibody. Immunoperoxidase staining for the proliferation‐related nuclear antigen Ki‐67 was performed using MIB‐1 monoclonal antibody.

RESULTS

Normal anterior pituitary cells did not contain immunoreactive hst protein. Lactotrophs in five of 14 prolactinomas (36%) stained strongly for hst compared with immunoreactive pituicytes in only one of 21 nonfunctioning, GH‐, and ACTH‐secreting adenomas (P = 0.05). Immunoreactive hst in adenoma cells was detected in 3 of 5 invasive prolactinomas, and in 2 of 9 noninvasive PRL‐cell adenomas. Immunostaining for the proliferation‐related antigen Ki‐67 showed a higher proliferation index in hst‐positive adenomas (3.94 + 0.85%) as compared with those immunonegative for hst (1.98 + 0.7%; P = 0.05).

CONCLUSIONS

hst protein may be directly involved in prolactinoma development or progression, particularly in invasive tumours, probably due to the growth promoting effects of FGF‐4.
Melatonin administration and pituitary hormone secretion
Authors: Kostoglou‐Athanassiou Ifigenia , Treacher David F. , Wheeler Michael J. , Forsling Mary L.
Published: March 2003

OBJECTIVE

The relationship between the pineal gland and pituitary function remains controversial, while the role of melatonin in the adaptation of the organism to the light‐dark cycle of the environment is becoming increasingly recognized. The aim of this study was to investigate the effect of a manipulation of the melatonin rhythm on pituitary hormone secretion in man.

DESIGN

Double‐blind controlled clinical study.

SUBJECTS

Ten adult healthy male volunteers, aged 21–33 years, were studied on two occasions: once after the administration of melatonin 5 mg orally for 4 days at 1700 hours and once after the administration of placebo, at similar times. On the day of each study the subjects undertook their normal duties but refrained from taking heavy exercise, from smoking and drinking alcohol.

MEASUREMENTS

Serum cortisol, growth hormone, prolactin and plasma vasopressin, oxytocin, melatonin, sodium, potassium, osmolality and packed cell volume were measured over the following 24 hours.

RESULTS

The cortisol peak was advanced and prolactin release increased after melatonin administration, while growth hormone was not affected. Vasopressin and oxytocin levels were found to increase during the night in the control study, but the period of the nocturnal increase in vasopressin concentrations was reduced after the administration of melatonin and the nocturnal increase of oxytocin was absent.

CONCLUSION

Altering the melatonin rhythm may affect neuroendocrine function, influencing the nocturnal pattern of neurohypophysial hormone secretion, augmenting prolactin release and advancing the peak of cortisol release.
L‐Arginine and insulin‐tolerance tests in the diagnosis of adult growth hormone deficiency: influence of confounding factors
Authors: Fisker  , Jørgensen  , Ørskov  , Christiansen
Published: March 2003

OBJECTIVE

In the attempt to define a GH stimulation test with high specificity and reproductibility, few studies have addressed the influence of potential interfering external factors on the test result. We therefore tested the influence of physical activity (admission to hospital on test morning) and mild heat exposure on the GH response to l‐arginine stimulation test (Arg) and insulin‐tolerance test (ITT).

DESIGN

One Arg stimulation test and one ITT were performed in all subjects during standard conditions (overnight hospital stay, 10 hours fasting). In addition, each subject was randomized to undergo either two additional Arg tests, or two ITTs, performed under two different conditions: admission to hospital on the morning of the test and during standard conditions except for heat exposure before testing. The four tests were performed in random order.

PATIENTS

Twenty‐two patients (six women, 16 men) (mean age ± SEM, 38.3 ± 5.3 years and 36.1 ± 2.7 years, respectively) presenting with pituitary disease and a group of healthy age and gender‐matched normal subjects (six women, 13 men) (age 38.3 ± 4.8 years and 35.7 ± 2.4 years, respectively) participated.

MEASUREMENTS

During the GH‐stimulation tests serum GH, cortisol, blood glucose, and plasma glucagon were measured and compared in the three different test conditions.

RESULTS

During standard conditions, peak GH response was higher in the ITT compared to the Arg test in the control group (23.4 ± 3.6 mU/l vs 11.6 ± 2.0 mU/l, P = 0.004), and the specificity of the ITT was higher (18/19 versus 13/19, P = 0.047). Minor heat exposure before the ITT (temperature rise 0.24 ± 0.05°C, range 0.0–0.5°C) did not change the GH response in the healthy adults whereas admission to hospital on the morning of the test reduced the GH response significantly (P < 0.05). The lowest blood glucose did not change in the three situations and did not correlate with peak GH during the ITT. In the patients there were no significant differences between the GH response during different conditions. Plasma glucagon did not significantly differ between the different test conditions in the control group (P = 0.88), but there was a significant decrease in the glucagon response to the test performed after hospital admission on the test morning in the patients (P < 0.025). Serum cortisol response in the control group did not differ in the three situations.

CONCLUSIONS

Since provocative GH responses are influenced by external factors, conditions should be standardized to optimize the reproductibility and specificity of the tests. Furthermore the higher specificity of the insulin‐tolerance test as compared to the arginine stimulation test was confirmed.
Corticosteroid metabolism in human granulosa‐lutein cells
Authors: Thomas  , Thomas  , Tetsuka  , Mason  , Hillier
Published: March 2003

OBJECTIVE

The aims of this study were to determine the type and level of 11β‐hydroxysteroid dehydrogenase (11βHSD) in human granulosa‐leutein cells (GLE) shortly before ovulation and to correlate activity with the outcome of treatment in patients undergoing in vitro fertilization and embryo transfer (IVF/ET).

