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Showing papers in "Scandinavian Journal of Immunology in 1993"


Journal ArticleDOI
TL;DR: Gastric synthesis of IL‐8 is likely to be an important factor in regulating mucosal neutrophil infiltration and activation in patients with H. pylori infection and the local production of IgA antibodies to IL‐9 may represent a down‐regulatory response of the host to limit mucosal damage associated with a chronic bacterial infection.
Abstract: Gastric infection with Helicobacter pylori is frequently characterized by neutrophil infiltration. The production of the neutrophil-activating peptide (NAP-1/IL-8) and mucosal IgA autoantibodies to IL-8 by human antral biopsies have been examined during short-term in vitro culture. Detectable IL-8 was secreted by 84% of H. pylori-negative patients with normal antral mucosa (range < 0.07-61.5 ng/mg biopsy protein, n = 19). Concentrations in 4 patients with reactive gastritis and 10 with inactive gastritis were not significantly different from subjects with normal mucosa. In H. pylori-positive patients with active gastritis and neutrophil infiltration into the epithelium (n = 17) IL-8 secretion was significantly increased relative to subjects with normal mucosa (P < 0.0001), inactive gastritis (P < 0.001) and reactive gastritis (P < 0.01). IL-8 concentrations in active gastritis were significantly correlated with the extent of epithelial surface degeneration (r = 0.64). IgA autoantibodies were present in 19 patients (13 active, 4 inactive gastritis) and concentrations were significantly correlated with IL-8 production (P < 0.001). Gastric synthesis of IL-8 is likely to be an important factor in regulating mucosal neutrophil infiltration and activation in patients with H. pylori infection. The local production of IgA antibodies to IL-8 may represent a down-regulatory response of the host to limit mucosal damage associated with a chronic bacterial infection.

255 citations


Journal ArticleDOI
TL;DR: Findings provide further evidence for the contention that, depending on defined stimuli, monocytes may develop either into accessory cells or into classical macrophages.
Abstract: To investigate the differentiation and activation of monocytes, the combined effects of 1,25-dihydroxyvitamin D3 (D3) and IL-4 on human blood monocytes were examined with respect to expression of MHC class-II antigens, accessory activity, and phagocytic capacity. IL-4 was reported to upregulate the expression of MHC class-II antigens and accessory activity of monocytes. The experiments described here demonstrate that D3 inhibits the expression of all three subtypes of MHC class-II antigens (HLA-DR, -DP and -DQ) as well as the accessory activity of monocytes, both in a dose- and time-dependent manner. However, D3 enhances the immunoglobulin- and complement-dependent phagocytosis by monocytes in a dose- and time-dependent manner. When monocytes are treated with both IL-4 and D3, the effects of D3 are reverted by IL-4, suggesting that IL-4 induces the development of monocytes into accessory cells, whereas D3 stimulates differentiation of monocytes into classical macrophages. These findings provide further evidence for the contention that, depending on defined stimuli, monocytes may develop either into accessory cells or into classical macrophages.

174 citations


Journal ArticleDOI
TL;DR: Whether T cells from rheumatoid synovial inflammation belong to the Th1‐ or Th2‐like functional subsets or the mycobacterial antigen‐specific group, all CD4+’αβ T‐cell clones produced IFN‐γ at high levels, while the production of IL‐4 was generally absent or low, consistent with a Thl‐like profile.
Abstract: This study was performed in order to characterize whether T cells from rheumatoid synovial inflammation belong to the Th1- or Th2-like functional subsets. Cytokine production was studied in 26 CD4+ alpha beta+ and 2 CD8+ alpha beta T-cell clones from the synovial fluid, the synovial membrane and peripheral blood of 5 patients. Fifteen of the CD4+ clones were raised against various mycobacterial antigens and 11 CD4+ clones and 2 CD8+ clones were raised unspecifically using PHA and/or IL-2. The specificities of these clones are not known. In the mycobacterial antigen-specific group, all CD4+ alpha beta T-cell clones produced IFN-gamma at high levels, while the production of IL-4 was generally absent or low (< 1 ng/ml), consistent with a Th1-like profile. Some of these clones, however, also produced various amounts of IL-10 which has been regarded as a Th2 product but can be produced also in lower amounts by Th1 cells. One HSP-65-specific clone produced levels of IL-4 and IL-10 in the same order as that of IFN-gamma, thus appearing to be Th0-like. Among the 11 unspecific CD4+ clones, 7 showed a Th1-like pattern but with lower levels of IFN-gamma than the antigen-specific clones. However, three clones did not produce any IFN-gamma activity but produced IL-4 and one of them also produced distinct amounts of IL-10, compatible with a Th2-like pattern. In addition, one of the clones also showed an almost equally strong IFN-gamma and IL-4 production, thus most likely representing a Th0-like clone.

136 citations


Journal ArticleDOI
TL;DR: The frequencies of two polymorphisms within the MHC‐linked heat shock protein (HSP) 70 genes in patients with insulin‐dependent diabetes mellitus (IDDM) and healthy control individuals are characterized.
Abstract: In the present study we characterized the frequencies of two polymorphisms within the MHC-linked heat shock protein (HSP) 70 genes in patients with insulin-dependent diabetes mellitus (IDDM) (n = 114) and healthy control individuals (n = 110). Significant differences in genotype and allelic frequencies were observed for both polymorphisms between randomly selected patients and controls. However, for the HSP70-2 polymorphisms this was solely due to linkage disequilibrium with DR3. The rate HSP70-Hom 2-allele was significantly more frequent in controls than in patients. It showed strong association with certain tumour necrosis factor (TNF) (class III) and HLA-B and -A (class I) alleles independent of HLA-DQ and -DR alleles. By typing 257 individuals from 55 IDDM multiple-case families two extended MHC-haplotypes, including class II-, TNF- and class I-markers, carrying the rare HSP70-Hom allele were defined. One was only transmitted to diabetic offspring, whereas the other was only transmitted to unaffected offspring. The functional implication of the polymorphism in the heat shock-inducible HSP70-2 gene was analysed by studying HSP70-2 mRNA expression after heat shock in peripheral blood mononuclear cells from individuals with different HSP70-2 genotypes. Preliminary data showed that individuals homozygous for the PstI 8.5-kb allele consistently had slightly lower expression than heterozygous and 9.0-kb homozygous individuals.

