bluuebunny/crossref_metadata_2025 huggingface dataset:
Spontaneousin vitroproduction of rheumatoid factor during infectious exacerbations of cystic fibrosis: correlation with circulating immune complex levels
Authors: KEOGAN M T , CALLAGHAN M , YANNI G , MULHERIN D , FEIGHERY C , BROWN D L , FITZGERALD M X , BRESNIHAN B
Published: July 2010

Rheumatoid factor (RF) production has been demonstrated during infections, including infectious exacerbations of cystic fibrosis (CF). The aim of this study was to evaluate the relationship of RF production to infection, and examine the mechanisms involved. Serial peripheral blood mononuclear cell (PBMC) cultures with measurement of spontaneous production of IgM RF, IgA RF, total IgM and IgA, and measurement of serum levels of immune complexes were carried out during exacerbations of CF. The percentage of B cells expressing CDS was examined in a second cohort of acutely infected CF patients, and related to IgM RF production. IgM RF production was significantly elevated during acute infection compared with convalescence (P<0·05), stable CF subjects (P<0·005) and normal controls (P<0·05). IgM RF production did not correlate with total IgM production in the majority of patients, but was closely related to circulating immune complex levels in 8/10 subjects. IgA RF production did not increase significantly during infection, and did not correlate with total IgA or IgM RF production, or with circulating immune complex levels. CDS+ B cells were not increased in the CF group, and the percentage of CD5+ B cells did not correlate with IgM RF synthesis. These observations suggest that RF production during infection is specifically induced, possibly by immune complex autoimmunization, and is not simply the result of polyclonal B cell activation. Different patterns of IgM RF and IgA RF synthesis suggest different mechanisms of induction.
Determination of the cytokine profile in American cutaneous leishmaniasis using the polymerase chain reaction
Authors: CACERES-DITTMAR G , TAPIA F J , SÉNCHEZ M A , YAMAMURA M , UYEMURA K , MODLIN R L , BLOOM B R , CONVIT J
Published: July 2010

The lymphokine profiles were determined in the skin lesions of the three distinct clinical forms of American cutaneous leishmaniasis (ACL), using a reverse transcriptase polymerase chain reaction (RT-PCR) and primers for various lymphokines. The message for interferon-gamma (IFN-y). tumour necrosis factor-beta (TNF–β) and IL-8 was expressed in the three clinical forms of ACL. IL-lβ in RNA was expressed in most localized (LCL) and mucoeutaneous (MCL) leishmaniasis. but in only few of the diffuse cutaneous leishmaniasis (DCL). IL-2 mRNA was detected in about half of the lesions, with more prominent values for MCL. IL-4 mRNA was present in most lesions from the three clinical forms, but markedly increased in DCL. IL-5 and IL-10 mRNAs were expressed in all MCL and in half of the DCL lesions and weakly expressed in LCL lesions. IL-10 mRNA was more abundant in MCL lesions. In contrast. IL-6 and TNF-s mRNAs were expressed in a Iarge number of LCL. In MCL, IL-6 mRNA was expressed in most cases and TNF–α mRNA in all the cases. In DCL. 11,–6 mRNA was absent and TNF–α mRNA was weakly expressed. These results suggest that most T cells present in the MCL and DCL lesions secrete a mixture of type 1 and type 2 cytokine patterns, but in DCL granulomas type 2 cytokines predominate. In LCL the cytokine patterns show a mixture of type l and type o with a preponderance of IFN–γ over IL-4, and low levels of IL-5 and IL-10. The Iack of IL-6 and TNF–α mRNAs, and the low expression of IL-lβ in DCL lesions suggest a defect in the anligcn-processing cells that may account for the state of unrcsponsiveness in these patients.
Interferon-gamma stimulates the secretion of IL-1, but not of IL-6, by glomerular mesangial cells
Authors: KAKIZAKI Y , KRAFT N , ATKINS R C
Published: July 2010

IL-1 activity in culture supernatant and cell lysate from rat mesangial cells stimulated with interferon-gamma (IFN-γ) was measured by a thymocytc proliferation assay. While IFN-γ alone had no effect on the secretion or the intracellular pool of IL-1, the enhancement by IFN-γ of IL-1 secretion in response to lipopolysaccharide (LPS) was observed. The stimulatory effect of culture supernatant on thymocyte proliferation was abrogated by preincubation with the anti-IL-l antibody. At least 4–h incubation with IFN-γ and LPS was required to detect enhancing effect of I FN-γ. The addition of as little as 1 U/ml IFN-γ significantly increased IL-1 secretion in the presence of 10 μg/ml LPS. The IL-6 activity in culture supernatants was determined by measurement of thymidine uptake in mouse IL-6-dependent cell line (MH60.BSF2). Mesangial cells secreted IL-6 in culture supernatant without additional stimuli and LPS distinctly increased it as described previously. However, in contrast to IL-1 production, no effect of IFN-γ on IL-6 secretion was observed in the presence or absence of LPS. Moreover, we determined whether enhanced IL-1 release is associated with la expression on mesangial cells. IFN-γ alone and the combination with LPS induced marked expression of la antigen, whereas LPS alone did not. We conclude that IFN-γ stimulates the production of IL-1, but not IL-6, by mesangial cells and suggest an important role of IFN-γ in the pathogenesis of glomerulonephritis by regulating the mesangial production of IL-1 and the accessory cell function of mesangial cells.
Tumour necrosis factor soluble receptors behave as acute phase reactants following surgery in patients with rheumatoid arthritis, chronic osteomyelitis and osteoarthritis
Authors: CHIKANZA I C , ROUX-LOMBARD P , DAYER J-M , PANAYI G S
Published: July 2010

