bluuebunny/crossref_metadata_2025 huggingface dataset:
Colony-stimulating factors and interferon-gamma differentially affect cell surface molecules shared by monocytes and neutrophils
Authors: BUCKLE A M , JAYARAM Y , HOGG N
Published: July 2010

Monocytes and neutrophils share a common progenitor and perform many similar functions, as reflected in the expression of several shared membrane molecules. We show here that such molecules can be independently regulated by the cytokines interferon-gamma (IFN-γ) and the colony-stimulating factors (CSF) in the context of the cell type on which they are present. Thus, IFN-γ causes neutrophils to express the high-affinity receptor for IgG, FcγRI, which has been considered to be a monocyte-specific receptor. Although neutrophil FcγRI never reaches the levels present on monocytes. it is induced more rapidly and by lower amounts of IFN-γ than monocyte FcγRI. Of the CSF, only macrophage (M) CSF has an effect which is to cause a decrease in expression of monocyte FcγRI. Secondly, after culture monocytes express FcγRIII which is constitutively expressed by neutrophils. Although ncutrophil FcγRIII can be readily modulated by IFN-γ. granulocyte (G) CSF and granulocyte macrophage (GM) CSF, these cytokines do not alter the low levels of monocyte FcγRIII. Thirdly, expression of the gp55 protein recognized by CD14 monoclonal antibodies is decreased after exposure of monocytes to IFN-γ. Neutrophils express low levels of CD14 and, in this case. IFN-γ causes an increase in the CD 14 antigen on these cells. Of all the molecules investigated, only HLA class II is confined to monocytes. with increases in expression induced by IFN-γ but not by the CSF.
The neuropeptide substance P stimulates the effector functions of platelets
Authors: DAMONNEVILLE M , MONTÉ D , AURIAULT C , CAPRON A
Published: July 2010

Sensory neuropeptides, such as substance P. appear as potent mediators of various immunological reactions, and inhibit or stimulate a wide range of functions of immune inflammatory cells. Platelets were recently shown to participate as effector cells in an IgE or lymphokine-dependent killing of parasites. Substance P and its carboxy-terminal fragment SP (4–11) induce the cytotoxic activity of platelets towards the larvae of Schistosoma mansoni, respectively, by 90% and 40%, whereas the modified C terminal SP, the SP-free acid, exhibits no effect on the platelets. The neuropeptide effects occur at low doses (10-8 M), are specific as shown by inhibition studies with a substance P antagonist, theD-SP. Binding data obtained after flow cytofluorometry with FITC-SP lead to the conclusion that SP binds specifically to about 20% of the homogenous population of platelets. Moreover, IgE could modulate the SP-dependent functions of platelets since the pre-incubation with myeloma human IgE or with AP2 monoclonal antibodies—known to inhibit the IgE-dependent killing of these cells leads to a dramatic decrease of the SP dependent cytotoxic activity of platelets towards the larvae. These findings identify a potent mechanism for nervous system regulation of host defence responses.
Relative increase of inflammatory CD4+ T cells in the cerebrospinal fluid of multiple sclerosis patients and control individuals
Authors: ROTTEVEEL F T M , KUENEN B , KOKKELINK I , MEAGER A , LUCAS C J
Published: July 2010

Of three patients with multiple sclerosis (MS) and two non-MS individuals a large number of CD4+ T cell clones was obtained from the cerebrospinal fluid and peripheral blood by direct limiting dilution. The CD4+ T cell clones from cerebrospinal fluid and peripheral blood lymphocytes were compared for their cytotoxic activity and lymphokine production. Cytotoxic capacity of cloned T cells was analysed with the use of anti-CD3 antibodies and target cells bearing Fc receptors for murine IgG. Recently, we demonstrated the existence of two different subsets of human CD4+ T cell clones by phenotypic and functional criteria. One type of CD4+ T cell with anti-CD3 mediated cytotoxic activity, in analogy with murine studies, is the inflammatory or TH1 subtype, the main producer of interleukin (IL-2), interferon-gamma (IFN-γ) and tumour necrosis factor (TNF)-α, -β, whereas the other type of CD4+ T cell clone lacked anti-CD3 mediated cytotoxicity and produced minimal amounts of IL-2 concomitant with reduced levels of IFN-γ and TNF-α, -β. The present study demonstrates that among three MS patients, relatively more inflammatory CD4+ T cell clones with cytotoxic activity and IFN-γ and TNF-α, -β production were derived from the cerebrospinal fluid as compared with peripheral blood lymphocytes. Also among control individuals more inflammatory CD4+ T cell clones could be obtained from the cerebrospinal fluid as from the peripheral blood. The enrichment of inflammatory CD4+ T cells, therefore, appears to be physiological rather than associated with the disease.
Anti-endothelial cell antibodies in patients withWegener's granulomatosis and micropolyarteritis
Authors: FERRARO G , MERONI P L , TINCANI A , SINICO A , BARCELLINI W , RADICE A , GREGORINI G , FROLDI M , BORGHI M O , BALESTRIERI G
Published: July 2010

Anti-endothelial cell antibodies (AECA) have been detected by cell surface radioimmunoassay innine out of 15 patients with micropolyarteritis (MPA) and in two out of five patients with Wegener'sgranulomatosis. AECA mostly belonged to the IgG isotype and were present in the active phase ofthe diseases. These antibodies were not detectable in 10 sera from patients with essential mixedcryoglobulinaemia. suggesting that they were not a mere epiphenomenon consequent to theinflammatory vascular injury. The binding activity was not related to ABH antigens or to HLA class Iantigens displayed by resting human endothelial cells in culture and was not influenced by removingimmune complexes. Absorption of the anti-neutrophil cytoplasmic antibodies (ANCA). present inMPA and Wegener's granulomatosis sera, did not affect the endothelial binding. AECA-positive seradid not display lytic activity against endothelial cells, neither alone nor after addition of freshcomplement or normal human peripheral blood mononuelear cells. Although AECA arc notcytolytic for endothelial cell monolayers in vitro, the reactivity against intact endothelial cells suggeststheir possible involvement in in vivo pathological processes affecting vascular structures in smallvessel primary vasculitides.
The affinity of anti-HBc antibodies in acute and chronichepatitis B infection
Authors: WEN Y M , DUAN S C , HOWARD C R , FREW A F , STEWARD M W
Published: July 2010