135 citations


Journal ArticleDOI
TL;DR: In addition to these activated T cells, a dominating population of CD25+ CD3‐CD4+ subepithelial pan‐HLA–class I+ macrophages (CD68+) with variable expression of the p75 β‐chain was often induced by gluten challenge.
Abstract: Jejunal biopsy specimens from 10 patients with treated coeliac disease and seven non-coeliac controls were challenged in vitro with peptic-tryptic gluten digest. Mucosal T cells were examined in situ by three-colour immunofluorescence staining for expression of the activation marker CD25 (the p55 alpha-chain of interleukin-2 receptor) and the nuclear proliferation marker revealed by monoclonal antibody Ki-67. Intraepithelial T cells expressed CD25 rarely whereas the proportion of activated lamina propria T cells increased (P 90%) were CD4+CD8-, co-expressed CD45R0 and the p75 beta-chain, and often also the integrin alpha E beta 7 but not HLA-DR. In addition to these activated T cells, a dominating population of CD25+CD3-CD4+ subepithelial pan-HLA-class II+ macrophages (CD68+) with variable expression of the p75 beta-chain was often induced by gluten challenge.

93 citations


Journal ArticleDOI
TL;DR: The EA‐hy‐926 cells used in this study showed responses similar to HUVEC when stimulated with TNF but not when stimulating with IL‐4 or IFN.
Abstract: EA-hy-926 is a cell line produced by hybridizing human umbilical vein endothelial cells (HUVEC) and the epithelial cell line A549. To establish whether EA-hy-926 could be used as a model for endothelial cells (EC) in leucocyte-EC adhesion interactions, the effect of interleukin-4 (IL-4), tumour necrosis factor (TNF) or interferon-gamma (IFN) stimulation on their adhesiveness and expression of E-selectin, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) was compared with that of HUVEC and A549. Although HUVEC exhibited increased adhesiveness and adhesion molecule expression with IL-4, TNF or IFN, EA-hy-926 exhibited these responses only with TNF. CD11/CD18-dependent binding accounted for a significant component of basal binding to HUVEC and EA-hy-926, but did not account for the increased binding of T cells, JY, J6, ICH-BJ or ICH-KM cell lines to TNF-stimulated monolayers. At least part of the CD11/CD18-independent adhesion was attributable to VCAM-1 induction on HUVEC and EA-hy-926. TNF-stimulation also induced E-selectin expression on EA-hy-926 and HUVEC and an accompanying increase in neutrophil (PMN) binding. The EA-hy-926 cells used in this study, therefore, showed responses similar to HUVEC when stimulated with TNF but not when stimulated with IL-4 or IFN.

91 citations


Journal ArticleDOI
TL;DR: It is shown that cholera toxin strongly potentiates antigen presentation by intestinal epithelial cells, probably by enhancing co‐stimulation, in an allogeneic system using cells from the IEC‐17 rat epithelial cell line as antigen presenting cells (APC).
Abstract: In the present study we show that cholera toxin (CT) strongly potentiates antigen presentation by intestinal epithelial cells, probably by enhancing co-stimulation. This was demonstrated in an allogeneic system using cells from the IEC-17 rat epithelial cell line as antigen presenting cells (APC). These cells were induced by optimal concentrations of IFN-gamma to express good amounts of Ia antigen and cultured for 24-48 h in the presence or absence of CT. Thereafter the cells were thoroughly washed and added to cultures containing MHC-incompatible spleen cells as responder cells. Epithelial cells exposed to CT demonstrated greatly enhanced ability to trigger allogen-specific T-cell proliferation as compared with IEC-17 cells treated with IFN-gamma alone. The mechanism for the enhanced APC function was investigated by analysing CT-treated IEC-17 cells for increased class II MHC antigen expression or enhanced production of cytokines with known co-stimulatory function. We found no significant increase in class II MHC antigen expression. By contrast, CT strongly promoted, in a dose-dependent fashion, the production of both IL-1 and IL-6 cytokines by IEC-17 cells as compared with untreated epithelial cells. This effect of CT was specific and not due to contaminating endotoxin because excess amounts of soluble toxin receptor, ganglioside GM1, added to the IEC-17 cultures completely abrogated the cytokine response to CT. These results together with our previous findings of enhanced antigen presentation by macrophages stimulated by CT suggest that the potent adjuvant function of CT for induction of mucosal immune responses might be attributed to an enhanced co-stimulating ability of several putative APC in the mucosal immune system: macrophages, B cells and epithelial cells.