Tumour necrosis factor-alpha (TNF-α) is involved in diverse biological processes including immune and inflammatory reactions and the response to surgical stress. Two soluble TNF receptor protein fragments. TNF-sR55 (from the p55 kD TNF receptor) and TNF-sR75 (from the p75 kD TNF receptor), are released by cells during inflammation and may modulate the effects of TNF-α. We have studied the kinetics of secretion of TNF-α, TNF-sR55 and TNF-sR75 in the sera of patients with rheumatoid arthritis (RA) and control subjects with osteoarthritis (OA) or chronic osteomyelitis (OM) before and after major surgery. Significantly higher pre-operative levels of TNF-sR55 and TNF-sR75 were found in RA and OM as compared with OA (P<0·02). Following surgery, TNF-sR55 increased within 24 h in RA, OM and OA (P<0·05), whereas TNF-sR75 increased significantly only in OM and OA patients (P<0·05). By contrast, no TNF-α was detectable before and after surgery in any of the subjects, but this may have been due to impaired detection (by ELISA) of TNF-α when it is bound to TNF-sR. These findings suggest that TNF-sR55 and TNF-sR75 may be further markers of the host's reaction to inflammatory insults. They may also play a role in modulating the immune and inflammatory reactions by inhibiting the systemic effects of TNF-α.
Cytokines involved in the progression of multiple myeloma
Authors: MERICO F , BERGUI L , GREGORETTI M G , GHIA P , AIMO G , LINDLEY I J D , CALIGARIS-CAPPIO F
Published: July 2010

We have investigated which of the cytokines that are relevant in the in vitro growth of multiple myeloma (MM) malignant plasma cells are actually produced in vivo by MM patients. To this end, we have measured the levels of IL-1β, IL-3, IL-4, IL-6, IL-7, IL-8 and tumour necrosis factor-alpha (TNF-α) both in sera and in the supernatant of bone marrow (BM) stromal cell cultures from patients with MM and monoclonal gammopathy of undetermined significance (MGUS). The significance of our findings is three-fold. First, IL-6 and IL-8 are produced by MM BM stromal cells, while IL-1β, TNF-α, IL-4 and IL-7 are not. Second, IL-3 is the only cytokine consistently raised in serum samples; we have also detected low levels of serum IL-6 in a minority of cases, usually in advanced stage of the disease. Third, MM BM stromal cells are active IL-6 and IL-8 producers, while both normal and MGUS BM stromal cells are low producers, thus suggesting that in the BM of MM a number of environmental cells, that would normally be quiescent, are instead activated and that, in MM, activated BM stromal cells play an active role in supporting the progressive expansion of the B cell clone.
A novel specificity of anticytoplasmic autoantibodies directed against eosinophil peroxidase
Authors: DOLMAN K M , DAMSMA I , TOOL A T J , SONNENBERG A , von dem BORNE A E G Kr , GOLDSCHMEDING R
Published: July 2010

Vasculitis associated anticytoplasmic autoantibodies (ANCA) are directed against enzymes in the granules of both neutrophils and monocytes. These autoantibodies can be detected by indirect immunofluorescence technique (IIFT) using ethanol-fixed cytospins. We here report the identification of a novel specificity of autoantibodies, present in the sera of eight patients, that reacted only with eosinophils in the IIFT. By immunoprecipitation and ELISA experiments it was shown that the autoantibodies in these sera were directed against eosinophil peroxidase (EPO). There was no apparent influence on initial substrate conversion rate, but reduced plateau levels suggested increased inactivation of the enzyme in the course of the peroxidase reaction. Flow cytometry studies demonstrated the presence of EPO on the surface of primed eosinophils. Anti-EPO sera and purified anti-EPO immunoglobulins significantly increased the release of reactive oxygen species from primed eosinophils. The patients with anti-EPO antibodies suffered from clinically diverse disorders, with more or less generalized manifestations involving the kidneys, blood vessels, lungs and/or joints.
Autoantibodies in malaria, tuberculosis and hepatitis B in a West African population
Authors: ADEBAJO A O , CHARLES P , MAINI R N , HAZLEMAN B L
Published: July 2010

Following reports of associations between autoantibodies and living in the tropics, we have studied the seroprevalence and nature of anti-nuclear antibodies, anti-cardiolipin antibodies, antibodies to extractable nuclear antigens and anti-neutrophilic cytoplasmic antibodies in 351 West Africans with malaria, tuberculosis or hepatitis B, or in good health. Amongst healthy West Africans we found a seroprevalence of 7% for anti-nuclear antibodies with several staining patterns, and of 30·3% for anti-cardiolipin antibodies. Among patients with tuberculosis and malaria there was twice that frequency of anti-nuclear antibodies (predominantly speckled in pattern), and anti-neutrophil cytoplasmic antibodies (predominantly IgM) were demonstrated in a few cases. A possible association beween IgG anti-cardiolipin antibodies and tuberculosis was observed (P<0·05), but antibodies to double-stranded DNA were not elevated and no antibodies to extractable nuclear antigens were found in any of the patients or healthy individuals studied. Our findings suggest the need for caution in the interpretation of autoantibody tests in subjects from or living in the tropics, as well as in patients with tropical infections.
IL-4 and interferon-gamma production in children with atopic disease
Authors: TANG M , KEMP A , VARIGOS G
Published: July 2010

