bluuebunny/crossref_metadata_2025 huggingface dataset:
Anti-neutrophil cytoplasmic antibodies (ANCA) against bactericidal/permeability-increasing protein (BPI) and cystic fibrosis lung disease
Authors: Mahadeva R , Dunn A C , Westerbeek R C , Sharples L , Whitehouse D B , Carroll N R , Ross-Russell R I , Webb A K , Bilton D , Lomas D A , Lockwood C M
Published: March 2003

Persistent infection with Pseudomonas aeruginosa and inflammatory mechanisms play an important role in cystic fibrosis (CF) lung disease. ANCA against BPI, a potent host defence protein with anti-bacterial and anti-endotoxin properties, have been described in CF. We have assessed the relationship of anti-BPI antibodies to pulmonary disease severity in 148 CF subjects. IgA and IgG anti-BPI antibodies were found in 55.4% and 70.3% of CF patients, respectively, and higher levels were strongly associated with colonization with P. aeruginosa (P = 0.001 and 0.039 for IgA and IgG antibodies, respectively). IgA and IgG anti-BPI antibodies were independently associated with more severe lung disease as assessed by chest radiograph score (P = 0.023) and a significantly lower forced expiratory volume in 1 s (FEV1)% (P = 0.01). The pathophysiological relevance of the autoantibodies was investigated further by determining their epitope specificity and their effect on bacterial phagocytosis in vitro. Both isotypes of anti-BPI antibodies were specific for the C-terminus of BPI shown recently to be important for BPI-mediated opsonization, and in vitro affinity-purified anti-BPI antibodies significantly reduced BPI-induced phagocytosis of Escherichia coli compared with controls. These data indicate that anti-BPI autoantibodies are associated with colonization with P. aeruginosa and worse lung disease in CF. The inhibition of bacterial phagocytosis suggests that these autoantibodies may contribute to the persistence of P. aeruginosa in the CF lung and so play a role in perpetuating CF lung damage.
Anti-lactoferrin antibodies and other types of anti-neutrophil cytoplasmic antibodies (ANCA) in reactive arthritis and ankylosing spondylitis
Authors: Locht H , Skogh T , Kihlström E
Published: March 2003

Fifty-five serum samples from patients with reactive arthritis (ReA), 40 from patients with ankylosing spondylitis (AS) and three from patients with chronic sacroiliac joint arthritis were analysed for the presence of ANCA of IgG class by means of enzyme immunosorbent assay using lactoferrin (Lf), myeloperoxidase (MPO) and antigen extracted from azurophil granules (‘α-antigen’) containing proteinase 3 (PR3) as substrate. IgG-ANCA were found in 31 (56%) patients with ReA. Twenty-three (42%) had anti-Lf antibodies, nine (16%) had anti-MPO and eight (15%) had anti-α-antigen antibodies, none of which reacted with PR3. Only six (14%) AS or sacroiliac joint arthritis patients had ANCA (P < 0.001). Three (7%) had anti-Lf, two (5%) anti-MPO and two (5%) anti-α-antigen antibodies. Yersinia and Salmonella bacteria were separated by SDS–PAGE and blots were incubated with serum from rabbits immunized with human Lf. The hyperimmune serum recognized a band of 78 kD from both bacteria which was not seen when preimmune serum was used. The reaction to the 78-kD antigen could be completely inhibited when anti-Lf antibodies were absorbed on Lf coupled to cyanogen bromide-activated Sepharose, possibly indicating cross-reacting epitopes in Lf and enterobacterial antigen.
Is an epitope on keratin 17 a major target for autoreactive T lymphocytes in psoriasis?
Authors: Gudmundsdottir A S , Sigmundsdottir H , Sigurgeirsson B , Good M F , Valdimarsson H , Jonsdottir I
Published: March 2003

Psoriasis is a T cell-mediated inflammatory skin disease that has been associated with infections by group A β-haemolytic streptococci. In a previous study of patients with active psoriasis we demonstrated an increased frequency of circulating Th1-like cells that responded to 20 amino acid (aa) streptococcal M-peptides sharing sequences with human keratin. These cells disappeared after ultraviolet B (UVB)-induced clinical remission. Using T cells from the blood of 17 psoriatic patients and 17 healthy controls we have now compared the numbers of interferon-gamma (IFN-γ)-producing cells induced by seven 18–20 aa keratin peptides and five corresponding M-peptides. The most frequent and strongest responses were observed to a peptide from keratin 17 that shares ALEEAN sequence with M-protein. The responses to this peptide were stronger than to the corresponding M-peptide containing the ALEEAN sequence. After UVB treatment T cell responses to all the M- and keratin peptides were abolished, while responses to the positive control antigen streptokinase/streptodornase (SK/SD) were not affected. These findings are consistent with the notion that aa sequences which keratin has in common with M-protein may be a major target for autoreactive T cells in psoriasis.
Qualitative and quantitative studies of autoantibodies to phospholipids in diabetes mellitus
Authors: Gargiulo P , Goldberg J , Romani B , Schiaffini R , Ciampalini P , Faulk W P , McIntyre J A
Published: March 2003