Antibodies to hepatitis B core antigen (anti-HBc) are found in the sera of all individuals infected withhepatitis B virus. A role for these antibodies has been suggested in determining the outcome ofinfection. In this study, the affinity of anti-HBc antibodies in asymptomatic virus carriers wascompared with that of antibodies present in the sera of patients with chronic liver disease. Persistentlyinfected individuals with no evidence of clinical disease were found to have anti-HBc antibodies ofgreater affinity, compared with the chronic liver disease group. Sera from patients with chronichepatitis contained high levels of low-affinity antibody whereas antibody levels in asymptomaticcarriers were significantly lower. These findings are discussed in relation to the predicted role of anti-HBc antibodies in mediating hepatitis B virus-related hepatocellular injury.
Complement activation and the prognostic value of C3a in patients at risk of adult respiratory distress syndrome
Authors: ZILOW G , STURM J A , ROTHER U , KIRSCHFINK M
Published: July 2010

In vivo and in vitro studies have shown that complement activation plays an important role in the pathogenesis of the adult respiratory distress syndrome (ARDS). In a prospective study of polytrauma patients at risk of ARDS (n= 38) complement parameters were determined over a period of 14 days in serial plasma samples (obtained every 6 h during the first 48 h). Polytrauma induced a rapid and remarkable complement activation. Low levels of the complement proteins C3, C4, C1 inhibitor (Cl INH) factor I and factor H during the first 48 h indicated complement consumption in all patients. Elevated C3a levels in the first few hours after injury were associated with the later development of ARDS. A more sensitive indicator than C3a alone was the calculated C3a:C3 ratio discriminating ARDS and non-ARDS patients. A second rise of C3a levels and C3a:C3 ratio from day 4 on paralleled the course of extravascular lung water. To assess the mode of complement activation, the activation-specific protein complexes Cl rCls-Cl INH and C3b(Bb)P were measured in some of the patients. We demonstrate that in the first 48 h complement activation occurred via the alternative pathway only and was later followed by an additional activation via the classical pathway. Our observations suggest that monitoring of C3a and C3 in plasma can identify polytrauma patients at high risk for ARDS at an early stage of the disease.
Thyroid peroxidase and the induction of autoimmune thyroid disease
Authors: MCLACHLAN S M , ATHERTON M C , NAKAJIMA Y , NAPIER J , JORDAN R K , CLARK F , SMITH B REES
Published: July 2010

Animal models of autoimmune thyroid disease are associated with thyroglobulin (Tg) as autoantigen whereas in man the autoimmune response to microsomal antigen/thyroid peroxidase (TPO) appears to play a major role in thyroiditis. Consequently, we have compared the ability of TPO and Tg to induce thyroid autoantibodies and thyroid damage in mice known to be susceptible (CBA/J) or resistant (BALB/c) to thyroiditis induced using murine Tg. Groups of three to five mice were immunized twice using Freund's complete adjuvant with 80–100μg highly purified porcine (p) TPO, pTg, rat (r) Tg, human Tg, bovine serum albumin (BSA) or BSA + 0.2 μg pTg (the level of Tg contamination of TPO). Four weeks after immunization with TPO, plasma from CBA/J (but not BALB/c) mice contained IgG class antibodies which bound to TPO-coated tubes in the presence or absence of excess Tg (and could therefore be clearly distinguished from Tg antibodies) but there was no evidence of thyroiditis in either strain of mice. In contrast, in CBA/J mice immunized with rTg and, to a lesser extent in mice that had received pTg, thyroid tissue was infiltrated with lymphoid cells and/or neutrophils and antibodies to pTg (but not pTPO) were present. Our observations demonstrate that induction of TPO antibody alone is insufficient to lead to thyroiditis in CBA/J mice. Further, these studies emphasize the complex interactions between MHC and different thyroid antigens in the processes leading to thyroid destruction.
Mechanism of a lymphocyte abnormality associated with HLA-B8/DR3: role of interleukin-1
Authors: HASHIMOTO S , MICHALSKI J P , BERMAN M A , MCCOMBS C
Published: July 2010

Lymphocytes from normal individuals with the histocompatibility antigens HLA-B8 and DR3 have impaired proliferative responses when stimulated with suboptimal concentrations of mitogens. We have previously shown that an important factor in the impaired response is a failure to produce normal quantities of interleukin-2 (IL-2). To examine the mechanism of decreased responsiveness further, we measured interleukin-1 (IL-1) production of low responder subjects compared with controls. The peripheral blood mononuelear cells of five low responder individuals with HLA-B8/ DR3 stimulated with 0.05 μg/ml of phytohaemagglutinin (PHA) accumulated only 0.036 U/ml of IL-I compared with 0.32 U/ml for normal respondcrs. There was a highly significant correlation between the PHA-stimulated IL-1 concentration at 12 h and the subsequent IL-2 concentration at 48 h (r=0.89, P<0.000l) suggesting a role of decreased IL-1 production in the impaired response. A study of unfractionated or column-fractionated culture supernatants revealed no evidence that the decreased IL-1 activity in the supernatants of low responder subjects was related to increased IL-1 inhibitor concentrations. These results suggest that impaired IL-2 production and lymphocyte proliferation in healthy subjects with HLA-B8/DR3 may be mediated at least in part by decreased IL-1 production, and implicates a defect of a very early event in lymphocyte activation.
Effects of a novel immunosuppressive agent, FK506, on human B cell activation
Authors: SUZUKI N , SAKANE T , TSUNEMATSU T
Published: July 2010