In vitro studies have implicated reciprocal roles for IL-4 and interferon-gamma (IFN-γ) in the regulation of IgE production. As elevated IgE is a major feature of atopic disease, an important question is whether an imbalance of IL-4 and IFN-γ is present in vivo. The production of IL-4 and IFN-γ in phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cell cultures from atopic children was examined to determine if there is an increased production of IL-4 and/or a reduced production of IFN-γ. Highly atopic children with IgE >600 U/ml produced significantly more IL-4 and less IFN-γin vitro than age-matched non-atopic controls. Production of IL-4 and IFN-γ in mildly atopic children was equivalent to controls. These findings indicate that highly atopic children have an imbalance of IL-4 and IFN-γ production and that the degree of imbalance relates to severity of the atopic state. The ratio of in vitro IL-4: IFN-γ production correlated positively with serum IgE, which suggests that the balance of these two cytokines is a factor in the regulation of IgE, in vivo. It remains to be determined whether this imbalance of IL-4 and IFN-γ in the highly atopic children is the cause or result of the disease process.
Effect of anti-IL-4, interferon-gamma and an antifungal triazole (SCH 42427) in paracoccidioidomycosis: correlation of IgE levels with outcome
Authors: HOSTETLER J S , BRUMMER E , COFFMAN R L , STEVENS D A
Published: July 2010

Paracoccidioidomycosis is characterized by depressed cellular but enhanced humoral immune responses, which suggests a Th2 type of response to infection. We investigated possible therapeutic roles for anti-IL-4, interferon-gamma (IFN-γ) and/or SCH 42427 (SCH), a new triazole antifungal agent, and their effect on serum IgE levels in a murine model of chronic Paracoccidioides brasiliensis infection. BALB/c mice infected by the pulmonary route were studied with three programmes. The subacute model and one acute model experiment investigated cytokine secretion by lymph node cells (LNC), and in a second acute experiment mice were given anti-IL-4, IFN-γ or nothing 24 h post infection, then killed at 4 weeks. In the chronic model, mice began treatment at 4 weeks post infection, receiving either SCH, IFN-γ alone, SCH + IFN-γ. or no treatment for 8 weeks. At 2-week intervals lung and spleen burdens of infection and serum polyclonal IgE levels were determined. In the subacute model (non-progressive infection), initially there was dual production of IL-4 and IFN-γ by antigen-stimulated LNC. In the acute progressive infection model IL-4, but not IFN-γ. was secreted. Anti-IL-4 treatment of the acute phase resulted in enhanced host resistance to infection, which correlated with decreased scrum IgE. The chronic model, in which the in vivo efficacy of SCH against P. brasiliensis was shown, suggests possible synergy between immunomodulation and antimicrobial chemotherapy (IFN-γ and SCH). Decreased organ burdens of infection in the chronic model after treatment with SCH, SCH plus IFN-γ, or anti-IL-4 correlated with decreased serum IgE. These promising novel approaches lo treatment of systemic fungal infections suggest a Th2 type of response to P. brasiliensis infection, which can be reversed with successful therapy.
In vitrocytotoxicity as a marker of hypersensitivity to sulphamethoxazole in patients with HIV
Authors: CARR A , TINDALL B , PENNY R , COOPER D A
Published: July 2010

Hypersensitivity to trimethoprim-sulphamethoxazole (TMP-SMX) in patients with HIV infection may be a result of either immune dysregulation, a direct cytotoxicity of the SMX-hydroxylamine metabolite (SMX-HA) (rather than SMX per se), or glutathione deficiency. We evaluated the in vitro cytotoxicity of SMX and SMX-HA to peripheral blood mononuclear cells (PBMC) of HIV-infected subjects to determine if the degree of in vitro cytotoxicity is associated with hypersensitivity, whether glutalhione inhibits cytotoxicity, and whether in vitro cytotoxicity is predictive for the development of hypersensitivity. Given that fever is often a prominent feature of hypersensitivity, we also assessed whether SMX or SMX-HA could induce the in vitro production of IL-lβ, IL-6 or tumour necrosis factor-alpha (TNF-α) by PBMC. Thecytotoxicitics of SMX and SMX-HA to PBMC were assessed in 45 HFV-infected patients with prior TMP-SMX therapy, and in eight HIV controls. Twelve HIV-infected subjects were studied prospectively before primary Pnvumocystis carinii pneumonia (PCP) therapy or reehallenge with TMP-SMX in previously hypersensitive subjects. Cylokine production was measured in four hypersensitive and two non-hypersensitive HlV-infccted subjects, and three HIV-uninfected controls. The cytotoxicity of SMX-HA to PBMC was significantly greater in the 22 HIV-infectcd patients with prior hypersensitiviiy than both the 23 HIV-infeeled paiients without hypersensitivity and the control group. Cytotoxicity was significantly reduced by glutalhione only in the hypersensitive group. SMX did not induce eylotoxicity in any group. In 12 subjects studied prospectively. SMX-HA cytotoxicity was also significantly greater in those with subsequent hypersensitivity. Exposure of PBMC to SMX-HA resulted in a modest increase in the production of IL-6, IL-1β and TNF-α, although no major difference was detected between subjects with or without hypersensitivity. These data suggest that SMX-HA and glutathione deficiency are involved in the pathogenesis of hypersensilivity to TMP-SMX in HIV-infected patients, and that in vit ro cytotoxicity could be useful in the diagnosis of hypersensilivity and predicting its likelihood.
Blockade of immunoregulatory Fc-signalling by HIV peptides: oligopeptides from HIV gpl20 and gp41 bind the Fc portion of IgG and increase thein vitroanti-ssDNA response
Authors: RAHIMPOUR R , ANDERSON C C , SINCLAIR N R StC
Published: July 2010

