bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of periodontal treatment on the serum antibody levels to heat shock proteins
Authors: YAMAZAKI K , UEKI-MARUAYAMA K , HONDA T , NAKAJIMA T , SEYMOUR G J
Published: February 2004

We have shown previously that both humoral and cellular immune responses to heat shock protein 60 (HSP60) are elevated in chronic periodontitis patients compared with non-diseased subjects. The aim of the present study was to determine whether periodontal treatment could influence the level of serum antibodies to human HSP60 and Porphyromonas gingivalis GroEL, a bacterial homologue of human HSP60. Sera were obtained from 21 patients with moderate to advanced chronic periodontitis at the baseline examination and again after completion of treatment. Antibody levels were determined using an enzyme-linked immunosorbent assay. The mean anti-P. gingivalis GroEL antibody levels were down-regulated significantly by periodontal treatment when recombinant P. gingivalis GroEL was used as an antigen, whereas antibody levels to P. gingivalis GroEL-specific peptide were significantly elevated following successful periodontal therapy. The mean level of anti-human HSP60 antibody remained unchanged although individual levels of antibody either increased or decreased after periodontal treatment, suggesting that synthesis of these antibodies might be regulated independently during the course of periodontal infection. Although their regulatory mechanisms in chronic infection are not understood, further study would provide insight not only into the role of these antibodies in the pathogenesis of periodontitis but also into the possible link between periodontitis and systemic diseases such as coronary heart disease.
Hepatitis B core antigen stimulates interleukin-10 secretion by both T cells and monocytes from peripheral blood of patients with chronic hepatitis B virus infection
Authors: HYODO N , NAKAMURA I , IMAWARI M
Published: February 2004

In chronic hepatitis B virus (HBV) infection, immune responses to hepatitis B core antigen (HBcAg) are weak. Interleukin (IL)-10 is a potent immunosuppressive cytokine which we reported recently to be secreted in response to HBcAg by peripheral blood mononuclear cells (PBMCs) from patients with chronic HBV infection or healthy controls. Using an enzyme-linked immunospot assay, we compared the ability of HBcAg to stimulate IL-10 production by PBMC with that of lipopolysaccharide (LPS), phytohaemagglutinin-P and hepatitis C virus-derived antigens in 16 patients with chronic HBV infection and six healthy controls. Frequencies of IL-10 spot-forming cells (SFC) in response to HBcAg were comparable to those obtained with LPS in patients with chronic HBV infection. Frequencies of IL-10 SFC in response to HBcAg or to LPS were significantly higher in patients with chronic HBV infection than in healthy controls. IL-10 SFC in response to HBcAg consisted of 26–35% T cells, 62–70% monocytes and less than 1% B cells in patients with chronic HBV infection. Only monocytes contributed to IL-10 production in controls. Frequencies of HBcAg stimulated IL-10 SFC representing T cells and monocytes were significantly higher in patients with elevated serum alanine aminotransferase (ALT) and detectable HBV DNA than in patients with normal ALT and undetectable HBV DNA. The potent ability of HBcAg to stimulate IL-10 production by PBMC may contribute importantly to immune tolerance toward HBV.
1α, 25-dihydroxyvitamin D3 increases IgA serum antibody responses and IgA antibody-secreting cell numbers in the Peyer's patches of pigs after intramuscular immunization
Authors: VAN DER STEDE Y , VERFAILLIE T , COX E , VERDONCK F , GODDEERIS B M
Published: February 2004

Pigs were injected intramuscularly (i.m.) twice with human serum albumin (HSA) with or without 1α,25-dihydroxyvitamin D3[1α,25(OH)2D3] with a 5-week interval. The supplementation of 1α,25(OH)2D3 enhanced the HSA-specific IgA serum antibody response but decreased the IgM, IgG, IgG1 and IgG2 responses. Furthermore, higher numbers of HSA-specific IgA antibody-secreting cells were obtained in systemic lymphoid tissues (local draining lymph node, spleen and bone marrow) as well as in Peyer's patches and lamina propria of the gut (GALT). In addition, the in vivo mRNA expression for Th1 [interferon (IFN)-γ, interleukin (IL-2)], Th2 (IL-4, IL-6 and IL-10) and Th3 [transforming growth factor (TGF)-β] cytokines as well as the percentage of different cell subsets (CD2+, CD4+, CD8+, IgM+, MHC II+, CD25+) of monomorphonuclear cells from the local draining lymph node were determined at different time-points after the i.m. immunizations. Cytokine profiles did not resemble a typical Th-cytokine profile using 1α,25(OH)2D3: higher levels of IL-10 and significantly lower levels of IL-2 were observed the first day after the primary immunization. However, significantly higher levels of IL-2 and significantly lower levels of IFN-γ were observed the first day after the second immunization. Furthermore, after the second immunization TGF-β mRNA expression decreased more quickly in the 1α,25(OH)2D3 group. This difference became significant 7 days after the second immunization. One week later a significantly higher percentage of CD25+ cells was observed in this group, indicating more activated T and B cells using the steroid hormone. These results suggest that in pigs the addition of 1α,25(OH)2D3 to an intramuscularly injected antigen can enhance the antigen-specific IgA-response and prime GALT tissues, but the relation with cytokines and cell phenotype in the local draining lymph node needs further clarification.
Chronic delayed-type hypersensitivity reaction as a means to treat alopecia areata
Authors: ZÖLLER M , FREYSCHMIDT-PAUL P , VITACOLONNA M , MCELWEE K J , HUMMEL S , HOFFMANN R
Published: February 2004

