bluuebunny/crossref_metadata_2025 huggingface dataset:
Anti-thyroglobulin autoantibodies in sera from patients with chronic thyroiditis and from healthy subjects: differences in cross-reactivity with thyroid peroxidase
Authors: NAITO N , SAITO K , HOSOYA T , TARUTANI O , SAKATA S , NISHIKAWA T , NIIMI H , NAKAJIMA H , KOHNO Y
Published: July 2010

A significant portion (about 12·7%) of healthy subjects was found to contain anti-thyroglobulin (anti-Tg) antibodies in their sera. We compared the binding activities of these antibodies and of anti-Tg autoantibodies from sera of patients with chronic thyroiditis with human thyroid peroxidase (TPO). The results obtained by ELISA indicated that out of 10 healthy subjects with anti-Tg antibodies, only four had anti-Tg antibodies capable of binding to TPO, whereas anti-Tg autoantibodies from almost all patients with chronic thyroiditis possessed high binding activities to TPO. By use of the immunoprecipitation method, it was also shown that although all anti-Tg autoantibodies from patients precipitated TPO, a majority of anti-Tg antibodies from healthy subjects could not precipitate TPO. Such findings cannot be ascribed to the differences in levels of anti-Tg autoantibodies and anti-TPO autoantibodies in sera and the differences in avidities of anti-Tg antibodies in sera between healthy subjects and patients with chronic thyroiditis. Thus, it can be concluded that anti-Tg antibodies from healthy subjects differ from those of patients with chronic thyroiditis with respect to TPO binding, probably due to difference in fine specificities of these anti-Tg antibodies.
Cells with dendritic morphology and bright interleukin-lα staining circulate in the blood of patients with rheumatoid arthritis
Authors: BARKLEY D E H , FELDMANN M , MAINI R N
Published: July 2010

Freshly isolated peripheral blood mononuclear cells (PBMC) from 10 healthy volunteers, 28 patients with rheumatoid arthritis (RA), eight patients with osteoarthritis, and five patients with ankylosing spondylitis were examined for interleukin-1α (TL-1α) and interleukin-1β (IL-1β) production using monoclonal antibodies and an indirect immunofluorescent method. In freshly isolated PBMC from healthy controls very few cells were stained for either IL-1 type. All 20 RA patients who were not receiving parenteral gold therapy had PBMC staining for IL-1α. In these patients, up to 7.5% of PBMC showed bright IL-1α staining (range 12·7·5%). No IL-1β staining was seen. These IL-1α staining cells had a dendritic morphology and the percentage of cells staining correlated well with levels of C-reactive protein, an index of disease activity in these RA patients. Significantly fewer IL-1α staining cells were present in the peripheral blood of RA patients receiving gold therapy and in the blood of patients with osteoarthritis and ankylosing spondylitis. These IL-1α-containing cells. circulating in the blood of RA patients and correlating with disease activity have not been previously described. These results support the idea that IL-1α plays an important role in the pathogenesis of rheumatoid inflammation.
Soluble histocompatibility antigens in synovial fluids of patients with rheumatoid arthritis
Authors: STEVENSON F K , DOUGLASS W A , SPELLERBERG M B , WALTERS M T , CAWLEY M I D
Published: July 2010

Soluble histocompatibility antigens of the class II region have been detected in synovial fluids obtained from patients with rheumatoid arthritis. A capture immunoassay involving two monoclonal antibodies was used; interference by rheumatoid factor, which is a feature of such assays, was overcome by mild pretreatment of fluids with 2-mercaptoelhanol. No HLA class II antigen could be detected in matched sera from patients, even when levels were high in synovial fluids. Released HLA-class II material was of high molecular weight (> 1000 kD) and was linked to HLA-class I antigen. However, no significant amounts of other common cell surface antigens were detected in the complex, suggesting a preferential release of MHC antigens from cells of the inflamed synovium. Attempts to induce production of similar material from a cell line which expresses HLA class II strongly at the cell surface, by stressing the cells in various ways did not succeed, indicating that release is an active process.
Parasite antigens recognized by patients with cutaneous leishmaniasis
Authors: JAFFE C L , SHOR R , TRAU H , PASSWELL J H
Published: July 2010

Humoral and cell-mediated responses to crude and purified parasite antigens were examined in patients with active cutaneous leishmaniasis caused by Leishmania major. The patients had serum antibody titres against parasite lysates ranging from 1/500 to 1/10000 and recognized multiple components by Western blotting with molecular weights between 5000 and >200000. Several components, particularly at 5 and 50 kD, were recognized by most of the patients. The lymphoproliferative responses to two pure antigens, promastigote surface protease and lipophos-phoglycan. both considered potential candidates for the development of a human vaccine were measured. No response to promastigote surface protease was observed even at 10 μg/ml; however, weak proliferation to lipophosphoglycan was consistently present. T cell blots examining peripheral blood lymphocyte proliferation, showed antigen-specific responses to a 72–82 kD component in promastigote lysates.
Cells and mediators which participate in immunoglobulin synthesis by human mononuclear cells. III. Null cells secrete a factor(s) (human immunoglobulin synthesis/secretion-facilitating factor) that can replace the null cells in the synthesis of immunoglobulin by cultured B cells
Authors: TAYLOR S , JODOUIN C A , RICHTER M
Published: July 2010