Concomitant ligation of antigen receptors with Fc-receptors negatively signals B cells. Antibodies to the Fc portion of IgG prevent this negative Fc-signalling, provided that these antibodies do not emit Fc signals. Prevention of Fc signals leads to augmented antibody responses to self and foreign antigens, and reduces the requirement for T cells by 10- to 100-fold in T cell-dependent antibody responses. In EL1SA assays, peptides from conserved portions of the glycoproteins. H1V-I gp120 or gp41 from HIV-1 and HIV-2 bind to the Fc portion of IgG, but do not bind the F(ab′)2 portion of IgG. HIV-derivcd peptides. which bind to the Fc portion of IgG. augment the antibody-forming cell response to single-stranded (ss)DNA. The spontaneous response to ssDNA using spleen cells from young mice, and the response in the presence of exogenous DNA using spleen cells from old mice, are augmented to the greatest extent. These results demonstrate that HIV peptides bind to the Fc portion of IgG and augment immune responses to DNA; they suggest the possibility that blockade of the Fc portion of IgG antibodies is associated with a reduction in Fc-mediated regulation of anti-self responses. Blockade of regulatory Fc-signalling may account for increased circulating immune-globulins and autoantibodies in clinical AIDS.
Dendritic cells infected in vitro with human T cell leukaemia/lymphoma virus type-1 (HTLV-1); enhanced lymphocytic proliferation and tropical spastic paraparesis
Authors: ALI A , PATTERSON S , CRUICKSHANK K , RUDGE P , DALGLEISH A G , KNIGHT S C
Published: July 2010

Evidence supporting a role of the dendritic cell (DC) in stimulating autologous T cell activity in tropical spastic paraparesis (TSP) was sought by studies of cells taken from healthy volunteers and exposed Vo HTLV-1 in vitro. DC were co-cultured with an HTLV-1 -producing cell line (MT-2) at 1:1 or 10:1 ratios. These DC stimulated high levels of proliferation in autologousT cells. This was similar to that seen in an autologous mixed leucocyte reaction (AMLR) using cells from TSP patients. The requirement for both DC and virus was confirmed. since neither DC co-cultured with uninfected MT-2 cells nor addition of infected MT-2 cells directly to T cells caused significant stimulation. DC exposed to the highest dose of HTLV-1 (1:1) for 24 h before addition of T cells caused strong stimulation that increased after 8 h but almost disappeared by 72 h. In situ hybridization showed that approximately 25% of DC became infected in cultured cells after preincubation for 24 h, and over 50% were infected with a 72-h preincubation. We suggest that infection of DC by HTL V-l may be an initial step in altering the immune system in seronegative patients, and that persistent T cell stimulation in those with genetic susceptibility may underlie the produclion of neurological disease.
Selective impairment of T lymphocyte activation through the T cell receptor/CD3 complex after cytomegalovirus infection
Authors: TIMóN M , ARNAIZ-VILLENA A , RUIZ-CONTRERAS J , RAMOS-AMADOR J T , PACHECO A , REGUEIRO J R
Published: July 2010

Cytomegalovirus(CMV) infection is reported to cause transient immunosuppression in man. In this Study we have analysed ihe effect of CMV on T lymphocyte function in 29 children diagnosed for acute CMV infection. Peripheral blood mononuclear cells (PBMC) obtained from the palients showed a significant specific impairment in their proliferative response to enterotoxins A and C1. to coneanavalin A and to the anti-CD3 MoAb OKT3. The impaired responses were corrected with exogenous IL-2. In conlrast. slimulation using phytohaemagglutinin. as well as activation signals delivered through the surface molecules CD26 or CD28. elicited normal proliferalive responses in CMV PBMC. The results indicate that the T cell anergy associated with CMV infection is restricted to the T cell receptor/CD3 activation pathway.
Novel human immunoglobulin heavy chain constant region gene deletion haplotypes characterized by pulsed-field electrophoresis
Authors: OLSSON P G , RABBANI H , HAMMARSTRÖM L , SMITH C I E
Published: July 2010

Fifteen patients with selective IgG1 deficiency were screened for immunoglobulin H chain C region locus (IGHC) gene deletions and three deletion haplotypes were found: del Gl, delGI G4 and del G4. These haplotypes, together with four deletion haplotypes described by us previously (del G1 (NY), del G1 (VIT) del G1-G2 (NY) and del G2-G4 (HJE)). were further characterized using pulsed-field gel electrophoresis (PFGE) to determine the physical extent of the deletions. The MluI fragment sizes confirmed the deletions, although the deduced sizes of the most extensive deletions indicated that material had been inserted into the locus.
Bronchoalveolar lavage cell analysis and lung function impairment in patients with systemic lupus erythematosus (SLE)
Authors: GROEN H , ASLANDER M , BOOTSMA H , van der MARK Th W , KALLENBERG C G M , POSTMA D S
Published: July 2010

We examined the relationship between peripheral blood and bronchoalveolar lavage (BAL.) lymphocyte phenotypes and lung function in 19 patients with SLE. and evaluated their association with disease activity. Lung function assessment showed a mildly restrictive pattern with frequent impairment of transfer factor for carbon monoxide (TL,CO) and diffusing capacity of the alveolocapillary membrane (Dm), of late-expiratory airflow rales and with a high prevalence of increased airway resistance. TL,CO, KCO and Dm correlated inversely with the numbers of CD+ cells and CD56+/CD16+/CD3− (NK) cells in BAL. Oxygen radical production, both by stimulated and unslimulated BAL cells and blood polymorphonuclear leucocytes (PMN) was significantly increased in SLE. In comparison with healthy controls, patients with SLE had a lower percentage of CD19+ B cells in the BAL versus an increased percentage of these cells in peripheral blood. HLA-DR expression on CD4+ and CDS+ lung lymphocytes was markedly increased in SLE. Current SLE disease activity was not associated with changes in BAL or peripheral blood lymphocyte phenotypes. Our data suggest that an ongoing cell-mediated immune response is present in the lungs in SLE, particularly involving activated CD8+ T cells and CD56+/CD16+/CD3− NK cells. It is associated with up-regulated local production of oxygen radicals and with impaired pulmonary diffusing capacity. This inflammatory process seems to be independent of general SLE disease activity.
Triggering of respiratory burst by phagocytosis in monocytes of patients with systemic lupus erythematosus
Authors: GYIMESI E , KÁVAI M , KISS E , CSIPÖ I , SZÜCS G , SZEGEDI G Y
Published: July 2010