The acute phase of alopecia areata (AA) is characterized by an increase in CD44v3+ and CD44v10+ skin-infiltrating leucocytes (SkIL). Induction of a contact eczema, one of the therapeutic options in AA, can be mitigated strongly by a blockade of CD44v10. The observation that induction of a delayed type hypersensitivity (DTH) reaction abrogates an autoimmune reaction, where both responses apparently use similar effector mechanisms, is surprising and prompted us to search for the underlying mechanisms. AA-affected C3H/HeJ mice were treated with the contact sensitizer SADBE (squaric acid dibutylester) and leucocyte subpopulations and their activation state was evaluated in SkIL and draining lymph nodes. AA-affected mice exhibited an increased number of SkIL with a predominance of T lymphocytes. After treatment with the contact sensitizer SADBE recovery of SkIL was reduced and monocytes predominated. However, a significantly increased number of leucocytes was recovered from draining lymph nodes. Draining lymph node cells from untreated and treated AA mice exhibited all signs of recent activation with high-level expression of co-stimulatory and accessory molecules and an increased percentage of CD44v3+ and CD44v10+ leucocytes. In contrast, SkIL of SADBE-treated AA mice contained relatively few activated T cells and reduced numbers of CD44v3+ and CD44v10+ cells. Thus, the activation state and the distribution of leucocyte subsets in SADBE-treated AA mice are consistent with a blockade of leucocyte extravasation. Accordingly, the therapeutic effect of long-term SADBE treatment may rely on impaired leucocyte traffic.
Anti-CD4 monoclonal antibody treatment in acute and early chronic antigen-induced arthritis: influence on T helper cell activation
Authors: POHLERS D , NISSLER K , FREY O , SIMON J , PETROW P K , KINNE R W , BRÄUER R
Published: February 2004

To examine the effects of anti-CD4 mAb treatment in acute and chronic antigen-induced arthritis (AIA), C57BL/6 mice were treated intraperitoneally either with the depleting anti-CD4 mAb GK1·5 or with rat-IgG (control) on Days −1, 0, 1, 3, 5, and 7. Arthritis was monitored by assessment of joint swelling and histological evaluation in the acute (Day 3) and the chronic phase (Day 21) of AIA. To determine the effects on cellular immune responses, in vivo T-cell reactivity (delayed type hypersensitivity; DTH) was measured, as well as protein levels of TH1- (IL-2, IFN-γ) and TH2-cytokines (IL-4, IL-10) in joint extracts and supernatants of ex vivo stimulated spleen and lymph node cells. The humoral immune response was analysed by measuring serum antibodies against methylated bovine serum albumine (mBSA) and extracellular matrix proteins. Treatment with GK1·5 reduced swelling, inflammation, and destruction of the arthritic joint. Unexpectedly, the effects were even more pronounced in the acute than in the chronic phase. The anti-inflammatory effect was accompanied by a diminished DTH against the arthritogen mBSA and a decrease of TH1-cytokine production in spleen and pooled body lymph nodes, whereas the TH2-cytokine production in these organs was unchanged and the humoral immune response was only moderately reduced. There was a failure of depleting CD4+ T-cells in the joint, reflected also by unchanged local cytokine levels. Therefore, systemic rather than local effects on the TH1/TH2 balance appear to underlie the therapeutic efficacy of anti-CD4 treatment in AIA.
Dysregulation of IL-2 and IL-8 production in circulating T lymphocytes from young cystic fibrosis patients
Authors: HUBEAU C , LE NAOUR R , ABÉLY M , HINNRASKY J , GUENOUNOU M , GAILLARD D , PUCHELLE E
Published: February 2004

It is well documented that patients with cystic fibrosis (CF) are unable to clear persistent airway infections in spite of strong local inflammation, suggesting a dysregulation of immunity in CF. We and others have reported previously that T lymphocytes may play a prominent role in this immune imbalance. In the present work, we compared the reactivity of CD3+ T cells obtained from young CF patients in stable clinical conditions (n = 10, aged 9–16·5 years) to age-matched healthy subjects (n = 6, aged 9–13·5 years). Intracellular levels of interferon (IFN)-γ, interleukin (IL)-2, IL-8 and IL-10 were determined by flow cytometry after whole blood culture. The data identified T lymphocyte subsets producing either low levels (M1) or high levels (M2) of cytokine under steady-state conditions. We found that the production of IFN-γ and IL-10 by T lymphocytes was similar between young CF patients and healthy subjects. In contrast, after 4 h of activation with PMA and ionomycin, the percentage of T cells producing high levels of IL-2 (M2) was greater in CF patients (P = 0·02). Moreover, T cells from CF patients produced lower levels of IL-8, before and after activation (P = 0·007). We conclude that a systemic immune imbalance is present in young CF patients, even when clinically stable. This disorder is characterized by the capability of circulating T lymphocytes to produce low levels of IL-8 and by the emergence of more numerous T cells producing high levels of IL-2. This imbalance may contribute to immune dysregulation in CF.
LTB4 is present in exudative pleural effusions and contributes actively to neutrophil recruitment in the inflamed pleural space
Authors: PACE E , PROFITA M , MELIS M , BONANNO A , PATERNÒ A , MODY C H , SPATAFORA M , FERRARO M , SIENA L , VIGNOLA A M , BONSIGNORE G , GJOMARKAJ M
Published: February 2004