We examined the effect of new immunosuppressive agent, FK506, on the human B cell function, in comparison with that of cyclosporin A (CyA) and tried to define the discrete activation step(s) which is selectively affected by FK506 and CyA. We used polyclonal B cell activators, Staphylococcus aureus Cowan I (SAC) and pokeweed mitogen (PWM). We found that (i) the initial B cell activation process by PWM, which is on the basis of T cell-dependent manner, is susceptible to the inhibitory effects of FK506 and CyA. while initial B cell activation on the basis of T cell-independent manner by SAC is resistant to these drugs: (ii) they also inhibit helper factor production by T cells; (iii) once they are activated, the B cells become resistant to inhibition by the drugs; and (iv) on an equimolar basis, FK506 exhibits 100-fold greater inhibitory activity than does CyA. Thus FK506 mainly interferes with interactions between T cells and other cells which are essential for B cell activation process, resulting in inhibition of B cell function.
Loss of natural killer activity as an indicator of relapse in acute leukaemia
Authors: TRATKIEWICZ J A , SZER J
Published: July 2010

A role for naturally occurring cytotoxic cells in immunosurveillance against malignancy has been presumed in several studies. The natural killer activity (NKA) of peripheral blood mononuclear cells was therefore measured at regular intervals in patients with acute leukaemia and expressed specific cytotoxicity. Sixty controls had a median NKA of 33·6% (range 15·4–71). Seventy-three patients with acute lymphoblastic leukaemia (ALL) and acute non-lymphoblastic leukaemia (ANLL) with untreated or relapsed disease had a median activity of 2·4% (range 0–13·4) (P< 0·001), while 57 patients who had achieved complete remission had a median activity of 22·7% (range 9·5–64·4). In 15 patients, reductions of NKA were seen prior to 16 episodes of relapse. In ten of these (nine with ANLL and one with ALL), 11 relapses were preceded within 10 weeks by drops in NKA to less than 30% of remission levels. The median NKA of the group prior to the drop in activity was 25·5% and the median first low value was 60%. Five patients who relapsed after allogeneic bone marrow transplantation had significant, sustained drops in NKA, 5–9 weeks earlier. The median NKA prior to the drop was 25·6% and the median first low value was 8·0%. We therefore conclude that there was a marked reduction in NKA in patients with active acute leukaemia when compared with healthy blood donors and that this activity substantially improved in complete remission. All patients who relapsed had significantly reduced NKA which in some, significantly preceded the time of relapse. These data suggest that the regular assessment of NKA in patients with acute leukaemia may be a useful diagnostic tool.
Murine monoclonal antibodies to DNA. A comparison of MRL/lpr NZB/W and chronically graft-versus-host-diseased mice
Authors: BRINKMAN K , VAN DAM A , VAN DEN BRINK H , TERMAAT R-M , BERDEN J , SMEENK R
Published: July 2010

Hybridomas producing monoclonal antibodies to DNA were prepared from NZB/W F1 (n= 20), MRL/lpr (n= 13), mice with a chronical graft versus-host-disease (GVHD) (n= 8) and polyclonally stimulated mice (n = 9). Screening was performed by means of an anti-DNA EL1SA. Reaction patterns in four different anti-DNA assays (anti-DNA ELISA, indirect immunofluorescence on Crithidia luciliae. PEG assay and Farr assay) as well as avidity and cross-reactivity of these monoclonals were studied in relation to anti-DNA (sub)class and murine origin of the clones. It was found that monoclonal anti-DNA derived from mice with chronic GVHD did not differ from monoclonal anti-DNA derived from NZB/W Fl or MRL/lpr mice, with respect to isotype distribution, avidity towards DNA, cross-reactivity and assay behaviour in the anti-DNA assays mentioned before. In contrast, monoclonal anti-DNA obtained from polyclonally stimulated mice were all of the IgM isotype and displayed a stronger cross-reactive behaviour than the other three models. Altogether, these results exclude the possibility that anti-DNA in the GVHD mice originates from the non-specific pool of natural autoantibodies and further emphasize the relevance of chronic GVHD as a murine model of systemic lupus erythematosus.
Immunomodulation in mineral dust-exposed lungs: stimulatory effect and interleukin-1 release by neutrophils from quartz-elicited alveolitis
Authors: KUSAKA Y , CULLEN R T , DONALDSON K
Published: July 2010

Quartz deposition in the rat lung causes an intense and persistent neutrophil alveolitis leading to parenchy mal fibrosis. Bronchoalveolar leucocytes (BAL) from quartz-exposed rat lungs were studied for their effects on splenic lymphocyte proliferation; titanium dioxide (TiO2) was used as a control, non-fibrogenic dust. Seven days after the intratracheal instillation of 1 mg of quartz or TiO2 suspended in phosphate-buffered saline (PBS), BAL were recovered by lavage; the effect of PBS alone was also studied. TiO2-elicited BAL (macrophages > 98%) inhibited splenocytes responding to suboptimal phytohaemagglutinin (PHA) more than PBS-elicited BAL (macrophages > 98%); the effect was dependent on the BAL:splenic lymphocyte ratio. Quartz-elicited whole BAL (macrophages 49%, neutrophils 51 %), and an alveolar macrophagc-enriched population with purity of 87% separated from it, were less inhibitory to splenocyte mitogenesis than PBS-elicited BAL. A neutrophil-enriched population, with a purity of 80%, markedly enhanced splenocyte response to PHA. In addition, whole quartz BAL and the macrophagc-enriched population obtained from it enhanced the mitogenesis of T cell-enriched lymphocytes at a much lower BAL:lymphocyte ratio. The neutrophil-enriched quartz BAL enhanced mitogenesis substantially more than the whole or macrophage-enriched population from quartz-exposed lung. Supernatants from normal macrophages, PBS BAL, TiO2 BAL, quartz BAL and both alveolar macrophage and neutrophil-enriched quartz populations were assessed for interleukin-1 (IL-1) activity. Quartz-BAL, quartz macrophages and quartz neutrophils all produced significantly higher IL-1 levels than PBS BAL; the supematants from quartz neutrophils, however, showed the highest IL-1 activity. These findings suggest that quartz-elicited bronchoalveolar leukocytes, especially neutrophils, enhance lymphocyte proliferation and that increased IL-1 secretion by these cells is likely to be the effector molecule involved. These findings have important implications for immune response in mineral dust-stimulated lung and for inflammatory lung disease in general.
Specificities of IgM and IgG anti-histone H1 autoantibodies in autoimmune mice
Authors: MONESTIER M , FASY T M , DEBBAS M E , BOHM L
Published: July 2010