88 citations


Journal ArticleDOI
TL;DR: This lecture attempts to establish an analogy between linguistics and immunology, between the descriptions of language and of the immune system, using the example of diphtheria toxin and tetanus toxin.
Abstract: Grammar is a science that is more than 2000 years old. whereas immunology has become a respectable part of biology only during the past hundred years. Though both sciences still face exasperating problems, this lecture attempts to establish an analogy between linguistics and immunology, between the descriptions of language and of the immune systetn. Let me first recall some of the essential elements of the immune system, with which I shall be concerned. In 1890, von Behring and Kitasato [12] were the first to discover antibody molecules in the blood serum of immunized animals, and to demonstrate that these antibodies could neutralize diphtheria toxin and tetanus toxin. They also demonstrated tbe specificity of antibodies: tetanus antitoxin cannot neutralize diphtheria toxin, and vice versa. During the first 30 years, or more, after these discoveries, most immunologists believed that all eells of our body are capable of producing antibodies, and it took until the 1950s before it became clear, and until I960 before it was demonstrated [13], that only white blood cells, named lymphocytes, can produce antibodies. The total number of lymphocytes represent a little more than \\\"/n ofthe body weight of an animal. Thus, it would not be wrong to say that our immune system is an organ consisting of about 10'lymphocytes:

81 citations


Journal ArticleDOI
TL;DR: The oligoclonal nature of the anti‐HIV‐1 antibodies suggested by skewed kappa/lambda expression was confirmed with isoelectric focusing of affinity purified antibodies to p24 and gp120.
Abstract: Antibodies in sera of HIV-1 infected individuals against the HIV-1 core protein (p24), HIV-1 envelope glycoprotein (gp120) and reverse transcriptase (RT) are characterized by a skewed light chain isotype expression The kappa/lambda ratios of antibodies to p24 and gp120 in infected individuals were found to be unique in each individual, but constant over several years independently from disease progression The oligoclonal nature of the anti-HIV-1 antibodies suggested by skewed kappa/lambda expression was confirmed with isoelectric focusing of affinity purified antibodies to p24 and gp120 Analysis of the utilization of V gene families in purified anti-p24 and anti-gp120 antibodies revealed a restricted and biased VH gene family usage In contrast, the utilization of VK gene families appeared to be random The finding of stable and restricted antibody responses in infected individuals could be one of the causes for the failure to produce antibodies to HIV-1 that are effective against escape virus variants

75 citations


Journal ArticleDOI
TL;DR: The hypothesis that TLTF and IFN‐γ have a erueial regulatory function in the parasite host interactions and that these moleeules influence the disease eourse during experimental African trypanosomiasis is supported.
Abstract: A protein factor that stimulates CD8+ lymphocytes to produce and secrete IFN-gamma has been purified from Trypanosoma brucei brucei (T.b. brucei). This was accomplished by raising monoclonal antibodies (MoAbs) against a fraction of T.b. brucei obtained by gel filtration, which contained high levels of material inducing rat mononuclear cells (MNC) to IFN-gamma production. MoAbs from four hybridomas strongly inhibited trypanosome-induced IFN-gamma production. One of them (MO1) was used for purification of the trypanosome-derived lymphocyte triggering factor (TLTF) by affinity chromatography. SDS electrophoresis of the purified TLTF displayed a band of 42-45 kDa MW. Gel filtration of homogenates of whole parasites yielded several peaks of IFN-gamma-inducing activity with a lowest MW of 41-46 kDa. Bioactivity of all peaks was blocked by MO1, suggesting that a single molecule, or a single epitope of additional molecules, is responsible for the different peaks with IFN-gamma-inducing activity. IFN-gamma released from MNC stimulates T.b. brucei growth. Blocking of TLTF in vitro with MO1 inhibited MNC-supported growth of the parasites. To study the in vivo relevance of TLTF in the course of experimental African trypanosomiasis, MO1 was used to treat rats and mice at different times after infection. Treatments instituted at different time-points after infection suppressed parasite growth, abrogated the IFN-gamma production by splenocytes induced by the infection and prolonged survival of the animals. The data support the hypothesis that TLTF and IFN-gamma have a crucial regulatory function in the parasite-host interactions and that these molecules influence the disease course during experimental African trypanosomiasis.

73 citations


Journal ArticleDOI
TL;DR: The data are suggestive of a cell‐lo‐cell‐mediated mechanism by which monocytes down‐modulate NK‐cell function and phenotype and its serotonergic regulation.
Abstract: Autologous monocytes irreversibly suppressed functions of human natural killer (NK) cells including baseline and lymphokine-induced cytotoxicity, antibody-dependent cellular cytotoxicity (ADCC), and interleukin-2 (IL-2)-induced proliferation. The suppression of these NK-cell functions was cell contact-dependent and could be evoked only by purified monocytes, recovered directly from peripheral blood by countercurrent centrifugal elutriation (CCE). The presence of monocytes also induced the disappearance of CD16 and CD56 antigen on CD3- NK cells (CD3-/16+/56+-->CD3-/16-/56-). By contrast, T-cell proliferation and the expression of CD3 on CD56- T cells were not susceptible to cell contact-mediated suppression by monocytes. The biogenic amine serotonin abrogated monocyte-induced suppression of NK-cell functions as well as down-modulation of CD16/56 NK-cell antigen. Serotonin thus markedly augmented baseline and lymphokine-induced NK-cell cytotoxicity, ADCC, and NK-cell proliferation, and maintained the expression of NK-cell surface antigens in the presence of elutriated monocytes. The effect of serotonin was mediated by 5-HT1A-type serotonin receptors (5-HT1AR) as indicated by mimicry exerted by 5-HT1AR agonists such as 8-OH-DPAT and (+)-ALK, partial antagonism by the 5-HT1AR antagonists pindolol and cyproheptadine, and lack of antagonism by the 5-HT2R antagonist ketanserin or the 5-HT3R antagonist ondansetron. Our data are suggestive of a cell-to-cell-mediated mechanism by which monocytes down-modulate NK-cell function and phenotype and its serotonergic regulation.

