bluuebunny/crossref_metadata_2025 huggingface dataset:
Porphyromonas gingivalis-specific serum IgG and IgA antibodies originate from immunoglobulin-secreting cells in inflamed gingiva
Authors: OGAWA T , KONO Y , MCGHEE M L , MCGHEE J R , ROBERTS J E , HAMADA S , KIYONO H
Published: July 2010

Patients with adult periodontitis (AP) exhibit elevated serum antibody levels to Porphyromonas (Bacteroides) gingivalis; however, it is not known whether these antibodies originate from plasma cells in the local disease site or From peripheral lymphoid tissues. We studied the isotype and subclass levels and origin of antibodies to P. gingivalis fimbriae, since elevated serum anti-fimbriae responses were seen when compared with sera of healthy controls. IgG unti-fimbriae litres were dominant and the subclass response was IgG3 ≫ IgG1 > IgG2 ≫ IgG4; however, some IgA anti-fimbriae antibodies were also seen. The IgA subclass fimbriae-specific response was mainly IgA1; however, significant IgA2 anti-fimbrae antibodies were seen. We also assessed numbers of anti-fimbriae antibody producing cells from peripheral blood mononuclear cells (PMBC) and from either healthy or inflamed gingiva of AP subjects. Gingival mononuclear cells (GMC) of AP patients exhibited high numbers of immunoglobulin-producing (spot-forming) cells (SFC) including fimbriae-specific antibody secreting cells in a pattern of IgG > IgA > > > IgM. However, low numbers of SFC were seen in GMC from healthy gingiva; further, no anti-timbriac SFC responses were noted in healthy GMC. Although no fimbriae-specific immunoglobulin-producing cells were seen in PBMC. low numbers of antigen-specific SFC were found in pokeweed mitogen-triggercd PBMC from AP subjects. Treatment of AP patients for plaque and surgical removal of inflamed gingiva resulted in significant reductions in serum anti-fimbriac responses. These studies show that AP patients exhibit brisk serum IgG and IgA subclass anti-fimbriac antibodies, whose origin appear to be the plasma cells present in the localized inflamed tissues.
Detection of human and murine common idiotypes of DNA antibodies in tissues and sera of patients with autoimmune diseases
Authors: WATTS R A , RAVIRAJAN C T , WILKINSON L S , WILLIAMS W , GRIFFITHS M , BUTCHER D , HORSFALL A T , STAINES N A , ISENBERG D A
Published: July 2010

The expression in tissue and serum of a panel of murine and human common DNA antibody idiotypes (Ids) (BEG 2. PR4, F-423, I-402, II-28. IV-228. V-88) has been investigated. The murine V-88 Id was detected in eight out of 10 and the human BEG 2 Id in five out of 10 labial biopsies from patients with Sjögren's syndrome. The murine F-423, I-402 and IV-228 Ids were identified in one out of 10 biopsies. In each case the pattern of staining was similar with staining of the acinar basement membrane and a cell population. Using double-labelling immunohistochemistry this cell population were identified as plasma cells. No staining was seen in Four normal labial biopsies. The V-88 Id was detected on the epithelial aspect of the thickened basement membrane in three out of nine renal biopsies from patients with systemic lupus erythematosus (SLE). None of the other Ids (BEG2, PR4, IV-228. F-423 or I-402) could be detected in renal tissue. None of the Ids were found in skin biopsies from SLE patients. Id V-88 may, like the 16/6 Id to which it is phenotypically related, play a role in the pathogenesis of renal lesions in SLE. The BEG 2 Id could be detected in the serum of patients with rheumatoid arthritis (RA) and active untreated tuberculosis. Ids II-28, V-88 and I-402 were elevated in serum from patients with Sjogren's syndrome and II-28 Id in serum from patients with myositis and RA. None of the Ids were elevated in serum from patients with SLE. Apart from the BEG 2 Id, none of the Ids were elevated in serum from patients with tuberculosis or Gram-negative infections. The presence of murine Ids in human tissue and serum suggests that they arc cross-species idiotypes and have been conserved through evolution.
Detection of interleukin-6 and interleukin-1 production in human thyroid epithelial cells by non-radioactivein situhybridization and immunohistochemical methods
Authors: ZHENG R Q H , ABNEY E , CHU C Q , FIELD M , GRUBECK-LOEBENSTEIN B , MAINI R N , FELDMANN M
Published: July 2010

Human endocrine thyroid epithelial cells have been described to produce cytokines in vitro. In order to determine whether they do so in vivo during thyroiditis. parallel studies on mRNA expression with a non-radioactive in situ hybridization technique and immunohislochemical detection for the protein were performed on frozen sections of thyroid samples from autoimmune thyroiditis (Graves' disease and Hashimoto's thyroiditis), non-toxic goitre and normal thyroid tissue. cDNA probes were sulphonated and their hybridization with mRN A was detected with a sulphonyl-specific monoclonal antibody. This signal was amplified and visualized with the alkaline phosphatase anti-alkaline phosphatase (APAAP) system. The protein products were detected with immuno-purified rabbit F(ab′)2 antibody fragments recognizing recombinant human cytokines, visualized by the immuno-peroxidase technique. Each sample was studied at the two levels. Both interleukin-6 mRNA and protein were found in the endocrine cells. There was no obvious difference between autoimmune thyroiditis and non-toxic goitre. However, normal thyroid epithelial cells produced less interleukin-6. Interleukin-1α mRNA and its protein were found in epithelial cells from Hashimoto's thyroiditis samples, but not in the others, except one Graves' disease sample, in which only mRNA was detected. Interleukin- 1β was not detected in these cells, its mRNA was only found in one of the Graves' disease samples. These cytokines were also detected in some infiltrating cells.
Serum immunomodulatory factors in gastrointestinal diseases. A 30-50-kD serum fraction in Crohn's disease capable of modulating lymphocyte activation
Authors: BIANCONE L , BOIRIVANT M , FAIS S , RICCI G L , PAGANELLI R , PALLONE F
Published: July 2010