Autoantibodies to histone H1 represent the most common specificity among anti-histone autoantibodies in systemic autoimmune diseases. Here we analyse anti-H1 autoantibodies in mice from the following autoimmune strains: MRL/Mp-lpr/Ipr. NZB and NZB × NZW/F1. Autoantibodies of the IgM isotype bind predominantly to epitopes located in the COOH-terminal domain of the H1 molecule, whereas IgG autoantibodies in the MRL/Mp-lpr/Ipr and NZB strains also recognize epitopes requiring the integrity of both the COOH-terminal and the central globular domains of H1. In both of these strains, the titre of these IgG anti-H 1 antibodies rises during the course of the disease. The importance of three-dimensional structure of histone H1 was attested by a significant decrease in IgG binding after cleavage of the H1 molecule within the folded globular domain. The binding of these sera to H1 variants from various species was also investigated and a strong binding of MRL/Mp-lpr/lpr sera to certain phylogenetically distant histone H1 variant molecules (sea-urchin sperm H1 and chicken erythrocyte H5) was observed. This cross-reacting binding can be explained by the presence in MRL/Mp-lpr/lpr sera of autoantibodies to H10, a variant found in non-dividing cells and exhibiting sequence homologies lo the above mentioned variants. The significance and the possible implications of these data for the pathogenesis of autoimmunity are discussed.
Anti-M9 antibodies in sera from patients with primary biliary cirrhosis recognize an epitope of glycogen phosphorylase
Authors: KLEIN R , BERG P A
Published: July 2010

Anli-M9 antibodies in sera from patients with primary biliary cirrhosis (PBC) were previously found to recognize two antigenic determinants at 98 and 59 kD, using a purified antigen fraction derived from rat liver mitochondria in the Western blot. Here we show that these antibodies are directed against an epitope of the enzyme glycogen phosphorylase. By Western blotting, a determinant at 98 kD was obtained testing anti-M9 positive sera against phosphorylase from skeletal muscle, and after plasmin treatment a degradation product appeared at 59 kD. Both determinants were identical to the M9-specific determinants 98 and 59 kD as shown by absorption studies. When these antibodies were eluted from the 98 and 59 kD determinants of the M9 antigen after immunoblotting. they again recognized the same epitopes on plasmin-treated phosphorylase. Furthermore, phosphorylase enzyme activity could be also demonstrated in the purified M9 fraction, and anti-M9-positive/anti-M2-negative but not anti-M9-negative/anti-M2-positive sera could be shown to stimulate phosphorylase activity. Testing sera from 1189 patients with different hepatic and non-hepatic disorders against M9 and phosphorylase from skeletal muscle by ELISA. 20% were positive with phosphorylase and only 2% with the M9 fraction. These data indicate that the commercially available phosphorylase from skeletal muscle cannot be recommended as M9 source. It may still contain non-PBC-specific epitopes which are probably recognized by naturally occurring antibodies directed against this highly conserved protein.
Kinetics and pathogenicity of autoantibodies induced by mercuric chloride in the brown Norway rat
Authors: PUSEY C D , BOWMAN C , MORGAN A , WEETMAN A P , HARTLEY B , LOCKWOOD C M
Published: July 2010

Repeated low-dose injections of mercuric chloride (HgCl2) in the brown Norway (BN) rat result in polyclonal activation which includes the induction of anti-glomerular basement membrane (GBM) autoantibodies. We examined the kinetics of various auloantibodies produced in vivo, general features of polyclonal activation such as total IgG levels and immune complex formation, and the relationship between organ specific autoimmunity and tissue injury in the kidney and thyroid. The production of immune complexes and autoanlibodies to GBM and thyroglobulin was short lived, and the increase in levels of total IgG and antibodies to ssDNA and dsDNA was prolonged; the antibody response to collagen types I and II was intermediate in duration. Auloantibodies induced by HgCI2 caused only mild and variable tissue injury in the kidneys and did not induce abnormalities in the thyroid. These studies demonstrate that immunostimulation by mercury may result in the formation of a range of autoantibodies. with variable kinetics and pathogenicity.
Differential expression of the calcium-binding proteins MRP8 and MRP14 in granulomatous conditions: an immunohistochemical study
Authors: DELABIE J , WOLF-PEETERS C DE , OORD J J VAN DEN , DESMET V J
Published: July 2010

MRP14 and MRP8 are well-characterized calcium-binding proteins present in myeloid cells and mononuclear phagocytes. These antigens can easily be visualized in paraffin-embedded tissue, making use of monospecific polyclonal antibodies. This study evaluates MRP14 and MRP8 expression in mononuclear phagocytes in various granulomatous conditions. MRP14 is strongly expressed in all granulomatous conditions. MRP8 is variably expressed. Mononuclear phagocytes in granulomas of foreign body type, cat-scratch discase and erythema nodosum strongly express MRP8. In contrast, MRP8 expression is weak or absent in mononuclear phagocytes of sarcoidosis and tuberculosis. These results show differences in immunophenotype between non-phagocytic mononuclear phagocytes in delayed hypersensitivity type granulomas and phagocytic mononuclear phagocytes in non-hypersensitivity and non-immunological granulomas.
Relationships between 2H4 (CD45RA) and UCHL1 (CD45RO) expression by normal blood CD4+CD8−, CD4−CD8+, CD4−CD8dim+, CD3+ CD4−CD8− and CD3−CD4−CD8− lymphocytes
Authors: RICHARDS S J , JONES R A , ROBERTS B E , PATEL D , SCOTT C S
Published: July 2010