Journal ArticleDOI
TL;DR: Comparisons between RA patients of radiological stages I and II and between stage I and other stages showed significantly higher levels of sICAM‐1 in the sera of patients in the latter stages than those without vasculitis or pneumonitis.
Abstract: Intercellular adhesion molecule-1 (ICAM-1), a member of the immunoglobulin supergene family, is known to play an important role in inflammatory diseases. Using a previously developed enzyme-linked immunosorbent assay (ELISA) with two monoclonal antibodies (MoAbs) against human ICAM-1, levels of soluble ICAM-1 (sICAM-1) were measured in sera from patients with collagen diseases and in synovial fluids (SF) from patients with rheumatoid arthritis (RA). Although the results did not demonstrate that RA and other collagen diseases, as a group, had significantly higher levels of sICAM-1 in sera as compared with healthy controls, 21 of 138 cases (15%) with collagen diseases and 11 out of 57 patients (19%) with RA clearly showed higher levels of sICAM-1 in the sera. Comparisons between RA patients of radiological stages I and II and between stage I and other stages showed significantly higher levels of sICAM-1 in the sera of patients in the latter stages. RA patients with vasculitis and/or pneumonitis showed significantly higher levels of sICAM-1 than those without vasculitis or pneumonitis. Significant correlations were demonstrated between sICAM-1 and the factors IgG-RF, IgM-RF, erythrocyte sedimentation rate (ESR) and TNF-alpha in sera of RA patients. In addition, it was noted that the levels of sICAM-1 in SF were as high as those in the sera of patients with RA.

Journal ArticleDOI
TL;DR: In non‐human species, the binding ofprotein L was also found to be mediated through Ig light chains, and the results demonstrate the potential value of protein L as an immunochemical tool.
Abstract: Protein L, a cell wall molecule of certain strains of the anaerobic bacterial species Peptostreptococcus magnus, shows high affinity for human immunoglobulin (Ig) light chains. In the present study protein L was tested against a panel of human myeloma proteins of the IgG, IgM, IgA and IgE classes, and strong binding was seen with antibodies carrying kappa light chains. A high degree of specificity for Ig was demonstrated in binding experiments with human plasma proteins. Apart from human Ig, strong protein L-binding activity was also detected in the serum of 12 out of 23 tested additional mammalian species, including other primates and rodents. Subsequent analysis with purified Ig samples demonstrated the binding of protein L to Ig of important laboratory animal species such as the mouse, the rat and the rabbit. The affinity constants for the interactions between protein L and polyclonal IgG of these species were 2.6 x 10(9), 3.9 x 10(8) and 7.4 x 10(7), respectively. In non-human species, the binding of protein L was also found to be mediated through Ig light chains, and the results demonstrate the potential value of protein L as an immunochemical tool.

Journal ArticleDOI
TL;DR: The results indicate that TGF‐β1 and PGE2 suppress NK activity by different mechanisms.
Abstract: The separate and combined effects of transforming growth factor-beta 1 (TGF-beta 1) and prostaglandin E2 on human natural killer (NK) activity were studied. Peripheral blood lymphocytes (PBL) and large granular lymphocytes (LGL, 70-90% purity) were used as effector cells and K562 as targets. Overnight incubation of the effector cells with TGF-beta 1 resulted in a significant inhibition of NK activity. TGF-beta 1 did not influence the expression of CD3, CD16, CD18 or CD56 antigens on PBL. Combination of TGF-beta 1 with indomethacin gave the same NK-suppressive effect as TGF-beta 1 alone, showing that the inhibition of NK activity by TGF-beta 1 is not due to an increase in PGE2 levels. TGF-beta did not influence cAMP level in PBL whereas PGE2 significantly increased it. On the other hand, TGF-beta 1 and PGE2 showed an additive inhibitory effect on NK activity. TGF-beta 1 did not reduce the binding of PBL and LGL to K562. PGE2 suppressed the binding and TGF-beta 1 did not influence this suppression. TGF-beta 1 also suppressed IL-2-induced activation of NK activity and increase of expression of the granule proteins granzyme A and perforin. PGE2 did not appear to affect granzyme A and perforin contents. The results indicate that TGF-beta 1 and PGE2 suppress NK activity by different mechanisms.

Journal ArticleDOI
S. Klaesson1, O Ringdén1, L. Markling1, Mats Remberger1, I. Lundkvist1 
TL;DR: Intravenous Immunoglobulin (IVIG) at a concentration of 5 mg/ml, significantly inhibited mitogenic responses to phytohaemagglutinin (PHA), concanavalin A (conA) and pokeweed mitogen (PWM) by peripheral blood cells from healthy donors.
Abstract: Intravenous Immunoglobulin (IVIG) at a concentration of 5 mg/ml, significantly inhibited mitogenic responses to phytohaemagglutinin (PHA), concanavalin A (conA) and pokeweed mitogen (PWM) by peripheral blood cells from healthy donors. No difference in inhibition by IVIG was seen when stimulating different T-lymphocyte cell subsets. Inhibition by IVIG was dose-dependent. An increased response was observed when IVIG was added more than 12 h after PHA compared to adding 1 h before [p= 0·05]. Intravenous immunoglobulin added to mixed lymphocyte cultures (MLC), reduced the median response by more than 60% (range 14–89%; P= 0·03) and almost completely abrogated the lymphocyte response to Staphylococcus aureus protein A (SPA), whose median inhibition was 94% (range 90–99%; P= 0·02). When comparing 12 different commercial IVIG preparations at a concentration of 2.5 mg/ml, the median inhibition of the PHA stimulation ranged from 4% to 35% and the MLC response from 0% to 66%. In the presence of IVIG the lymphocyte response to different herpes virus antigens was reduced by <50%. No difference in inhibitory effect was seen when comparing IVIG and cytomegalovirus (CMV) hyper Ig, but CMV negative Ig resulted in lower inhibition (P= 0·05]. Three out of five IgG preparations (2.5 mg/ml) made from single donors inhibited PHA stimulation signifiantly more than commercial IVIG [P<0·05]. Mean inhibition was 61% compared to 35% Inhibition by pooled IgG from five donors was 56%. F(ab‘)2 fragments of IVIG inhibited the MLC response by more than 50% (range 34–75%), SPA stimulation by 97% (83–104%) and PHA stimulation by more than 30% (26–37%). One of two Fe preparations tested had an inhibitory effect, but the inhibition was less than that obtained with the F(ab')2 fragments [P= 0.04]. These results further strengthen the notion that IVIG exerts its immune modulatory effect by binding to leukocyte surface receptors. A clear inhibition was obtained with concentrations corresponding to the serum levels obtained when IVIG is given 250–500 mg/kg bodyweight. F(ab')2 fragments have the same inhibitory effect as intact IgG molecules but the role of Fc fragments still remains unclear. Differences in the immunosuppressivc effect of various IVIG preparations may be associated with the method of preparation.