In the accompanying communication, it was demonstrated that the null cells, the TM cells, monocytes and PWM are all obligatory participants in the synthesis and secretion of immunoglobulins by human B cells in culture. Here we demonstrate that the null cells secrete a factor, referred to as human immunoglobulin synthesis/secretion-facilitating factor (HISFF) that can replace the null cells in the cultures. HISFF is distinct from the known T cell-derived interleukins. HISFF functions in an HLA-unrestricted fashion since it can facilitate the synthesis and secretion of immunoglobulins by allogeneic B cells. The null cells cultured with TM helper cells and PWM required monocytes in the culture in order to secrete HISFF. Furthermore, B cells cultured with TM cells in medium containing HISFF, monocyte-derived factors and PWM nevertheless required monocytes in order to respond to the HISFF signal. Thus, the monocyte plays a pivotal role in the secretion of and response to HISFF. Normal levels of immunoglobulin were synthesized even when HISFF was added to the cultures of B cells, TM cells and monocytes, in the presence of PWM, as late as day 6 of the 7 day culture. We conclude that the null cells participate in immunoglobulin synthesis by the B cells by secreting a soluble mediator, HISFF, capable of replacing the null cells in the culture; and that the HISFF signal is the last signal received by the B cell before it begins to synthesize and secrete immunoglobulins.
Bispecific antibody: a tool for diagnosis and treatment of disease
Authors: SONGSIVILAI S , LACHMANN P J
Published: July 2010

Antibodies with two distinct binding specificities have great potential for a wide range of clinical applications as targeting agents for in vitro and in vivo immunodiagnosis and therapy, and for improving immunoassays. They have shown great promise for targeting cytotoxic effector cells, delivering radionuclides, toxins or cytotoxic drugs to specific targets, particularly tumour cells. We discuss potential applications of bispecific antibodies, the theoretical basis and problems associated with their production and purification. Cell fusion and chemical conjugation techniques, and propose a new manufacturing strategy by genetic engineering. This approach has enormous potential applications for producing tailor-made bispecific antibodies, and will enable widespread clinical uses of these antibodies both for diagnostic purposes and therapy.
Neutrophil-potentiating factors released from stimulated lymphocytes; special reference to the increase in neutrophil-potentiating factors from streptococcus-stimulated lymphocytes of patients with Behçet's disease
Authors: NIWA Y , MIZUSHIMA Y
Published: July 2010

The potentiating effect of the soluble factors released from normal or diseased lymphocytes on neutrophil functions were investigated in the presence or absence of mitogens and wall preparations of Streptococcus pyogenes. When normal T lymphocyte populations were stimulated with T cell mitogens or with streptococcal preparations, the supernatants from these cultures potentiated neutrophil chemotaxis, phagocytosis and O2- generation. Upon gel-filtration of these stimulated lymphocyte supernatants, the neutrophil-potentiating activity was inactivated by trypsin or by a 30-min incubation at 130°C, but was not affected by acid treatment at pH 2 or heat treatment at 56°C for 60 min. Its activity was almost not affected by antisera against human interleukin-1, interleukin-2, interferon-gamma or tumour necrosis factor. With the stimulation of T cell mitogens, the supernatants released from the lymphocytes of not only the patients with Behçet's disease but also healthy and diseased controls enhanced neutrophil functions. However, supernatants from streptococcal preparation-stimulated lymphocytes from patients with Behçet's disease had a higher potentiating effect on neutrophil functions. Our study suggests that the enhanced neutrophil functions in patients with Behçet's disease may be related to an abnormally high level of circulating activated T cells in these patients.
The inductive effect of interleukin-4 on IgG4 and IgE synthesis in human peripheral blood lymphocytes
Authors: ISHIZAKA A , SAKIYAMA Y , NAKANISHI M , TOMIZAWA K , OSHIKA E , KOJIMA K , TAGUCHI Y , KANDIL E , MATSUMOTO S
Published: July 2010

Using murine monoclonal antibodies against human IgG subclasses, specific and sensitive ELISAs assay to quantify the four human IgG subclasses in cell culture supernaUints were established. The effect of human recombinant interleukin-4 (IL-4) on the regulation of IgG subclasses by normal peripheral blood lymphocytes was investigated. In addition to the enhancement of lgE synthesis. IL-4 preferentially induced IgG4 synthesis in vitro, whereas IL-4 had no effect on IgGl, IgG2, and IgG3 synthesis. IL-4-induced IgG4 production was blocked in a dose-dependent manner by recombinant interferon-gamma and anti-human IL-4 monoclonal antibody. Collectively, this data indicates that IL-4 plays an important regulatory role in both IgG subclass and IgE synthesis.
Absence of Epstein-Barr virus-specific, HLA class II-restricted CD4+ cytotoxic T lymphocytes in infectious mononucleosis
Authors: ENSSLE K H , FLEISCHER B
Published: July 2010

