bluuebunny/crossref_metadata_2025 huggingface dataset:
The effect of gold sodium thiomalate and auranofin on lipopolysaccharide-induced interleukin-1 production by blood monocytesin vitro:variation in healthy subjects and patients with arthritis
Authors: DANIS V A , KULESZ A J , NELSON D S , BROOKS P M
Published: July 2010

The anti-rheumatic gold compounds gold sodium thiomalate (GST) and auranofin (AF) have variable and often unpredictable effects in patients treated for arthritis As inhibition of interleukin-1 (IL-1) production may be an important effect of these drugs, we investigated their effect on IL-1 production by lipopolysaccharide (LPS) stimulated monocytes in a serum-free. non-adherent culture system. A bi-modal effect was observed: low concentrations (GST 10–250 ng/ml and AF 1–100 ng/ml) potentiated IL-1 production, and higher concentrations (GST 200–1000 ng/ml and AF10–500 ng/ml) inhibited This bi-modal effect was observed for both secreted and cell-associated IL-1 activity with the exception that GST failed to inhibit cell-associated IL-1 generation. The potentiating effect was dependent on the continuous presence of gold for at least the first few hours after LPS stimulation. The inhibitory effect of GST was dependent on its presence after LPS Stimulation while that of AF was evident even if cells were pretreated with AF and washed before exposure to LPS. There was considerable individual variation in IL-1 production in response to LPS as well as in the effects of gold on cells from both healthy individuals and patients with arthritis. There was also some overlap in the range of concentrations of gold that potentiated and inhibited IL-1 production, and there was relative insensitivity to the inhibitory effects of gold in certain individuals. These results may explain some of the variability in the response of patients to chrysotherapy and support further studies to see if these in vitro effects might predict clinical response to gold.
Differential augmentation ofin vivonatural killer cytotoxicity in normal primates with recombinant human interleukin-1 and granulocyte-macrophage colony-stimulating factor
Authors: DAVIS T A , MONROY R L , SKELLY R R , DONAHUE R E , MACVITTIE T J
Published: July 2010

The effect of recombinant human interleukin-1 (IL-1) alpha, granulocyte-macrophage colony-stimulating factor (GM-CSF), and combined factor therapy (CFT) on Rhesus monkey peripheral blood natural killer (NK) activity in vivo was compared. During a 14-day treatment period, IL-1-treated animals demonstrated a 170% increase in NK activity against K562 target cells by day 4, reaching maximal levels (300%) at day 16, and returning to baseline by day 30. NK activity of GM-CSF-treated monkeys increased slightly (60–100%) during days 4–12. as did saline-treated monkeys, but returned to baseline values by day 16. A delayed increase in NK activity resulted after GM-CSF treatment, reaching a peak (260%) on day 23 and remaining elevated through day 39. CFT resulted in a bimodal response pattern, with two peaks of NK activity, one at day 16 and a second at day 39, The first peak of activity (223%) was significantly less than the activity attained with IL-1 alone; the second peak (300%) was of greater duration and occurred later than the peak observed in GM-CSF-treated monkeys. Unlike IL-1, GM-CSF treatment did not lead to a immediate stimulation of NK activity; augmentation was delayed by more than 7 days post treatment. CFT results suggest that GM-CSF reduced the direct NK response to IL-1; while IL-1 led to an enhanced delayed NK response. Therefore, IL-1 and GM-CSF augment NK activity through different but interrelated pathways.
Urinary excretion of CD23 antigen in normal individuals and patients with chronic lymphocytic leukaemia (CLL)
Authors: LING N R , STEVENSON F K , BROWN B
Published: July 2010

A soluble form of CD23 (sCD23) was found in the urine from 12 normal individuals but was not present in 20 normal sera, suggesting that sCD23 produced by cells in tissues is eliminated in the urine. The sCD23 from urine differed in physicochemical properties from the sCD23 found in supernales from B-lymphoblastoid cell lines (B-LCL) and in the sera of patients with B type chronic lymphocytic leukaemia (B-CLL). On SDS-PAGE analysis under reducing conditions urinary sCD23 showed two bands corresponding to molecular weights of 45 60 kD and 2S 35 kD indicating that sCD23may be excreted in combination with another molecule. When subjected to gel filtration in its native state, sCD23 from urine showed a major peak at approximately 150 kD and a minor peak (probably a breakdown product) at 21 kD. Urinary sCD23 was more strongly held by DEAE-cellulose and required 0–5 m buffer pH 8·0 for elution, suggesting that it is more anionic than sCD23 from culture supernates. Five MoAbs recognizing different epitopes on sCD23 from B-LCL supernates were tested on urinary sCD23. Four of the MoAbs were reactive but one (EBVCS-1) was not. Urinary sCD23 did not bind to IgE. The level of sCD23 found in normal urine (approximately 0·02–0·05 μg/ml) was exceeded in 17 of 24 cases of B-CLL. In one case with a high cell count and a serum concentration of 10 μg/ml. the urine contained 80 μg/ml sCD23. In another case a high serum sCD23 was not matched by a high urinary level. In this case the gel filtration pattern was closer to that found with urine sCD23 rather than the B-LCL pattern found with sera of other B-CLL patients.
Anti-human thyroid peroxidase and anti-human thyroglobulin antibodies present no cross-reactivity on recombinant peptides
Authors: HENRY M , ZANELLI E , MALTHIERY Y
Published: July 2010

Thyroglobulin (Tg) and thyroid peroxidase (TPO) are two antigens largely recognized by the sera from patients with autoimmune thyroid disease (AITD). Recently, the complete mapping of both antigens was established with rabbit polyclonal antibodies by the use of recombinant proteins expressed in prokaryotic vector. Several investigators have argued for the existence of a crossreactivity of some hetero- and autologous antibodies versus these two proteins. In the present study, using rabbit polyclonal antibody, mouse polyclonal antibody and autoimmune antibody (aAb). we observed no common epitope on human Tg (hTg) and human TPO (hTPO).
Evidence that X-linked severe combined immunodeficiency is not a differentiation defect of T lymphocytes
Authors: GOODSHIP J , MALCOLM S , LEVINSKY R J
Published: July 2010