Diabetes mellitus is associated with vascular and neurological complications. We have investigated the presence of antibodies to phospholipids and to phospholipid binding plasma proteins in blood samples collected from 68 clinically and biochemically characterized type I and type II diabetic patients and from 252 healthy blood donor controls. Each sample was analysed for antibodies to three phospholipids (cardiolipin, phosphatidylserine and phosphatidylethanolamine), the antibody isotypes (IgA, IgG and IgM), and whether antibody activity was plasma protein-dependent. Patients were considered to have anti-phospholipid antibodies when one or more of these 18 tests was found above predetermined control values. The results of these experiments revealed an increased incidence of anti-phospholipid antibodies in diabetic patients compared with control subjects. The incidence of IgA isotype to phosphatidylethanolamine was higher than the incidence of other isotypes to other phospholipids, and their reactivities were independent of phospholipid-associated proteins. In addition, these antibody findings were studied for associations with prothrombin degradation products, activated factor VII and activated protein C, and with the incidence of diabetic complications. The anti-phosphatidylethanolamine antibody association with proliferative retinopathy was significant.
γδ T cells in Behçet’s disease (BD) and recurrent aphthous stomatitis (RAS)
Authors: Freysdottir J , Lau S-H , Fortune F
Published: March 2003

The immunopathogenesis of BD is believed to be T cell-mediated. The objective of this study was to characterize the activation stage and cytokine profile of peripheral blood lymphocytes (PBL), with particular emphasis on γδ T cells. Venous blood was collected from 20 patients with BD, and for comparison, from 11 patients with RAS and from 15 healthy controls. Both the expression of activation markers (CD25, CD29, CD40 ligand, CD69 and HLA-DR) on freshly isolated PBL and T cell subsets, and the expression of intracellular cytokines (IL-4, IL-10, interferon-gamma (IFN-γ) and tumour necrosis factor-alpha (TNF-α)) on mitogen-stimulated PBL and T cell subsets were analysed by double immunofluorescent staining and flow cytometry. Significantly decreased proportion of αβ T cells and increased proportion of γδ T cells, CD56+ cells and CD8+γδ T cells were found in BD patients compared with healthy controls. This was also seen to a lesser extent in patients with RAS. Furthermore, in BD a significantly increased proportion of the γδ T cell population expressed CD69 and high levels of CD29 and were induced to produce IFN-γ and TNF-α compared with healthy controls. In contrast, an increased percentage of γδ T cells from RAS patients was induced to produce IFN-γ, but not TNF-α. These results indicate that in BD, activated γδ T cells, capable of producing IFN-γ and TNF-α, are present in peripheral blood, suggesting that γδ T cells are dynamic and may be regulating immunopathogenic events.
Contribution of serotype-specific IgG concentration, IgG subclasses and relative antibody avidity to opsonophagocytic activity against Streptococcus pneumoniae
Authors: Anttila M , Voutilainen M , Jäntti V , Eskola J , Käyhty H
Published: March 2003

The contribution of serotype-specific IgG concentration, subclasses, and avidity to opsonophagocytic activity (OPA) against Streptococcus pneumoniae (Pnc) was evaluated in sera of adults and infants immunized with different pneumococcal vaccines. Antibody concentrations and avidities were measured by enzymeimmunoassay (EIA) and OPAs by killing assay of Pnc. The most important factor contributing positively to OPA was the specific IgG level. In infants, a tendency to negative correlation was found between the concentration needed for killing of bacteria and avidity, suggesting that less antibodies of high rather than low avidity were required for killing. No such correlation was seen in adults. However, in adults the avidity was high already before vaccination and the variation was narrow. Thus, avidity was probably not a limiting factor influencing OPA. The effect of IgG2/IgG1 ratio on OPA was mostly negative but insignificant.
Elevated levels of β-chemokines in bronchoalveolar lavage fluid (BALF) of individuals infected with human T lymphotropic virus type-1 (HTLV-1)
Authors: Seki M , Kadota J-I , Higashiyama Y , Iida K , Iwashita T , Sasaki E , Maesaki S , Tomono K , Kohno S
Published: March 2003

Pulmonary complications are known to develop in HTLV-1 carriers, including T lymphocytic alveolitis, and increased IL-2 receptor α (CD25)-bearing T cells have been found in BALF. Several chemokines may contribute to accumulation of T lymphocytes in the lungs of HTLV-1 carriers. Here, we compared the distribution of T lymphocyte subsets and β-chemokines, such as macrophage inflammatory peptide-1α (MIP-1α), regulated on activation normal T expressed and secreted (RANTES), and macrophage chemoattractant protein-1 (MCP-1), in BALF and peripheral blood between HTLV-1 carriers and non-infected healthy normal subjects. Flow cytometric analysis with MoAbs to cell surface antigens was used to identify T lymphocyte subsets in BALF samples from HTLV-1 carriers (n = 13) and non-infected healthy controls (n = 10). The levels of different β-chemokines were estimated by ELISA. High percentages of CD3+ cells, CD3 expressing HLA-DR antigen and CD3+CD25+ cells were detected in BALF of HTLV-1 carriers compared with non-infected controls. The concentration of MIP-1α in BALF of patients was significantly higher than in non-infected healthy controls and correlated well with the percentage of CD3+CD25+ cells. The level of RANTES in BALF was also significantly high in HTLV-1 carriers, but did not correlate with the percentage of CD3+CD25+ cells. On the other hand, the level of MCP-1 in BALF of HTLV-1 carriers was not different from that of controls. Our results suggest a possible interaction between activated T cells bearing CD25 and β-chemokines, especially MIP-1α, which may contribute to the pulmonary involvement in HTLV-1 carriers.
Development of dextran sulphate sodium-induced experimental colitis is suppressed in genetically mast cell-deficient Ws/Ws rats
Authors: Araki Y , Andoh A , Fujiyama Y , Bamba T
Published: March 2003