The pleural space is a virtual compartment between the lung and chest wall that becomes filled with fluid and inflammatory cells during a variety of respiratory diseases. Here, we study the potential role of the eicosanoid metabolite leukotriene B4 (LTB4) in disparate diseases leading to acute (pneumonia) or chronic (tuberculosis, cancer) inflammation of the pleural space. LTB4 concentrations were significantly higher in pleural fluid due to pneumonia, tuberculosis and cancer with respect to congestive heart failure and correlated with neutrophil elastase, which is used as an indication of state of activation of neutrophils in the pleural space. Moreover, pleural LTB4 was biologically active, as an anti-LTB4 antibody partially neutralized the chemotactic activity of parapneumonic, tuberculous and cancer effusions. Macrophages, neutrophils, lymphocytes, mesothelial cells and cancer cells all expressed mRNA for 5-lipoxygenase, the enzyme that initiates leukotriene synthesis leading to the production of LTB4, in exudative pleural effusions. Upon stimulation in transudative pleural effusions, pleural macrophages produced, in a time-dependent fashion, a significantly higher concentration of LTB4 than mesothelial cells. These studies demonstrate that different cell types are capable of producing LTB4 in the inflamed pleural space and that this mediator may play a crucial role in the recruitment of neutrophils into the pleural space.
Dendritic cell subsets in childhood and in children with cancer: relation to age and disease prognosis
Authors: VAKKILA J , THOMSON A W , VETTENRANTA K , SARIOLA H , SAARINEN-PIHKALA U M
Published: February 2004

Dendritic cells (DC) are a heterogeneous group of uniquely well-equipped bone marrow-derived antigen-presenting cells. They circulate in blood as precursor cells (preDC). In humans, two blood-borne subtypes of preDC can be distinguished by their differential expression of CD11c (CD11c+ preDC; monocytoid DC) and CD123 (CD123+ preDC; plasmacytoid DC). We studied the incidence of monocytoid and plasmacytoid DC in peripheral blood samples from 39 children of various ages (0·4–16·8 years) by flow cytometry, and found a significant negative correlation between the number of plasmacytoid DC and age (r = 0·421, P = 0·012). Monocytoid DC counts did not change significantly with age. Similarly, we analysed DC subsets in 19 children with cancer at the time of diagnosis prior to initiation of any myelosuppressive or antiproliferative treatment and compared the results with those obtained from gender- and age-matched control children. Patients with cancer had significantly less circulating monocytoid DC than controls (medians 13·2 versus 21·4 cells/µl, respectively, P = 0·042) at diagnosis, whereas absolute plasmacytoid DC counts did not differ significantly between the study groups. However, clinical outcome of the children with cancer (2·9–5 years follow-up after diagnosis) correlated with plasmacytoid DC count. Children with high plasmacytoid DC counts at diagnosis (above median) survived significantly worse (6/10 deceased) than those with low counts (1/9 deceased) (P = 0·034). Thus, circulating plasmacytoid DC counts are related to age during childhood, and development of cancer is associated with low number of monocytoid DC. A low circulating plasmacytoid DC count at diagnosis was a good prognostic sign.
Biologically active neutrophil chemokine pattern in tonsillitis
Authors: RUDACK C , JÖRG S , SACHSE F
Published: February 2004

To gain an insight into the mechanisms of chronic and acute inflammation, the production of neutrophil chemokines in different types of tonsillitis − hyperplastic tonsillitis (HT), recurrent tonsillitis (RT) and peritonsillar abscesses (PA) − was investigated. The chemokines interleukin-8 (IL-8), growth-related oncogene-α (GRO-α), epithelial cell-derived neutrophil attractant-78 (ENA-78) and granulocyte chemotactic protein-2 (GCP-2) were detected and shown to have different biological activities. With respect to the biological properties of CXC chemokines, the biological activity of the chemokines was identified using a three-step high-performance liquid chromatography (HPLC) technique, a bioassay involving measurement of neutrophil chemotaxis in a single Boyden chamber in tissue of HT, RT and PA. Using reverse transcription-polymerase chain reaction (RT-PCR), the chemokine concentrations were determined in the different tonsillitis entities. The chemokine pattern was dominated in PA by IL-8 and GRO-α and in RT by GRO-α. Hyperplastic tonsils of patients without a history of infection generated about five times lower IL-8 than PA. A protein concentration of GCP-2 was induced in PA and RT, whereas ENA-78 remained the same in all entities. In conclusion, it would appear that IL-8 was up-regulated in acute inflammation, whereas GRO-α dominated in chronic inflammation. ENA-78 seems not to play a pivotal role in inflammatory processes in tonsils. GCP-2 may serve as a substitute chemokine in certain inflammatory conditions as its quantity of mRNA and protein was higher in RT and PA than in HT.
Prolonged disturbances ofin vitrocytokine production in patients with severe acute respiratory syndrome (SARS) treated with ribavirin and steroids
Authors: JONES B M , MA E S K , PEIRIS J S M , WONG P C , HO J C M , LAM B , LAI K N , TSANG K W T
Published: February 2004