We tested the hypothesis that serum factors present in Crohn's disease interfere with the process of lymphocyte activation. The mitogen-induced proliferation and the expression of early activation antigens by normal lymphocytes cultured in the presence of either Crohn's disease sera or sera from different controls were evaluated. The mitogen-induced proliferation was significantly impaired in the presence of Crohn's disease sera. These sera markedly inhibited the mitogen-induced interleukin-2 receptor (IL-2R) expression (48% inhibition), while the effect of sera on the expression of the transferrin receptor and the 4F2 antigen was much less pronounced. Diafiltration experiments showed that the inhibitory effect was confined to a 30–50-kD serum fraction. Such a serum property was not related to the patients' disease activity and disappeared after surgical removal of the affected bowel. The capability of inhibiting the mitogen-induced IL-2R expression was not restricted to Crohn's disease and was observed with sera from other inflammatory and neoplastic gastrointestinal disorders. This study indicates that a marked inhibition of the IL-2R is a mechanism underlying the immunosuppressive property of the serum in Crohn's disease and in other gastrointestinal conditions.
Clonotypic analysis of human antibodies specific forNeisseria meningitidispolysaccharides A and C in adults
Authors: LE MOLI S , MATRICARDI P M , QUINTI I , STROFFOLINI T , AMELIO R D
Published: July 2010

Serum antibodies to the capsular polysaccharides A and C (PSA and PSC) of N. meningitidis in healthy adults before and after vaccination with the sole polysaccharides were analysed by isoelectric focusing (IEF). Before vaccination, 49% and 28% had naturally acquired antibodies against PSA and PSC, respectively, whereas 18 days after vaccine administration 84% and 91%, respectively, showed a detectable spectrotypic pattern. Oligoclonality appeared to be the main feature of naturally acquired and vaccine-induced antibodies for both polysaccharides. In all subjects the anti-PSA response, showing dominant bands at the same pH position, was more homogeneous than anti-PSC one. Most subjects with naturally acquired antibodies (25 out of 38 for PSA and 20 out of 22 for PSC) showed a spectrotypic pattern after vaccination, similar to that observed before vaccination (any differences were just related to band intensity), suggesting that PSA and PSC are able to recruit the same B cell clones previously primed with a T-dependent form of the antigen, i.e. the whole bacterium. However, in one-third of subjects with naturally acquired anti-PSA antibodies, the appearance of new alkaline bands after vaccination was observed. Furthermore, in subjects with absence of detectable natural antibodies, the vaccine-induced antibody response started in correspondence of alkaline pH areas, subsequently extending to neutral and acidic areas. Therefore, it may be hypothesized that alkaline antibody-secreting B cell clones are the first to be recruited. The final spectrotype in these subjects was similar to that observed in subjects with naturally acquired antibodies. This observation, together with the above reported data, allow us to conclude that natural (T-dependent pathway) and vaccine (T-independenl pathway) immunization induce the expression of the same antibody repertoire, for both meningococcal PSA and PSC.
Effect of recombinant human erythropoietin on human IgE productionin vitro
Authors: KIMATA H , YOSHIDA A , ISHIOKA C , MIKAWA H
Published: July 2010

Recombinant human erythropoietin enhanced spontaneous IgE production 200–300% enhance ment) in cultures of peripheral blood mononuclear cells (MNC) from atopic patients. In contrast, IgG and IgA production were only slightly enhanced (30–50% enhancement), and IgM production was not affected by erythropoietin. The enhancement of IgE production by erythropoietin was indirect since it required T cells and monocytes. However, erythropoietin effect was specific since enhancement was blocked by anti-erythropoietin antibody but not by control antibody. Interleukin-4 (IL-4) also enhanced spontaneous IgE production from atopic MNC. However, the enhancing effect by erythropoietin is different from that by IL-4, since the erythropoietin effect was not blocked by anti-IL-4 antibody, and conversely IL-4 effect was not blocked by anti-erythropoietin antibody. In contrast to the enhancing effect on atopic MNC, erythropoietin failed to induce IgE production in cultures of MNC from normal donors while IL-4 induced IgE production from normal MNC. However, when normal MNC were pre-incubated with IL-4. erythropoietin enhanced IgE production from IL-4-pre-incubated MNC. Moreover. B cells separated from IL-4-pre-incubated MNC produced IgE which was enhanced by erythropoietin. However, this effect required T cells and monocytes. These results indicate that erythropoietin could regulate ongoing IgE production in vitro by T cell- and monocyte-dependent mechanisms.
Human T cell responses to β-galactosidase
Authors: OXLEY J D , BROOKES R H , RAYFIELD L S , SHEPHERD P S
Published: July 2010