Cytotoxic T lymphocytes (CTL) with the CD4+ phenotype that recognize major histocompatibility complex (MHC) class II antigens are detectable very frequently in cultures of human alloreactive or virus-specific T cells. The significance of these CD4+ CTL for an immune reaction in vivo is not clear, Since Epstein-Barr virus (EBV) transformed B cells express HLA-class I and class II antigens equally well both CD8+ and CD4+ CTL should be stimulated during an acute EBV infection. We analysed the MHC specificity and the phenotype of EBV-specific CTL from patients with infectious mononucleosis (IM). When tested directly without any previous culture, T cells from patients in the acute phase of IM showed specific MHC-restricted cytotoxicity against the autologous B cell line. Addition of a HLA class I specific monoclonal antibody (MoAb) but not of a HLA class II specific MoAb resulted in a complete blocking of the lytic activity. Cell sorting revealed that the entire cyloloxic activity was present in the CD8+ fraction whereas no specific CTL were detectable in the CD4+ fraction. The absence of cytotoxicity in CD4+ cells was not due to a lack of activation of these cells since both CD8+ and CD4+ cells were activated in situ, showing spontaneous growth in interleukin-2 (IL-2) and expressing the activation marker TP103. Frequency estimation revealed that 1/300-1/600 CD8+ but only 1/2000-1/4000 CD4+ T cells gave rise to a specific CTL colony after 10 days. If CD4+ colonies were tested repeatedly for cytotoxicity we found that CD4+ CTL acquired their cytotoxicity during in vitro culture. In addition, we isolated EBV-specific CD4 T cell clones able to lyse their stimulator cells in the presence but not in the absence of leclin even after a long period of culture. Taken together our results show that cytotoxicity mediated by CD4+ T cells does not play a role in an anti-viral immune response.
Peptide inhibitors of C3 breakdown
Authors: PEAKE P , SZELKE M , JONES D M , SINGLETON A , SUEIRAS-DIAZ J , LACHMANN P J
Published: July 2010

We have investigated the development of substrate-based inhibitors of complement enzymes. Sequences around the scissile Arg77-Ser78 bond of C3 have been synthesized and tested as inhibitors of C3 convertase. The best inhibition was found with the tetrapeptide Ac-Arg-Ser-Asn-Leu-OH (H-576); extending this sequence in cither direction reduced inhibitory activity. Preliminary experiments with peptides in which the scissile bond —CO—NH— was replaced with non-hydrolysable moieties such as— CO—CH2— (H-497) and —CH2—NH— (H-336) failed to show enhanced inhibition. One of the longer chain inhibitors H-416 containing DArg77-Ser78 was unexpectedly found to potentiate iC3 cleavage by factors I and H but did not inhibit the intact alternative pathway. The same peptide also bound to factor H. Tt is concluded that the binding requirements of the C3 convertase are more sophisticated than can be satisfactorily imitated simply by linear sequences around the scissile bond of C3.
Human milk contains proteins that stimulate and suppress T lymphocyte proliferation
Authors: MINCHEVA-NILSSON L , HAMMARSTRÖM M-L , JUTO P , HAMMARSTRÖM S
Published: July 2010

The modulatory effect of human milk proteins from colostrum and late milk on the proliferative response of human T lymphocytes activated by mitogens (OKT3 and leucoagglutinin from Phaseolus vutgaris) and alloantigens was studied. High concentrations (10-100 μg/ml) of crude colostral milk proteins had an inhibitory effect on T cell growth while low concentrations (0.1-1 μg/ml) enhanced T cells growth. In contrast, proteins from late milk did not inhibit T lymphocyte proliferation while the enhancing effect was retained. Colostrum was fractionated by ammonium sulphate precipitation and gel filtration on sepharose 6B. The inhibitory activity was recovered in a protein fraction containing Iactoferrin as its major component. Lactoferrin was. however, not responsible for the observed inhibition. On the contrary. lactoferrin in most cases augmented the proliferative response induced by polyclonal activators. The inhibitory activity was found to bind coneanavalin A-sepharose suggesting an association with glycoprotein. Inhibitory fractions contained glycoproteins of the following molecular sizes26, 74/76 (doublet),84, 145 and 160 kD under reducing conditions. The inhibitory effect appeared to be lymphocyte specific since the active fraction did not inhibit the growth of tissue culture cells (I-leLa cells and human fibroblasts) or bacteria. Furthermore, the fraction was not toxic for lymphocytes. The inhibitory colostrum factor may prevent the newborn from overreacting immunologically against the environmental antigens encountered at birth.
Detection of IL-2 at mRNA and protein levels in synovial infiltrates from inflammatory arthropathies using biotinylated oligonucleotide probes in situ
Authors: HOWELL W M , WARREN C J , COOK N J , CAWLEY M I D , SMITH J L
Published: July 2010

A non-radioactive in situ hybridization method for IL-2 mRNA detection based on the use of four biotinylated oligonucleotide probes, plus appropriate positive and negative control probes was developed and applied to synovial surgical and needle biopsies from rheumatoid arthritis (RA), spondyloarthropathy (SpA), psoriatic arthritis (PsA) and juvenile chronic arthritis (JCA) patients. In eight surgical biopsies (six RA, one SpA, one PsA) this non-radioactive system showed similar sensitivity to that of a previously described 32P-labelled probe system, and in addition detected IL-2 mRNA in five out of seven biopsies from SpA and PsA patients and in two out of two JCA needle biopsies. IL-2 mRNA was found in the absence of IL-2 protein in RA biopsies (six surgical, 12 needle), but variable amounts of IL-2 protein were detected in six out of seven needle biopsies from SpA. PsA and JCA patients, where CD3+ lymphoid infiltrates were present. These data suggest differences in IL-2 regulation and expression in RA and non-RA inflammatory arthropathies.
Functional characterization of skin-infiltrating lymphocytes in atopic dermatitis
Authors: REINHOLD U , KUKEL S , GOEDEN B , NEUMANN U , KREYSEL H W
Published: July 2010