Ws/Ws rats have a small deletion of the c-kit gene, and are deficient in both mucosal-type mast cells (MMC) and connective tissue-type mast cells (CTMC). In the present study we investigated the role of intestinal MMC in the development of dextran sulphate sodium (DSS)-induced experimental colitis using Ws/Ws rats. Ws/Ws and control (+/+) rats were given a 3% DSS aqueous solution orally for 10 days, and the subsequent mucosal damage was evaluated macroscopically and histologically. The mucosal myeloperoxidase (MPO) activities and histamine levels were also measured. (i) DSS induced severe oedema and hyperaemia with sporadic erosions in the control (+/+) rats, but these changes were significantly attenuated in the Ws/Ws rats (P < 0.01). (ii) The microscopic mucosal damage score was lower in the Ws/Ws rats than in the control (+/+) rats (P = 0.06). (iii) There were no significant differences in mucosal MPO activity between the Ws/Ws and control (+/+) rats (P = 0.46). (iv) The mucosal histamine levels in the colon were significantly reduced in the Ws/Ws rats compared with the control (+/+) rats (P < 0.05). (v) Significant positive correlations were observed between mucosal histamine levels and the degree of mucosal oedema (calculated as colonic wet weight/protein content) (r = 0.778, P < 0.01), and between histamine levels and the macroscopic damage (r = 0.623, P < 0.05), respectively. (vi) DSS induced a local recruitment of MMC in the colonic mucosa of Ws/Ws rats, and mucosal damage gradually increased in accordance with this MMC recruitment. These results indicate that MMC play an important role in the development of DSS colitis.
Expression of costimulatory molecules CD80 and CD86 and their receptors CD28, CTLA-4 on malignant ascites CD3+ tumour-infiltrating lymphocytes (TIL) from patients with ovarian and other types of peritoneal carcinomatosis
Authors: Melichar B , Nash M A , Lenzi R , Platsoucas C D , Freedman R S
Published: March 2003

Costimulation of T lymphocytes by the leucocyte surface molecules CD80 and CD86 expressed on antigen-presenting cells (APC) is required for the development of T cell responses. The CD28 and CTLA-4 molecules on T cells serve as receptors for the CD80 and CD86 costimulatory antigens. We have examined the frequency of expression of CD80 (B7.1), CD86 (B7.2), CD28 and CTLA-4 surface antigens on TIL isolated from malignant ascites or peritoneal washings of 26 patients with ovarian carcinoma and five patients with non-ovarian peritoneal carcinomatosis. Expression of CD80 and CD86 antigen was detected by reverse transcription-polymerase chain reaction (RT-PCR), and by FACS analysis. Significantly higher proportions of intraperitoneal CD3+ cells expressed CD86 antigen than the CD80 antigen (14 ± 9% versus 3 ± 3%, P < 0.05). Moreover, CD3+CD86+ cells were significantly more frequent in the peritoneal fluid (14 ± 9%) than in the peripheral blood (3 ± 0.4%, P < 0.05) of ovarian patients or normal controls (3 ± 1%). CTLA-4 and CD28 antigen were expressed, respectively, on 9 ± 4% and 86 ± 14% of ascitic CD3+ cells of ovarian cancer patients. Both CD80 and CD86 antigens were expressed primarily on HLA-DR+ ascites TIL and were present in a very low proportion of HLA-DR− ascites TIL. These HLA-DR+ cells may represent a population of lymphocytes that have been activated in vivo, and function as APC. An anti-CD86 MoAb or a combination of anti-CD86 and anti-CD80 MoAbs significantly inhibited the proliferation of cultured intraperitoneal TIL. We have shown that in addition to CD28 and CTLA-4, CD3+ intraperitoneal TIL express the costimulatory molecules CD80 and CD86. The expression of these molecules on T cells could be dependent upon certain factors in the tumour microenvironment that could determine the outcome of in vivo immune responses.
Treatment with anti-CD86 costimulatory molecule prevents the autoimmune lesions in murine Sjögren’s syndrome (SS) through up-regulated Th2 response
Authors: Saegusa K , Ishimaru N , Yanagi K , Haneji N , Nishino M , Azuma M , Saito I , Hayashi Y
Published: March 2003

Intraperitoneal administration with anti-CD86 (B7.2) MoAb into the murine model for primary SS in NFS/sld mutant mice resulted in dramatically inhibitory effects on the development of autoimmune lesions, while no significant effects were observed when the mice were administered with anti-CD80 (B7.1) MoAb. We found that spleen cells in the murine SS model treated with anti-CD86 MoAb showed a significant impairment of autoantigen-specific T cell proliferation. T cell activation markers (CD44high, CD45RBlow, Mel-14low) were significantly down-regulated in the spleen cells gated on CD4 in anti-CD86-treated mice. We detected a higher level of cytokine production of IL-4 from splenic T cells in anti-CD86-treated mice, but not of IL-2, and interferon-gamma (IFN-γ), compared with those in the anti-CD80- and PBS-treated SS model. Moreover, serum autoantibody production against α-fodrin autoantigen was almost entirely suppressed in anti-CD86-treated mice. These data provide strong evidence that in autoimmune exocrinopathy resembling SS in NFS/sld mutant mice, the CD86 costimulatory molecule plays a crucial role in the initiation and subsequent progression of Th1-mediated autoimmunity in the salivary and lacrimal glands.
CD28 costimulation and CD28 expression in T lymphocyte subsets in HIV-1 infection with and without progression to AIDS
Authors: Choremi-Papadopoulou H , Panagiotou N , Samouilidou E , Kontopidou F , Viglis V , Antoniadou A , Kosmidis J , Kordossis T
Published: March 2003