The triggering of the respiratory burst by phagocytosis via different receptors in monocytes of patients with systemic lupus erythematosus (SLE) was investigated. The superoxide anion synthesis was assayed by reduction of fcrricytochrome C that was inhibited by superoxide dismutase. The mononuclear cell suspensions were triggered by IgG-coated latex. C3 complement fragment-coated and uncoated yeast (Saccharomyces cerevisiae). Superoxide generation induced by phagocytosis via FcγR was decreased in monocytes of patients with SLE. On the other hand. MoAbs against FcγRI, FcγRII and especially CR3 could also induce superoxide anion synthesis. At the same time, superoxide generation induced by anti-CR3 could be inhibited with C3-coated yeast.
Effect of low-dose ultraviolet-B radiation on the function of human T lymphocytesin vitro
Authors: TEUNISSEN M B M , SYLVA-STEENLAND R M R , BOS J D
Published: July 2010

Purified peripheral blood human T lymphocytes, derived from normal individuals, were assayed for their susceptibility to low doses of ultraviolet B (UVB) In vitro. Exposure of T cells to graded single doses (range 0–8 mJ/cm2) of UVB resulted in a dose-dependent reduction of viability. This phototoxic effect was not immediately apparent, however, but became manifest 48–72 h subsequent to irradiation. A dose as little as 0·5–1 mJ/cm2 was sufficient to cause 50% mortality. Irradiated T cells showed a reduced ability to proliferate, irrespective of the stimulus used, and a reduced ability to produce cytokines IL-2, IL-4, IL-5, interferon-gamma (IFN-γ) and tumour necrosis factor-alpha (TNF-α). This decreased ability was UVB-dose related and, remarkably, was exactly correlated to phototoxicity. UVB had no effect on CD4 and CD8 expression or their ratio, whereas the expression of IL-2R (CD25) was only slightly reduced. Our data suggest that UVB radiation neither selectively affects Th1 or Th2 nor CD4 or CD8 T cell subsets. The high susceptibility of T cells to UVB might explain, al least in part, the beneficial effect of phototherapy during treatment of certain immunodermatological diseases.
Transforming growth factor-beta (TGF-β) in human milk
Authors: SAITO S , YOSHIDA M , ICHIJO M , ISHIZAKA S , TSUJII T
Published: July 2010

The amount of TGF-β contained in human whey was studied by the colony formation of NRK47F cells. It was noted that a factor inducing colony formation did exist in human whey, and its action was neutralized when anti-TGF-β antibodies were introduced. This suggests that TGF-β does exist in human whey. In colostrum, the total amount of TGF-β was 1365·7 ± 242·9 ng /ml, of which the active form comprised 728·1 ± 248·7 ng/ml (n=21). In late milk, the total TGF-β was 952·5 ± 212·6 ng/ml, with an active form of 178·7 ± 157·3 ng/ml. Thus human milk contains a large amount of active TGF-β. Furthermore, it was revealed by the reverse transcriptase polymerase chain reaction that mRNAs coding TGF-β1 and TGF-β2 exist in human milk cells. These results suggest that both TGF-β1 and TGF-β2 exist in human milk.
IL-5 in post-traumatic eosinophilic pleural effusion
Authors: SCHANDENÉ L , NAMIAS B , CRUSIAUX A , LYBIN M , DEVOS R , VELU T , CAPEL P , BELLENS R , GOLDMAN M
Published: July 2010

Thoracic trauma or pneumothorax can result in pleural fluid eosinophilia. In this study we investigated the role of the eosinophilopoietic cytokine IL-5 in three cases of post-traumatic eosinophilic pleural effusions (EPE). Using a specific immunoenzymatic assay, significant levels of IL-5 were found in EPE (range 100–3000 pg/ml), while IL-5 was undetectable (<25 pg/ml) in corresponding serum samples and in non-eosinophilic pleural fluids. IL-5 present in pleural fluids was found bioactive in a proiiferative assay using a mouse CTLL-2 cell line transfected with the cDNA corresponding to the a chain of the human IL-5 receptor. Using a reverse polymerase chain reaction (PCR) method, we found IL-5 mRNA expression within pleural mononuclear cells from patients with EPE, but not in corresponding peripheral blood mononuclear cells (PBMC), confirming that IL-5 is synthesized locally in the pleural cavity. In the two cases in which pleural CD4+ cells were purified, these cells were identified as the major source of IL-5. Taken together, these data indicate that the development of post-traumatic EPE is related to a local secretion of IL-5 by CD4+ cells present in the pleural cavity.
Limiting dilution analysis of the allo-MHC anti-paternal cytotoxic T cell response I: normal primigravid and multiparous pregnancies
Authors: MANYONDA I T , PEREIRA R S , PEARCE S M , SHARROCK C E M
Published: July 2010