The peripheral blood of most normal individuals has been shown to contain T cells that respond to β-galactosidase (β-Gal), presumably as a result of natural priming. Three T cell clones (clones 1,2,4) specific for β-Gal were isolated from peripheral blood mononuclear cells (PBMC) after pretreatment with leucine methyl ester (LeuOMe); a fourth clone from the same individual was isolated from untreated cells. All four clones were CD4+ CD8- aβTcR+ and clone 1 was additionally shown to be cytotoxic. Epstein-Barr virus (EBV) transformed B cell lines were derived from LeuOMe-treated or untreated PBMC and used to study the efficiency of presentation of β-Gal to one of the clones. The results indicated that B cells transformed after LeuOMe treatment presented β-Gal at lower concentrations than untreated controls. β-Gal would therefore appear to be a highly suitable model antigen for studies of immunoregulation in humans.
Japanese encephalitis virus latency in peripheral blood lymphocytes and recurrence of infection in children
Authors: SHARMA S , MATHUR A , PRAKASH V , KULSHRESHTHA R , KUMAR R , CHATURVEDI U C
Published: July 2010

In a study group of 40 children who had been admitted to hospital with acute encephalitis, the disease was due to infection with Japanese encephalitis virus (JEV). Three children developed recurrence of disease 8-9 months later. No virus had been isolated from these three patients during the acute stage of their illness, but virus was recovered from all during the recurrence phase by co-cultivation of their peripheral blood mononuclear cells in primary mouse embryo fibroblast cultures. Virus was also recovered by co-cultivation of peripheral blood mononuclear cells collected 8 months after their acute disease from three out of eight randomly selected asymptomatic children within the study group but not from similar cultures set up from JEV-seronegative children used as controls. Virus was also isolated by co-cultivation of T lymphocytes of asymptomatic children as detected by indirect immunofluorescence or by inoculation in mice.
Gamma/delta T cells and the diagnosis of coeliac disease
Authors: SPENCER J , ISAACSON P G , MACDONALD T T , THOMAS A J , WALKER-SMITH J A
Published: July 2010

Gamma/delta T cells are increased in the gut epithelium of patients with coeliac disease compared with normal controls. The aim of this study was to determine whether the increase in yd intraepithelial lymphocytes (IEL) is specific for coeliac disease, in which case it could be of diagnostic importance. Biopsies were obtained from children with no intestinal disease, coeliac disease, cow-milk-sensitive enteropathy/post-enteritis syndrome (CMSE/PES) and miscellaneous other enteropathies (n = 67). Intraepithelial CD3+ and γδ T cells were identified in frozen sections using peroxidase immunohisto-chemistry. In normal biopsies there were 0-7 γδ IEL/100 cells in the epithelium. In untreated coeliac patients this increased to 9-22 γδ IEL/100 cells in the epithelium (P=0.000004). Of 27 patients with morphologic intestinal damage which was not due to coeliac disease, four with CMSE/PES had γδ IEL/100 cells in the epithelium in the same range as the patients with coeliac disease. Of these, two had high densities of CD3+ IEL in the epithelium and were indistinguishable from patients with untreated coeliac disease. The other two could be excluded as possible coeliacs because their CD3f IEL/100 epithelial cells were in the normal range. Thus an increase in γδ IEL is not specific for coeliac disease. However, enumeration of both of γδ IEL and CD3+ IEL densities will be useful in the exclusion of coeliac disease as a diagnosis in some children.
Expression of CD34 on human B cell precursors
Authors: SCHMITT C , EAVES C J , LANSDORP P M
Published: July 2010

CD34 is a 110-kD glycoprotein previously shown by a variety of monoclonal antibodies (MoAbs) to be expressed selectively on immature hematopoietic cells. However, more detailed characterization of CD34+ cells has been hampered by lack of anti-CD34 MoAbs that can be labelled directly with fluorochromes to facilitate subpopulation analysis by multi-parameter flow cytometry. We have recently isolated a murine anti-CD34 MoAb, designated as 8G12, that can be directly labelled with fluorochromes such as FITC. In this study, we have exploited this property of 8G12 to compare the reactivity of 8G12 and My10 with normal and leukaemic human marrow cells and to characterize normal early human B cell precursors by two- and three-colour immunofluorescencc analysis. Comparison of three-colour staining profiles of normal bone marrow cells incubated with both 8G12 and MY10, and either anti-CD10 or anti-CD19 MoAb revealed the reactivity patterns of 8G12 and MY10 to be indistinguishable. This conclusion was confirmed by a similar comparative analysis of 8G12 and MY10 staining of blood and bone marrow cells from 4 patients with B lineage acute lymphoblastic leukaemia (ALL). Of interest, both 8G12and MY 10 detected a CD34+ CD10+ CD19+ population in normal adult bone marrow. To determine whether a CD34+ CD10+ CD19- precursor population previously reported by others to exist in fetal liver could also be identified, CD10+ CD16-marrow cells were first isolated by FACS and the sorted cells then re-analysed for expression of CD19 and CD34. These studies showed that all of the sorted CD10+ cells that expressed CD34 appeared to coexpress CD19. No CD34+ CD10+ CD19- cells were detected (at a sensitivity of 0.1%). Further studies will be required to determine whether a very minor population of CD34+ CDl0+ CD19- cells may still be generated in the normal development of B cells in adult human marrow.
Immunophenotypic analysis of histiocytes involved in AIDS-associatedMycobacterium scrofulaceuminfection: similarities with lepromatous lepra
Authors: DELABIE J , DE WOLF-PEETERS C , BOBBAERS H , BILBE G , DESMET V J
Published: July 2010