Skin-infiltrating lymphocytes (SIL) were isolated from skin biopsies of patients with hyperimmunoglobulin E (IgE) atopic dermatitis (AD) and expanded in vitro in the presence of IL-2 in combination with IL-4. Phenotypic analysis of skin-derived cells revealed the predominance of CD4+ T helper/inducer phenotype in SIL populations. In 3H-thymidine incorporation assays, SIL showed proliferation in response to IL-2, IL-3, IL-4, ionomycin (Io)+ 12-o-tetradecanoyl-phorbol-13-acetate (TPA) and OKT3 + TPA. OKT4 with and without TPA did not induce proliferation. Tumour necrosis factor alpha (TNF-α) did not block proliferative responses of SIL to IL-2 and IL-4. Cultured SIL showed no cytotoxic activity against K562 and Jurkat target cells. Expanded skin-derived T cells were tested for their capacity to secrete several cytokines in vitro. SIL secreted significant amounts of IL-4, GM-CSF and TNF-α upon stimulation with mitogens but failed to secrete IFN-γ. Io in combination with phorbolester induced the secretion of larger amounts of IL-4, GM-CSF, TNF-α and low amounts of IFN-γ. The data indicate that SIL derived from AD lesions were defective in their capacity to secrete IFN-γ but were enriched in T cells capable of producing IL-4 upon stimulation. The results support the possibility of a predominant‘TH2-like’ cell-mediated immune response in lesional skin of AD patients.
Surface charge distribution is a determinant of antigen deposition in the renal glomerulus: studies employing ‘charge-hybrid’ molecules
Authors: BATSFORD S R , MIHATSCH M J , RAWIEL M , SCHMIEDEKE T M , VOGT A
Published: July 2010

The deposition of antigens and immune complexes (IC) in the renal glomerulus is charge-dependent. The demonstration that molecules of net anionic charge, but with discrete positively charged regions, exhibit affinity for the glomerular basement membrane (GBM) extends this concept. Charge hybrid (polar) molecules were constructed by covalently coupling small polycations (lysozyme or linear poly-l-lysinc chains with a mean of 17 and 20 residues) to larger polyanions (ovalbumin or human scrum albumin (HSA)). Although the products were of overall net anionic charge they still bound to glomerular structures. Immunofluorescence studies performed after i.v. injection of the samples into rats revealed that HSA:poly-l-lysine had the highest affinity. Radioisotopic measurements showed uptake of HSA: poly-l-lysinc to be a function of the number of lysinc residues; binding of HSA: poly-l-lysine20 was 2.5 times higher than HSA:poly-l-lysine17 (P<0.01). Prior injection of a small competing polycation (polyethyleneimine 1200) reduced uptake of HSA:poly-l-lysine by 75%, indicating the charge-based nature of the interaction. HSA:poly-l-lysine20 alone was effectively eliminated from the glomeruli within 72 h. Administration of HSA:poly-l-lysine followed by anti-HSA antibody induced immune complex formation in the capillary wall giving rise to a granular immunofluoresccnce pattern and discrete subendothelial and subepithelial deposits. Molecules with polar structure do occur naturally and may contribute to immune complex formation in glomerulonephritis.
Autoantibodies against Forssman glycolipids in Graves' disease and Hashimoto's thyroiditis
Authors: ARIGA T , YOSHIDA T , MIMORI T , YU R K
Published: July 2010

Sera from patients with Graves’ disease and Hashimoto's thyroiditis have been shown to react with the Forssman glycolipid antigen (Gb5) using the techniques of high performance thin-layer chromatography (HPTLC) immunostaining and ELISA. Human monoclonal antibodies (MoAbs) have been prepared by fusion of human myeloma with peripheral lymphocytes from patients with Graves’ disease. A MoAb, TRMo-4, reacted strongly and specifically with Gb5. These results suggest that anti-Forssman antibody may be involved in the pathogenesis of these autoimmune diseases. The detection of anti-Forssman glycolipid antibody may provide a useful means for clinical diagnosis and therapy.
Human colostrum contains an activity that inhibits the production of IL-2
Authors: HOOTON J W L , PABST H F , SPADY D W , PAETKAU V
Published: July 2010

The effect of human colostrum on T cell immune function was investigated. Colostrum inhibited the proliferation of human T cells activated by allogeneic, concanavalin A (Con A) or phytohaemagglutinin (PHA) stimulation. Colostrum also inhibited the production of IL-2 by Con A-activated human peripheral blood T cells and by Con A-activated Jurkat cells, a human T lymphoma line. Similarly, human colostrum inhibited the production of IL-2 by EL4 cells, a murine thymoma line, when stimulated with phorbol myristate acetate. The inhibitory activity was not cytotoxic and could not be neutralized by antibody to transforming human growth factor β.
Isotype distribution of mucosal IgG-producing cells in patients with various IgG subclass deficiencies
Authors: NILSSEN D E , SÖDERSTRÖM R , BRANDTZAEG P , KETT K , HELGELAND L , KARLSSON G , SÖDERSTRÖM T , HANSON L Å
Published: July 2010

The subclass distribution of IgG-producing immunocytes was examined by immunohistochemistry in nasal and rectal mucosa of infection-prone patients with untreated IgG subclass deficiencies. Biopsy specimens from the two sites were obtained in 18 clinically and serologically well-characterized adult subjects; only a nasal or rectal sample was available from nine similar patients. Chronic lung disease was common in the patient groups with selective serum IgG1 deficiency and combined IgG1 and IgG3 deficiency, whereas the other categories of patients had mainly upper airway and other mild infections. Serum IgG2 or IgG3 deficiency was usually expressed also at the cellular level in rectal mucosa, and the proportion of rectal IgG1 cells was significantly correlated with the IgG1 level (r= 0.90, P < 0.001). Likewise, there tended to be a decreased expression of the actual subclass at the cellular level in nasal mucosa of patients with serum IgG1 or IgG2 deficiency. Conversely, the median nasal proportion of IgG3 celts was remarkably unaffected by a deficiency of this subclass in serum and reclal mucosa. Interestingly, these patients rather tended to have raised IgG3 and reduced IgG2 cell proportions in their nasal mucosa, although this apparent local IgG3 compensation was nevertheless strongly correlated with the serum IgG3 level (r = 0.87, P < 0.002). These disparities may reflect different antigenic and mitogenic exposure of the two tissue sites; for example, a persistent protein bombardment of the nasal mucosa that could conceivably override locally a B cell maturation defect. The possible clinical consequences of such variable mucosal expression of IgG subclass deficiencies remain to be studied.
Measurement of interleukin-6 in bronchoalveolar lavage fluid by radioimmunoassay: differences between patients with interstitial lung disease and control subjects
Authors: JONES K P , REYNOLDS S P , CAPPER S J , KALINKA S , EDWARDS J H , DAVIES B H
Published: July 2010