Anti-paternal cytotoxic T lymphocyte precursor frequencies (CTLpF) were determined by limiting dilution analysis (LDA) in the peripheral blood of eight primigravid and seven multiparous women during the three trimesters of pregnancy. In five of these women the responses to cord blood lymphocytes (CBL) and paternal lymphocytes were also determined at the time of delivery and at 6 weeks post delivery. As controls, CTLpF against unrelated third party donors were determined. A wide range of CTLpF against unrelated third groups of targets was found in both the primigravid and multiparous women, reflecting the wide range of frequencies found in random populations. These frequencies remained fairly constant during and 6 weeks after the pregnancy. Splitwell analysis demonstrated that the responses generated in our culture system were specific to the stimulator. The LDA data conform to single-hit kinetics, indicating that only cytotoxic T cells were limiting in the assay. Proliferative responses of maternal lymphocytes to paternal, cord blood and third party MHC antigens also remained unchanged as determined by time-course mixed lymphocyte reactions (MLR). Our data suggest that there is no significant allo-stimulation or suppression of the maternal immune system during normal pregnancy. The mother remains immunocompetent and is capable of both cytottjxic and proliferalive responses lo paternally-derived fetal MIC antigens. Our findings confirm that in normal pregnancy the trophoblast. which is devoid of classical MHC antigens, forms an effective immune barrier which prevents interaction of the maternal and fetal immune systems.
Studies on the mechanism of complement-mediated inhibition of antibody binding to HIV gp41
Authors: HIDVÉGI T , PROHÁSZKA Z , UJHELYI E , THIELENS N M , DIERICH M P , HAMPL H , ARLAUD G , NAGY K , FÜST G
Published: July 2010

We have previously demonstrated that HIV envelope gp41 binding to specific antibodies decreases after preincubation of fluid-phase gp41 in normal human serum. This inhibition is proven to be mediated by the classical complement pathway. In this study recombinant gp41 (rgp41) and/or synthetic peptides were preadsorbed to solid phase, and then complement (normal human serum heated human serum, purified Clq/heated Clq) and anti-gp41 antibodies were added either after each other or simultaneously, and the amounts of bound antibody, and deposited C3b, C4b and Clq were measured. Complement-dependent inhibition of antibody binding to solid-phase rgp41 was found, and Clq seems to be at least partially responsible for this phenomenon. Heating of Clq did not affect this process. Higher amounts of anti-gp41 antibodies significantly and dose-dependently enhanced C4b and C3b fixation to solid-phase rgp41. In the case of synthetic peptides corresponding to the immunodominant region of gp41, significant antibody binding to the solid-phase peptides was also detected, and pretreatment of peptides preadsorbed to solid phase with normal human serum almost totally abolished the antibody binding.
Variation in interferon-gamma responses toCoxiella burnetiiantigens with lymphocytes from vaccinated or naturally infected subjects
Authors: IZZO A A , MARMION B P
Published: July 2010

Previous work in our laboratory has shown that lymphocytes from persons vaccinated with a formalin-inactivated Phase I Q fever vaccine(Q-VaxCSL Ltd) show a mitogenic response to Coxiella burnetii antigens. The mitogenic response is the sum of that from various subsets of CD4+, T helper cells, CD8+ T cells and probably B cells. It does not distinguish between T helper cell responses leading to formation of interferon-gamma (IFN-γ)—a cytokine responsible for clearing intracellular infection with C. burnetii organisms—and responses of other T cell subsets which may produce disease-enhancing cytokines. The present study analyses (i) the capacity of Q-Vax to induce T cell sensitization which leads to IFN-γ responses on antigen stimulation, and (ii) the immunomodulatory, (down-regulatory) effects of the Phase I lipopolysaccharide (LPS) of the organism, which interacts with monocyte/macrophages to limit IL-2 production and production of IFN-γ by sensitized T lymphocytes.
Circulating adhesion molecules in tuberculosis
Authors: LAI C K W , WONG K C , CHAN C H S , HO S S , CHUNG S Y , HASKARD D O , LAI K N
Published: July 2010

Leucocyte-endothelial adhesion molecules have been implicated in the pathogenesis of inflammatory diseases. To evaluate their role as markers of disease activity in tuberculosis, we have used an antigen capture ELISA to measure the serum concentrations of circulating intercellular adhesion molecule-1 (cICAM-1), E-selectin (cE-selectin) and vascular cell adhesion molecule-1 (cVCAM-1) in 34 patients with active tuberculosis (27 with pulmonary disease and seven with lymph node disease) before the commencement of standard chemotherapy, 15 subjects who had previously completed treatment for pulmonary tuberculosis, and 27 healthy volunteers. Circulating ICAM-1 and E-selectin levels were significantly elevated in patients with active tuberculosis when compared to those with treated disease (P⩽0.01), and healthy controls (P< 0.02). Circulating VCAM-1 was raised in patients with active or old pulmonary tuberculosis (P<0.02 versus healthy controls) but not in those with tuberculous lymphadenitis. Significant correlations were observed between the levels of cICAM-1 and cE-selectin (ρ= 0-63, P=0 0001), and between cICAM-1 and cVCAM-1 (ρ = 0.28, P=0.016). Taking the mean +2 s.d. of the serum level in healthy controls as the upper limit of normal range, circulating ICAM-1 had the best discriminative power in identifying active tuberculosis, being elevated in about 80% of patients but was raised in only 6.7% of subjects with treated disease and in 3.7% of normal subjects. Our data support the possibility that three adhesion molecules may be involved in the pathogenesis of tuberculosis and cICAM-1 may be a useful marker of disease activity.
Spontaneous cytotoxicity of intestinal intraepithelial lymphocytes: clues to the mechanism
Authors: ROBERTS A I , O'CONNELL S M , BIANCONE L , BROLIN R E , EBERT H C
Published: July 2010