The present study reports a rare case of systemic M. scrofulaceum infection in an AIDS patient and analyses the inflammatory infiltrate in a lymph node by immunohistochemistry. Special emphasis is put on the histiocytes. The diffuse infiltrate consists mainly of large histiocytes that contain numerous bacilli. These cells display the phenotype of mature histiocytes and in addition coexpress the antigens recognized by RFD7 and RFD9, both markers of different subsets of histiocytes which have been reported to be co-expressed by the infected histiocytes in the infiltrate of lepromatous lepra. Interdigitating reticulum cells are rare as well as T cells which are mainly of the suppressor/cytotoxic type. These findings arc similar to those reported for lepromatous lepra and might indicate common deficiencies in T cell macrophagc interactions in both conditions. Superimposed on the diffuse infiltrate of large histiocytes we observed ‘monocytic granulomas’, the presence of which might be related to a reactional state comparable to erythema nodosum leprosum, a reactional state of lepromatous lepra.
An abnormal T cell repertoire in hypergammaglobulinaemic primary Sjogren’s syndrome
Authors: KAY R A , HAY E M , DYER P A , DENNETT C , GREEN L M , BERNSTEIN R M , HOLT P J L , PUMPHREY R S H , BOYLSTON A W , OLLLER W E R
Published: July 2010

T cell antigen specificity is determined by the products of the genes which encode the variable regions of their receptors. Of the T cell receptor (TCR) variable region gene products examined, only Vβ6.7a TCR-positive lymphocytes were reduced in primary Sjogren’s syndrome patients with IgG 1 hypergammaglobulinaemia compared with an age-, sex- and HLA-matched control population. The levels of Vβ6.7a T cells were also significantly reduced when these patients were compared with an age- and sex-matched but HLA-unmatched control group and non-tissue typed normal people of both sexes. Since published studies show no such abnormality in rheumatoid arthritis, systemic lupus erythematosus or other autoimmune diseases, this abnormality may reflect a pathogenic process specific to primary Sjogren’s syndrome.
Abnormalities within CD4 and CD8 T lymphocyte subsets in type 1 (insulin-dependent) diabetes
Authors: ILONEN J , SURCEL H M , KÄÄR M-L
Published: July 2010

Abnormalities in the proportions of various T lymphocyte subpopulations have been found in a number of autoimmune diseases. Monoclonal antibodies labelled with various fluorochromes were used here to define the percentages of subsets, and especially to divide CD4+ (helper/inducer) and CD8+ (suppressor/cytotoxic) cells into phenotypic subgroups. Blood samples were analysed from 25 patients (age 10.1±3.7 years) with recently diagnosed insulin-dependent diabetes mellitus (IDDM) and 25 age- and sex-matched control subjects. The percentages of CD4+ cells and CD4+CD45RA+ cells described as naive T helper cells or suppressor/inducers were increased in the IDDM patients (P<0.05 and P<0.05, Student’s t-test, respectively), whereas the percentage of CD4+CD45RA-cells (memory T-helper cells, helper/inducers) was similar in the patients and controls. The percentage of CD8+CD11b+ cells containing suppressor/effector lymphocytes was decreased in the IDDM patients as compared with the controls (P<0.01) but no significant difference was seen in total CD8+ cells. The percentages of CD3+ cells and the proportions of these simultaneously positive for HLA-DR antigen (activated T cells) were also increased in the recent IDDM patients (P<0.001 and P<0.05, respectively), while the proportion of CD20+ B cells was decreased (P<0.05). The findings support the view that disturbed immune regulation occurs in IDDM and indicate that further division of T cell subpopulations may clarify our understanding of the disease process.
Soluble IL-2 receptor and CD25 cells in psoriasis: effects of cyclosporin A and PUVA therapy
Authors: DUNCAN J I , HORROCKS C , ORMEROD A D , POWLES A V , WHITING P H , FRY L , THOMSON A W
Published: July 2010

A study was conducted to quantify soluble IL-2 receptor (sIL-2R) levels in sera of 57 chronic plaque psoriasis patients and correlate these measurements with disease activity and the numbers of IL-2R-positive (CD25+) lymphocytes in lesional biopsies of 11 cyclosporin A (CsA) and 13 psoralen plus ultraviolet radiation (PUVA) treated patients. Levels of sIL-2R showed a strong correlation with the psoriasis area and severity index (PASI). CsA and PUVA significantly reduced the PASI and sIL-2R levels to a similar degree after 4 weeks of treatment. Although the majority of CsA-treated patients who were biopsied showed reductions in lesional CD25+ cells, these did not reach statistical significance; in five patients biopsied who had PUVA treatment, no consistent effect on the numbers of CD25+ cells was observed. A significant correlation was found between CD25+ cells in lesional biopsies and the PASI score.
A study of immune responses to myelin and cardiolipin in patients with systemic lupus erythematosus (SLE)
Authors: KHALILI A , COOPER R C
Published: July 2010

Sera from 39 patients with SLE. 20 patients with cerebrovascular disease with no evidence of SLE, and 20 normal controls were tested for antibodies to cardiolipin (CL), brain total upper (UPG) and lower phase (LPG)glycolipids, myelin basic protein (MBP), myelin, and single strand DN A (ssDNA) by ELISA. Binding to the glycolipids and MBP was negative or negligible in all the groups, but significant binding was observed against CL, myelin and ssDNA in some of the SLE patients. Many sera from SLE patients with cerebral disorders and high CL binding also demonstrated high binding to myelin. These sera also labelled cell surface antigens on neonatal mouse neurons and astrocytes by immunofluorescence in tissue culture. A correlation was found to exist between anti-CL and antimyelin antibodies in SLE patients with cerebral lesions, but not between anti-ssDNA and anti-CL antibodies. As much as 80–90% of the specific activity of these antibodies could be absorbed out by the relevant antigens but only partially by the other antigens. In the control groups binding was low and no specific absorption could be demonstrated.
Immune adherence and clearance of hepatitis B surface Ag/Ab complexes is abnormal in patients with systemic lupus erythematosus (SLE)
Authors: MADI N , STEIGER G , ESTREICHER J , SCHIFFERLI J A
Published: July 2010