Bronchoalveolar lavage fluid (BALF) from subjects with a variety of interstitial lung diseases (active sarcoidosis, pigeon breeders' disease (PBD), asymptomatic pigeon breeders, patients with idiopathic pulmonary fibrosis) and from control subjects were assayed for interleukin-6 (IL-6) using a novel radioimmunoassay system. IL-6 was detectable in BALF from all groups, with disease groups showing significantly increased IL-6 levels compared with controls (P < 0.01 in all cases). When these results were standardized, using urea to compensate for dilution effects in the BALF, only the asymptomatic pigeon breeders had significantly higher IL-6 levels than the controls (P < 0.025), with all other groups showing no difference. When albumin was used for standardization, both the PBD group (P < 0.001) and the sarcoidosis patients (P < 0.01) had considerably lower levels of IL-6 than the control subjects. Using either albumin or urea for standardization, the PBD patients had significantly lower levels of IL-6 than do their asymptomatic counterparts (P < 0.001 in both cases). This is contrasted by the finding of greatly elevated levels of IgG in the BALF of the PBD patients compared with asymptomatics (P < 0.001). There was, however, no relation between IL-6 and IgG in any patient group, although the PBD patients had the lowest IL-6 and highest IgG as a group. These findings may suggest a mechanism by which asymptomatic subjects remain free from clinical complaints.
Immunogenicity of surfactant. II. Porcine and bovine surfactants
Authors: STRAYER D S , HALLMAN M , MERRITT T A
Published: July 2010

Protein-containing surfactants of human and animal origin are being used increasingly to treat neonatal and adult respiratory distress syndromes. This trend led us to examine the antigenicity of two important preparations of animal surfactant, cow lung surfactant extract (CLSE) and a porcine surfactant preparation, Curosurf. We describe here 15 monoclonal antibodies against Curosurf and four against CLSE. Antibodies were studied by Western blot analysis to determine their ability to recognize protein components of their respective surfactant preparations. They were also tested for their ability to inactivate surfactant in vitro, assayed using the pulsating bubble surfactometer. Several antibodies directed against CLSE or Curosurf functionally inactivate the surfactant to which they were raised. We determined the degree of immunologic cross-reactivity between antibodies directed to CLSE and Curosurf against the other surfactant and also against human surfactant, both by Western blot and by examining functional inactivation in vitro. Antibodies to these animal surfactants that are commonly used therapeutically may inactivate the specific animal surfactant to which they were raised, as well as human and other surfactants. Generally, when antibodies inactivate surfactant from more than one animal species, they inactivate heterologous surfactants comparably to the extent to which they inactivate the surfactant to which they are directed. Immune complexes between anti-surfactant antibodies and surfactant have been described in the course of neonatal respiratory distress syndrome. The potential pathophysiological importance of anti-surfactant antibodies may therefore lie in their ability to inactivate administered surfactant, other similar surfactants and endogenous surfactant. In so doing, these antibodies may potentiate surfactant deficiency or pulmonary injury initiated by other stimuli.
Molecular analysis of HLA-DQ A alleles in coeliac disease lack of a unique disease-associated sequence
Authors: MANTOVANI V , CORAZZA G R , ANGELINI G , DELFINO L , FRISONI M , MIRRI P , VALENTINI R A , BARBONI P , GASBARRINI G , FERRARA G B
Published: July 2010

Susceptibility to coeliac disease is strongly associated with some HLA class IF antigens, encoded by the HLA-D region. Since the HLA-DQ locus seems to be primarily involved, we have analysed by polymerase chain reaction amplification and allele-specific oligonucleotide hybridization the most polymorphic region of the HLA-DQ Al gene. No difference was observed between the 20 coeliac patients and 20 HLA-D-matched healthy controls who took part in the study. Furthermore, in patients and controls, the restriction fragment length polymorphism analysis of the HLA-DQ A gene using the restriction enzyme BglII did not disclose any specific disease-associated fragment. Our results are not consistent with a unique DQ A coeliac disease-associated sequence, but rather with the hypothesis that some polymorphic residues or allelic hypervariable regions, although found also in the normal population, can predispose to coeliac disease due to their higher frequency in this condition.
14C1, an antigen associated with human ovarian cancer, defined using a human IgG monoclonal antibody
Authors: GALLAGHER G , AL-AZZAWI F , WALSH L P , WILSON G
Published: July 2010