In order to gain information about the nature of the defect in X-linked severe combined immunodeficiency (XSCID). we investigated gene expression in different lymphoid and haematopoietic cells of female carriers by looking for non-random X chromosome usage. We have shown non-random X chromosome usage in T lymphocyte enriched (E+) fraction in all carriers. E cells and monocytes showed non-random X chromosome usage in three carriers tested. In the B cell series one carrier showed non-random inactivation in all EBV lines tested (10) and the same X chromosome was shown to be active in all cells, In other carriers there was a preference for use of the normal X chromosome but some B cell lines used the mutant X as well as the normal X. Similar results were found with granulocytes. In two female carriers DNA made directly from whole blood showed a non-random pattern of X chromosome usage. In fibroblast cultures from two female carriers more cells had the normal gene on the active X chromosome than had the defective gene on the active X chromosome. Within families there was heterogeneous expression of the gene. The gene that is defective in XSCID is expressed in all the cell types studied and. therefore, is not a T lymphocyte differentiation gene. The results are consistent with the gene being in a metabolic pathway as in certain autosomal recessive forms of SCID i.e. adenosine deaminase deficiency and purine nucleoside phosphorylase deficiency.
Asbestos fibres inhibit thein vitroactivity of ymphokine-activated killer (LAK) cells from healthy individualsand patients with malignant mesothelioma
Authors: MANNING L S , DAVIS M R , ROBINSON B W S
Published: July 2010

Asbestos exposure is associated with an increased incidence of several malignances. including malignant mesothelioma (MM). This study evaluates the relationship between asbestos exposure and the in vitro generation and function of LAK cells, an immune effector cell population with powerful lytic activity against MM cells. Both serpentine (chrysotile) and amphibole (amosite and crocidolilc) forms of asbestos fibres suppress LAK cell generation, viability (by 5–11%, P < 0.02) and cell recovery (by 13.15%, P < 0.02). However, the LAK cells generated in the presence of the amphiboles were as effective as unexposed cells in lysing both standard tumour cell targets (K562, 56.4% lysis versus 61.5%. respectively, P > 0.5; NS; Daudi. 60.5% lysis versus 64.5%P > 0.5; NS). and MM tumour cell targets (mean of three MM cell lines 48.3%versus 46.3%. P > 0.5; NS), whereas the function of LAK cells generated in the presence of chrysotile was significantly reduced against three out of the five tumour cell targets tested (P < 0.03). In the presence of asbestos fibres, LAK cell function was reduced against all five tumour cell targets (P < 0.0l). irrespective of whether the cell donors were healthy individuals or patients with MM. NK cell activity was also suppressed (P < 0.0l). The serpentine form of asbestos, chrysotile, was significantly more suppressive of both effector cell functions than either of the amphiboles (P < 001). These findings suggest that asbestos exposure may suppress the function and in some instances the generation of immune effector cell mechanisms, thereby increasing the risk of disease and malignancy.
A new murine model of autoimmune orchids induced by immunization with viable syngeneic testicular germ cells alone. I. Immunological and histological studies
Authors: ITOH M , HIRAMINE C , HOJO K
Published: July 2010

Experimental autoimmune orchitis (EAO) was produced in C3H/He mice with as high as 100% incidence by two or three s.c. injections of 1 × 107 viable syngeneic testicular germ cells (TC) without resorting to adjuvants, Bordetella pertussis vaccine, or other immunological manipulations. On day 40 after the first injection of TC. the lesions induced were characterized by interstitial infiltration of inflammatory cells and severe hypospermatogenesis in the testis with resulting whole organ atrophy and. in contrast, by a complete lack of epididymitis. Immunological studies revealed that this form of immunization caused both delayed-type hypersensitivity and humoral antibody responses to syngeneic TC. We compared the susceptibilities to the induction of this type of EAO among six different strains of inbred mice comprising A/J, AKR, BALB/C, C3H/He, C57BL/6 and DBA/2 mice. All strains except for DBA/2 mice developed lesions of EAO to a greater or lesser extent, and severe disease was induced with high frequency in two strains, C3H/He and A/J. As this murine model of EAO can be induced without the use of Freund's complete adjuvant and B. pertussis vaccine, it is simply ‘autoimmune’ in nature and may provide new ways for further investigation into the immunological mechanisms which regulate deleterious autoimmune reactions to germ cell antigens leading to the male infertility.
Immunoglobulin and cytokine production by neonatal lymphocytes
Authors: WATSON W , OEN K , RAMDAHIN R , HARMAN C
Published: July 2010

Growth and differentiation of cord blood B cells were studied using T cell-depleted populations. In the absence of in vitro activation, cord blood B cells proliferated in response to cytokines including interleukin-2 (IL-2) and interleukin-4 (IL-4); anti-μ-stimulated cord B cells had a lesser response to IL-2 than adult cells. IgM synthesis by cord blood B cells was enhanced by interleukin-6 (IL-6) and decreased by IL-2. In cultures activated by Staphylococcus aureus Cowan I (SAC), cord blood B cells produced lesser increases in IgM than adult B cells regardless of the cytokine added. Cord blood B cells produced no IgG or IgA with any cytokine preparation with or without SAC activation. Supernatants of cord blood T cells pulse-stimulated with phytohaemagglutinin and phorbol myristate acetate contained less IL-2 and IL-6 and had less growth and differentiation activity than adult T cell supernatants. The results confirm a limited cord blood B cell response and also suggest a limitation in production of B cell stimulatory lymphokines by cord blood T cells.
Granulocyte-macrophage colony-stimulating factor augments neutrophil killing ofTorulopsis glabrataand stimulates neutrophil respiratory burst and degranulation
Authors: KOWANKO I C , FERRANTE A , HARVEY D P , CARMAN K L
Published: July 2010

The effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the interaction between the fungus Torulopsis glabrata and human neutrophils was examined. Pre-incubation of neutrophils with GM-CSF increased the neutrophil fungal killing. The cytokine also increased the oxygen-dependent respiratory burst in response to opsonized fungi, measured by the lucigenin-dependent chemiluminescence assay and superoxide release. Under the same conditions the cytokine augmented release of constituents from both specific and azurophilic granules. Besides these priming effects, GM-CSF was a weak stimulus of the neutrophil respiratory burst and degranulation. The priming and stimulatory effects of GM-CSF were observed at 10–1000 U/ml with an optimal concentration of 100 U/ml.
Detection of circulating intercellular adhesion molecule-1 antigen in malignant diseases
Authors: TSUJISAKI M , IMAI K , HIRATA H , HANZAWA Y , MASUYA J , NAKANO T , SUGIYAMA T , MATSUI M , HINODA Y , YACHI A
Published: July 2010

A new monoclonal antibody (MoAb) H A58 (IgG 1) was prepared, which recognizes the binding site on the intercellular adhesion molecule-1 (ICAM-1) antigen to the lymphocyte function-associated antigen-1 (LFA-1). The double-determinant immunoassay (DDIA) was established with use of MoAb HAS8 and another anti-ICAM-1, MoAb CL207, to detect the soluble, shedding ICAM-1 antigen. Human recombinanl interferon-gamma (IFN-γ) induced not only the expression of cell surface ICAM-1, but also the shedding ICAM-I antigen in an IFN-y concentration-dependent and incubation-time-dependent manner. DDIA was applied to detect the shedding ICAM-1 antigen in the sera of patients with malignant or benign diseases. The incidence of posilivity for ICAM-I antigen in malignant diseases was higher than that in benign diseases or in healthy controls. Furthermore, the sera of cancer patients with liver metastasis showed higher levels of the shedding ICAM-1 antigen. These findings suggest that serum ICAM-1 antigen may be a useful marker to monitor tumor burden in cancer patients.
Naturally occurring anti-interferon-α2a antibodies in patients with acute viral hepatitis
Authors: IKEDA Y , TODA G , HASHIMOTO N , UMEDA N , MIYAKE K , YAMANAKA M , KUROKOWA K
Published: July 2010

The occurrence of antibodies against recombinant human interferon-α2a (IFN-α2a) in patients with acute viral hepatitis (AVH) was examined by ELISA. Naturally occurring IgG anti-IFN-α2a were found in 50% of patients with type A, 50% of those with type B and in 8.3% of those with non-A, non-B AVH. The corresponding frequencies of IgM antibodies were 80%, 30% and 33.3%, respectively. IgM anti-IFN-α2a were found more frequently in patients with AVH type A than in normal control subjects (P < 0.01). Anti-IFN-α2a were detectable at the highest frequency 3 weeks after acute onset and then became negative. An absorption experiment revealed that IgM anti-IFN-α2a did not cross-react with recombinant human IFN-α2b. Immunoblotting analysis confirmed the binding of antibodies to IFN-α2a. Sera positive for IgG and/or IgM anti-IFN-α2a were unable to neutralize IFN-α2a. The appearance of anti-IFN-α2a was not correlated with disease severity. There was no evidence to suggest that anti-IFN-α2a impaired the elimination of hepatitis virus. This is the first study to demonstrate the occurrence of anti-IFN-α2a in patients with AVH. Detection of anti-IFN-α2a may be useful for clarifying any underlying immune events in various diseases.
Cytokine production by peripheral blood mononuclear cells in IgA nephropathy
Authors: LAI K N , LEUNG J C K , LI P K T , LUI S F
Published: July 2010

The regulation of cytokine production and T cell proliferation by other cytokines is mandatory in mediating inflammatory responses but the full understanding is far from complete. We have previously reported increased production of IL-2 and IL-2 receptors (IL-2R) in IgA nephropathy. The present study was undertaken to examine other cytokine production during T cell activation in IgA nephropathy. Peripheral blood mononuclear cells (PBMC) from 17 IgA nephritic patients and 14 controls were cultured with phytohaemagglutinin and phorbol myristate acetate for 48 h for maximal cytokine production. IL-2Rs and IL-4 receptors (IL-4Rs) expressed on cultured PBMC were studied by a radioimmunoassay using monoclonal antibodies against these receptors. Although the total cellular IL-2R expression and percentages of T helper and T suppressor cells did not differ between the patients and controls, there was a significant increase in activated T helper cells expressing IL-2R in patients with IgA nephropathy. The total cellular IL-4R expression was elevated in IgA nephritic patients (P<0.005). IL-2 production by PBMC was raised in IgA nephritic patients compared with controls (P<0.05) but no difference in IL-4 or IL-6 production was observed. The interferon-gamma production by PBMC was significantly increased in patients with IgA nephropathy (P< 0.025). No correlation was observed between individual cytokine levels. Our data suggest there are selective increases in cytokine production in IgA nephropathy.
Recombinant interferon-gamma inhibits the expression of IL-4 receptors on human lymphocytes
Authors: BYRON K A , VARIGOS G A , WOOTTON A M
Published: July 2010

Inlerferon gamma (IFN-γ) has been shown to inhibit many of the activities of IL-4, including the induction of lgE synthesis and the proliferation of T cell clones. Here we demonstrate that IFN-γ is able to inhibit the expression of IL-4 receptors on peripheral blood lymphocytes from both normal healthy donors and from patients with chronic lymphocytic leukaemia. Inhibition was shown to be dose-dependent and did not affect the binding affinity of the receptor as shown by Scatchard analysis. IFN-γ was unable to displace labelled IL-4 from its membrane receptor, which demonstrates that IFN-γ and IL-4 do not compete for the same membrane binding protein. The ability of IFN-γ to down-regulate IL-4 receptors may be important in controlling certain immune responses.
Identification of a putative shared epitope between Coxsackie virus B4 and alpha cardiac myosin heavy chain
Authors: BEISEL K W , SRINIVASAPPA J , PRABHAKAR B S
Published: July 2010