Severe acute respiratory syndrome (SARS) is a new disease which has spread rapidly and widely. We wished to know whether evaluation of in vitro cytokine production could contribute to improved understanding of disease pathogenesis and to better patient management. Numbers of unstimulated and mitogen-stimulated cytokine-secreting peripheral blood mononuclear cells were measured repeatedly during and after hospitalization in 13 patients with SARS using enzyme-linked immunospot technology. Numbers of interferon-gamma, interleukin (IL)-2, IL-4, IL-10 and IL-12 secreting cells induced by T cell activators were below normal in many or most patients before and during treatment with corticosteroids and ribavirin but returned essentially to normal after completion of treatment. Staphylococcus aureus Cowan 1 (SAC)-stimulated IL-10 secreting cells were increased in early SARS but fell during treatment. SAC-induced IL-12 secreting cells were deficient before, during and long after treatment. Numbers of cells induced to produce IL-6 and tumour necrosis factor-alpha by T cell or monocyte activators were higher than normal in many early SARS patients and were still increased in some during and after treatment. We conclude that prolonged dysregulated cytokine production occurs in SARS and that future studies should be directed at improving anti-inflammatory and antiviral therapies in order to limit cytokine impairment.
Anti-Saccharomyces cerevisiae antibodies (ASCA) in Crohn's disease are associated with disease severity but not NOD2/CARD15 mutations
Authors: WALKER L J , ALDHOUS M C , DRUMMOND H E , SMITH B R K , NIMMO E R , ARNOTT I D R , SATSANGI J
Published: February 2004

Anti-Saccharomyces cerevisiae antibodies (ASCAs) have been proposed as serological markers, which may differentiate Crohn's disease (CD) from ulcerative colitis (UC) and predict disease phenotype. Their importance in pathogenesis is unproven. We investigated the relationship between ASCAs, disease phenotype and NOD2/CARD15 genotype in CD and whether ASCAs were related to antibodies to other fungal proteins. Serum from 228 patients [143 CD, 75 UC, 10 with indeterminate colitis (IC)] and 78 healthy controls (HC) were assayed for ASCA. Antibodies (IgA, IgG) to other fungal proteins (Fusarium species ATC20334, Mycoprotein) were measured in the same samples using an in-house enzyme-linked immunosorbent assay (ELISA) assay. ASCAs were present in 57% of CD, 19% of UC, 30% of IC and 8% of HCs. ASCA-positive status was a predictor for CD with sensitivity of 57%, specificity of 87%, positive predictive value of 78% and negative predictive value of 68%. ASCA was associated with proximal (gastroduodenal and small bowel involvement) rather than purely colonic disease (P < 0·001) and with a more severe disease phenotype and requirement for surgery over a median follow-up time of 9 years (P < 0·0001). No associations with NOD2/CARD15 mutations were seen. There was no association between ASCA and antibodies to MP (IgA or IgG). These data implicate ASCA as a specific marker of disease location and progression in CD, emphasizing the heterogeneity within IBD.
T cell help is required to induce idiotypic–anti-idiotypic autoantibody network after immunization with complementary epitope 289–308aa of La/SSB autoantigen in non-autoimmune mice
Authors: PAPAMATTHEOU M G , ROUTSIAS J G , KARAGOUNI E E , SAKARELLOS C , SAKARELLOS-DAITSIOTIS M , MOUTSOPOULOS H M , TZIOUFAS A G , DOTSIKA E N
Published: February 2004

Immunotherapies against autoimmune diseases have been of limited success. Preventive vaccines could be developed on the basis to abrogate unwanted immune responses to defined autodeterminants. In this study it is shown that immunization of BALB/c mice with two linear T and B cell epitopes of the human La/SSB autoantigen (spanning the regions 289–308aa and 349–364aa) and their complementary forms specified by the complementary mRNA, results in characteristic B and T cell responses. Mice immunized with the 289–308aa epitope or its complementary peptide elicited specific antibodies against both epitopes. In contrast, mice immunized with the 349–364aa epitope or its complementary peptide mounted antibody titres against the immunizing peptide only. According to these data, the 289–308aa epitope and its complementary form were capable to generate an idiotypic–anti-idiotypic response, which were cross-regulated. Peptide-specific T cell proliferation and cytokine production in vitro revealed the induction of a two-stage T helper response (Th1→Th2 type) after immunization with either the epitope 289–308 or its complementary peptide. IgG1 was the predominant subclass after immunization with the two forms of epitopes 289–308 and 349–364, while a response of the IgG2b > IgG2a was obtained after the immunization with the complementary form of 349–364 epitope reflecting the TH2/TH1 polarization, respectively. Our data suggest that the complementary peptides of two immunodominant epitopes of human LaSSB can mimic the autoantibodies against these epitopes and establish an active idiotypic–anti-idiotypic network.
Serum and salivary IgA antibody responses to Saccharomyces cerevisiae, Candida albicans and Streptococcus mutans in orofacial granulomatosis and Crohn's disease
Authors: SAVAGE N W , BARNARD K , SHIRLAW P J , RAHMAN D , MISTRY M , ESCUDIER M P , SANDERSON J D , CHALLACOMBE S J
Published: February 2004