In a prospective study of 152 HIV-1 patients (with and without progression to AIDS) we examined CD28 MoAb costimulation and CD3 MoAb response using whole blood culture at baseline and up to either the time of AIDS diagnosis or the end of the observation period. CD28 antigen expression on both CD4+ and CD8+ T lymphocytes was also studied in both groups of patients. In patients who progressed to AIDS, CD28 MoAb costimulation was found to be decreased. Univariate time-dependent analysis showed that decreases in (i) absolute numbers of either CD4+, CD4+CD28+, CD8+CD28+ T cells, (ii) CD28 MoAb costimulation, and (iii) CD3 MoAb response, and an increase in CD8+CD28− %, are significant predictors for progression to AIDS. In addition, multivariate time-dependent analysis demonstrated that a decrease in CD28 MoAb costimulation (but not a decrease in CD3 MoAb response) was predictive for progression to AIDS, as were decreases in the percentage of CD4+ T cells and the absolute number of CD4+CD28+ T cells. Thus, CD28 MoAb costimulation can be considered a useful assay for monitoring HIV-1 infection. Furthermore, apart from the early increase in the percentage of CD8+CD28− T cells and an increase in the percentage of CD28− on CD8+ T cells in both groups of patients at baseline compared with normal controls, a negative correlation was found to exist between the percentages of CD4+ or CD4+CD28+ T cells and the percentage of CD8+CD28− T cells; this suggests that these cells are probably mutually regulated.
Anti-MHC autoimmunity in Behçet’s disease: T cell responses to an HLA-B-derived peptide cross-reactive with retinal-S antigen in patients with uveitis
Authors: Kurhan-Yavuz S , Direskeneli H , Bozkurt N , Özyazgan Y , Bavbek T , Kazokoglu H , Eksioglu-Demiralp E , Wildner G , Diedrichs-Möhring M , Akoglu T
Published: March 2003

Immune response to retinal autoantigens plays a central role in the pathogenesis of uveitis. A synthetic peptide (B27PD) from a common sequence of various HLA-B molecules associated with uveitis, such as HLA-B27 and 51, which shares amino acid homologies with a retinal-S antigen (S-Ag)-derived peptide (PDSAg), was shown to be immunogenic in human and experimental uveitis in the rat. In this study we investigated T cell responses to B27PD and PDSAg in patients with Behçet’s disease and posterior uveitis (BD-posterior uveitis; n = 33) in comparison with non-Behçet anterior uveitis (AU, n = 14), Behçet’s patients without uveitis (BD, n = 15) and healthy controls (HC, n = 32) in a 6-day proliferation assay. Patients with BD and posterior uveitis had significantly higher responses (stimulation index (SI) 2·8 ± 1·3) than those with AU (SI 1·5 ± 0·4), BD without uveitis (SI 1·1 ± 0·4) and HC (SI 1·1 ± 0·6) for B27PD (P < 0·0001). Responses to PDSAg were also higher in BD with posterior uveitis patients (SI 3·3 ± 1·6) than AU (SI 1·5 ± 0·4), BD without uveitis (SI 1·2 ± 0·3) and HC (SI 1·1 ± 0·6) (P < 0·0001). A significant correlation between the responses to PDSAg and B27PD (r = 0·56, P < 0·001) was observed. Elevated levels of IL-2 and tumour necrosis factor-alpha were also observed in culture supernatants obtained from peripheral blood mononuclear cells after stimulation with the peptides, but no correlation was found between the proliferative responses and cytokine levels. These results suggest that cellular immunity to cross-reactive HLA-B and S-Ag-derived peptides might play a role in the pathogenesis of posterior uveitis in BD.
The proportion of circulatingγδT cells increases after the first week of onset of tularaemia and remains elevated for more than a year
Authors: Kroca M , Tärnvik A , Sjöstedt A
Published: March 2003

In various human intracellular bacterial diseases, an increase of the proportion of circulating Vγ9Vδ2 T cells has been observed. The prevalence of the finding among infected subjects and the time course of the elevation remain to be investigated. In the present study, comprising blood samples from a large number of cases of ulceroglandular tularaemia, the percentage of Vγ9Vδ2 T cells within the first week of onset of disease (5·3 ± 0·7% (mean ± s.e.m.)) did not differ from that of control subjects (5·3 ± 0·8%). Thereafter, percentages increased rapidly and within the interval of 8–40 days mean levels were > 20% (P < 0·001). Of 45 individuals sampled within 3 months of onset, 42 showed a percentage of Vγ9Vδ2 T cells of > 10%. Significantly increased levels were still recorded at 18 months (13·8 ± 2·4%; P < 0·05) but not at 24 months (10·2 ± 2·1%; P > 0·10). Thus, a consistent increase of circulating Vγ9Vδ2 T cells was demonstrated in tularaemia. The initial delay and the prolonged course of elevation may suggest a role in immunoregulation and/or immunological memory. Furthermore, the percentage of γδ T cells expressing tumour necrosis factor-alpha in response to phorbol myristate acetate was decreased during the first week and up to 40 days after onset, possibly reflecting the modulation of an inflammatory response.
Elevated humoral immune response to heat shock protein 60 (hsp60) family in periodontitis patients
Authors: Tabeta K , Yamazaki K , Hotokezaka H , Yoshie H , Hara K
Published: March 2003