We characterized and established relationships between the expression of membrane 2H4 (CD45R A) and UCHL1 (CD45RO) by enriched lymphocyte fractions prepared by selective immunomagnetic depletion of monoclonal antibody-defined populations. Cell fractions analysed in this study could be divided into two broad groups according to the presence (CD.3+VCD4+CD8−, CD3+CD4−CD8+ CD3+ CD4− CD8dim+ and CD3+CD4− CD8−) or absence (CD3−CD4−CD8dim+ and CD3− CD4− CD8−) of the CD3 antigen. Preliminary studies confirmed a reciprocal relationship for CD45RA and CD45RO expression by major lymphoid components and further showed that the level or intensity of membrane 2H4 staining (2H4+, 2H4int and 2H4−) could be directly related to UCHLI expression. As a reflection of their differential functions, the various CD3+ populations examined showed much greater heterogeneity in 2H4 and UCHLI expression. CD3+CD4+CD8 cells generally showed significant proportions of 2H4+, 2H4int and 2H4− components, whereas the CD3+CD4−CD8+ population was characterized by a predominance of 2H4+ cells. The results of this current investigation further suggested a higher proportion of dual-positive (2H4+ UCHL1+) cells and a much greater degree of inter-individual variation than previously suspected. In contrast to CD3* lymphocytes, natural killer (NK) associated CD3−CD4− CD8dim+ and CD3−CD4−CD8−populations were mostly 2H4+ with only minor 2H4int components and very low expression of UCHL1. An additional observation of note was that the proportions of 2H4+ and 2H4− cells comprising the CD4+ CD8− fraction in any given individual was highly correlated (P= 0·002) with the distributions of 2H4+ and 2H4− components within the CD4− CD8+ fraction. This suggests the possible existence of a common control mechanism for the acquisition of immunological memory by distinct lymphocyte populations and further indicates that individual variations in the distribution of 2H4/UCHLI lymphocyte subpopulations may be a direct consequence of immunological experience’ rather than age alone.
Lack of association of T cell receptor beta-chain constant region polymorphism with insulin-dependent diabetes mellitus in Finland
Authors: REIJONEN H , SILVENNOINEN-KASSINEN S , JLONEN J , KNIP M
Published: July 2010

Allelic polymorphism in the T cell receptor constant beta-chain gene region has been reported to be associated with autoimmune diseases, including insulin-dependent diabetes mellitus (IDDM). The present analysis of 164 children and adolescents with IDDM and 193 controls for Bq/II polymorphism using a TcR-Cβ cDNA probe revealed two allelic restriction fragments with sizes of 10.5 kb (U) and 9.6 kb (L). No particular association was observed between the RFLP genotypes and IDDM (UU 27%versus 31%; UL 53%versus 52%; and LL 20%versus 17%, in diabetic subjects and controls, respectively), nor were any differences found between patients with various HLA risk antigens. The frequency of heterozygotes was 52% in 63 DR3-positive diabetic subjects and 53% in 73 DR3-negative ones. The results do not support any involvement of the TcR constant region genes in susceptibility to IDDM.
Role of interleukin-2 and interleukin-6 in the mitogen responsiveness of T cells from patients with ‘common-variable’ hypogammaglobulinaemia
Authors: NORTH M E , WEBSTER A D B , FARRANT J
Published: July 2010

We have assessed the ability of interleukin-2 (IL-2) and interleukin-6 (IL-6) to augment the proliferative response of T lymphocytes from ‘common-variable’ hypogammaglobulinaemia (CVH) patients and from normal controls, to the mitogens phytohaemagglutinin (PHA) and OKT3. We show that with cells from the control group and from those patients whose T cells respond to PHA within the control range, both IL-2 and IL-6 will significantly augment the response to OKT3. However, in those patients with a T cell defect in which the PHA response is below the control range, neither IL-2 nor IL-6 could restore the PHA or OKT3 response to normal. Responses to IL-2 or IL-6 alone were always in or above the control range.
IgG subclass antibodies toPseudomonas aeruginosain sera from patients with chronicPs. aeruginosainfection investigated by ELISA
Authors: PRESSLER T , PEDERSEN S S , ESPERSEN F , HØIBY N , KOCH C
Published: July 2010

ELISAs using subclass-specific monoclonal antibodies were developed for the quantification of human IgGl, IgG2, IgG3 and IgG4 antibodies to Ps. aeruginosa. We investigated the pattern of IgG subclass antibodies against Ps. aeruginosa in serum from patients with cystic fibrosis (CF). other patients with chronic Ps. aeruginosa infection, and healthy controls. Healthy controls and patients with CF but without Ps. aeruginosa infection showed no or very low litres of antibodies against Ps. aeruginosa. In the early stage of chronic Ps. aeruginosa infection, antibody litres in all four subclasses were significantly higher than either normals or CF patients without infection. Other patients with Ps. aeruginosa infection showed the same increased level of IgG subclass antibodies as CF patients in an early stage of infection. Sixteen patients (eight in good and eight in poor clinical condition) have been followed for an average of 13 years with multiple serum samples covering the pre-infection, early and late stages of chronic infection. Patients in a poor clinical condition showed significantly higher levels of IgG3 antibodies in the first year of infection and 2 years later also had significantly higher IgG2 antibody levels. We conclude that elevated levels of IgG2 and lgG3 antibodies to Ps. aeruginosa are a sign of poor prognosis in CF.
Reduced oxidative burst responses in monocytes and monocyte-derived macrophages from HIV-infected subjects
Authors: MÜLLER F , ROLLAG H , FRØLAND S S
Published: July 2010