Orofacial granulomatosis (OFG) is a condition of unknown aetiology with histological and, in some cases, clinical association with Crohn's disease (CD). However, the exact relationship between OFG and CD remains uncertain. The aim of this study was to determine whether OFG could be distinguished immunologically from CD by comparing non-specific and specific aspects of humoral immunity in serum, whole saliva and parotid saliva in three groups of patients: (a) OFG only (n = 14), (b) those with both oral and gut CD (OFG + CD) (n = 12) and (c) CD without oral involvement (n = 22) and in healthy controls (n = 29). Non-specific immunoglobulin (IgA, SigA, IgA subclasses and IgG) levels and antibodies to whole cells of Saccharomyces cerevisiae, Candida albicans and Streptococcus mutans were assayed by enzyme-linked immunosorbent assay (ELISA) in serum, whole saliva and parotid saliva. Serum IgA and IgA1 and IgA2 subclasses were raised in all patient groups (P < 0·01). Salivary IgA (and IgG) levels were raised in OFG and OFG + CD (P < 0·01) but not in the CD group. Parotid IgA was also raised in OFG and OFG + CD but not in CD. The findings suggest that serum IgA changes reflect mucosal inflammation anywhere in the GI tract but that salivary IgA changes reflect involvement of the oral cavity. Furthermore, the elevated levels of IgA in parotid saliva suggest involvement of the salivary glands in OFG. Serum IgA antibodies to S. cerevisiae were raised markedly in the two groups with gut disease while serum IgA (or IgG) antibodies to C. albicans were elevated significantly in all three patient groups (P < 0·02). No differences were found with antibodies to S. mutans. Whole saliva IgA antibodies to S. cerevisiae (and C. albicans) were raised in the groups with oral involvement. These findings suggest that raised serum IgA antibodies to S. cerevisiae may reflect gut inflammation while raised SIgA antibodies to S. cerevisiae or raised IgA or IgA2 levels in saliva reflect oral but not gut disease. Analysis of salivary IgA and IgA antibodies to S. cerevisiae as well as serum antibodies in patients presenting with OFG may allow prediction of gut involvement.
Antibody responses to human rotavirus (HRV) in gnotobiotic pigs following a new prime/boost vaccine strategy using oral attenuated HRV priming and intranasal VP2/6 rotavirus-like particle (VLP) boosting with ISCOM
Authors: GONZÁLEZ A M , NGUYEN T V , AZEVEDO M S P , JEONG K , AGARIB F , IOSEF C , CHANG K , LOVGREN-BENGTSSON K , MOREIN B , SAIF L J
Published: February 2004

Safer and more effective human rotavirus (HRV) vaccines are needed. We evaluated oral priming with attenuated WaHRV (AttHRV) followed by boosting with two intranasal (IN) doses of VP2/6 virus-like particles (2/6 VLP) with immunostimulating complexes (ISCOM) to determine if this regimen induces protection against diarrhoea and viral shedding in the gnotobiotic pig model. IgM, IgA and IgG antibody titres in serum and intestinal contents were quantified by enzyme-linked immunosorbent assay (ELISA) and serum neutralizing antibody titres were measured by a virus neutralization (VN) test. Seven groups of neonatal gnotobiotic pigs were vaccinated at post-inoculation days (PID) 0, 10 and 21 and challenged with virulent WaHRV at PID 28. The vaccine groups included: (1, 2) oral priming with AttHRV and boosting with two IN immunizations with 2/6 VLP–ISCOM (Att + 2/6 VLP–ISCOM) at VLP concentrations of 250 µg or 25 µg; (3, 4) three IN immunizations with 2/6 VLP–ISCOM at VLP concentrations of 250 µg or 25 µg (2/6 VLP–ISCOM); (5) three oral immunizations with AttHRV (3×AttHRV); (6) one oral immunization with AttHRV (1×AttHRV); (7) controls (ISCOM matrix and/or diluent). The pigs that received 3×AttHRV or Att + 2/6 VLP250–ISCOM had the highest protection rates against diarrhoea upon challenge at PID 28 with virulent WaHRV. The IgA antibody titres to HRV in intestinal contents were significantly higher in the Att + 2/6 VLP250–ISCOM group than in all other groups prechallenge (PID 28). Serum VN antibody titres were statistically similar after the first inoculation among the groups given AttHRV, but at PID 28 VN antibody titres were significantly higher for the 3×AttHRV and Att + 2/6 VLP250–ISCOM groups than for the 1×AttHRV group suggesting that boosting with 2/6 VLP also boosted VN antibody responses. In humans, intestinal IgA antibodies have been correlated with protection against symptomatic reinfection. Thus the vaccine regimen of one oral dose of AttHRV and two IN immunizations with 2/6 VLP250–ISCOM may be an alternative to multiple-dose live oral vaccines in humans.
The effect of eosinophils on collagen gel contraction and implications for tissue remodelling
Authors: ZAGAI U , SKÖLD C M , TRULSON A , VENGE P , LUNDAHL J
Published: February 2004

Asthma is characterized by an eosinophilic inflammation and a subepithelial fibrosis in the airways. Eosinophils contain several cytotoxic substances, such as eosinophil cationic protein (ECP), which can promote inflammation and cause tissue damage. This has generated the hypothesis that eosinophils may drive remodelling of extracellular matrix (ECM). To investigate the role of eosinophils we used an in vitro model for remodelling, the three-dimensional collagen gel contraction assay. Two sources of eosinophils were used in this study, isolated human peripheral eosinophils (purity > 95%) and stimulated [interleukin (IL)-5, IL-3 and granulocyte macrophage–colony stimulating factor (GM-CSF)] HL-60 clone 15 cells. Human eosinophils or HL-60 cells were cast together with human lung fibroblasts (HFL1) in type I collagen gels. Both types of eosinophils augmented fibroblast-mediated collagen gel contraction in a time and concentration-dependent manner. At 48 h, the gel area in HFL1/eosinophil co-culture was 46·5% ± 0·5 (mean ± s.e.m.) of initial area and in HFL1 culture 52·3% ± 0·1 (P < 0·001). Respective figures for HFL1/stimulated HL-60 co-culture and HFL1 culture only were 44·1% ± 0·5 and 52·4% ± 0·4 (P < 0·001). The release of ECP was increased when fibroblasts were cultured with eosinophils compared to eosinophils cultured alone. In addition, native ECP added to fibroblast gel cultures also augmented contraction. Our results suggest that eosinophils may interact with mesenchymal cells, promoting remodelling of ECM and that ECP constitutes one potential eosinophil-derived mediator driving this process. We conclude that this may be one important mechanism by which eosinophil–ECM interactions will lead to airway tissue remodelling in asthma.
PCT-233, a novel modulator of pro- and anti-inflammatory cytokine production
Authors: BISSONNETTE E Y , PROULX L-I , TURMEL V , DROUIN R , PURCELL M
Published: February 2004