Complement levels and complement receptor 1 (CR1) on erythrocytes (E) are reduced in systemic lupus erythematosus (SLE). To see whether these abnormalities are responsible for defective transport and elimination of immune complexes (IC) from the circulation, patients with active SLE (14) and normal volunteers (14) were injected with preformed IC (hepatitis B surface Ag/Ab). Two minutes after injection only 25±9 ± 19±1% (mean ± 1 s.d.) of the circulating IC were bound to Ein the SLE patients as compared to 63 ± 3±7% in the normal subjects (P= 0±0001). For SLE patients, the reduced immune adherence was best explained by a combination of complement depletion and low CRI binding capacity (±= 0±80, P = 0±0001).

The disappearance of IC as estimated from the area under the elimination curve was faster in SLE lhan in controls (P= 0±02), and correlated with CRI (±= 0±54, P= 0±0001) and immune adherence observed in vivo (±= 0±33, P= 0±013). Finally, immune adherence was absent and IC disappeared very rapidly in a patient with C2 deficiency and an SLE-like disease. These observations suggest that in SLE the defective immune adherence reaction might be responsible for the accelerated disappearance of IC from the circulation.
The effect of anti-neutrophil cytoplasm autoantibodies on the signal transduction in human neutrophils
Authors: LAI K N , LOCKWOOD C M
Published: July 2010

The effect of anti-neutrophil cytoplasm autoantibodies (ANCA) on neutrophil activation was studied by pre-incubating neutrophils with IgG and F(ab')2 prepared from ANCA patients with Wegener’s granulomatosis or microscopic polyarteritis. We measured the generation of inositol triphosphate (IP3) (a product of hydrolysis of membrane phospholipid which acts as a second intracellular messenger), and the translocation of protein kinase C (PKC) upon stimulation by a chemotactic peptide, fMet-Leu-Phe (fMLP). ANCA+ F(ab')2 did not induce a significant increase in IP3 generation. Nonetheless, ANCA+ F(ab')2 and ANCA+ IgG pretreatment of human neutrophils reduced the production of inositol phosphates upon subsequent fMLP stimulation compared with experiments performed when cells were pretreated with F(ab')2 and IgG prepared from ANCA healthy subjects. A significantly reduced generation of IP3 and inositol biphosphate (IP2) was observed. ANCA+ F(ab')2 pretreatment of neutrophils inhibited fMLP-stimulated IP3 generation in a dose-dependent manner. The membrane-bound PKC activity upon stimulation by FMLP and PMA was reduced in neutrophils pretreated with ANCA+ F(ab')2 and IgG. These results indicate that ANCA affect in vitro signal transduction (IP3 generation, and translocation of PKC) in human neutrophils. Apparently, further activation of signal transduction by chemotactic peptide is significantly blunted in cells pre-incubated with ANCA+ F(ab')2 but not with F(ab')2 from healthy controls.
CD5+ B cells and naturally occurring autoantibodies in cancer patients
Authors: STEIN R , WITZ I P , OVADLA J , GOLDENBERG D M , YRON I
Published: July 2010

We have determined the percentage of CD5+ B lymphocytes in the peripheral blood of cancer patients and healthy controls, using antibodies directed at the CDS and CD 19 (pan-B) markers. The frequencies of CD5+ B cells, expressed as a percentage of total B cells, ranged from 14.3 to 57.5% in the controls and from 14.8 to 82.8% in the patient population. One-third of the cancer patients had frequencies > 2 s.d. above the mean of the control population. The CD5+ B cell fraction expressed as a percentage of total lymphocytes was also significantly elevated in this group of cancer patients. These results suggest that the CD5+ B cell compartment may be affected by the malignant process or by the therapy modality employed. The plasma levels of several naturally occurring autoantibodies, the products of the CD5+ B cells, were also assessed in cancer patients and controls. No significant differences were observed when reactivity to several autoantigens was measured. These included nuclear components and phospholipids.
Identification of the major fibroblast growth factors released spontaneously in inflammatory arthritis as platelet derived growth factor and tumour necrosis factor-alpha
Authors: THORNTON S C , POR S B , PENNY R , RICHTER M , SHELLEY L , BREIT S N
Published: July 2010

Rheumatoid arthritis is characterized by chronic inflammation and proliferation of a number of important elements within the joint including the synovial fibroblasts. Elevated levels of a number of cytokines such as II-1, IL-2, IL-6, interferon-gamma (IFN-γ), transforming growth factor-β and tumour necrosis factor-alpha (TNF-α) have been detected in the synovial fluid of patients with rheumatoid arthritis and other inflammatory arthritides. It seems likely that local release of such mediators may be responsible for the proliferation and overgrowth of connective tissue elements in these disorders. In order to ascertain whether there was evidence to suggest local production or release of fibroblast growth factors in the joint in inflammatory arthritis, and to determine their identity, cells were obtained from the synovial fluid of 15 patients with chronic inflammatory arthritides, All subjects’ synovial fluid cells spontaneously released growth factor activity for fibroblasts. This was present in large amounts, being detectable in culture supernatants diluted to a titre of at least 1/625. By a series of depletion experiments using solid-phase bound antibodies to cytokines, it was possible to demonstrate that this activity was due to TNF-α and platelet-derived growth factor (PDGF). Thus, this study showed for the first time that functionally active PDGF was released from synovial fluid cells. Both PDGF and TNF-α appeared to contribute in approximately equal amounts to this fibroblast growth factor activity, and were synergistic in effect. Thus this study provides evidence for the local production and release of these two cytokines and suggests that together they are the dominant factors in fibroblast proliferation within the synovial cavity.
Effects of interferon-gamma and tumour necrosis factor-alpha on the development of cytotoxic T lymphocytes in autologous mixed lymphocyte tumour cultures with human melanoma
Authors: ROTH A D , HORNICEK F J , GERSTNER C G , KIRKWOOD J M
Published: July 2010