Molecular mimicry is an important postulated mechanism for autoimmunity in viral myocarditis. The 356-1 monoclonal antibody neutralizes Coxsackie virus B4 by binding to the VPI protein and cross-reacts with mouse alpha cardiac myosin heavy chain. We used this monoclonal antibody to screen a λgt11 expression library made from CD-1 mouse hearts. Of the 48 positive plaques/106 recombinant phages examined. 14 of the strongest-reacting clones were purified for additional studies. The inserts were amplified by polymerase chain reaction and the amplified products ranged from about 150 to 1400 bp in size. Northern hybridization using these inserts demonstrated that 11 out of 14 reacted with a message equivalent to that of cardiac myosin in size. Additional Southern hybridization studies suggested that these 11 inserts contained overlapping sequences in the light meromyosin fragment of cardiac myosin. Sequence analysis confirmed that these 11 independent, recombinant clones contained a common sequence representing amino acid residues 1299-1647. Within this fragment only one isoform-specific site matched the observed reactivity pattern of 356-1 among hearts from various species. Thus, we were able to identify a putative shared epitope represented by residues 1632-1647.
Fibronectin and basement membrane components in renal amyloid deposits in patients with primary and secondary amyloidosis
Authors: WESTERMARK G T , NORLING B , WESTERMARK P
Published: July 2010

Kidney biopsies from one patieni with primary (AL) and three with secondary (AA) amyloidosis were used for an ultrastructural study of the collocalization of basement membrane proteins and the extracellular matrix protein fibronectin within amyloid deposits. Antibodies against amyloid P component, laminin. and heparan sulphate proteoglycan core protein all reacted with the basement membranes and the amyloid depositions in AA and AL amyloidosis. Monoclonal and polyclonal antibodies against collagen type IV reacted only with the basement membranes. Anti-fibronectin reaction was found in association with the basement membranes in all four cases, while labelling of amyloid depositions was found only in one of the AA amyloid cases and in the AL amyloid depositions. It is concluded that basement membrane components may be of importance for the formation of amyloid fibrils.
Kupffer cell depletionin vivoresults in preferential elimination of IgG aggregates and immune complexes via specific Fc receptors on rat liver endothelial cells
Authors: BOGERS W M J M , STAD R -K , JANSSEN D J , VAN ROOIJEN N , VAN ES L A , DAHA M R
Published: July 2010

In the present study we have investigated the clearance kinetics and tissue distribution of monomeric (m) IgG and soluble aggregates of IgG (AlgG) and immune complexes (IC) in normal and Kupffer cell (KC) depleted rats. In normal rats, clearance of mIgG occurred in a biphasic manner with a first half-life (T) or 36±3 ± 6±3 min and a second T±(T2) of 168±4±4±7 min. AlgG composed of 20-27 IgG molecules per aggregate were cleared significantly faster than mIgG with a TI of 2±5±0±1 min and a T2 of 32±5±5±6 min. KC depletion did not have a significant effect on the clearance rate of mIgG (TI: 33±4 ± 8±9 min; T2: 159±5± 12±5 min), while clearance of AlgG was delayed significantly wilh TI 4±8 ± 0±7 min and T2 41±2 ± 3±2 min. Eight minutes after injection. 77% of AIgG was found in the liver in normal rats while 62% was found in the liver of KC-depleted rats. Double immunofluorescence studies indicated that AIgG in the liver was associated with KC and endothelial cells (EC) in normal rats. In KC-depleted rats, AIgG was strongly associated with EC. A similar staining pattern was observed when IgG-immune IC were administered. The clearance of AIgG in KC-depleted rats was inhibited fully by pre-administration of high concentrations of IgG but not by pretreatment with IgA. asialofetuin (ASFe) or ovalbumin (OVA). Aggregated F(ab')2IgG was cleared with a comparable rate to mIgG from the circulation, again suggesting Fcγ receptor-mediated elimination of AIgG by EC. There was a reduced degradation of AIgG in rats depleted of KC as compared with normal rats. These data suggest binding and degradation of AIgG by EC in vivo.
Studies on the humoral immune response to a synthetic vaccine against Plasmodium falciparum malaria
Authors: SALCEDO M , BARRETO L , ROJAS M , MOYA R , COTE J , PATARROYO M E
Published: July 2010

A synthetic vaccine against the asexual blood stages of P. falciparum, the SPF 66 synthetic hybrid polymer, composed of peptides derived from three merozoite membrane proteins as well as one peptide from the sporozoite CS protein, has been developed by our group and tested in different protection assays in Aotus monkeys as well as in human volunteers. This study evaluates the humoral immune response induced by the SPf 66 protein vaccination in adult human volunteers from the Colombian Pacific coast as follows: determination of specific IgG antibody levels against SPf 66 by FAST-ELISA after each immunization; analysis of antibody reactivity with P. falciparum schizont lysates by immunoblots; and determination of the in vitro parasite growth inhibition. A clear boosting effect, dependent on time and dose, was observed in the antibody production kinetics. These antibodies also specifically recognize three proteins of the P. falciparum schizont lysate corresponding to the molecular weights of the proteins from which the amino acid sequence was derived. These sera were also capable of markedly inhibiting in vitro parasite growth.
Reduced secreted μ mRNA synthesis in selective IgM deficiency of Bloom's syndrome
Authors: KONDO N , OZAWA T , KATO Y , MOTOYOSHI F , KASAHARA K , KAMEYAMA T , ORII T
Published: July 2010