Journal ArticleDOI
TL;DR: Comparison of reactivity on a panel of self antigens of affinity‐purified anti‐DNA and anti‐thyroglobulin IgG autoantibodies from the serum of patients with systemic lupus erythematosus and autoimmune thyroiditis indicates that polyreactivity of autoantsibodies is a feature that does not allow one to distinguish between natural and disease‐associated autoantIBodies as well as between V‐region‐connected and unconnected autoant
Abstract: Polyreactivity was earlier recognized as a feature of naturally expressed autoantibodies in serum. In the present study, we have compared the reactivity on a panel of self antigens of affinity-purified anti-DNA and anti-thyroglobulin (TG) IgG autoantibodies from the serum of patients with systemic lupus erythematosus (SLE) and autoimmune thyroiditis with their affinity-purified counterparts isolated from the serum of healthy individuals. Anti-DNA autoantibodies exhibited a similar degree of polyreactivity whether originating from patients or from healthy adults. Natural anti-TG autoantibodies were also found to be polyreactive. Anti-TG autoantibodies from patients with Hashimoto's thyroiditis showed little or no polyreactivity. Natural anti-TG autoantibodies were equally polyreactive whether or not they belonged to a fraction of normal IgG that is connected through V regions with other IgG molecules from the same source. These results indicate that polyreactivity of autoantibodies is a feature that does not allow one to distinguish between natural and disease-associated autoantibodies as well as between V-region-connected and unconnected autoantibodies.

Journal ArticleDOI
TL;DR: The purpose of the present work was to compare complement activation and binding of C3 to unmodified and heparin‐treated polystyrene surfaces of microtitre plates and found that the modified surface bound lower amounts of protein than the unmodified.
Abstract: In recent years, conjugation of heparin to biomaterials has been shown to improve its biocompatibility. The purpose of the present work was to compare complement activation and binding of C3 to unmodified and heparin-treated polystyrene surfaces of microtitre plates. When polystyrene was incubated with human serum, C3 was deposited on the surface by both adsorption and binding dependent on activation of the classical (CPW) and alternative (APW) pathways. After end-point attachment of heparin, significant C3 deposition, although at reduced levels, occurred only by CPW-mediated mechanisms, while adsorption and APW-mediated binding were strongly reduced. Generally, the modified surface bound lower amounts of protein, e.g. serum albumin and IgG, than the unmodified. By contrast, it had increased affinity for C1q which leads to binding of C1 and activation of complement via the CPW. Nevertheless, the net effect of the surface modification on the complement reaction was an overall reduction of C3 binding due to obliteration of APW. This can be related to an enhanced factor H/I-dependent down-regulation of C3b and to the lowered protein-adsorbing property of the surface, both of which have inhibitory effects on APW and on the C3 shunt-dependent activation of the complement system.

Journal ArticleDOI
TL;DR: Protection was reflected by statistically significant increases in survival rate of mice immunized with MoAb KT4 which showed variable serum levels of yeast killer toxin‐like anti‐Id antibodies.
Abstract: Anti-Id antibodies were raised in mice against a monoclonal antibody (MoAb KT4) that neutralized the in vitro activity of a Pichia anomala yeast killer toxin. Monoclonal antibody was administered to BALB/C syngeneic mice with different schedules of immunization before intravenous challenge with increasing amounts of yeast killer toxin-sensitive Candida alhicans cells. The course of candidosis was studied in comparison with mice non-immunized and immunized with an isotypc-matched unrelated MoAb subdivided into control groups. Protection was reflected by statistically significant increases in survival rate of mice immunized with MoAb KT4 which showed variable serum levels of yeast killer toxin-like anti-Id antibodies. MoAb KT4 affinity chromatography purified mouse anti-Id antibodies were capable of killing in vitro the yeast ceils of the Candida albicans strain used for the experimental infection. This is the first report of antimicrobial protection that exploits the role of anti-idiotypic antibodies presumably acting in vivo as antibiotics (idiotypic vaccination).

Journal ArticleDOI
TL;DR: The stimulation of peripheral blood mononuclear cells by purified autologous and/or allogeneic monoclonal IgG was studied in five patients with multiple myeloma, nine patients with monOClonal gammopathy of undetermined significance (MGUS) and six healthy individuals.
Abstract: The stimulation of peripheral blood mononuclear cells by purified autologous and/or allogeneic monoclonal IgG was studied in five patients with multiple myeloma (MM), nine patients with monoclonal gammopathy of undetermined significance (MGUS) and six healthy individuals. Single cells secreting IFN-γ or IL-2 were identified using an enzyme-linked immunospot assay. Patients' cells were preferentially stimulated by autologous monoclonal IgG at low concentrations (1–100 pg/ml). while l00 ng/ml or higher stimulated T cells both from patients and, to a lesser degree, healthy individuals. This biphasic dose-response of T-cell stimulation by autologous monoclonal IgG was reproduced in all patients. The numbers of cells secreting IFN-γ and IL-2 in response to allogeneic IgG were significantly lower than the numbers obtained using autologous IgG in patients with MM and MGUS. Cells from healthy individuals were stimulated by allogeneic monoclonal IgG, but to a lesser extent. The results of this study support the presence of idiotype-reactive T cells in patients with MM and MGUS and also may suggest a general but less pronounced T-cell reactivity to monoclonal IgG among these patients.