Plant extracts have been implicated in various immunoregulatory effects that are poorly understood. Thus, we investigated the modulatory activity of PureCell Complex (PCT)-233, an active molecular complex from mesophyll tissue of Spinacia oleacea on the inflammatory process. Alveolar macrophages (AM) were treated with PCT-233 and/or budesonide, a well-known anti-inflammatory agent, before or after being stimulated with lipopolysaccharides (LPS). Pro- and anti-inflammatory cytokine production, tumour necrosis factor (TNF) and interleukin (IL)-10, respectively, were measured in cell-free supernatants at different times after the treatment. PCT-233 increased unstimulated AM release of both TNF and IL-10, whereas heat- and light-inactivated PCT-233 stimulated only the release of TNF without affecting IL-10 production, suggesting that different mechanisms are involved in the modulation of TNF and IL-10 release by PCT-233. The presence of LPS did not modify PCT-233-stimulated TNF production, but the ratio TNF/IL-10 production by LPS-stimulated AM was reduced significantly in the presence of PCT-233. Pretreatment of AM with PCT-233 and budesonide before LPS stimulation reduced TNF production at both protein and mRNA levels, whereas IL-10 production was increased. Moreover, TNF/IL-10 ratio was reduced further with the combination PCT-233/budesonide. Interestingly, AM treatment with PCT-233 and budesonide 18 h after LPS stimulation did not modulate TNF release significantly but it did increase IL-10 production, and a synergistic effect was observed with the combination PCT-233/budesonide. These exciting data suggest that PCT-233 possesses some anti-inflammatory properties, even when added during the inflammatory process, and could potentiate the effect of other anti-inflammatory agents.
Retinoic acid enhances the gene expression of human polymeric immunoglobulin receptor (pIgR) by TNF-α
Authors: TAKENOUCHI-OHKUBO N , ASANO M , CHIHAYA H , CHUNG-HSUING W U , ISHIKASA K , MORO I
Published: February 2004

In this study, the detailed mechanisms for the effects of vitamin A on the expression of polymeric immunoglobulin receptor (pIgR) were examined. Expression of the pIgR by tumour necrosis factor (TNF-α) was enhanced by the addition of all-trans retinoic acid (ATRA) or 9-cis retinoic acid (9CRA). This enhancement was mediated mainly by RARα, and regulated at the transcriptional level. Transcription factor nuclear factor-κB (NF-κB) binding and activation were not influenced by addition of ATRA. These data imply that RA, in combination with TNF-α, could up-regulate the expression of pIgR. In addition, we hypothesize that up-regulation of pIgR by RA is controlled through the RAR-dependent signalling pathway and that it plays a role in enhancement of mucosal immunity.
Evidence that factors other than particular thyrotropin receptor T cell epitopes contribute to the development of hyperthyroidism in murine Graves’ disease
Authors: PICHURIN P N , CHEN CHUN-RONG , NAGAYAMA Y , PICHURINA O , RAPOPORT B , MCLACHLAN S M
Published: February 2004

Immunization with thyrotropin receptor (TSHR)-adenovirus is an effective approach for inducing thyroid stimulating antibodies and Graves’ hyperthyroidism in BALB/c mice. In contrast, mice of the same strain vaccinated with TSHR-DNA have low or absent TSHR antibodies and their T cells recognize restricted epitopes on the TSHR. In the present study, we tested the hypothesis that immunization with TSHR-adenovirus induces a wider, or different, spectrum of TSHR T cell epitopes in BALB/c mice. Because TSHR antibody levels rose progressively from one to three TSHR-adenovirus injections, we compared T cell responses from mice immunized once or three times. Mice in the latter group were subdivided into animals that developed hyperthyroidism and those that remained euthyroid. Unexpectedly, splenocytes from mice immunized once, as well as splenocytes from hyperthyroid and euthyroid mice (three injections), all produced interferon-γ in response to the same three synthetic peptides (amino acid residues 52–71, 67–86 and 157–176). These peptides were also the major epitopes recognized by TSHR-DNA plasmid vaccinated mice. We observed lesser responses to a wide range of additional peptides in mice injected three times with TSHR-adenovirus, but the pattern was more consistent with increased background ‘noise’ than with spreading from primary epitopes to dominant secondary epitopes. In conclusion, these data suggest that factors other than particular TSHR T cell epitopes (such as adenovirus-induced expression of conformationally intact TSHR protein), contribute to the generation of thyroid stimulating antibodies with consequent hyperthyroidism in TSHR-adenovirus immunized mice.
Elevated expression and release of tissue-type, but not urokinase-type, plasminogen activator after binding of autoantibodies to bullous pemphigoid antigen 180 in cultured human keratinocytes
Authors: SCHMIDT E , WEHR B , TABENGWA E M , REIMER S , BRÖCKER E-B , ZILLIKENS D
Published: February 2004