The presence of antibodies to the 60-kD human and Porphyromonas gingivalis GroEL hsp60 in the sera and inflamed gingival tissues of periodontitis patients was examined. In order to obtain the antigens, recombinant plasmids carrying human hsp60 and P. gingivalis GroEL genes were constructed and expressed as histidine-tagged recombinant proteins. Immunoreactivities of these proteins were confirmed by MoAbs specific to mammalian hsp60 and cross-reactive with both mammalian and bacterial hsp60. Western blot analysis clearly demonstrated that the number of periodontitis patients showing a positive response to P. gingivalis GroEL was higher than the number of periodontally healthy subjects. Furthermore, anti-P. gingivalis GroEL antibody was detected in all samples of gingival tissue extracts. For human hsp60, a higher frequency of seropositivity was found in the periodontitis patients than in the healthy subjects. In addition, the periodontitis patients demonstrated stronger reactivity compared with the healthy subjects. Quantitative analysis of serum antibodies by ELISA also demonstrated that the levels of antibodies in the sera of patients were significantly higher than those of control subjects. In the gingival tissue extracts, seven out of 10 patients demonstrated a positive response to human hsp60 and tso of these demonstrated strong positivity. Affinity-purified serum antibodies to human hsp60 and P. gingivalis GroEL from selected patients reacted with P. gingivalis GroEL and human hsp60, respectively, suggesting cross-reactivity of antibodies. These results suggest that molecular mimicry between GroEL of the periodontopathic bacterium P. gingivalis and autologous human hsp60 may play some role in immune mechanisms in periodontitis.
Intra-articular IL-10 gene transfer regulates the expression of collagen-induced arthritis (CIA) in the knee and ipsilateral paw
Authors: Lubberts E , Joosten L A B , Van Den Bersselaar L , Helsen M M A , Bakker A C , Xing Z , Richards C D , Van Den Berg W B
Published: March 2003

We studied the effects of local IL-10 application, introduced by a recombinant human type 5 adenovirus vector, in the mouse knee joint during the early phase of CIA. One intra-articular injection with the IL-10-expressing virus (Ad5E1mIL-10) caused substantial over-expression of IL-10 in the mouse knee joint, using virus dosages which did not induce distracting inflammation. High expression of IL-10 was noted for a few days, being maximal at day 1. One intra-articular injection of Ad5E1mIL-10 in the knee joints of collagen type II (CII)-immunized mice, before onset of CIA was noted, reduced the incidence of collagen arthritis in that knee. Of high interest, the protective effect of local IL-10 expression by Ad5E1mIL-10 was not restricted to the knee joint alone. The arthritis incidence in the ipsilateral paw was highly suppressed. In contrast, local IL-10 over-expression was not effective when treatment was started after onset of CIA. Further analysis in the acute streptococcal cell wall-induced arthritis model revealed that local IL-10 over-expression markedly suppressed the production of tumour necrosis factor-alpha (TNF-α) and IL-1α, but had no significant effect on IL-1β and IL-12 production in the inflamed synovium. These data indicate that local over-expression of IL-10 in the knee joint of mice regulates the expression of collagen arthritis, probably through down-regulation of TNF-α.
IVIG-bound IgG and IgM cloned by phage display from a healthy individual reveal the same restricted germ-line gene origin as in autoimmune thrombocytopenia
Authors: Hoffmann M , Uttenreuther-Fischer M M , Lerch H , Gaedicke G , Fischer P
Published: March 2003

Intravenous immunoglobulin preparations (IVIG) have shown positive effects in the treatment of immune defects and autoimmune diseases. It is not clear how IVIG interacts with the components of the immune system. To investigate this, we cloned previously a large number of phage displayed IgG Fab fragments derived from three patients with autoimmune thrombocytopenia (AITP) that were specifically bound by IVIG molecules. Many of these Fabs reacted with platelets. Sequencing revealed that the most frequently used germ-line gene segments of all IVIG-bound Fabs were identical to those observed for many other autoantibodies. Particularly, the loci 3–30 or 3–30/3–30.5, 3–23 and 3r, 3l, and 2a2 represented the most abundant genes used for the heavy (VH) and light chain V region (VL), respectively. This suggested a specific interaction of IVIG molecules with B cells that present B cell receptors derived from these germ-line genes. In the current study we determined the genetic origin of IVIG-reactive IgG and IgM cloned from a healthy person. A favoured selection of antibodies derived from the same germ-line origins as in AITP was observed. Because 3–30 and 3–23 are the most frequently rearranged VH germ-line gene segments among human B cells, our results suggest that this favoured anti-idiotypic interaction may have an important role for the development and control of the normal B cell repertoire.
Cytokine dysregulation in activated cystic fibrosis (CF) peripheral lymphocytes
Authors: Moss R B , Hsu Y-P , Olds L
Published: March 2003

Recent studies demonstrate in vivo and in vitro cytokine dysregulation in CF epithelial cells. To see if these abnormalities may be generalized to other cells expressing cystic fibrosis transmembrane conductance regulator (CFTR) but not directly exposed to local inflammation, we studied mRNA transcription, intracellular protein production and extracellular secretion of IL-2, IL-4, IL-5, IL-10 and interferon-gamma (IFN-γ) from freshly isolated blood mononuclear and CD4+ T cells from CF patients and controls. Cells were activated by phorbol myristate acetate (PMA) and anti-CD3, PMA–ionomycin, or lipopolysaccharide (LPS) and assessed for cytokine mRNA transcription by semiquantitative reverse transcriptase-polymerase chain reaction, intracellular protein production by flow cytometry, and secretion by supernatant ELISA. Cytokine expression was highly stimulus-dependent. CF cells showed higher IL-10 transcription than control cells after maximal activation by LPS (P = 0·01); despite this, cytokine production and secretion were equivalent to controls. CF cells showed lower cellular IL-10 production after PMA-anti-CD3 activation (P = 0·002). CF cells secreted less IFN-γ than control cells after maximal activation by PMA-anti-CD3 (1836 ± 273 pg/ml versus 9635 ± 3437 pg/ml, P = 0·04). IL-2, IL-4 and IL-5 regulation was similar to controls. We conclude that CF mononuclear cells show selective cytokine dysregulation after maximal activation, namely reduced IFN-γ secretion and increased IL-10 mRNA without increased production or secretion. These findings extend defects described in respiratory epithelial cells to circulating immunoregulatory cells, suggesting a link between CF genotype and cytokine dysregulation.
The endothelium is an extrahepatic site of synthesis of the seventh component of the complement system
Authors: Langeggen H , Pausa M , Johnson E , Casarsa C , Tedesco F
Published: March 2003