Oxidative burst responses of monocytes and monocyte-derived macrophages (MDM) were studied in 40 subjects with HIV infection of different clinical stages. Oxidative burst was assessed as reduction of nitroblue tetrazolium (NBT) with or without stimulants. Results were determined as oxidative burst responses per cell and as a stimulatory ratio between stimulated and unstimulated NBT reduction. Cells from 12 HIV-seronegative homosexual men and 38 blood donors served as control groups. In patients with asymptomatic HIV infection, monocyte oxidative burst responses were reduced compared with the blood donors. In MDM from the same patients, stimulatory ratios were reduced. In AIDS patients, stimulatory ratios of both monocytes and MDM were reduced compared with controls. In contrast to the progressive deterioration of CD4+ lymphocyte counts as well as other immune functions in HIV infection, monocyte oxidative burst responses are impaired already in the asymptomatic phase of the infection, almost to the same extent as in patients with AIDS.
Epitope mapping of an HLA-B27 monoclonal antibody that also reacts with a 35-kD bacterial outer-membrane protein
Authors: TOUBERT A , HAMACHI M , RAFFOUX C , PARK M S , YU D T Y
Published: July 2010

Ye-3 is an HLA-B27-specific murine monoclonal antibody recognizing the heat-modifiable protein of Enterobacteriaceae. Here we used recombinant hybrid molecules between the HLA-B7 and HLA-B27 antigens to delineate the epitope recognized by Ye-3. Results of these experiments indicated that the segment of HLA-B*2705 spanning residues 77–81 was critical to the reactive epitope. It is known to be a major serological and functional recognition site of HLA-B*2705 and our data give support for its involvement also in the serological cross-reactivity with bacterial antigens.
Changes in plasma levels of interleukin-2 receptor in relation to disease exacerbations and levels of anti-dsDNA and complement in systemic lupus erythematosus
Authors: TER BORG E J , HORST G , LIMBURG P C , KALLENBERG C G M
Published: July 2010

Interleukin-2 receptor (IL-2R) is expressed and released predominantly by activated T cells. In order to investigate whether disease exacerbations of systemic lupus erythematosus (SLE) are preceded by T cell activation, we prospectively measured levels of IL-2R once a month, from 6 months prior to exacerbations until 1 month afterwards. To assess the temporal relation between T cell activation and B cell activation, we measured, in addition, levels of anti-dsDNA, complement C3JC4, and total IgG. During a mean follow-up period of 23 months, 40 exacerbations occurred in 21 out of the 71 participating patients. For the present study one exacerbation per patient was evaluated. During exacerbation levels of IL-2R were increased in 18 out of the 21 cases and correlated with levels of anti-dsDNA (P < 0.02). C3 (P < 0.02), and C4 (P < 0.01), but not with the score of the disease activity index. Levels of IL-2R rose prior to the excerbation (P < 0.02) and fell afterwards following treatment (P < 0.05). Even in the absence of disease activity or during minor disease symptoms IL-2R levels were higher (P < 0.01) than in healthy controls. Sixteen out of the 21 exacerbations (76%) were preceded by a significant increase in IL-2R. Changes in levels of anti-dsDNA and complement C3JC4 tended to precede changes in levels of IL-2R. We conclude that increased levels of IL-2R, compatible with T cell activation, are present in SLE already during inactive disease. These levels further increased prior to exacerbations of disease. As such, IL-2R is an indicator of disease activity in SLE. Serial measurement of IL-2R is a sensitive test for predicting disease exacerbations of SLE.
Accelerated expression of anti-Sm and Y2 idiotype differ in dependence on T cell subsets in MRL/+ mice
Authors: MILLER M L , DANG H , TALAL N
Published: July 2010

We investigated the role of MRL T cells in the induction of anti-Sm antibodies and Y2 idiotype. Four injections of Sm antigen in Freund's complete adjuvant were required to induce peak amounts of specific anti-Sm antibody in young BALBJc and MRL/+ mice. The Y2 idiotype was expressed in MRL/+ mice but not in BALBJc mice. Expression of both anti-Sm, predominantly IgG2a heavy chain, and Y2 idiotype was augmented in MRL/+ mice after two injections of Sm if. prior to immunization, mice received splenicTcells from naive MRL/lpr or immunized, but not naive MRL/+ mice. These results suggest that the lpr gene contributes to the ability of autoimmune T cells to augment the anti-Sm antibody response. Treatment of primed M RL/+ donor T cells with anti-CD4, but not anti-CD8, antibodies and complement removed the ability to augment anti-Sm antibody production. In contrast, augmentation of Y2 idiotype production was abrogated by pretreatment of donor T cells with either anti-CD4 or anti-CD8. These results suggest that, while MRLJ+ CD4 + T cells play an important role in anti-Sm antibody production, additional interaction between CD4+ and CD8+ T cells augments Y2 expression.
Sera with high levels of anti-smooth muscle and anti-mitochondrial antibodies frequently bind to cytoskeleton proteins
Authors: DIGHIERO G , LYMBERI P , MONOT C , ABUAF N
Published: July 2010