Journal ArticleDOI
TL;DR: The high numbers of MBP and MBP peptide‐reactive T cells could play a role in the pathogenesis of ON via secretion of effector molecules, one of them being IFN‐γ, as well as in the transfer of ON to MS.
Abstract: The cause of multiple sclerosis (MS) is unknown. Recently reported abnormal T-cell responses to several myelin proteins and myelin basic protein (MBP) peptides in peripheral blood constitute one line of evidence that autoimmune mechanisms could be involved in the pathogenesis of the disease. Monosymptomatic unilateral optic neuritis (ON) is a common first manifestation of MS and important to examine for a possible restriction of the T-cell repertoire early in the disease. T-cell activities to MBP and the MBP amino acid sequences 63-88, 110-128 and 148-165 were examined by short-term cultures of mononuclear cells from cerebrospinal fluid (CSF) and blood in the presence of these antigens, and subsequent detection and counting of antigen-specific T cells that responded by interferon-gamma (IFN-gamma) secretion. Most patients with MS and ON had MBP and MBP peptide-reactive T cells in CSF, amounting to mean values of between about 1 per 2000 and 1 per 7000 CSF cells and without immunodominance for any of the peptides. Numbers were 10-fold to 100-fold lower in the patients' blood. Values were similar in ON and MS, and no evidence was obtained for a more restricted T-cell repertoire in ON. The MBP peptide-recognizing T-cell repertoire was different in CSF than in blood in individual patients with ON and MS, thereby giving further evidence for an autonomy of the autoimmune T-cell response in the CSF compartment. No relations were observed between numbers of autoreactive T cells and presence of oligoclonal IgG bands in CSF or abnormalities on magnetic resonance imaging of the brain in ON or clinical variables of MS. The high numbers of MBP and MBP peptide-reactive T cells could play a role in the pathogenesis of ON via secretion of effector molecules, one of them being IFN-gamma, as well as in the transfer of ON to MS.

Journal ArticleDOI
TL;DR: Different serum IgG and IgG subclass levels were found among Gm phenotypes of a normal population, implying the importance of Gm(f,″,b/f,”,b) and GM(a,′,g/a,″-g) phenotypes causing lower amounts of IgG antibodies.
Abstract: Different serum IgG and IgG subclass levels were found among Gm phenotypes of a normal population. One hundred and fifty-seven Caucasian blood donors were investigated for the reciprocal Gm allotypes on IgG subclass loci namely: for IgG1, G1m(f) and G1m(a); for IgG2, G2m(n) and G2m("); and for IgG3, G3m(b) and G3m(g), and subgrouped in the seven most common Gm phenotypes. The frequencies of Gm phenotypes and haplotypes were given, including numbers of the previously little known G2m(n,") heterozygous individuals. Mean serum quantities +/- SD and range of IgG, IgG1, IgG2, IgG3 and IgG4 were given for different Gm phenotypes. The IgG content was significantly lower in the Gm(f,",b/f,",b) phenotype in which the IgG2 levels were also significantly lower, compared with values of the other phenotypes. IgG3 levels were significantly lower in the Gm(a,",g/a,",g) phenotype compared with other phenotypes. These data imply the importance of Gm(f,",b/f,",b) and Gm(a,",g/a,",g) phenotypes causing lower amounts of IgG antibodies. In evaluating IgG subclass deficiency, the range for the low responding Gm(f,",b/f,",b) and Gm(a,",g/a,",g) phenotypes should be considered.

Journal ArticleDOI
TL;DR: The data obtained suggest that alternative pathway activation is, in some cases, a major factor determining the effectiveness of a particular complement source in the lysis of xenogeneic tumour cells.
Abstract: Sera from 20 species of mammals were tested for their ability to lyse erythrocytes from 18 species of mammals and birds by the alternative complement pathway. Erythrocytes were not lysed by homologous complement, with one minor exception, but all erythrocytes tested were lysed by at least one complement source, and all sera tested except that of the horse lysed at least one type of erythrocyte. Control experiments indicated that lysis was via the alternative complement pathway and that antibodies were not involved. Complement from the various species could be ranked from most active to least active, and erythrocytes could be ranked from most susceptible to least susceptible. There was an inverse correlation between complement activity and erythrocyte susceptibility. The ranking of the orders of placental mammals, from strongest to weakest complement, was carnivore > artiodactyl (ruminants and swine) > primate = armadillo > rodent > rabbit > horse. Opossum serum had activity that placed it in the centre of this range. Ferret complement, the most potent tested, lysed all erythrocytes tested except for homologous erythrocytes, with APCH50 titres as high as 4000. Although the overall reactivity pattern was clear, there were several striking exceptions. For example, the only complement source which lysed ferret erythrocytes was sera of the mouse. The amount of sialic acid present on erythrocytes of 14 mammals was determined, and was, in general, directly correlated with resistance to alternative complement pathway lysis, although there were prominent exceptions to this correlation, involving erythrocytes of the horse, burro and human. All 20 types of complement were also tested for their ability to lyse antibody-coated human tumour cells, under conditions in which both the classical and alternative complement pathways were functional. The data obtained suggest that alternative pathway activation is, in some cases, a major factor determining the effectiveness of a particular complement source in the lysis of xenogeneic tumour cells.