We have studied the influence of tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) on the development of cytotoxic T lymphocytes (CTL) against melanoma in mixed lymphocyte tumour cultures (MLTC). In these MLTC, TNF-α at 104 U/ml increased the expansion of the CTL up to 104-fold over recornbinant IL-2(rIL-2) alone. IFN-γ at 104 U/ml and combinations of TNF-α plus IFN-γ at 102-103 U/ml promoted the proliferation more variably. MLTC generated with rIL-2 showed a predominance of CD8+ cells, while 2 weeks of culture in the presence of IFN-γ at 104 U/ml, or with IFN-γ and TNFα at 1 ± 102-103 U/ml, favoured the emergence of CD4+ cell populations. The cytotoxic activity of the lymphocytes generated in these MLTC showed a consistent decline of K562 cytotoxic activity following exposure to the combination of IFN-γ and TNF-α. Despite the altered T cell subset distribution with different combinations of cytokines. no consistent alteration in the specific anti-tumour cytotoxicity against melanoma was detected. These results suggest that TNF-α and IFN-γ influence the activation, phenolypic. and functional outcome of MLTC-generated CTL, and may account for the phenotypic variations observed in T cell populations generated in vitro.
Eosinophils adhere to and stimulate replication of lung fibroblasts ‘in vitro’
Authors: SHOCK A , RABE K F , DENT G , CHAMBERS R C , GRAY A J , CHUNG K F , BARNES P J , LAURENT G J
Published: July 2010

Eosinophils have been implicated in several disorders associated with the development of fibrosis. This led us to investigate the interactions between eosinophils and fibroblasts in vitro. Adhesion between purified guinea pig peritoneal eosinophils and monolayers of human fetal lung fibroblasts was assessed using the rose bengal dye staining assay. Fibroblast replication was assessed using a colorimetric assay based upon the uptake and subsequent release of methylcnc blue. Addition of phorbol myristate acetate induced a rapid, time-dependent increase in cosinophil adhesion (127% and 328% over basal adhesion after 10 and 30 min, respectively). Phorbol myristate acetate-induced adhesion was inhibited by the peptides RGDS and GRGDS (48% and 42%, respectively using 1 mM peptide) and by nordihydroguaiaretic acid, an inhibitor of the lipoxygenase pathway of arachidonic acid metabolism (46% inhibition at 15 μm), In addition. 24 h culture of fibroblast monolayers with interleukin 1α (IL-lα) or tumour necrosis factor α (TNFα) resulted in enhanced adhesion (10 U/ml IL-lα stimulated adhesion by 55% of control, 500 U/ml TNFα by 75% of control). Conditioned media from cultured eosinophils stimulated fibroblast replication in a time-dependent fashion with maximal stimulation at 3 h. In contrast, media from guinea pig peritoneal macrophages in culture did not show such an effect. This study indicates that eosinophils are capable of both adhering to and releasing mitogens for fibroblasts in vitro. These observations suggest that eosinophils have the capacity to play a role in the development of fibrosis in disorders where they have been shown to be present.
Cytokines in sera from insulin-dependent diabetic patients at diagnosis
Authors: CAVALLO M G , POZZILLI J P , BIRD C , WADHWA M , MEAGER A , VISALLI N , GEARING A J H , ANDREANI D , THORPE R
Published: July 2010

Cytokines are known to play an important role in autoimmunity and have been suggested to be involved in the pathogenesis of insulin-dependent diabetes (IDDM). In the present study we have measured IL-1, IL-2, IL-4, IL-6, interferon-gamma(IFN-γ)and tumour necrosis factor (TNF) (using both immunoassays and bioassays) in sera from 50 patients affected by IDDM at the time of clinical diagnosis and 51 age and sex matched controls. Detectable levels of IL-I, IL-2, IL-6 and IFN-μgK were found in the serum of a small percentage of subjects and were not significantly different between patients and controls. IL-4 was detectable in a higher number of both patients and controls and circulating TNF-α (> I U/ml) was found in a percentage of patients (24%) significantly higher than controls (P<0.01). Raised levels of TNF-α were detectable using an immunoenzymatic assay whereas TNF bioactivity in these samples was negligible. We conclude that the presence of immunoreactive TNF-α in the patient’s sera may reflect an increased localized production of this cytokine at pancreatic level. However, the measurement in serum of other cytokines does not add information on the role that they may play in the pathogenesis of IDDM.
Sputum eosinophils from asthmatics express ICAM-1 and HLA-DR
Authors: HANSEL T T , BRAUNSTEIN J B , WALKER C , BLASER K , BRUIJNZEEL P L B , VIRCHOW J-C , VIRCHOW C
Published: July 2010