Using ELISA methods, 54 sera from chronic active hepatitis (CAH) patients displaying high levels of anti-smooth muscle antibodies (SMA) and 18 sera from primary biliary cirrhosis (PBC) patients with high levels of anti-M2 antibodies were examined for the presence of high antibody levels against actin, tubulin, myosin, tropomyosin, troponin. vimenlin and desmin. Our results showed that: (i) in CAH with high SMA activity, increased antibody levels were found in 51-9% of sera for actin, 31.5% for myosin, 35.2% for tubulin, 34.0% for tropomyosin, 11.3% for troponin. 22.6% for vimentin and 43.4% for desmin, compared with natural antibody levels in 21 normal sera; (ii) Similar high levels of these antibodies were found in the case of PBC; (iii) in most cases, sera simultaneously bound to several antigens of the panel; and (iv) approximately 26% of the CAH sera were found to be negative with the seven antigens examined while 22% were reacted with a cytoskeleton protein (CP) other than actin. These results indicate that current opinion associating SMA with anti-actin activity in CAH is confirmed for only 50% of cases and that although a good correlation between SMA and anti-CP antibodies can be obtained, there is still a significant percentage of SMA for which the putative antigen recognized needs to be determined.
Plasma and tissue interleukin-2 receptor levels in inflammatory bowel disease
Authors: MAHIDA Y R , GALLAGHER A , KURLAK L , HAWKEY C J
Published: July 2010

Plasma and tissue interleukin-2 receptor (IL-2R) levels were determined in patients with active ulcerative colitis and Crohn's disease. Compared with healthy controls (median 440 U/ml; range 240–900), significantly higher levels of plasma IL-2R were present in patients with active ulcerative colitis (median 1180 U/ml; range 580–7150; P < 0.002) and Crohn's disease (median 1340 U/ml; range 480–9000; P < 0002). Compared with other laboratory parameters, plasma IL-2R levels were related most closely to clinical score of disease activity in Crohn's disease. Plasma IL-2R levels also reflected the clinical course and may provide a more accurate assessment of disease activity in Crohn's disease. In plasma of patients undergoing intestinal resection of active inflammatory bowel disease, raised levels of IL-2R were present in samples from mesenteric vein (draining inflamed intestine) compared with those from peripheral vein. In tissue homogenates of colonic biopsies, significantly higher levels of IL-2R were present in specimens from colons with active ulcerative colitis compared with healthy controls (median 230.2, range 20.7–581.5 versus 77.9, range 34.2–291.3; P < 0.02).
Signal requirements for activation of leukaemic T cells from a chronic lymphocytic leukaemia (T-CLL)
Authors: ZOCCHI M R , POGGI A , HELTAI S , VILLA A , INVERARDI L , VICARI A , SABBADINI M G , FERRARINI M
Published: July 2010

In order to define the signal requirements Tor leukaemic T cell activation, the proliferation and interleukin-2 (IL-2) production of peripheral lymphocytes from a patient with a HTLV-I., CD4+, CD45RA+ CD45RO+ CD25- T-CLL were evaluated after the delivery of different stimuli. Unlike resting CD4+ normal T lymphocytes that can be activated only by a two-signal stimulation. T-CLL cells proliferated and released IL-2 in response to a pair of anti-CD2 monoclonal antibodies (MoAbs) or concanavalin A (Con A) in the absence of both accessory cells (AC) and phorbol myristate acetate (PMA). The two stimuli were also able to induce CD25 expression within 12–20 h on the majority of T-CLL cells. A response to anti-CD3 and anti-CD28 MoAbs was detected only in the presence of PMA, similar to that observed in normal resting T lymphocytes matched for phenotype. Both Con A-and CD2-induccd proliferation were strongly inhibited by the addition of anti-CD2S MoAb. Furthermore, T-CLL lymphocytes acquired anti-tumour lytic activity after culture in the presence of PMA and ionomycin. We conclude that HTLV1 CD25 T-CLL can be characterized not only by morphological and phenotypical studies but also on the basis of signal requirements for cell activation.
NADP+ reduction by human lymphocytes
Authors: KLEIN A , CHAN A W-L , CAPLAN B U , MALKIN A
Published: July 2010

The hexose monophosphate shunt (HMPS) is known to be responsible for the reduction of NADP+ by lymphocytes. We tried to find other enzymatic systems that might provide the lymphocytes with NADPH. By measuring the absorbance at 340 nm we noted that the addition of NADP+ to a preparation of disrupted lymphocytes resulted in the formation of NADPH at a rate of 4 nmol/106 cells per min. This phenomenon could not be changed by negative feedback inhibition of HMPS, and could not be attributed to the low concentration of glucose, glucose-6-phosphate (G-6-P) and isocitrate found in the cell preparation (NADP+ -dependent isocitratc dehydrogenase in addition to HMPS NADP+ reducing enzymes was found to be present in lymphocytes). Because of the activity of a NADP+ -dependent lactate dehydrogenase, pyruvate oxidized the NADPH as it was being formed. Here we demonstrate the presence of an unknown NADP+ reducer in lymphocytes which seems to play an additional role to HMPS in NADP+ reduction by lymphocytes. NADP+-dependent lactate dehydrogenase may play a role in regulating the NADP+/NADPH ratio.
Phase I trial with recombinant interleukin-2 (rIL-2): immune activation by rIL-2 alone or following pretreatment with recombinant interferon-gamma
Authors: FARACE F , MATHIOT C , BRANDELY M , TURSZ T , DORVAL T , POUILLART P , TRIEBEL F , HERCEND T , FRIDMAN W H
Published: July 2010