Human intestinal intraepithelial lymphocytes (IEL) demonstrate target cell-restricted spontaneous cytotoxic (SC) activity that is due to CD2+CD3+CD8+CD16−CD56– effector eells: they kill epithelial cell (EC) tumours (such as DLD-1 colon cancer eells), but not natural killer (NK)-sensitive K-562 cells. The present study shows that the measured levels of SC activities by IEL correlated with those of autologous lamina propria lymphocytes (LPL), but not with those of peripheral blood lymphocytes (PBL). Also, the susceptibilities of DLD-1 cell clones to lysis by IEL and PBL effector cells did not correlate, suggesting different mechanisms of lysis. Antibody blocking experiments showed that the main surface molecules involved in lysis depended on the effector cell type: αEβt (HML-l) on IEL and CDI6 on PBL. No antibody-dependent cell-mediated cytotoxicity (ADCC) was demonstrated by IEL, even after stimulation with inerferon-gamma (IFN-γ). Few IEL expressed Fc receptors for IgG. This study describes further differences between the SC activities of IEL and PBL.
Antigenic structure of the hepatitis C virus envelope 2 protein
Authors: ZHANG Z-X , SÖNNERBORG A , SÄLLBERG M
Published: July 2010

The antigenic structure of the envelope 2 (e2) protein of the hepatitis C virus (HCV) was characterized by the use of 70 synthetic peptides and 131 human sera from persons with antibodies to HCV, Among 34 overlapping peptides spanning the e2 protein of HCV, two major antigenic regions were located to residues 484–499 and residues 554–569. The sequence of the two major antigenic regions of the e2 protein are partly well conserved within the described types of HCV. Both regions contain two Cys residues in close proximity, and the region at residues 554–569 contains a putative N-glycosylation site, which are factors that previously have been suggested to affect the immune recognition of the e2 protein. Using substitution peptide analogues where each position within residues 484–499 and 554–569 were sequentially substituted by Ala or Gly, the most essential residues for antibody binding were found to be the Pro-498, Ala-499, Ala-566, Pro-567, and Pro-568. All of these, except for the Pro-498 and Ala-566, are conserved among different HCV strains. Also, according to previous studies, position 496 often shows variations, which could be explained by position 496 being contained within the antigenic region at residues 484–499. Interestingly, none of the Cys residues at positions 486, 494, 564 and 569 were found to be essential for antibody binding, indicating that these are not essential in maintaining the e2antigenicity of the peptides. In a material of 114 confirmed anti-HCV positive sera, derived from patients during the acute or the chronic phase of HCV infection, the prevalence of antibodies to the two major linear antigenic regions of the e2 protein was found to be 55% among HCV RNA-positive sera, and 53% among HCV RNA-negative sera. In conclusion, we have identified and characterized two major linear antigenic regions outside the two hypervariable regions of the e2 protein. Since these regions are accessible to the B cells of the infected host, these two regions are likely to be surface exposed either on the precursor polyprotein or the native e2 protein. Also, we could confirm that antibodies to the e2 protein co-exist with HCV viraemia.
Cross-regulatory role of interferon-gamma (IFN-γ), IL-4 and IL-10 in schistosome egg granuloma formation:in vivoregulation of Th activity and inflammation
Authors: CHENSUE S W , WARMINGTON K S , RUTH J , LINCOLN P M , KUNKEL S L
Published: July 2010

This study examined the relationship ofIL-4, IL-10 and IFN-γ with regard to the local granuloma (GR) and draining lymph node (LN) response to Schistosoma mansoni eggs. Synchronized GR were induced in naive and schistosome-infected mice at the vigorous (8 weeks) and late chronic (20 weeks) stages. In LN cultures, IL-10 and IFN production peaked on day 4 and was greatest for 8 week-infected mice. All GR cultures contained IFN, but compared with naive mice IL-10 production was accelerated at 8 weeks and abrogated at 20 weeks, consistent with expansion and abatement of Th2 activity, Cytokine neutralization was performed in egg-challenged, naive mice that were adoptively sensitized with lymphoid cells from 8 week-infected donors. GR size, GR macrophage tumour necrosis factor (TNF) production and egg antigen-elicited IL-2, IL-4, IL-5, IL-10 and IFN were examined on day 4 of GR formation, Anti-IFN augmented GR area by 40%, increased local IL-4 and IL-10, but decreased IFN and TNF production. In corresponding LN cultures, IFN decreased by about 50%, while IL-2, IL-4, IL-IO and lL-5 increased by nearly two-, four-, five- and six-fold, respectively, Anti-IL-10 did not affect GR size or GR cytokines, but increased IFN levels in LN cultures four-fold and decreased IL-2, IL-4, lL-5 and IL-10. Anti-IL-4 abrogated GR area by 40%, along with a reduction in local IL-4 and TNF production. In LN, IL-4 depletion reduced IL-4 and IL-5 by 60–70% and increased IFN levels. These results support the notion of a cross-regulatory network in which IFN inhibits Th2 and IL-10 inhibits ThI cells. IL-4 fosters Th2 cell differentiation in LN, but also performs a critical recruitment function in the eosinophil-rich schistosome egg-induced GR, whereas IFN contributes to enhanced GR macrophage function.
Evaluation of Onchocerca volvulus-specific IgG4 subclass serology as an index of onchocerciasis transmission potential of three Gabonese villages
Authors: EGWANG T G , DUONG T H , NGUIRI C , NGARI P , EVERAERE S , RICHARD-LENOBLE D , GBAKIMA A A , KOMBILA M
Published: July 2010

The major objective of this study was to evaluate the usefulness of IgG4 ELISA and Western blot analysis, using a crude extract of Onchocerca volvulus adult worms as antigens, for diagnosing onchocerciasis in a Gabonese paediatric population with mixed filarial infections. The subjects had loaisis, streptocercosis or mansonellosis in addition to onchocerciasis. Control sera from loaisis or mansonellosis subjects residing outside the endemic zone were used to provide the cut-off point for positive results. The IgG4 ELISA had a specificity of 96% but a lower sensitivity of 78·7%. It detected 25 onchocerciasis cases out of 65 individuals who were negative on parasitological examination. Furthermore, the ELISA provided a more accurate picture of onchocerciasis transmission in a village with very low skin microfilartal load. A 27·5-kD antigen was identified on Western blots as a marker of onchocerciasis. The paediatric population provided a reliable window for assessing the parasitologic and serologic parameters in the three villages with disparate levels of onchocerciasis transmission.
Suppression of experimental systemic lupus erythematosus (SLE) with specific anti-idiotypic antibody–saporin conjugate
Authors: BLANK M , MANOSROI J , TOMER Y , MANOSROI A , KOPOLOVIC J , CHARCON-POLAK S , SHOENFELD Y
Published: July 2010