The level of the terminal complement components secreted by human umbilical vein endothelial cells (HUVEC) was measured by a sensitive ELISA which allows the detection of 30–50 pg/ml of these components. C7 was the only terminal component detected in measurable amounts in the cell supernatant. The mean value was 11 ng/106 cells at 96 h and was slightly higher than that of C3 (9 ng/106 cells). HUVEC and serum C7 analysed by SDS–PAGE and immunoblot exhibited the same electrophoretic mobility. A proportion of C7 secreted by HUVEC was incorporated into the terminal complement complex (TCC) assembled spontaneously in the supernatant of cells cultured in C7-deficient human serum, and was not detected by the standard ELISA for C7 measurement. By adding the amount of C7 present in the TCC to that of free C7, the total amount of the component released by HUVEC was calculated to be approximately 35 ng/106 cells. Further TCC was produced following complement activation of the cell supernatant through the alternative pathway. Synthesis of C7 by HUVEC was confirmed by inhibition experiments in the presence of cycloheximide and by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of C7 mRNA expression. Addition of IL-1α and tumour necrosis factor-alpha to the cell culture stimulated the secretion of C3, but had no effect on the synthesis of C7. By contrast, interferon-gamma had only a marginal effect on the production of C3, but markedly down-regulated the synthesis of C7 as assessed both by ELISA and RT-PCR.
Effects of secretory leucocyte protease inhibitor on the production of the anti-inflammatory cytokines, IL-10 and transforming growth factor-beta (TGF-β), by lipopolysaccharide-stimulated macrophages
Authors: Sano C , Shimizu T , Sato K , Kawauchi H , Tomioka H
Published: March 2003

We studied the effects of secretory leucocyte protease inhibitor (SLPI) on the production of the anti-inflammatory cytokines, IL-10 and TGF-β, by lipopolysaccharide (LPS)-stimulated macrophages, using half-sized SLPI (1/2 SLPI) containing the C-terminal domain (Arg58-Ala107). ELISA testing of macrophage culture fluids showed a temporary production of IL-10 by the macrophages in the early phase (24 h) after LPS stimulation at low (1 ng/ml) or high (10 μg/ml) concentrations. On the other hand, TGF-β production was initiated after day 3 and progressively increased. 1/2 SLPI significantly increased IL-10 and TGF-β production by macrophages in response to a low dose as well as a high dose of LPS. Reverse transcription-polymerase chain reaction analysis showed that 1/2 SLPI caused a significant increase in the expression of both IL-10 and TGF-β mRNAs by LPS-stimulated macrophages. Thus, although the profile of macrophage TGF-β production by LPS-stimulated macrophages is markedly different from that of their IL-10 production, SLPI causes an up-regulation of the production of these anti-inflammatory cytokines by LPS-stimulated macrophages.
Expression of MHC class II in T cells is associated with increased HIV-1 expression
Authors: Saifuddin M , Spear G T , Chang C-H , Roebuck K A
Published: March 2003

HIV-1 replicates in activated T cells at significantly higher levels than in resting cells. Thus, certain molecules up-regulated during T cell activation appear to be important for HIV-1 replication. In this study, we present evidence suggesting that expression of MHC class II (class II) molecules on CD4+ T cells facilitate HIV-1 replication. T cells that expressed class II supported greater virus replication than T cells lacking class II. The class II+ cells, when either infected with HIV-1 or transfected with an env-minus HIV-1 provirus plasmid, produced 10–20-fold greater virus expression than class II− cells. Anti-class II antibody markedly inhibited virus expression in class II+ cells (but not class II− cells) and also decreased the nuclear binding activity of AP-1, an inducible transcription factor important in T cell activation and HIV-1 expression. Most importantly, the induction of class II expression by transfection of the MHC class II transactivator (CIITA) stimulated HIV-1 replication in Jurkat T cells. Taken together, these data suggest that expression of MHC class II molecules and/or CIITA in T cells enhances HIV-1 transcription.
Changed colonic profile of P-selectin, platelet-endothelial cell adhesion molecule-1 (PECAM-1), intercellular adhesion molecule-1 (ICAM-1), ICAM-2, and ICAM-3 in inflammatory bowel disease
Authors: Vainer B , Nielsen O H
Published: March 2003

Cell adhesion molecules (CAM) are essential for the capture and migration of leucocytes. Ulcerative colitis (UC) and Crohn's disease (CD) are characterized by a continuous infiltration of leucocytes into intestinal tissue, and the colonic contents of P-selectin, PECAM-1, ICAM-1, ICAM-2, and ICAM-3 were therefore studied. Concentrations of these cell adhesion molecules were measured by an ELISA technique in sonicated colonic tissue from patients with UC and CD and controls with non-inflammatory disease and compared with the diagnosis and disease activity. P-selectin, PECAM-1, and ICAM-1 concentrations were elevated in UC patients compared with controls (P = 0.034, P = 0.014, P = 0.017, respectively), whereas that of ICAM-2 was not. The concentrations of these CAM did not differ in CD. In contrast, higher concentrations of ICAM-3 were found in the CD patients than in either UC (P = 0.001) or controls (P = 0.004). The CAM concentrations increased with disease activity, although only ICAM-1 was significantly elevated (P = 0.017). As considerable differences were found between UC and CD with comparable stages of inflammation, the mere presence of inflammation cannot solely explain the results. The observed differences in the CAM concentrations in UC and CD support the hypothesis that UC and CD are two distinct disease entities with separate pathogenic mechanisms.
Angiogenesis in inflammatory joint disease: a target for therapeutic intervention
Authors: Brenchley P E C
Published: March 2003