Alterations of imniunological parameters were analysed in patients with advanced malignancies during a phase I trial with rIL-2. Five-day infusions of rIL-2 at doses from 1 × 106 to 24 × 106 biological response modifiers program (BRMP) U/m2 per day were given to 29 patients, with a minimum of three patients per dose. The dose of 24 × 106 U/m2 per day was the maximal tolerated dose (MTD). Immunological parameters were analysed at days 0, 8 and 11 of the rIL-2 courses. Following a leucopenia during rlL-2 infusion, a lymphocytosis was found in all patients except one. The lymphocylosis peaked at day 8 and was detected at doses of rIL-2 as low as 1 × 106 U/m2 per day, reaching a plateau at a dose of 16 × 106 U/m2 per day. Although all lymphocyte subsets were increased in patients receiving rIL-2. some patients had predominant T cells (CD3+, NKH1(CD56)-), others had predominant natural killer (NK) cells (CD3-. NKH1(CD56)+), and yet others showed a mixed profile. A strong induction of cells cytotoxic for K.562 targets was found in all patients at days 8 and 11. Eighteen patients received, 1 month later, a second treatment in which infusion of rIL-2 was preceded by a course of 5 days infusion of 2 × 106 U/m2 per day recombinant interferon-gamma (rIFN-γ)- The infusion of rIFN-γ prior to rIL-2 had no effect on the rIL-2-induced alterations of immunological parameters. Taken together, our results suggest that immune stimulation by rIL-2 occurs even at low doses and is maximal at a dose below the MTD; and that pretreatment with low-dose rlFN-γ does not modify the immune stimulation by rIL-2.
Interferon-gamma potentiates antibody affinity in mice with a genetically controlled defect in affinity maturation
Authors: HOLLAND G P , HOLLAND N , STEWARD M W
Published: July 2010

Interferon-gamma (IFN-γ), the tetrapeptide tuftsin and the synthetic nonapeptide from interleukin-1 beta (IL-1 β) (amino acids 163–171) have previously been shown to act on macrophages and/or T cells and to enhance antibody titres to T cell-dependent antigens. The ability of these immunomodulatory agents to potentiate antibody affinity in addition to antibody titre has been studied in a line of mice that fail to demonstrate normal maturation of antibody affinity (low N/M mice). The results presented here confirm that each of the agents potentiate antibody levels following simultaneous injection with a T cell-dependent antigen but demonstrate that only IFN-γ is able to enhance antibody affinity in these mice. The observation that IFN-γ can enhance both antibody affinity and antibody levels suggests that it could be an important adjuvant for vaccine use.
Thyroid autoantigens and human T cell responses
Authors: FUKUMA N , MCLACHLAN S M , RAPOPORT B , GOODACRE J , MIDDLETON S L , PHILLIPS D I W , PEGG C A S , SMITH B REES
Published: July 2010

We investigated the ability of T cells from patients with Hashimoto's thyroiditis and with Graves' disease as well as control donors to proliferate in response to thyroid peroxidase (TPO) and thyroglobulin using (i) lymphoid cells from different lymphoid organs; (ii) unfractionated or CD8-depleted lymphoid suspensions or T cells+autologous low density cells (LDC); (iii) 200-μI well cultures and 20-μl hanging-drop microcultures; and (iv) intact TPO and thyroglobulin, denatured thyroglobulin and 12 synthetic peptides predicted on the basis of the amino acid sequence of TPO to be T cell epitopes. In 200-μl well cultures, proliferative responses (assessed in terms of 3H-thymidine uptake) to intact TPO or thyroglobulin, digested thyroglobulin or synthetic TPO peptides were not significantly different in unfractionated or CD8-depleted lymphoid suspensions from blood, thyroid or lymph nodes of TPO/thyroglobulin autoantibody-positive patients, autoantibody-negative patients or control donors. In contrast, blood T cells from some high titre patients with Hashimoto's thyroiditis (but not from healthy individuals) proliferated in response to intact thyroglobulin or TPO presented by autologous LDC in hanging-drop microcultures. Heat denatured thyroglobulin (with which thyroglobulin autoantibodies do not interact) did not stimulate proliferation and this observation, together with the ability of T cells from some patients to respond to intact thyroglobulin in the absence of LDC, indicated that thyroglobulin-specific B cells may be involved in antigen presentation. As we were unable to demonstrate proliferation by blood T cells + LDC from all thyroglobulin antibody-positive patients with Hashimoto's thyroiditis, our studies suggest that the presence of sufficient precursor T cells, as well as the number and type of antigen-presenting cells, are critical for T cell proliferative responses to human TPO and thyroglobulin.
The effect of recombinant interferon-alpha on lymphocyte subpopulations and HLA-DR expression on liver tissue of HBV-positive individuals
Authors: YOO Y K , GAVALER J B , CHEN K , WHITESIDE T L , VAN THIEL D H
Published: July 2010

Interferon-alpha (IFN-α) has been reported to be beneficial in the treatment of chronic active hepatitis occurring as a result of hepatitis B virus (HBV) infection. Treatment with IFN-a has been proposed as a means of reducing the high rate of allograft infection in clinical liver transplantation in patients transplanted for HBV-related chronic active hepatitis and cirrhosis who are positive for hepatitis B surface antigen (HBsAg). We obtained resected whole livers from two groups of patients who received liver transplants. Group A consisted of 11 patients who were HBsAg+ but were not treated with IFN-α, and group B consisted of 10 patients who were also HBsAg+ but received IFN-α therapy for 29.4 ± 5.6 days prior to orthotopic liver transplantation. No differences between the two groups existed in terms of a variety of demographic and clinical characteristics. The liver tissue was stained with monoclonal antibodies to cell surface antigens unique to different mononuclear cell populations by the avidin-biotin-immunoperoxidase technique to determine the effect of IFN-α on the lymphocyte subsets as well as HLA antigen expression on liver-infiltrating mononuclear cells, The number of HLA-DR+ lymphocytes in the liver was significantly increased (P < 0.005) within the portal areas in group B compared with that found in group A (84 ± 14 versus 33 ± 5 per one high-power field). Moreover, the intensity of the HLA-DR antigen expression on lymphocytes in the portal areas (P < 0.02) and in the hepatic lobule (P < 0.05) was greater in group B than in group A. The number of natural killer (NK) cells was increased in the portal areas (P < 0.05) of group B compared with group A. These alterations in the lymphocyte and NK cell populations present in the liver in response to IFN-α therapy presumably reflect an IFN-α-induced enhancement of the immune response to virus-infected cells.