We have selected a human EBV-transformed cell line from the involved lymph nodes of an ovarian cancer patient which secretes an IgG 1 k antibody, able to recognize an antigen present on the surface of ovarian cancer cells. The antigen, termed ‘14C1,’ has previously been shown by immunohistological techniques to be present on the surface of the malignant cells within tumour specimens. Western blotting analysis has shown that the majority of primary ovarian cancer specimens and three continuous cell lines derived therefrom express 14C1; other tissue types were negative. Preliminary biochemical characterization has been carried out, which shows that the 14C1 antigen has a molecular weight range of 25–32 kD and an isoelectric point from pf 6.3 to 6.8. We believe that the 14C1 antigen is immunologically relevant to ovarian cancer patients and may therefore represent a novel target for both active and passive immunotherapy.
Antigen(s)-specific tumour-infiltrating lymphocytes from tumour induced by human herpes virus-6 (HHV-6) DNA transfected NIH 3T3 transformants
Authors: PURI R K , LELAND P , RAZZAQUE A
Published: July 2010

Tumour infiltrating lymphocytes (TIL) have recently been shown to mediate potent therapeutic effects in certain malignancies in mice and in humans. To understand the mechanism of TIL immunolherapy it would be advantageous to generate tumour-specific TIL and to study a defined system of TIL and target cells in which the tumour epitope(s) recognized by TIL might be identified. We have established tumourigenic cell lines by transfection of NIH 3T3 cells with the entire genome of human herpcsvirus-6 (HHV-6) and its small fragment (about 5% of the viral DNA sequence). Injection of these cells into nude mice produced tumours termed G-2T and 14-2T. respectively. Cell lines derived from these tumours when injected in NIH Swiss mice produced tumours. G-2TS and 14-2TS. respectively. We have generated TIL from G-2TS tumour that can kill G-2TS tumour cells in vitro but not other related tumours (14-2TS or MCA-106). These TIL can be expanded between 2–6.5 every 3–5 days. The TIL proliferated in tissue culture in response to recombinant intcrleukin-2 and interlcukin-4 and maintained their tumour specificity for up to 6 months in vitro. Their phenotypc was Thy 1.2+. Lyt-2+ and L3T4. The availability of such tumour-specific stable TIL lines and specific viral-transformed targets will provide an opportunity to characterize the tumour-associated antigen critical for the specific cytotoxicity in this system and thereby to clarify the mechanism of this promising immunological approach to cancer therapy.
T cell proliferative responses to molecular fractions of periodontopathic bacteria
Authors: IVANYI L , NEWMAN H N , MARSH P D
Published: July 2010

Soluble antigenic preparations of Veillonella parvula and Bacteroides gingivalis were separated by SDS-PAGE and used after electroblotting and solubilization for in vitro lymphocyte stimulation in 13 patients with severe periodontitis and 12 controls. The cellular responses of controls and patients to V. parvula antigens were represented by four main proliferation-inducing fractions with 74–66, 52–46, 22–19 and 12 kD mol. wt. These fractions induced slightly enhanced DNA synthesis in lymphocytes from eight patients who failed to respond to whole antigenic extract. Lymphocyte samples from Veillonella whole extract unresponsive patients were also examined for in vitro proliferation by B. gingivalis fractions. Almost all stimulatory activities could be classified into five regions of 84–74, 35–31, 28–25, 17–15 and 12 kD.
Production by cultured human monocytes of mesangial cell proliferation factor(s) differing from interleukin-1 and interleukin-6
Authors: MORIOKA T , NARITA I , SHIMIZU F , OITE T
Published: July 2010

Conditioned media from human peripheral blood leucocytes treated with lipopolysaceharide (LPS) induced a marked increase in the 3H-thymidine incorporation of cultured mesangial cells at low scrum concentration (four to six times higher than control). Two sizes 100–70 and 8–12 kD) of monocyte-derived mesangial cell proliferating factors (MDF) were separated by column chromatography. Their peaks were distinct from those of thymocytc proliferating activity. The addition of anti-human interleukin-1 (IL-1) or anti-recombinant human interleukin-6 (IL-6) antibody to the fractionated MDF failed to have any effect on the mitogenic activity toward mesangial cells. The addition of anti-human platelet-derived growth factor (PDGF) antibody to the low molecular weight fraction decreased mesangial cell mitogenic activity 40–60% of control), but addition to the higher fraction did not 80–100% of control). From these data it seems that a large portion of the monocytedcrived mesangial cell growth factor was not comprised of IL-1 or IL-6 but of PDGF-likc molecules; and that there is an unknown mesangial cell proliferating factor (or factors) besides IL-1. IL-6and PDGF.
Effect of a recombinant HIV gpl60 vaccine on monokine production
Authors: DOOLEY D P , COX R A , LOONEY D J
Published: July 2010

An investigation was undertaken to determine whether a recombinant gp 160 envelope protein, which is currently being evaluated as a vaccine for AIDS, induces or modulates the production of tumour necrosis factor-alpha (TNF-α) or interleukin-1β (IL-1β). Incubation of monocytes from healthy, HIV-seronegative persons with 0.0001–1.0 μg of the recombinant vaccine did not result in the secretion of TNF-α or IL-1β, nor did the recombinant product augment or suppress monokine production by lipopolysaccharide (LPS) stimulated monocytes. The vaccine was also without a stimulatory or modulatory effect upon TNF-α or IL-1β secretion by monocytes from a patient with the AIDS-related complex (ARC) and from the monocytic THP-1 cell line. The lack of effect of gp 160 on monokine production has important implications for its efficacy as a vaccine for AIDS.
Individual germinal centres of myasthenia gravis human thymuses contain polyclonal activated B cells that express all the VH and VK families
Authors: GUIGOU V , EMILIE D , BERRIH-AKNIN S , FUMOUX F , FOUGEREAU M , SCHIFF C
Published: July 2010