The evidence reviewed here clearly supports the concept that pathological angiogenesis is an important component in inflammatory joint erosion. Of the primary angiogenic factors, VEGF-A is clearly a key participant in this mechanism and a range of anti-VEGF strategies is being developed to neutralize its biological function [54,55]. Currently available drugs are also being screened for VEGF antagonistic effects. In a study of the effects of existing disease-modifying anti-rheumatic drugs on cultured synovial cells, bucillamine and dexamethasone showed significant inhibition of VEGF production [56]. In addition, COX-1 and COX-2 non-steroidal anti-inflammatory drugs have been shown to inhibit angiogenesis by blocking VEGF-induced signal transduction [57]. Modulation of the immune network in RA using TNF-α antagonists is producing promising results, but as outlined in this review, this treatment on its own is unlikely to control joint angiogenesis. It is possible that combination therapy, e.g. TNF-α antagonist and a VEGF signal transduction inhibitor, will be more effective by using antagonists that block different but key control points in the disease pathology.
Distribution of age-related thymulin titres in normal subjects through the course of life
Authors: Consolini R , Legitimo A , Calleri A , Milani M
Published: March 2003

The thymus has a dominant immunological role in utero and in early childhood, being a primary source of T lymphopoiesis, and its investigation may be particularly relevant for the immunological study of paediatric patients. Thymulin, a nonapeptide secreted by the thymus, is an essential hormone for T lymphocyte differentiation and function. As thymulin values in the normal population have not been well documented, especially for children under the age of 1 year, we detail thymic endocrine function by presenting age-related plasma thymulin levels in a large series (n = 93) of healthy individuals, ranging from birth to old age. We demonstrate that thymulin is already detectable at birth; it then gradually increases with age, reaching the highest level in children aged 5–10 years. Starting at adolescence, thymulin titres gradually start to fall, reaching the lowest value at 36 years of age and remaining steady until 80 years (the oldest person tested).
The effect of testosterone replacement treatment on immunological features of patients with Klinefelter's syndrome
Authors: Koçar I H , Yesilova Z , Özata M , Turan M , Sengül A , Özdemir I Ç
Published: March 2003

Although the effects of androgen deficiency in the immune system have long been appreciated, little is known about the immunological features of patients with Klinefelter's syndrome (KS). On the other hand, interest in androgens as a possible treatment for some autoimmune diseases is growing. In the present study, some immunological parameters were evaluated in 26 patients with KS prior to androgen replacement treatment (ART) and the results were compared with those in 19 healthy control subjects. Patients were then treated with testosterone for 6 months and the pre- and post-treatment findings were compared. Serum levels of IgG, IgA, IgM, C3c and C4 were measured by nephelometry and lymphocyte subsets and CD4+/CD8+ ratios were examined by flow cytometry. IL-2 and IL-4 levels were measured by ELISA. Pretreatment levels of the serum IgA, IgG, IgM, IL-2 and IL-4 of the patients were higher than those of the controls and were all decreased significantly following ART. The pretreatment absolute numbers and percentages of CD3+, CD4+, CD19+ cells and CD4+/CD8+ ratios of patients with KS were higher than those of the controls and were all decreased with ART. Percentages of CD8+ cells were increased significantly, while C3 and C4 levels were both significantly decreased after ART. It is concluded that the lack of testosterone in patients with KS enhances cellular and humoral immunity and that ART may suppress this.
TNFA promoter polymorphism and susceptibility to brucellosis
Authors: Caballero A , Bravo M J , Nieto A , Colmenero J D , Alonso A , Martín J
Published: March 2003

The aim of this study was to investigate the possible influence of the tumor necrosis factor alpha (TNFA) gene promoter polymorphisms and HLA class II genes on the susceptibility to or development of human brucellosis. TNFA genotypes (at positions −308 and −238) were determined in 59 patients with brucellosis and 160 healthy controls by polymerase chain reaction-restriction fragment length polymorphism. There were no significant differences between the patients and the controls for the TNFA-238 genotypes. However, when the overall TNFA-308 genotype distribution of the brucella patients was compared with that of the control subjects, a significant skewing was observed (P = 0·02). The TNFA-308.1/2 genotype was present at significantly higher frequency in the total patient as a whole compared with control subjects (30% versus 15%; P = 0·01, odds ratio (OR) 2·49, 95% confidence interval (CI) 1·16–5·33). No statistically significant differences in the distribution of HLA-DRB1 or DQB1 alleles were observed between brucella patients and control subjects. Stratification to correct for interdependence of TNFA-308.2 and HLA-DR3 alleles confirmed that, in spite of their strong linkage disequilibrium, the association of TNFA-308.2 with brucellosis was independent of HLA-DR3.
Macrophage inflammatory protein-1α(MIP-1α), MIP-1β, and RANTES mRNA semiquantification and protein expression in active demyelinating multiple sclerosis (MS) lesions
Authors: Boven L A , Montagne L , Nottet H S L M , De Groot C J A
Published: March 2003