Serum IgM concentrations were low although serum IgG and IgA concentrations were normal in both our patients with Bloom's syndrome. Although the percentages of surface Ig-earing cells were not reduced, the numbers of Ig-ecreting cells were markedly reduced. The membran-ound μ(μm) and secreted μ (μs) mRN As arc produced from transcripts of a single immunoglobulin μ gene by alternative RNA processing pathways. The control of μs mRNA synthesis depends on the addition of poly(A) to μs -erminal segment. In both patients, μ mRNA was well detected but μs -erminal mRNA was scarcely detected, suggesting that μ mRNA was well transcribed but μs mRNA was not. There was, at least, no mutation or deletion in the μs -erminal coding sequence, the RNA splice site (GG/TAAAC) at the 5’ end of μs -erminal segment and the AATAAA poly(A) signal sequence in both patients. Our results suggest that selective IgM deficiency in Bloom's syndrome is due to an abnormality in the maturation of surface Ig-earing B cells into Ig-ecreting cells and a failure of μs mRNA synthesis. Moreover, reduced μs mRNA synthesis may be due to the defect on developmental regulation of the site at which poly(A) is added to transcripts of the μ gene.
Increased CD11/CD18 expression on peripheral blood leucocytes of patients with sarcoidosis
Authors: SHAKOOR Z , HAMBLIN A S
Published: July 2010

Sarcoidosis is a multisystem disease of unknown etiology characterized by non-caseating granulomata, formed mainly from macrophages surrounded by lymphocytes and plasma cells. Using a novel method for the preparation of blood leucocytes for flow cytometry, we report increased expression of LeuCAMs (CD11/CD18) on peripheral blood leucocytes of 11 Caucasian and 10 Afro-Caribbean patients with sarcoidosis compared with age-, sex- and race-matched controls. Whilst the percentages of the cells expressing CD11/CD18 were no different, the density, expressed as mean fluorescence intensity (M FI), was greater for all leucocytes in sarcoids than in normal individuals. The expression of intercellular adhesion molecule-1 (ICAM-1), a ligand for LFA-1 which is expressed on all leucocytes, was not significantly different from normal, whereas HLA-DR was expressed more intensely on sarcoid monocytes (P< 0·01) and blood lymphocytes (P< 0·005) than control cells. Our findings are consistent with leucocyte activation although we were unable to confirm reports of elevated tumour necrosis factor-alpha (TNF-α) in the patients' plasma using an ELISA. Increased expression of adhesion molecules on peripheral blood leucocytes may play a role in the cellular extravasation, aggregation, and granuloma formation seen in sarcoidosis.
Intravenous immunoglobulin in the treatment of paediatric cerebral malaria
Authors: TAYLOR T E , MOLYNEUX M E , WIRtMA J J , BORGSTEIN A , GOLDRtNG J D , HOMMEL M
Published: July 2010

Hyperimmune globulin can inhibit and reverse the cytoadherence between Piasmodium falciparum- infected erythrocytes and melanoma cells in vitro. Cytoadherence is believed to mediate disease in cerebral malaria. Therefore we studied the efficacy of i.v. immunoglobulin, purified from the plasma of local semi-immune blood donors, as an adjunct to standard treatment for cerebral malaria in Malawian children.

The immunoglobulin preparation (IFAT antimalarial antibody titre 1:5120) recognized erythrocyte-associated antigens of each of 22 Malawian P. falciparum isolates studied, and reversed binding of Malawian isolates to melanoma cells. Immunoglobulin did not reverse binding to human monocytes or to cells of the human hystiocytic lymphoma cell line U937.

Thirty-one children with P. falciparum parasitaemia and unrousable coma were enrolled. All were treated with i.v. quinine dihydrochloride; in addition patients were randomized to receive either immunoglobulin (400 mg/kg by i.v. infusion over 3 h) or placebo (albumen and sucrose by similar infusion) in a double blind trial with sequential analysis. Of 16 patients receiving immunoglobulin, five (31%) died and five survivors had neurological sequelae. Of 15 patients receiving placebo, one (7%) died and two had sequelae. Parasite clearance, fever clearance and coma resolution times in survivors were similar in the two groups.

Although the difference in outcome between the two groups was not significant, the trial was stopped because immunoglobulin was demonstrated not to be superior to placebo.
Thyroid autoimmunity and hypothyroidism during long-term treatment with recombinant interferon-alpha
Authors: GISSLINGER H , GILLY B , WOLOSZCZUK W , MAYR W R , HAVELEC L , LINKESCH W , WEISSEL M
Published: July 2010

Forty-five patients with myeloproliferative or myelodysplastic syndromes, treated with recombinant interferon-alpha (rIFN-α) for a minimum of 1 up to 4 years, were examined for the occurrernce of thyroid autoimmunity. During treatment, the rate of thyroid autoimmunity rose to more than 20%. The deerease in severity and frequency of thyroid autoimmunity after withdrawal of IFN shows that this is a potentially reversible side effect. The key determinant for the manifestation of this IFN-related autoimmune phenomenon seems to be a predisposition for autoimmunity, since patients with initially detectable thyroid antibodies are prone to exacerbations of thyroid autoimmunity. Concurrent with thyroid autoimmunity. hypothyroidism occurred but did not correlate with the levels of thyroid antibodies, although severe hypothyroidism in two patients was accompanied by increased levels of thyroid antibodies. This investigation shows that thyroid autoimmunity and consecutively hypothyroidism must be expected in certain patients treated with rIFN-a during long periods. Furthermore, it may be assumed that IFN-α does not induee the development of autoimmunity, but rather enhances the levels of pre-existent thyroid antibodies.
CD4+ T cells clear virus but augment disease in mice infected with respiratory syncytial virus. Comparison with the effects of CD8+ T cells
Authors: ALWAN W H , RECORD F M , OPENSHAW P J M
Published: July 2010