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TL;DR: It is demonstrated that CR1 mediates the interaction between opsonized virus and T cells in addition to its ability to serve as a cofactor for the cleavage of C3b into smaller fragments that interact with CR2.
Abstract: Complement activation by HIV results in the binding of C3 fragments to the gp160 complex and enhanced infection of C3 receptor-bearing target cells. We have studied complement-mediated enhancement of infection of the human CD4-positive T-cell line HPB-ALL which expresses the CR1 (CD35) and CR2 (CD21) receptors for C3. CR1 and CR2 are present on 15% and 40% of normal peripheral blood CD4-positive T lymphocytes respectively. Opsonization of the virus with complement resulted in a 3- to 10-fold enhancement of infection of HPB-ALL cells, as assessed by measuring the release of p24 antigen in culture supernatants throughout the culture period. Blockade of CR2 with cross-linked anti-CR2 monoclonal antibodies decreased infection to the level observed with unopsonized virus. Blocking CR1 reduced complement-mediated infection by 50-80%. Experiments using serum deficient in complement factor I demonstrated that CR1 mediates the interaction between opsonized virus and T cells in addition to its ability to serve as a cofactor for the cleavage of C3b into smaller fragments that interact with CR2. A requirement for CD4 in complement-mediated enhancement of infection was observed with HIV-1 Bru but not with HIV-1 RF. Thus, CR1 and CR2 contribute in an independent and complementary fashion to penetration of opsonized virus into complement receptor-expressing T cells. Involvement of CD4 in infection with opsonized virus depends on the viral strain.

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Matsuda S, Oka S1, Mitsuo Honda, Takebe Y, Takemori T 
TL;DR: IgA antibodies were analysed in sera and saliva from 40 HIV‐1 seropositive individuals to determine the carrier and removal status of IgA in serum and saliva of individuals infected with HIV.
Abstract: IgA antibodies were analysed in sera and saliva from 40 HIV-1 seropositive individuals. The level of total IgA in serum was elevated according to the progress of the disease. IgA antibodies against p24 and gp160 were detected in the asymptomatic phase of infection. However, they declined in the symptomatic phases in contrast with IgG antibodies. Interestingly, three patients in the symptomatic phase who showed high levels of lgA antibodies were all in relatively good clinical condition. The IgG and IgA antibodies in saliva declined in the symptomatic phase. The level of IgG anti-p24 antibodies in saliva correlated with that in serum, suggesting that IgG anti-p24 antibodies in saliva originated from those in the serum. These results indicate that IgA antibodies are regulated independently from IgG antibodies and that the mucosal immune system is impaired early in the symptomatic phase of HIV infection, which starts with mucosal impairment. Detection of IgA antibodies may be useful for prognosis of the disease in HIV-infected individuals. The results indicate also that treatment for the impaired IgA mucosal immune system should be taken into consideration.

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TL;DR: The present results suggest that various populations of macrophages, newly recruited (ED1+) as well as resident Kupffer cells (ED2+), are involved in the immune response against tumour cell deposits in the liver.
Abstract: Macrophages play a role in the host defence against cancer. Little is known about changes in macrophage populations during early metastatic growth. To evaluate the distribution, number and phenotype of macrophages in the development of hepatic metastases in a rat model (Wag/Rij rats and syngeneic CC531 colon carcinoma cell line), an immunohistochemical study was performed with the monoclonal antibodies ED1 (monocytes, and all macrophages), ED2 (resident tissue macrophages, like Kupffer cells) and ED3 (a subpopulation of macrophages which may play a role in the recruitment of lymphocytes). OX19 and Hisl4 were used to identify lymphocytes. In this study a new monoclonal antibody CC52 is described, which recognizes the CC531 tumour cell line. Liver metastases were induced by injection of CC53I colon carcinoma cells into a mesenteric vein. Rats were killed at various intervals. Results show three major macrophage populations during hepatic tumour growth: (1) on day 3, infiltrates are observed around the micrometastases, which contain mainly newly recruited macrophages (ED1+ and ED2−); (2) after 7 days, ED3-positive (ED3 +) macrophages together with T lymphocytes are found in the infiltrates; (3) an increase in the number of ED2-positive (ED2+) Kupffer cells is observed in the liver parenchyma after 14 days. In conclusion, the present results suggest that various populations of macrophages, newly recruited (ED1+) as well as resident Kupffer cells (ED2+), are involved in the immune response against tumour cell deposits in the liver.

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TL;DR: Opsonic activity of hyperimmune rabbit IgG against fibronectin‐binding proteins of Staphylococcus aureus is described and it was shown that mice passively immunized with hyperimmune IgG anti‐FnBP (one or two doses, intravenously) before challenge with S. a Aureus was more effective than observed in control groups.
Abstract: In this report, opsonic activity of hyperimmune rabbit IgG against fibronectin-binding proteins (gal-FnBP A and ZZ-FnBP B) of Staphylococcus aureus is described. Moreover, the action of IgG purified from serum of rabbits immunized with 'combined vaccine' (fibronectin-binding protein A+collagen-binding protein+alpha-toxoid) is shown. The opsonic activity has been studied in an in vitro phagocytosis assay as well as in vivo. Mice which had been infected intraperitoneally with S. aureus strain Cowan 1 pretreated (opsonized in vitro) with specific anti-FnBPs IgG were able to eliminate the staphylococci from the peritoneal cavity and liver more rapidly than controls. Also, clearance from the bloodstream of intravenously injected S. aureus Cowan 1 as well as S. aureus U320, opsonized with IgG anti-FnBPs or anti-FnBP+CnBP+alpha-toxoid, was more effective than observed in control groups. In other in vivo experiments it was shown that mice passively immunized with hyperimmune IgG anti-FnBP (one or two doses, intravenously) before challenge with S. aureus Cowan I eliminated the bacteria better than controls injected only with preimmune IgG.