The importance of the idiotypic network is represented in experimental SLE induced by active immunization of naive mice with an anti-DNA idiotype (Ab1) emulsified in adjuvant. The mice after 4 months of incubation generate Ab3 having anti-DNA activity. In addition, the mice develop other serological markers for SLE associated with clinical and histopathological manifestations characteristic of the disease. To confirm further the etiological role of the idiotype in this experimental model the mice were treated with specific anti-idiotypic antibodies (anti-id) which were also conjugated to a toxin-saporin (Immunotoxin (IT)). Pretreatment of hybridoma cell line producing the anti-anti-Id (anti-DNA = (Ab3)) for 48 h with the anti-Id MoAb (Ab2) reduced the production of anti-DNA by 58%, while pretreatment with the IT resulted in 86% decrease in anti-DNA secretion (saporin alone had only 12% effect). The anti-Id MoAb had no effect on the production of immunoglobulin by an unrelated cell line. In vivo treatment of mice with experimental SLE led to a significant decrease in titres of serum autoantibodies, with diminished clinical manifestations. The results were more remarkable when the IT was employed. These suppressive effects were specific, since an anti-Id treatment of experimental anti-phospholipid syndrome was of no avail. The anti-Id effect was mediated via a reduction in specific anti-DNA antibody-forming cells, and lasted only while anti-Id injections were given. Discontinuation of the anti-Id injection was followed by a rise in titres of anti-DNA antibodies. No immunological escape of new anti-DNA Ids was noted. Our results point to the importance of pathogenic idiotypes in SLE and to the specific potential of implementing anti-idiotypic therapy, enhanced by the conjugation of the anti-Id to an immunotoxin, in particular one with low spontaneous toxicity.
T cell reactivity to proteinase 3 and myeloperoxidase in patients with Wegener's granulomatosis (WG)
Authors: BROUWER E , STEGEMAN C A , HUITEMA M G , LIMBURG P C , KALLENBERG C G M
Published: July 2010

T cell-mediated immunity is hypothesized to play an important role in the pathogenesis of granulomatous inflammation and vasculitis as found in patients with WG. The antigenic specificities of those T cells remain, however, unknown. Anti-neutrophil cytoplasmic antibodies (ANCA) present in patients with WG are directed to proteinase 3 (PR3) and myeloperoxidase (MPO). In the present study we investigated the proliferative capacity of peripheral Wood mononuclear cells (PBMC) from patients with WG and age-and sex-matched controls in response to the WG autoantigens PR3 and MPO. Possible mitogenic effects of active PR3 and toxic effects of active MPO were excluded by using heat-inactivated PR3 and MPO. Antigen-specific stimulation induced by these autoantigens was studied by using processed PR3 and MPO in the lymphocyte stimulation test (LST). Proliferation induced by processed antigen correlated with that by heat-inactivated free antigen. The general capacity to proliferate in response to mitogens and recall antigens did not differ between patients and controls. However, patients with WG who were or had been positive for PR3-ANCA (n = 17) responded more strongly to PR3 than to MPO and showed higher responses to PR3 compared with controls (n = 13). Within the PR3-ANCA group T cell proliferation did not correlate with ANCA titre. In a small group of patients with MPO-ANCA (n=5) no differences were observed compared with controls for MPO-specific proliferation. The data presented demonstrate that autoreactive PR3-specific T cells are present in patients with WG. Their fine specificity and possible role in the pathogenesis of WG have to be defined in further studies.
T cell receptor V-gene usage in oral lichen planus; increased frequency of T cell receptors expressing Vα2 and Vβ3
Authors: SIMARK-MATTSSON C , BERGENHOLTZ G , JONTELL M , TARKOWSKI A , DAHLGREN U I
Published: July 2010

In order to analyse the clonality of T cells in the inflammatory infiltrate of oral lichen planus (OLP), mucosal biopsies were obtained from seven patients with manifest disease. The biopsies were stained with MoAbs directed against 11 different T cell receptor (TCR) V-gene families, anti-CD4, anti-CD8 and lL-2 receptor (1L-2R). For comparison, the frequencies of the different TCR V-families were determined in biopsies from five patients with oral candidosis as well as in peripheral blood from three patients with OLP and from six healthy blood donors (HBD), The occurrence of the investigated TCR V-families varied between 0% and 7% in venous blood obtained from both HBD and OLP patients, T lymphocytes expressing the TCR Vβ3 and Vα2 in OLP biopsies were, however, detected in frequencies ranging between 18% and 40% of the total fraction of lymphocytes, a consistent finding for all the OLP infiltrates studied. The other nine TCR V-families examined appeared in low frequencies both in biopsies and in peripheral blood, Vα2+ and Vβ+β3+ cells were often localized adjacent to the basal membrane. In contrast, T cells in Candida-induced lesions did not express a biased TCR distribution, and most V-families studied appeared in frequencies of 0–6%. Thus, T lymphocytes in OLP lesions express a substantially higher frequency of TCR Vα2 and Vβ3 than expected from the distribution in blood. The clonal expansion of T cells observed in OLP suggests that a superantigen is involved in the pathogenesis of the disease. Whether this superantigen is of exogenous or endogenous origin needs to be investigated.