Respiratory syncytial (RS) virus-specific T cell lines were derived from the spleens of BALB/c mice primed by intranasal infection with RS virus. The lines were expanded by repealed antigenic stimulation in vitro, and separated into CD4+ and CD8+ T cell-enriched fractions by immunomagnetic adhesion. The effects of passive transfer of these fractions into RS virus infected mice were observed. The most severe immunopathological changes were seen in mice receiving CD4+ cells. Transfer of CD4+, CD8+ or both cell fractions caused RS virus-infected mice to become ill and lose weight. Both cell lines caused an increase in the severity of lung pathology (as monitored by bronchoalveolar lavage) with the appearance or lung haemorrhage and polymorphonuclear cell efflux. In addition, recipients of CD4+ cells developed striking pulmonary eosinophilia. In CD4+ cell recipients. 5×105 cells were sufficient to decrease lung virus titr, whereas 2×106 CD8+ cells were needed to produce a similar effect. The unseparated T cell line and the CD4+ cell fraction secreted significant amounts of IL-3, IL-4 and IL-5 (P < 0·001). High levels of IL-2 were produced only by the unseparated T cell line. The CD8+ cell fraction secreted IL-3 only. The results show that, cell-for-cell CD4+ cells are more anti-viral and more immunopathogenic than CD8+ cells in RS virus infected mice. Such effects may have contributed to the augmented disease seen in some infants vaccinated against RS virus.
Lymphotropic viruses in ‘common variable’ immunodeficiency—PCR analysis of lymphocyte DNA for HIV-1 and HHV-6
Authors: SA'ADU A , THOMSON B J , BOUNTIFF L , WEBSTER A D B
Published: July 2010

DNA of circulating lymphocytes obtained from 17 patients with ‘common variable’ immunodeficiency (CVID) was tested for the presence of HIV-1 sequences, using a variety of primers complementary to conserved regions of HIV-1 genomes. None of the patients was positive. The DNA from 12 CVID patients was tested for the presence of human herpes virus 6 (HHV-6), using appropriate primers for this virus which is known to remain latent following primary infection in the majority of people during childhood. Similar numbers of patients and normal subjects were positive (about 30%), suggesting that CVID patients are not particularly prone to reactivation of this virus. Despite previous anecdotal reports of an unexplained association between HIV-1 and CVID, we conclude that there is no evidence that this virus is implicated in the pathogenesis of CVID. A role for HHV-6 also seems unlikely.
T cells are responsible for the enhanced synovial cellular immune response to triggering antigen in reactive arthritis
Authors: SIEPER J , BRAUN J , WU P , KINGSLEY G
Published: July 2010

In reactive arthritis (ReA) there is specific proliferation of synovial fluid (SF) mononuclear cells (MNC) to the triggering bacterial antigen; comparatively little or no response is seen in peripheral blood (PB). To investigate the mechanism of this elevated local immune response, we examined patients with typical ReA who showed an enhanced antigen-specific synovial immune response in bulk culture. Using separated fractions of T cells and antigen-presenting cells (APC) from PB and SF we showed that the synovial T cells rather than SF APC are responsible for the specific proliferation. By limiting dilution analysis, the frequency of T cells responding to the specific antigen was found to be significantly increased compared with the frequency of irrelevant antigen-specific T cells. Furthermore, the frequency of T cells responding to the specific antigen was higher in SF (between 1/619 and 1/4846, mean 1/2389) than in PB (between 1/1286 and 1/16279, mean 1/7350). We conclude that the specific synovial cellular immune response in ReA is mainly due to an expansion of antigen-specific T cells within the joint. However, the non-specific hyper-reactivity of SF T cells and differences between SF and PB APC may make a more minor contribution.
Defective IL-6 secretion in HIV-infected haemophilia patients
Authors: WEIMER R , ZIPPERLE S , DANIEL V , ZIMMERMANN R , OPELZ G
Published: July 2010

To study the role of IL-6 in HIV-induced B cell defects, in vitro B cell responses and IL-6 secretion were determined simultaneously in 67 haemophilia patients, Twenty-three palients were HIV (Group 1). 27 HIV+ stage CDC II, III (Group 2). and 17 vi-ere HIV4 stage CDC IV (Group 3). Pokeweed mitogen (PWM) was used for TeelI-dependent and Staphylococcus aureus Cowan 1 (SAC I) for T cell-independent B cell stimulalion. B cell differentiation was assessed in a reverse haemolytic plaque assay and by ELISA determination of IgG and IgM in culture supernatants. An ELISA was used to measure IL-6 in plasma and culture supernatants, HIV+ patients showed impaired immunoglobulin-secreting cell (ISC) responses after T cell-independent and T cell-dependent stimulation (P < 0·000l and P<0·01, respectively), whereas IL-6 secretion. IgM and IgG responses were comparable to those in hcallhy controls. HIV+ patients at stage CDC II, Ml or IV demonst rated significantly reduced mitogen-stimulated IL-6 secretion (P < 0·05. PWM; P < 0·001. SAC lj as well as impaired ISC and IgG responses (P < 0·01. PWM; P≤ 0·0001, SAC I), CDC IV patients showed reduced IgM responses in addition (P < 0·02, PWM; P < 0·0005. SAC I). Plasma iL-6 levels were elevated both in HIV+ patients (CDC II, III patients: 165 ± 73 pg/ml. P < 0·005; CDC IV patients: 58 ± 18 pg/ml, P < 0·001) and in HIV patients (283 ± 65 pg/ ml. P<0·0001) which appeared to be a T cell effect induced by treatment with haemophilia factor concentrates. Our data provide evidence for different types of B cell deficiencies in HIV patients (impaired ISC response only) and HIV+ patients (impaired ISC as well as IL-6 and IgM /lgG responses). The defective IL-6 secretion in HIV+ patients is likely to affect terminal B cell differentiation and this may explain the reduced immunoglobulin secretion in these patients in response to antigenic challenge.
Differential cytokine gene expression and secretion after phagocytosis by a human monocytic cell line ofToxoplasma gondiicompared withMycobacterium tuberculosis
Authors: FRIEDLAND J S , SHATTOCK R J , JOHNSON J D , REMICK D G , HOLLIMAN R E , GRIFFIN G E
Published: July 2010