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E. Rakasz, A. Gal, J. Biro, G. Balas, András Falus 
TL;DR: Present data suggest that local imbalance in the ratio of these three cytokines in different pathological cases might influence the glucocorticosteroid sensitivity of the lymphocytes, monocytes and hepatocytes as target cells.
Abstract: In order to elucidate the role of the inflammatory cytokines in regulating glucocorticosteroid binding (GCSB) and glucocorticosteroid receptor (GR) level we incubated a B-cell line (CESS), a promonocytic cell line (U937) and a hepatoma cell line (HepG2) in the presence of varying concentrations of IL-1 beta, IL-6 and TNF-alpha for 24 h. Glucocorticosteroid binding was determined by the method of 'whole cell uptake', and the cellular appearance of the glucocorticosteroid receptor was detected by immunocytochemistry. A rise in the glucocorticosteroid binding was induced by all three cytokines. The increase in level of glucocorticosteroid receptors in the cells shown by immunocytochemistry was much more pronounced. However, antagonistic effects were demonstrated by both methods between IL-6 and TNF-alpha, and between IL-1 beta and TNF-alpha when they were applied simultaneously, in U937. Present data suggest that local imbalance in the ratio of these three cytokines in different pathological cases might influence the glucocorticosteroid sensitivity of the lymphocytes, monocytes and hepatocytes as target cells.

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TL;DR: In the U‐937 cell line but not in normal PB monocytes, the tryptase expression was upregulated 3–50 fold following phorbol ester (PMA)‐induced differentiation, but no such induction was seen after retinoic acid, interferon‐γ or vitamin D3 exposure.
Abstract: Expression of a mast cell tryptase mRNA was detected in two human monocytic cell lines, the U-937 and the Mono Mac 6, and in normal human peripheral blood (PB) monocytes. In the U-937 cell line but not in normal PB monocytes, the tryptase expression was upregulated 3-50 fold following phorbol ester (PMA)-induced differentiation, but no such induction was seen after retinoic acid, interferon-gamma or vitamin D3 exposure. The tryptases expressed in PMA-induced and non-induced U-937 and in Mono Mac 6 were characterized by PCR amplification and nucleotide sequence analysis. The U-937 cell line was found to express a tryptase identical to one of the previously cloned mast-cell beta tryptases (Tryptase I), and the tryptase expressed in Mono Mac 6 was found to be nearly identical to the previously cloned alpha tryptase. By northern blot analysis with oligonucleotide probes specific for the alpha and beta tryptases both cell lines were found to express only one type of tryptase. Densitometric quantifications of tryptase mRNA levels, in the two cell lines, showed approximately 80 times higher mRNA levels in Mono Mac 6 compared to non-induced U-937. Immunohistochemical staining for tryptase showed a marked heterogeneity in the Mono Mac 6 cell line. Only one out of 10 cells were positive for the protein but the levels in these cells were very high, equivalent, or even higher than the levels seen in the human mast cell line HMC-1. This shows that the expression of a single tryptase, in this case the alpha tryptase, is sufficient for the production of a stable protein and probably also a stable proteolytically active tetramer. The family of human mast-cell tryptases has been considered to represent a class of proteases specifically expressed in mast cells and basophilic leucocytes. The expression of tryptases in two monocytic cell lines and in normal PB monocytes indicate that in humans, the lineage specificity of these serine proteases is less restricted than earlier expected. The cloning of a full length cDNA for the murine counterpart to the human mast cell tryptases, the MMCP-6, is presented. No expression of the MMCP-6 was detected in a panel of mouse monocyte or macrophage cell lines indicating a species difference in the lineage specificity of the 'mast cell tryptases'.

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TL;DR: Results indicate a novel class of T‐cell‐definable tumour antigens recognized by tumour‐reactive CTL on human tumours and may be significant for understanding of cellular immunity in ovarian cancer, identification of CTL‐defined tumourAntigens and future adoptive specific immunotherapeutic approaches in ovariancancer.
Abstract: CTL-TIL lines have been developed from tumour infiltrating lymphocytes (TIL) from the ascites of patients with ovarian carcinoma, and used to investigate whether common tumour antigens are expressed on allogeneic ovarian tumours epithelial tumour lines derived from colon and pancreatic carcinoma. Three CTL lines expressed preferential cytolytic activity against autologous tumour cells and against certain allogeneic ovarian tumour cells that shared HLA-A2 molecules. Analysis of the target specificity of these CTL lines indicated that they also lysed human colon and pancreatic tumour lines sharing HLA-A2. CTL-TIL clones isolated from these lines were found to lyse HLA-A2+ ovarian, colon and pancreatic tumours, and to recognize clonally distributed common epitopes on pancreas and colon tumour clones. These results indicate that shared tumour antigens can be found among tumours of common epithelial cell origin. These results indicate a novel class of T-cell-definable tumour antigens recognized by tumour-reactive CTL on human tumours and may be significant for understanding of cellular immunity in ovarian cancer, identification of CTL-defined tumour antigens and future adoptive specific immunotherapeutic approaches in ovarian cancer.

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TL;DR: The results show that both Th l and Th2 clones can protect against tumour development and the anti lympboma properties of both the Thl and Th 2 clones appear to involve as yet undefined cytotoxic and growth inhibiting molecules.
Abstract: Idiotypes (Id) can serve as individual markers on B cells; therefore, cytotoxic Id-specific T cells may play a significant role in immunological surveillance of Id+ B-cell tumours. We have investigated the anti-tumour activity of CD4+ BALB/c Th1 and Th2 clones which recognize a processed Id of the syngeneic lambda 2(315) L chain in the context of the class II MHC molecule I-Ed. Id-specific T cells and A20/46 B lymphoma cells transfected with the lambda 2(315) gene were injected s.c. into the same site of BALB/c mice (Winn assay). The results show that both Th1 and Th2 clones can protect against tumour development. The protection was Id-specific because T cells did not influence tumour development by an A20/46 B lymphoma cell line transfected with the pSV2neo expression vector alone. In vitro studies showed that the Th1 clones were cytotoxic to lambda 2(315)-transfected B lymphoma cells; by contrast, the Th2 clone was not cytotoxic in 51Cr-release assay even though the Th2 cells inhibited the growth of lambda 2(315) B lymphoma cells. The anti-lymphoma properties of both the Th1 and Th2 clones appear to involve as yet undefined cytotoxic and growth inhibiting molecules.