Using in situ hybridization, we analysed the immunoglobulin repertoire expressed by the B cells present in myasthenia gravis thymuses from four patients. B cells, mostly in activated state, were clustered in germinal centres, in which multiple isotypes were identified. A majority of cells expressed IgG as compared with IgM. with a roughly similar contribution of k and δ chains. Hybridization with the six VH and the 4 Vk human family probes was observed in serial sections, providing additional evidence that individual germinal centres were polyclonal. The thymic B cell repertoire closely reflected the VH and the Vk family usage of normal peripheral blood lymphocytes with the preferential utilization of VH3, Vk1 and Vk3.
LDL-mediated interaction of DNA and DNA-anti-DNA immune complexes with cell surface
Authors: KABAKOV A E , SAENKO V A , POVERENNY A M
Published: July 2010

The effect of low-density lipoprotein (LDL) on the binding of DNA and DNA-anti-DNA immune complexes to cultured human skin fibroblasts was examined. Using radioisotope analysis. ELISA and indirect immunofluorescent staining, a correlation between plasma membrane-bound DNA or DNA-anti-DNA immune complexes and cell-associated LDL was established. It was demonstrated that cytotoxicity and internalization of DNA-anti-DNA immune complexes may be LDL mediated. The data obtained suggest that the binding of the major part of DNA and immune complexes bound to surface of normal skin fibroblasts is due to the formation of a DNA-LDL-LDL receptor linkage. A possible role of LDL-containing immune complexes in the pathogenesis of systemic lupus erythematosus is discussed.
Expression of decay-accelerating factor is reduced on hyperplastic synovial lining cells in rheumatoid synovitis
Authors: ITOH J , NOSE M , FUJITA T , KATO M , OHYAMA A , KYOGOKU M
Published: July 2010

Decay-accelerating factor (DAF), a membrane inhibitor of homologous complement activation, is present in synovial cells lining joint space and detected in synovial fluid. DAF is considered to protect synovial membrane from complement-mediated injury associated with articular inflammation. We studied the immunohistopathological features of DAF molecules in synovial membrane of rheumatoid synovitis using a DAF-specific monoclonal antibody, IC6. Reacting molecules with the 1C6 antibodies in synovial tissue extracts formed a 70-kD band in Western blot analysis. DAF was strongly detected on the flat synovial lining cells, but weakly on the hyperplastic and multi-layered lining cells in rheumatoid synovitis. The latter cells reacted with anti-Lcu-M3 antibodies specific for a cell surface marker of activated macrophages, sometimes accompanied by C3 and IgM deposition on the superficial synovial membrane. These results suggest that active rheumatoid synovitis characteristically with hyperplastic synovial lining cells is out of control by DAF, thereby permitting further complement-mediated injury.
Human hybridomas derived from CDS+ B lymphocytes of patients with chronic lymphocytic leukemia (B-CLL) produce multi-specific natural IgM (kappa) antibodies
Authors: JAHN S , SCHWAB J , HANSEN A , HEIDER H , SCHROEDER C , LUKOWSKY A , ACHTMAN M , MATTHES H , KIESSIG S T , VOLK H D , KRUEGER D H , VON BAEHR R
Published: July 2010

Great numbers of CD5+ B lymphocytes were detected in the peripheral blood of patients with B-CLL. To study the antibody repertoire of this immune cell subpopulation on a monoclonal level, we fused the lymphocytes derived from five different donors to a highly efficient HAT-sensitive heteromye-loma line (CB-F7). A fusion frequency of up to 10-5 allowed us to analyse hundreds of initial hybridoma lines per fusion. In all culture supernatants in three out of five fusions IgM lambda antibodies were detected, in two experiments only IgM kappa was measured, suggesting monoclonality of the primary hybridoma cell lines. The later fusions resulted in hybridomas producing multi-specific antibodies against both an autoantigen and an infectious agent: (i) dsDNA/influenza virus haemagglutinin; (ii) dsDNA/class V outer membrane protein type C from Neisseria meningitidis. However, no antibodies of the described specificity were detected in blood sera of patients, indicating a ‘switch-on’ of the immunoglobulin secretion capacity of malignant B cells during fusion to a myeloma partner. We discuss the results as further evidence for the natural multi-reactive antibody repertoire of CD5+ B cells.
Expression of the neural cell adhesion molecule (CD56) by human myeloma cells
Authors: DRACH J , GATTRINGER C , HUBER H
Published: July 2010

Recent studies in multiple myeloma indicate that molecules associated with different haematopoietic lineages may be expressed aberrantly by myeloma cells. In order to investigate this phenomenon further, we studied the immunophenotype of bone marrow cells from 21 patients with multiple myeloma using a panel of monoclonal antibodies against T.B. myelomonocytic, and natural killer (NK)-cell antigens. Leu-19/NK.H1 (CD56), a molecule identical to N-CAM, which is normally expressed by neuroectodermal and NK cells, was found in 13 patients (62%). Dual-parameter flow cytometry was used to correlate N-CAM positivity with DNA aneuploidy or cytoplasmic immunoglobuiin expression as markers of myeloma cells. When N-CAM was found positive, other haematopoietic antigens were expressed only in three out of 13 cases (23%). In contrast, myeloma cells not expressing N-CAM frequently exhibited pre-B cell markers, myeloid antigen, and HLA-DR, respectively (seven out of eight cases, 88%). Six out of eight N-CAM-negative myelomas were of the IgG lamba isotype, otherwise no clearcut association with basic clinical and laboratory parameters was noted. We conclude that N-CAM expression is a common finding in multiple myeloma. Whether its expression and the observed antigenic heterogeneity is just a manifestation of malignancy or N-CAM may play a role in the biology of multiple myeloma regarding tumour cell spread, remains to be explained.
Differential control of mesangial cell proliferation by interferon-gamma
Authors: KAKIZAKI Y , KRAFT N , ATKINS R C
Published: July 2010