MS is a demyelinating disease characterized by infiltration of monocytes and lymphocytes into the brain parenchyma, destruction of oligodendrocytes and loss of myelin. Since chemokines play a major role in the migration of monocytes and T cells, we here investigated the expression of the CC chemokines MIP-1α, MIP-1β, and RANTES in brain tissue from MS patients using reverse transcriptase-polymerase chain reaction techniques. Both MIP-1β as well as RANTES were found to be significantly elevated in brain tissue of MS patients. In addition, MIP-1α was also increased, although not significantly. Immunohistochemistry revealed that, whereas RANTES was mainly localized in reactive astrocytes, MIP-1α and MIP-1β immunoreactivity was predominantly found in perivascular and parenchymal macrophages, containing myelin degradation products. Thus, chemokines appear to be associated with MS and an increased chemokine expression may further enhance disease progression by attracting more leucocytes into the brain parenchyma and by activation of effector functions of astrocytes and microglial cells.
Atopic eczema is associated with delayed maturation of the antibody response to Pneumococcal vaccine
Authors: Arkwright P D , Patel L , Moran A , Haeney M R , Ewing C I , David T J
Published: March 2003

The aim of this study was to investigate a previously undocumented observation, that children with atopic eczema under 9 years of age tended to have a poor antibody response to Pneumococcal vaccination. Thirty-five children (mean age 8·8 years, range 3–16 years) with moderate to severe atopic eczema but no history of systemic infection were studied retrospectively. Pneumococcal antibody responses after immunization with Pneumovax II were compared with a hospital control group consisting of 36 children (mean age 6·0 years, range 3–16 years) with recurrent upper respiratory tract infections. Only 17% of children with atopic eczema aged 3–8 years responded to Pneumovax. This response was significantly poorer than that of the controls (57%) (odds ratio 0·20, 95% confidence interval (CI) 0·05–0·84, P = 0·03). There were no significant differences in the levels of total IgG2, the component of IgG associated with protective antibody responses to Pneumococcus between the two groups. Delay in maturation of the total IgG and IgG2 antibody response to Pneumococcus is a feature in this group of children with moderately severe atopic eczema.
Cardiolipin on the surface of apoptotic cells as a possible trigger for antiphospholipid antibodies
Authors: Sorice M , Circella A , Misasi R , Pittoni V , Garofalo T , Cirelli A , Pavan A , Pontieri G M , Valesini G
Published: March 2003

This study provides evidence that cardiolipin (CL) molecules are expressed on the surface of apoptotic cells and are recognized by antiphospholipid antibodies, purified from patients with the antiphospholipid antibody syndrome (APS). CL expression on cell surface was demonstrated by high performance thin layer chromatography analysis of phospholipids from plasma membrane purified fractions and by the positive staining with the CL-specific dye nonyl-acridine orange. This finding was complemented with the observation that aCL IgG purified from patients with APS bind to the surface of apoptotic cells. This staining shows a clustered distribution mostly localized on surface blebs. Interestingly, CL exposure on the cell surface preceded the DNA fragmentation, as shown by cytofluorimetric analysis. These findings demonstrate that exposure of CL molecules on the cell plasma membrane is an early event of the apoptotic cellular program that may represent an in vivo trigger for the generation of aCL.
Enhanced production of IL-8 in chronic but not in early ileal lesions of Crohn's disease (CD)
Authors: Brandt E , Colombel J-F , Ectors N , Gambiez L , Emilie D , Geboes K , Capron M , Desreumaux P
Published: March 2003

Distinct Th1/Th2 patterns have been observed during the evolution of CD. The aim of this study was to compare neutrophil involvement and IL-8 mRNA and protein expression during early recurrent lesions and chronic phases of CD. Twenty-nine patients with CD having ileocolonic resection with anastomosis were studied. Biopsies were obtained during surgery from the non-inflamed ileal mucosa and from chronic ileal lesions. Endoscopic ileal biopsies were also taken from early recurrent ileal lesions occurring 3 months after surgery. Neutrophil counts were performed and mucosal IL-8 levels were evaluated by competitive reverse transcriptase-polymerase chain reaction and immunohistochemistry. Early recurrent ileal lesions were characterized by low neutrophil counts and IL-8 production at the mRNA and protein levels compared with the ileal chronic lesions. The main cellular sources of IL-8 in the early recurrent lesions were neutrophils, while in chronic lesions the majority of IL-8-stained cells were CD3+ T cells and macrophages. These results confirmed that the nature of the inflammatory infiltrate and the expression of cytokine profiles may differ between the acute and chronic phases of CD.
Blood fetal microchimerism in primary biliary cirrhosis
Authors: Invernizzi P , De Andreis C , Sirchia S M , Battezzati P M , Zuin M , Rossella F , Perego F , Bignotto M , Simoni G , Podda M
Published: March 2003

The autoimmune nature of primary biliary cirrhosis (PBC) is well established. We tested the hypothesis that fetal microchimerism indicated by the persistence of circulating fetal cells in women years after pregnancy might contribute to the aetiopathogenesis of PBC through a graft-versus-host-like response. We extracted DNA from the peripheral blood cells of 36 women carefully selected from 173 consecutive PBC patients, who were matched with 36 healthy women by age, age of last son, and number of children. Both patients and controls had to have male offspring, and no history of miscarriages or blood transfusions; they could not be twins. We tested all of the samples for the presence of two specific Y-chromosome sequences (SY154 and SRY) by amplifying DNA in a nested polymerase chain reaction. Y-chromosome-specific DNA was detected in the peripheral blood cell DNA of 13 (36%) of the 36 women with PBC and in 11 (31%) of the 36 healthy controls. The two groups of PBC patients with and without male DNA sequences were similar in terms of their clinical, biochemical, and serological features. Y-chromosome sequences were found in three of the four PBC women with associated systemic sclerosis. All of the 24 Y-positive samples contained SY154 sequences, but only three PBC patients and six controls showed the presence of both SY154 and SRY sequences. This discrepancy may suggest that not only fetal cells but also fragments of fetal DNA are present in maternal circulation. Overall, our data do not support the hypothesis that fetal microchimerism plays a significant role in the onset or progression of PBC.