In bullous pemphigoid (BP), the binding of BP180-specific antibodies to their hemidesmosomal target antigen is not sufficient for blister formation, but must be accompanied by the release of proteases. Using plasminogen activator (PA) knock-out mice, the PA system has previously been shown to be a prerequisite for blister formation in experimental murine BP. Here, we found elevated levels of plasmin and tPA, but not of uPA, in blister fluid from BP patients (n = 7) compared to blisters from patients with toxic epidermal necrolysis (n = 4) and suction blisters in healthy controls (n = 7). Subsequently, we addressed the question whether keratinocytes release PA in response to the binding of anti-BP180 antibodies. Treatment of cultured normal human keratinocytes with BP IgG, but not with control IgG, led to both increased protein and mRNA levels of tPA, but not of uPA, as determined by ELISA and RT-PCR, respectively. The specificity of this finding was confirmed using BP180-deficient keratinocytes from a patient with generalized atrophic benign epidermolysis bullosa, where no tPA release was observed after stimulation with BP IgG. Our results show the elevated expression and release of tPA from normal human keratinocytes upon stimulation with antibodies to human BP180. Keratinocytes, by secreting tPA, may thus play an active role in blister formation of BP.
The role of B7 molecules in the cell contact-mediated suppression of T cell mitogenesis by immunosuppressive macrophages induced with mycobacterial infection
Authors: SHIMIZU T , SANO C , TOMIOKA H
Published: February 2004

We found previously that immunosuppressive macrophages (Mφs) induced by Mycobacterium intracellulare infection (MI-Mφs) transmitted their suppressor signals to target T cells through cell contact with target T cells. In this study, we examined what kinds of Mφ surface molecules are required for such cell–to–cell interaction. First, it was found that a B7-1-like molecule (B7–1LM) recognizable with one of three test clones of anti-B7-1 monoclonal antibodies (mAbs) was required for expression of the Mφ suppressor activity. Neither anti-B7-2, anti-ICAM-1, nor anti-VCAM-1 mAb blocked the Mφ suppressor activity. Second, MI-Mφs increased the expression of B7–1LM in parallel with the acquisition of the suppressor activity. Moreover, MI-Mφs bound with target T cells in a B7–1LM-dependent fashion. Third, mAb blocking of CTLA-4 on target T cells did not reduce the suppressor activity of MI-Mφs, suggesting the role of a putative molecule on target T cells other than CTLA-4 as the receptor for B7–1LM of MI-Mφs. Fourth, concanavalin A (Con A) stimulation of MI-Mφs was needed for effective cell contact with target T cells and subsequent expression of the suppressor activity of MI-Mφs. Fifth, the Con A-induced increase in the suppressor activity of MI-Mφs was inhibited by KN-62 but not by herbimycin A, H-7, nor H-88, indicating that Con A-induced up-regulation of MI-Mφ function is mediated by calmodulin-dependent protein kinase II or ATP/P2Z receptors, but independent of protein tyrosine kinase, protein kinase C, and protein kinase A. These findings indicate that a B7/CTLA-4-independent mechanism is needed for the transmission of the suppressor signals from MI-Mφs to target T cells.
Differential expression of mRNA for Th1 and Th2 cytokine-associated transcription factors and suppressors of cytokine signalling in peripheral blood mononuclear cells of patients with atopic dermatitis
Authors: ARAKAWA S , HATANO Y , KATAGIRI K
Published: February 2004

Atopic dermatitis is characterized by Th2-dominant immunity. Recently many intracellular molecules have been reported to regulate cytokine expression and T cell differentiation. GATA-3 and T-box expressed in T cells (T-bet) are transcription factors that play a critical role in the development of Th2 and Th1 immunity, respectively. Suppressor of cytokine signalling (SOCS)-3 and SOCS-5, are negative regulators of the cytokine signalling induced by IL-12 and IL-4, respectively. Txk is a transcription factor that activates IFN-γ gene directly. The present study was designed to identify intracellular molecules that are responsible for the pathogenesis and the imbalance of cytokines in atopic dermatitis. Semi-quantitative RT-PCR revealed that in peripheral blood mononuclear cells without any stimulation the levels of mRNA for GATA-3 and SOCS-3 were elevated, the levels of mRNA for Txk were depressed and the levels of mRNA for T-bet and SOCS-5 were comparable in patients with atopic dermatitis as compared with healthy controls. In addition, successful therapy normalized levels of mRNA for GATA-3 and Txk, although those for the others including IL-4, IL-5, IL-10, IL-13 and IFN-γ did not change. Levels of Txk mRNA correlated with those of IFN-γ, while the mRNA levels of the other regulators did not correlate with those of any of the cytokines. These results suggest GATA-3 and Txk might be involved in skin lesions, while SOCS-3 might be associated with an imbalance of cytokines that is difficult to normalize in atopic dermatitis.
An analysis of canine hair re‐growth after clipping for a surgical procedure
Authors: Diaz Sandra F. , Torres Sheila M. F. , Dunstan Robert W. , Lekcharoensuk Chalermpol
Published: February 2004

Abstract  Hair growth and replacement have been studied extensively in humans, sheep and laboratory rodents, but in dogs and other mammalian species few studies have been published. The objectives of this study were: (1) to determine the time required for the hair to re‐grow in dogs after clipping for a surgical procedure; (2) to define whether the season of the year influenced the period of time required for re‐growth and; (3) to determine if season might influence the telogen: anagen ratio. Eleven Labrador retrievers were recruited during spring, 10 during summer, six during autumn and 10 during winter. Hairs re‐grew to their preclipped length in 14.6 weeks, 14.5 weeks, 13.6 weeks and 15.4 weeks when shaved in the spring, summer, autumn and winter, respectively. The differences in these values were not significant suggesting that season has no effect on the rate of hair re‐growth in Labrador retrievers housed indoors (P = 0.12). The mean values for the telogen: anagen ratio in each season were: 5.2 (spring), 6.1 (summer), 9.5 (autumn), and 5.3 (winter). The differences in these values also were not significant (P = 0.89). The percentage of hairs in telogen was over 80% in all four seasons.
In vitrometabolism of dehydroepiandrosterone and testosterone by canine hair follicle cells
Authors: Bamberg Elmar , Aichinger Amanda , Mitteregger Gerda
Published: February 2004