Toxoplasma gondii infection may be clinically silent in immunocompetent individuals but may cause fatal disease in immunocomipromised patients such as those with HIV infection. Proinflammatory cytokines are known to be important in murine resistance to T. gondii but there are no data from human models of infection. We have investigated whether phagocytosis of T. gondii, of Mycobacterium tuberculosis (a pathogen which elicits a granulomatous host immune response) and of men latex particles by THP-I cells, a human monocytic line, caused gene expression and secretion of tumour neerosis factor (TNF), IL-6 and IL-8. These cytokines are important in recruitment and activation of T lymphocytes, and both TNF and IL-6 may have direct antitoxoplasmacidal and antimycobacterial activity. Phagocytosis of T. gondii by THP-1 cells resulted in minimal gene expression and secretion of TNF, IL-6 and IL-8 similar to that following phagocytosis of inert latex particles. In contrast, phagocytosis of M. tuberculosis resulted in increased gene expression of TNF and IL-8 as well as increased secretion of all three cytokines. particularly IL-8. These observations may partially explain the frequency of non-inflammatory host responses to T. gondii in immunocompetent individuals.
Complement activation in septic baboons detected by neoepitope-specific assays for C3b/iC3b/C3c, C5a and the terminal C5b-9 complement complex (TCC)
Authors: MOLLNES T E , REDL H , HØGÅSEN K , BENGTSSON A , GARRHD P , SPEILBERG L , LEA T , OPPERMANN M , GÖTZE O , SCHLAG G
Published: July 2010

We have investigated the cross-reactivity of various species in ncocpitope-specific methods for quantification of human complement aclivatlon products. In contrast to most other species examined, baboon showed a substantial cross-reactivity supporting a high degree of homology between human and bahoon complement. An assay for C3b, iC3b and C3c (MoAh bH6) showed moderately good reactivity, in contrast to a C3a assay which did not cross-react. Excellent reactivity was found for C5a using MoAbs C17/5 and G25/2. The reactivity of an established TCC assay (MoAb aEU to a C9 neoepitopc and polyclonal antibody to C5) was improved substantially by replacing the anti-C5 antibody with a new MoAb to C6 particularly selected on the basis of baboon cross-reactivity. Plasma samples from baboons receiving 2·5 · 109 and 1·0 ± 1·010 live Escherichia coli bacteria/kg were examined with the assays described. In vivo complement aetivation with the lowest dose was moderate and kept under control, in contrast to the highest dose, where an uncontrolled increase in all activation products continued throughout the infusion period. These results support the hypothesis that sufficiently high amounts of endotoxin lead to uncontrolled activation of complement as seen in irreversible septic shock. The results arc discussed with particular emphasis on activation of the terminal complement pathway.
Short-term administration of anti-L3T4 MoAb prevents diabetes in NOD mice
Authors: KURASAWA K , SAKAMOTO A , MAEDA T , SUMIDA T , ITO I , TOMIOKA H , YOSHIDA S , KOIKE T
Published: July 2010

We treated 2-week-old and 8-week-old non-obese diabetic (NOD) mice with 1 mg of anti-L3T4 MoAb weekly for 4 weeks. This short-term treatment of anti-L3T4 MoAb prevented the development of overt diabetes in NOD mice, in both groups, even after cessation of the therapy. However, there were overt mononuclear cell infiltrations in the majority of islets, and no appreciable differences in the degree of insulitis between treated and control mice. There were also no significant differences in the percentage of L3T4+ T cells expressing Vβ5, Vβ8 and Vβ11 antigens between the treated and the control group. In contrast, most of male NOD mice injected with 200 mg/kg of cyclophosphamide did not become diabetic when the spleen cells from the MoAb-treated female NOD mice were transferred to these animals 48 h before the cyclophosphamide injection. Thus, the tolerance induced by the short-term administration of anti-L3T4 MoAb to NOD mice may not be due to clonal deletion, but rather to newly generated suppressor cells in the animals.
Control of human thyroid autoantibody production in SCID mice
Authors: MACHT L M , CORRALL R J , BANGA J P , ELSON C J
Published: July 2010

In order to determine the phenotype of the cells required for thyroid autoantibody production, peripheral blood mononuclear cells (PBMC) from patients with autoimmune thyroid disease (AITD) were transferred to severe combined immunodeficient (SCID) mice. The production of human IgG. thyroglobulin (Tg) antibody and thyroid peroxidase (TPO) antibody in the SCID recipients was monitored for up to 4 months. PBMC from 10 of 13 AITD patients produced substantial IgG(× 100 μ/ml) and detectable Tg and TPO antibodies in recipient mice. PBMC pretreated to deplete or enrich T cells produced low or undetectable thyroid-specific antibody in SCID mice. Depletion of CD4+ T cells resulted in much lower or undetectable IgG, Tg and TPO antibodies compared with levels seen in recipients of control PBMC. By contrast, depletion of CD8+ T cells from the PBMC had no overall effect on autoantibody production, although with PBMC from some patients CD8+ depletion possibly enhanced both IgG and autoantibody production. In eight of 10 experiments, autoantibody levels reached maximal titres before total IgG levels peaked. It is considered that thyroid autoantibodies are produced from memory B cells activated in SCID mice and that this activation is T cell- and CD4+ T cell-dependent.
Characterization of cytokine production in infectious mononucleosis studied at a single-cell level in tonsil and peripheral blood
Authors: ANDERSSON J , ANDERSSON U
Published: July 2010