DESIGN

GLC from 32 patients undergoing IVF/ET were tested for type and level of 11βHSD activity in relation to treatment outcome.

PATIENTS

Periovulatory follicles were aspirated by ultrasound guided transvaginal puncture following a standard controlled ovarian stimulation protocol, ∼36 h after administering an ovulation‐inducing dose of human chorionic gonadotrophin (HCG). GLC were separated from follicular fluid by density‐gradient centrifugation and taken for measurement of 11βHSD activity in vitro; oocytes were used for IVF/ET.

MEASUREMENTS

Interconversion of cortisol (F) and cortisone (E), and dexamethasone (D) and 11‐dehydrodexamethasone (DHD) was measured in standardized assays comprising incubation of GLC with 3H‐labelled substrate, with separation of substrate and product by thin‐layer radiochromatography.

RESULTS

Conversion of F to E varied from 10.5 to 30.9% while that of E to F was between 2.4 and 44.6%. In the GLC of 25 patients in whom both activities were measured, dehydrogenase (F to E) activity predominated in 13 and reductase (E to F) in 12. By contrast, D (substrate for 11βHSD2 but not 11βHSD1) showed less than 1% metabolism in this system while DHD (substrate for 11βHSD1 and 11βHSD2) was converted significantly (65.6–90.5%) to D in the four patients tested. There was no significant difference in the interconversion of F and E between patients who became pregnant and those who did not.

CONCLUSIONS

The dehydrogenase and oxoreductase reactions catalysed by 11βHSD both occur in granulosa‐lutein cells at the time of follicular rupture, probably due to 11βHSD1. A lack of measurable conversion of dexamethasone to 11‐dehydrodexamethasone suggests that dehydrogenation due to 11βHSD2 is low or absent. Neither type nor level of 11βHSD activity measured under the present assay conditions correlates with IVF outcome.
A study of the association between a polymorphism in the CTLA‐4 gene and postpartum thyroiditis
Authors: Waterman E. A. , Watson P. F. , Lazarus J. H. , Parkes A. B. , Darke C. , Weetman A. P.
Published: March 2003

OBJECTIVE

Postpartum thyroiditis (PPT) is an autoimmune thyroid disease which shares immunological and clinical features with autoimmune hypothyroidism and Graves' disease, and is believed to be caused by a combination of genetic and environmental factors. Recently, an association has been described between Graves' disease or autoimmune hypothyroidism and a polymorphism in the CTLA‐4 gene, which encodes a T cell receptor for the B7 family of ligands, and we wished to test whether a similar association exists with PPT.

DESIGN

A population‐based case‐control study of a CTLA‐4 gene microsatellite polymorphism was performed, to look for an association with PPT.

PATIENTS

Caucasoid women (n = 122) were studied; 58 had thyroid antibodies (against thyroglobulin or thyroid peroxidase) alone during the postpartum period (PPT−) and 64 had thyroid antibodies and some form of postpartum thyroid dysfunction (PPT+).

RESULTS

There was no significant difference between this whole group of women and 161 local Caucasoid thyroid antibody‐negative women for the CTLA‐4106 base pair (AT)n microsatellite polymorphism (relative risk = 1.3; P = 0.3), nor did the PPT+ group differ from controls when analysed separately (P = 0.2). When the postpartum women were subdivided in groups according to clinical pattern of PPT and the type of thyroid antibodies, there were no associations within the subgroups.

CONCLUSIONS

No significant association exists between postpartum thyroiditis and a polymorphism in the CTLA‐4 gene. Furthermore, a natural variation in the prevalence of the polymorphism in healthy UK populations underscores the need to select appropriately matched normal subjects in future case‐control studies.
Growth hormone induced increase in serum IGFBP‐3 level is reversed by anabolic steroids in substance abusing power athletes
Authors: Karila Tuomo , Koistinen Hannu , Seppälä Markku , Koistinen Riitta , Seppälä Timo
Published: March 2003

OBJECTIVE

Body builders may use growth hormone (GH) and anabolic steroids to increase fitness and muscle mass. GH increases serum concentrations of IGF‐I and IGFBP‐3. The combined effects of GH and anabolic androgenic steroids on IGFBP‐3 and IGF‐I levels are not known under authentic substance‐abusing conditions. The aim of this study was to investigate this in substance‐abusing power athletes.

DESIGN AND PATIENTS

Nine healthy, non‐obese and non‐competing male power athletes, all aggressive substance abusers, used GH and/or anabolic steroids independently of this study. Blood samples were taken both during and between the drug intake. Sixteen substance non‐abusing wrestlers served as controls.

MEASUREMENTS

Serum IGF‐I concentration was measured by radioimmunoassay and the IGFBP‐3 concentration was measured by two immunofluorometric assays, one detecting proteolytic fragments of IGFBP‐3. The capacity of serum to proteolytically cleave IGFBP‐3 was studied by the proteolysis assay.

RESULTS

While growth hormone increased the IGFBP‐3 and IGF‐I concentrations, anabolic steroids decreased the same. Concomitant use of growth hormone and anabolic steroids decreased the IGFBP‐3 concentration in five out of six abuse periods in spite of the fact that the IGF‐I concentration remained elevated in four of them. However, in two men who were on low calorie diet both the IGF‐I and IGFBP‐3 concentrations decreased during combined GH/anabolic steroid abuse. No proteolytic fragmentation of IGFBP‐3 was observed.

CONCLUSION

Massive abuse of anabolic steroids decreases both the basal and GH‐stimulated IGFBP‐3 concentrations, whereas its effects on serum IGF‐I concentration are variable and affected by low calorie diet. This study demonstrates that detection of GH doping by measuring the IGF‐I and/or IGFBP‐3 levels has notable confounding factors.