Sputum from symptomatic asthmatics is a rich source of eosinophils from the respiratory tract. Following liquefaction of sputum with dithioerythritol (DTE), a cell suspension suitable for indirect double immunofluorescence with flow cytometry was obtained. Eosinophils were identified using anti-CD9 fluorescein conjugate, and particular surface markers measured with the relevant mouse MoAb followed by goat anti-mouse immunoglobulin phycoerythrin conjugate. Blood and sputum eosinophil surface markers were determined in parallel from asthmatics not receiving steroid therapy. Sputum eosinophils were found to have considerably elevated levels of CDl1b, a reflection of eosinophil activation. Sputum but not blood eosinophils were found to express ICAM-I (nine out of 11 cases) and HLA-DR (eight out of 11 cases). Furthermore, following culture of normal blood eosinophils with pooled T cell supernatants, ICAM-l and HLA-DR could be induced in vitro. The induction of eosinophil adhesion molecules such as ICAM-l and HLA-DR may influence eosinophil localization and function in asthma.
Neutrophil chemotactic factor produced by Japanese encephalitis virus stimulated macrophages
Authors: KHANNA N , AGNIHOTRI M , MATHUR A , CHATURVEDI U C
Published: July 2010

The mechanism of neutrophil leucocytosis in cases of Japanese encephalitis is not known. We here report that during Japanese encephalitis virus (JEV) infection in mice the splenic macrophages secrete a chemotactic factor that attracts the neutrophils. The peak activity of macrophage derived factor (MDF) was observed on day 7 following infection. The MDF acted in a dose-dependent manner. This chemoattractant was purified by low pressure liquid chromatography and gave a single band of 10 kD on silver stained polyacrylamide gel. The MDF was found to be heat resistant and sensitive to prolonged incubation with proteases.
Inhibitory effect of 11β-hydroxypregna-l,4-diene-3,20-dione (ΔHOP) on lymphocyte proliferation
Authors: RONDINONE C M , SCHILLACI R , CASTILLO M B , ROLDÁN A
Published: July 2010

We have compared the immunosuppressive effect of ΔHOP and glucocorticoids on lymphocyte proliferation and IL-1 secretion. The new synthetic steroid only inhibited proliferation of phytohaemagglutinin (PHA)-stimulated human lymphocyte at high concentrations and the effect did not persist after washing out the steroid- In contrast, glucocorticoids produced the classical dose-response inhibition and the effect persisted when they were removed from the cultured medium. Although both steroids decreased IL-1 secretion from human monocytes stimulated with lipopolysaccharide (LPS), they exert the same effect through a different mechanism. The experiments we report suggest that the decrease of IL-1 synthesis produced by ΔHOP could be caused by inhibition of LPS phagocytosis. These results support our hypothesis that ΔHOP exerts its immunosuppressive and anti-inflammatory effect by a non-genomic mechanism.
Piroxicam, indomethacin and aspirin action on a murine fibrosarcoma. Effects on tumour-associated and peritoneal macrophages
Authors: VALDÉZ J C , PERDIGÓN G
Published: July 2010

Growth of a methylcholanthrene-induced fibrosarcoma in BALB/c mice was accompanied by an increase in the activation state of tumour-associated macrophages (TAM). as measured by their FclgG receptor expression, phagocytic index and β-glucuronidase levels. All of these parameters were markedly higher in TAM than in peritoneal macrophages (PM) derived from the same animal. On the other hand, PM from tumour-bearing mice showed lower activation parameters than PM from normal animals. We also studied the effect on tumour development of three inhibitors of prostaglandin synthesis: indomethacin. piroxicam and aspirin. Intraperitoneal administration of these drugs during 8 d was followed by the regression of palpable tumours. Indomethacin (90 mg/d) induced 45% regression, while with piroxicam (two 400 mg/d doses and six 200 mg/d doses) and aspirin (1 mg/d) 32% and 30% regressions, respectively, were observed. The growth rate of non-regressing tumours, which had reached different volumes by the end of the treatment, was delayed to a similar extent by the three anti-inflammatory non-steroidal drugs (NSAID). With respect to TAM, the treatment did not induce any significant change in their activation state, though both piroxicam and indomethacin increased slightly the TAM number. In contrast, NSAID administration was followed by a remarkable increase in the activation parameters of PM when compared with PM from tumour-bearing mice receiving no treatment. Indeed, these parameters were in some cases higher than those of PM from normal mice. The leukocytosis (60 000/μl) with neutrophilia (80%) induced by tumour growth on peripheral blood leukocytes (PBL) was reversed by the treatment to values close to normal, in parallel with the reduction of tumour size. A drop in haematocrit was also noted which was most probably a consequence of tumour growth rather than of the treatment. This study reveals that the three NSAID tested have a remarkable antitumour activity, which correlates with the restoration of PM activity and PBL values.
Immune complex binding efficiency of erythrocyte complement receptor 1 (CR1)
Authors: MADI N , PACCAUD J-P , STEIGER G , SCHIFFERLI J A
Published: July 2010