Cytokine profile and production was studied at a single-cell level in cells obtained from 14 patients with acute infectious mononucleosis (IM), with less than 7 days of symptomatic disease, by use of cytokine-specific MoAbs and indirect immunofluorescence technique. In producer cells, all the studied cytokines, except IL-1, accumulated in the Golgi system, which resulted in a characteristic morphology of the staining. Less than one in a thousand mononuclear cells obtained directly from IM blood and stained within 2 h of sampling produced IL-2, interferon-gamma (IFN-γ), IL-4, IL-5, IL-6, IL-10, GM-CSF, tumour necrosis factor-alpha (TNF-α) or TNF-β, spontaneously. However, these cells were induced to cytokine synthesis by T cell receptor ligation in vitro using immobilized anti-CD3 MoAbs for 2–3 h restimulalion under conditions which did not activate normal cells. By this approach 168±120 cells/10000 peripheral blood mononuclear cells produced IFN-γ as compared with 10±8 cells/10000 non-stimulated cultured cells obtained from IM patients (P < 0.001) and 1 /10000 cells obtained from healthy controls, respectively. No induced production of IL-2, IL-3, IL-4, IL-5, IL-10, GM-CSF or TNF-β was detected in IM cells obtained from peripheral blood by this restimulalion. In contrast, a spontaneous cylokine production was evident in tonsil material obtained from four IM patients tonsilectomized because of respiratory obstruction. From this site 160±40 cells/10000 cells produced IL-2, 40±30 cells IL-6. 30±30 cells TNF-β and 35±25 cells IFN-γ, respectively. No such spontaneous IL-2, IL-6, TNF-γ or IFN-γ production was evident in control cells obtained from patients tonsilectomized because of chronic tonsil hyperplasia.
HIV induces deletion of T cell receptor variable gene product-specific T cells
Authors: BANSAL A S , GREEN L M , KHOO S H , PUMPHREY R S H , HAENEY M R , MANDAL B K
Published: July 2010

Using a flow cytometrie method, CD4+, CD8+, αβ TCR+ and TCR variable region gene product (TVRGP)-specific T cells were analysed in heallhy heterosexual males (HHeM). HIV-seronegative homosexual males (SNHM). asymptomatic seropositive homosexual males (ASPH) and homosexual males with AIDS who were either well (AIDS-A). or unwell in hospilal (AIDS-B). Total CD4+ and CD8+ T cell numbers were similar in HHeM and SNHM. CD4+ T cells were significantly reduced in ASPH relative to both HHeM and SNHM and in AIDS-A and AIDS-B relative to SNHM. TVRGP-specific T cells expressed as a percentage of TCR αβ+ cells showed no significant difference in HHeM. SNHM and AIDS-B. The proportion of αβ+ cells expressing the Vβ5·1, Vβ12 and Vα2 gene product (GP) was. however, significantly reduced in ASPH and AIDS-B relative to HHeM. SNHM and AIDS-A. Possible causes of TVRGP-specific T cell deletion arc discussed.
Increased expression and occupancy of receptors for tumour necrosis factor on blood monocytes from tuberculosis patients
Authors: CADRANEL J , PHILIPPE C , PHILIPPE B , MILLERON B , FOUQUERAY B , MAYAUD C , BAUD L
Published: July 2010

Blood monocytes from tuberculosis patients release high amounts of tumour necrosis factor-alpha (TNF-α). Because the biological efficiency of TNF-ot would depend on the expression of TNF-α receptors on target cells, we thought to analyse the capacity of blood monocytes from a group of patients with pulmonary tuberculosis to bind 125I-TNF-α. We report a slight but not significant enhancement in specific binding of 125I-TNF-α on monocytes of 15 consecutively studied patients compared with 10 controls. Per cent cell surface bound and internalized 125I-TNF-α was identical in the two groups. To evaluate the receptor occupancy by endogenously generated TNF-α, similar experiments were performed after cell exposure to low-pH glycine buffer. Under these conditions, specific binding of 125I-TNF-α was significantly higher on tuberculosis monocytes compared with control monocytes. Moreover, the occupancy of TNF-α receptors by endogenously generated TNF-α that was found to be significantly higher on tuberculosis monocyles than on control monocytcs. was directly related to the enhanced capacity of mononuclear cells to generate TNF-αIn vitro, It normalized after 3 months of antituberculous therapy. Scatchard analysis of the binding data revealed that tuberculosis infection caused a significant increase in high affinity 125I-TNF-α binding to monocyles without any significant change in the dissociation constant. Collectively, these results indicate an up-regulation of TNF-α generation and binding to blood monocytes in patients with pulmonary tuberculosis. They provide support to the hypothesis that TNF-α is of critical importance in the pathogenesis of this infection.
Lymphoproliferative responses to a merozoite surface antigen of Plasmodium falciparum: preliminary evidence for seasonal activation of CD8+/HLA-DQ-restricted suppressor cells
Authors: RILEY E M , MORRIS-JONES S , TAYLOR-ROBINSON A W , HOLDER A A
Published: July 2010

We have investigated the phenotype of human lymphocytes responding to a defined Plasmodium falctparum malaria antigen in vitro. Cells were obtained from the peripheral blood of malaria-immune donors from an endemic area of West Africa and were tested for proliferation in response lo cloned fragments of a merozoite surface protein (PfMSPI). Depletion and inhibition studies indicated that the majority of proliferating cells were CD4+ and restricted by HLA-DR or -DQ. A proportion of responding cells appeared to be CD8+, but their response was dependent on help from CD4+ cells. In two donors there was evidence that low responses could be enhanced by removal of CDS+ cells and/or blocking of antigen presentation by anti-HLA-DQ antibodies. This phenomenon was observed in cells collected during the wet (malaria transmission) season but not in cells collected from the same individual during the dry season.