Rat mesangial cells were shown to be sensitive to recombinant interferon-gamma (IFN-γ). IFN-γ reduced thymidine uptake by these cells and inhibited cell proliferation. Incubation of the cells with 1000 U/ml IFN-μ decreased thymidine uptake by up to 64% and cell numbers were decreased by 17%. The effects of IFN-y were dose and time dependent and were partially reversible by the anti-IFN-γ monoclonal antibody DB-I. This lymphokine did not reduce incorporation of RNA and protein precursors however. Measurements of 3H-uridine and 3H-leucine incorporation indicated significant increases in RNA and protein synthesis (37% and 45%, respectively) on a per cell basis. The mitogenic effects of IL-1 and platelet-derived growth factor (PDGF) were also susceptible to IFN-γ-mediated inhibition but the mitogenic response to epidermal growth factor (EGF) was much less sensitive. Weconclude that while IFN-γ may act to modulate the mitogenic signals provided by some factors such as IL-1 and PDGF, the response to EGF appears to be unaffected.
The gene corresponding to the putative Goodpasture antigen is present in Alport’s syndrome
Authors: SAVIGE J A
Published: July 2010

Alport’s syndrome is a heterogeneous group of inherited abnormalities of basement membranes that may result in progressive renal failure, defective hearing and lens abnormalities. The glomerular basement membrane (GBM) characteristically has areas of reduplication, lamellation and attenuation on electron microscopic examination. In the majority of affected males and some females, there is reduced or variable binding of serum from patients with anti-GBM disease (Goodpasture’s syndrome) to these basement membranes. These sera contain antibodies directed against the Goodpasture antigen which has been thought to be located in the non-collagenous domain of the alpha3 chain of type IV collagen and is presumed to be important in cross-linking of the collagen molecules. The reduced staining for the Goodpasture antigen suggests that this structure is either absent or masked in Alport’s syndrome. We have tested DNA from six unrelated individuals with Alport’s syndrome. All had been transplanted for renal failure. The diagnosis of Alport’s syndrome was made on the characteristic electron microscopic appearance of the renal basement membranes (n= 4), the presence of sensori-neural deafness (n=4), a family history of Alport’s syndrome (n=5) and the presence of circulating inhibitable anti-GBM antibody activity post-transplant (n=2), Oligonucleotides (20mers) corresponding to the 5′ and _3′ ends of the known 25 amino acid sequence for the putative Goodpasture antigen were used as primers for amplification of genomic DNA. The products were then blotted and probed with an intermediate 19-mer DNA. All Alport’s patients contained a 75-bp fragment corresponding to the published peptide sequence for the non-collagenous domain of the alpha3 chain of type IV collagen, suggesting that a large deletion of this region, the putative Goodpasture antigen, is unlikely to account for the defect in Alport’s syndrome.
Polymorphism of insulin antibodies in six patients with insulin-immune hypoglycaemic syndrome
Authors: DOZIO N , SODOYEZ-GOFFAUX F , KOCH M , ZIEGLER B , SODOYEZ J C
Published: July 2010

Insulin antibodies in six patients with immune hypoglycaemic syndrome were studied. The antibodies displayed a higher affinity for bovine insulin in two patients, were specific for human insulin in one patient and non-species specific in the other three patients. The predominant IgG subclass of the insulin antibodies was IgG4 in two patients, IgG3 in two and IgG 1 in two. In one of these, the other three subclasses were also detectable. Insulin autoantibodies of four patients were homogeneous with regard to light chains (k), and those of the other two contained both k and γ light chains. Analysis of insulin immune complex size by fast protein liquid chromatography was possible in three patients and demonstrated immune complexes with elution profile close to that of IgG, although not exactly superimposable to the one obtained with a mouse monoclonal insulin antibody. In two patients, avidity was too low to permit chromatography of the immune complexes, and, moreover, in these two cases insulin antibodies were of the IgG3 isotype and spontaneously formed aggregates independently of insulin binding. We conclude that insulin antibodies of the insulin immune syndrome are polymorphic but different from those generated by insulin therapy.
Ciprofloxacin treatmentin vivoincreases theex vivocapacity of lipopolysaccharide-stimulated human monocytes to produce IL-1, IL-6 and tumour necrosis factor-alpha
Authors: BAILLY S , FAY M , FERRUA B , GOUGEROT-POCIDALO M A
Published: July 2010

Because in vitro treatment with quinolones, at pharmacological concentrations, modifies lipopolysac-charide (LPS) induced production of cytokines by monocytes, we studied the effect of orally administered ciprofloxacin (25 mg/kg) on the capacity of peripheral blood monocyles of healthy volunteers to produce tumour necrosis factor-alpha (TNF-α), IL-1 activity, IL-1 a, IL-1β and IL-6 ex vivoin response to endotoxin stimulation. After 7 days of ciprofloxacin, the extracellular and cellular production of TNF-α, the cellular production of IL-1 activity, the extracellular and cellular production of IL-1α, and the cellular production of IL-6 increased significantly. Seven days after the end of the treatment, values returned to basal levels or even lower. To our knowledge, this is the first demonstration that ciprofloxacin can modulate in vivo the capacity of human monocytes to react to an inflammatory stimulus such as endotoxin.