C3b-coated immune complexes adhere to the complement receptor I (CR1. CD35) on human erythrocytes. This multi-valenl binding might be favoured by the known clustering of CR1 and by the multiple C3b-binding sites on each CR1. The size of the CR1 clusters correlates directly with the number of CR1 /erythrocytes, and the different structural CR1 alleles bear between two and five C3b-binding sites. Using radiolabelled hepatitis B surface antigen-antibody complexes, we investigated whether CR1 numbers and structural alleles modulate the ability of erythrocytes to bind immune complexes, and assessed if any reorganization of immune complexes takes place at the erythrocyte surface after the initial binding reaction. The binding efficiency (immune complexes/CR1) correlated with CR1 number as determined by the maximal binding at 4°C, the kinetics of binding at 37°C, and the binding in the presence of excess immune complexes and of immune complexes of small size. Binding efficiencies were similar for erythrocytes with low CR1 from normal subjects and patients with AIDS or SLE. A monoclonal antibody blocking the C3b-binding sites (3D9) of CR1 interfered with binding efficiency at a lower concentration on cells bearing low CR1 numbers, suggesting that CR1 clustering is essential. The larger alleles of CR1 (DD and BB) were more efficient than AA alleles. The distribution of immune complexes, visualized by immunofluorescence, was heterogeneous on erythrocytes: about two out of three cells bore between one and 12 immune complexes. No visible immune complex reorganization took place after initial binding, as prefixed erythrocytes displayed the same immune complex distribution and number/erythrocytes as unfixed erythrocytes. The contribution of CR1 alleles in immune complex binding efficiency was confirmed by morphological analysis. These results demonstrate that immune adherence efficiency is the resultant of the CR1 clustering, as well as the particular alleles carried by erythrocytes. Moreover, there is little or no immune complexes surface reorganization after the initial binding reaction.
Cellular origins of serum complement receptor type 2 in normal individuals and in hypogammaglobulinaemia
Authors: LING N , HANSEL T , RICHARDSON P , BROWN B
Published: July 2010

A soluble form of complement receptor 2 (sCR2) is found in normal human serum. Amounts present arc about 30 90 ng/ml, which is of the same order as reported tor soluble CR1. Although B cells express surface CR2 and are the main peripheral blood source of sCR2 they do not appear to be the major tissue source of serum sCR2. Scrum levels of sCR2 of patients with hypogammaglobulinaemia were not significantly different from those of normal individuals even in the case of two brothers with Bruton's X-linked agammaglobulinaemia (XLA) lacking (CD 19+) B cells. On gel filtration through Sephacryl S-300 the sCR2 from XLA serum behaved exactly like sCR2 from normal serum or sCR2 affinity purified from cell supernates of a B lymphoblastoid line or from the T-ALL line MOLT-4. In all cases a single peak appeared at the same point in the chromatogram. Possible alternative sources of serum sCR2 are follicular dendritic cells (FDC) which are known to express CR2 strongly and T6+ lymphocytes within the thymus. Peripheral T cells from adults have not been reported to express CR2. However, investigation showed that cells from the Bruton's XLA cases produced small amounts of sCR2 in culture and although no CD21 was detected on the surface of the mononuclear cells by flow cytometry, the more sensitive direct antibody rosette test readily detected CD21. Further studies showed that non-B cells from control samples of cord blood or blood of young children also weakly expressed CD21.
Severe combined immunodeficient (SCID) mice: a model for investigating human thyroid autoantibody synthesis
Authors: MACHT L , FUKUMA N , LEADER K , SARSERO D , PEGG C A S , PHILLIPS D I W , YATES P , MCLACHLAN S M , ELSON C , SMITH B REES
Published: July 2010

We have studied the ability of lymphocytes from the blood, thyroid and lymph nodes of patients with autoimmune thyroid disease (AITD) to produce auloantibodies to thyroglobulin (Tg) and/or thyroid peroxidasc (TPO) in SCID mice. Human IgG class Tg and/or TPO antibodies were detectable in plasma from SCID mice 7 days after transfer of 15–25 × I06 cells mouse and the highest levels were recorded 2–3 weeks later. In contrast, Tg and/or TPO antibodies were undctcctablc in recipients of lymphocytes from thyroid antibody negative controls. AITD thyroid lymphocytes produced the most antibody in recipient mice and lower levels were observed in recipients of AITD blood and lymph node lymphocytes. The amounts of Tg and or TPO antibody detected were in accordance with the ability of thyroid and lymph node lymphocytes to secrete these autoantibodies spontaneously in culture (indicating the presence of cells activated in the patient) and with the capacity of blood lymphocytes (probably B memory cells) to secrete Tg and or TPO antibodies in culture in response to pokeweed mitogen. Tg antibodies in plasma from SCTD recipients of thyroid lymphocytes were of subclasses IgG1, IgG2 and IgG4 and the proportions closely resembled those of the donor's scrum Tg antibodies. Blood lymphocytes transferred to SCiD recipients were also able to produce Tg antibodies of subclasses1, 2 and 4 but the subclass distribution varied between mice and the reason for this is not clear at present. Since SCID mice provide an environment in which B lymphocytes from patients with AITD can be activated without mitogen to secrete thyroid antibodies, this model will provide a powerful system for elucidating the mechanisms regulating the secretion of human antibodies to Tg and TPO.
Peripheral blood and synovial fluid T cells differ in their response to alloantigens and recall antigens presented by dendritic cells
Authors: STAGG A J , HARDING B , HUGHES R A , KEAT A , KNIGHT S C
Published: July 2010

Properties of T cells from inflammatory lesions were analysed by comparing the response of peripheral blood (PB) and synovial fluid (SF)T cells from 19 patients with a range of arthropathies to enriched allogeneic dendritic cells (DC) in a primary mixed leucocyte reaction (MLR). In 17 patients the proliferative response of SF T cells was significantly (P < 0.05) less than that of PB lymphocytes. The reduced response of SF T cells was observed in all disease categories studied and could not be attributed to differences in cell number requirements or response kinetics. Addition of recombinant interlcukin-2 enhanced the response of SF T cells in a dose-dependent manner. Cell mixing experiments suggested that active suppression was not the underlying mechanism of the poor MLR response of SF T cells. In contrast to the MLR response, SF T cells were able to mount vigorous proliferative responses to recall antigen presented by autologous antigen-presenting cells. The possibility is discussed that T cells compartmentalized at inflammatory lesions are a unique population with a diminished ability to interact with DC and respond to primary stimuli but an ability to respond to secondary antigenic challenge.