Abstract The metabolism of radioactive dehydroepiandrosterone (DHEA) and testosterone was studied in dermal papilla cells (DPC) and dermal fibroblasts (DFB) derived from hair follicles from two different body sites (head, flank) of four male, castrated beagle dogs. Thin layer chromatography was used for separation, and autoradiography for identification of the radioactive metabolites. DHEA was metabolized mainly to 11α‐OH‐testosterone and only to a minor extent to 11α‐OH‐androstenedione and another unidentified metabolite. The highest percentage of metabolization of DHEA was found in DFB of the head. Testosterone was metabolized only to a minor extent (less than 10%) to 5α‐dihydrotestosterone and epiandrosterone and there was no significant difference between either the two cell types or the two locations. These results clearly show that the metabolization of androgens in canine DPC and DFB is different from that observed in cells from the human hair follicle.
The tear film and ocular mucins
Authors: Davidson Harriet J. , Kuonen Vanessa J.
Published: February 2004

The trilaminar tear film, composed of the lipid, aqueous and mucin layers, has many functions including defending the ocular surface. The aqueous layer has several soluble antimicrobial factors that protect the ocular surface. Ocular mucins have recently been studied with regard to their role in the defense of the eye as well as in dry eye syndromes. To date, 15 mucin genes have been identified, and six of these mucin genes are localized to or secreted by ocular glands or epithelia. Understanding the production, secretion and function of ocular mucins will aid in the treatment of dry eye syndromes and ocular surface microbial infections.
Basic concepts in statistics for veterinary ophthalmologists
Authors: Montiani‐Ferreira F. , Cardoso F. F. , Petersen‐Jones S.
Published: February 2004

This review article provides a comprehensive coverage of basic statistical procedures commonly used in biologic experiments and is intended to be a basic guide to the selection of appropriate tests and also to aid the reader in the evaluation of published studies. The focus is on the use, interpretation and presentation of the main statistical tests and their key concepts for the practicing veterinary ophthalmologist. Several examples derived from actual ophthalmic research are presented in the text. Lastly, the article discusses computer‐based statistical analysis, provides a list of different software available and gives a practical example of a ‘real life’ statistical analysis.
Change in intraocular pressure during maturation in Labrador Retriever dogs
Authors: Mughannam Albert J. , Cook Cynthia S. , Fritz Curtis L.
Published: February 2004

Objective To measure intraocular pressure (IOP) in a group of dogs as puppies and young adults to determine if there is any change during maturation.

Animals studied Thirty‐two healthy Labrador Retriever dogs.

Procedures Intraocular pressure was measured using a Tonopen XL initially at approximately 6 weeks of age (T1), then again approximately 1 year later (T2). Exact ages were known based on whelp date.

Results The dogs had marginally higher IOP OU at T2 (mean = 14.9 mmHg) compared to T1 (mean = 13.4 mmHg). However, the difference was not statistically significant. No differences were seen based on sex and litter. Intraocular pressure OD was statistically greater than OS at T1 but not at T2.

Conclusions Normal values for intraocular pressure are the same in puppies and adults. The results of this study do not support the previously suggested theory that younger dogs have sustained increased IOP as a requirement to drive growth of the globe. However, it does not rule out the possibility that a dynamic relationship between intraocular pressure and expansion of the globe may exist.
Lens morphometry determined by B‐mode ultrasonography of the normal and cataractous canine lens
Authors: Williams David L.
Published: February 2004

Purpose To determine axial lens thickness, anterior chamber depth and axial globe length in canine eyes with normal lenses and in eyes with immature, mature, congenital, posterior polar and diabetic cataract.

Methods B‐mode ultrasonography was performed in 50 normal dogs and, as a prephacoemulsification screening procedure, in 100 dogs with cataract. Axial B‐mode ultrasonograms were used to determine lens thickness, anterior chamber depth and globe diameter. Statistical comparisons between groups were made by analysis of variance and multivariate analysis, with a significance level ofP< 0.05.

Results Axial globe lengths were not statistically significantly different between groups apart from the smaller globes in younger dogs with congenital cataract. Axial lens thickness in diabetics (8.4 ± 0.9 mm) was statistically significantly different from the lens thickness in normal eyes (6.7 ± 1.0 mm), eyes with immature cataract (6.4 ± 0.8 mm) and eyes with mature cataract (7.4 ± 0.9 mm) although these groups, while varying in thickness, were not statistically significantly different from each other. Anterior chamber depth was statistically significantly reduced in eyes with diabetic cataract (2.9 ± 0.1 mm) from that in normal eyes (3.8 ± 0.1 mm), eyes with immature cataract (3.5 ± 0.1 mm) and eyes with mature cataract (3.2 ± 0.6 mm) although these groups, while varying in chamber depth, were not statistically significantly different from each other.

Conclusions Lenses with diabetic cataracts were significantly increased in axial thickness compared to lenses in other eyes, although lenses with mature cataracts showed a trend towards increased axial thickness and immature cataracts demonstrated a trend towards reduced thickness. While previous studies on cataract pathobiology have suggested a reduction in lens thickness in immature cataract through lens protein loss and an increase in thickness in mature cataracts through intumescence, this study is the first to document these changes in the canine lens.