bluuebunny/crossref_metadata_2025 huggingface dataset:
Serum CD14 levels in poly traumatized and severely burned patients
Authors: KRÜGER C , SCHÜTT C , OBERTACKE U , JOKA T , MÜLLER F E , KNÖLLER J , KÖLLER M , KÖNIG W , SCHÖNFELD W
Published: July 2010

Recently it has been demonstrated that the CD14 molecule which is expressed on monocytes and macrophages serves as a receptor for lipopolysaccharide (LPS) bound to LPS-binding protein (LBP) and thus mediates LPS-induced tumour necrosis factor (TNF) production. Here we report that CD14 is found as a soluble (s) molecule in serum. In healthy volunteers sCD14 levels (mean ± s.e.m.) were 3.7±0.05 μg/ml (n= 30, 25–50 years of age) as determined by ELISA (detection limit 20 μg/ml serum) using two monoclonal antibodies in a sandwich technique. In polytraumatized patients (n= 16) significantly decreased levels (1.7 ± 0.3) were detected immediately after the trauma, which increased to 4.9±0.3 μg/ml within the first 6 days post trauma. sCD14 remained elevated during the first 14 days post trauma in patients with the most severe injuries (injury severity score > 45 points), whereas a return to normal levels was observed in patients with an injury score of < 45 points. In addition, the levels of the high-density lipoproteins that partially inactivate free endotoxin are significantly decreased post trauma. No correlation between parameters of inflammation (C3a and neopterin levels, leucocyte counts, amount of band cells), liver function and sCD14 levels was established. Comparable to polytraumatized patients, increased sCD14 serum levels were observed in five patients with burn trauma (burned area > 35%) within the second week post trauma when clinical signs of septicaemia were evident.
Increased spontaneous secretion of IL-6 from B cells of patients with B chronic lymphatic leukaemia (B-CLL) and autoimmunity
Authors: LAHAT N , AGHAI E , MAROUN B , KINARTY A , QUITT M , FROOM P
Published: July 2010

We studied B cells from 18 patients with B-CLL, six of them with autoimmune haemolytic anaemia, for spontaneous secretion of IL-6. Our aim was to determine whether the increased incidence of autoimmune disease found in B-CLL patients is associated with enhanced spontaneous IL-6 secretion. IL-6 was measured by the effect of B cell supernatants on the proliferation of an IL-6 dependent plasmacytoma cell line T1165. The highest IL-6 values (7.4±1.8 U/ml) were measured in supernatants derived on day 3 of culture from lymphocytes of the six patients with B-CLL and concomitant autoimmune disease. The maximal IL-6 values for 10 patients with B-CLL only were 2.8±0.3 U/ml and for 10 age-matched controls, 0.8±0.3 U/ml (P < 0.01, each group compared with the other). We conclude that there is an association between B-CLL, autoimmune disease and the spontaneous in vitro secretion of IL-6. Further studies are needed to determine whether the IL-6 secretion plays a role in the pathogenesis of autoimmune disease in patients with B-CLL.
In vivotreatment with interferon causes augmentation of IL-2 induced lymphokine-activated killer cells in the organs of mice
Authors: PURI R K , LELAND P
Published: July 2010

Interferon-alpha (IFN-α) has been shown to synergize with IL-2 in the regression or a variety of established murine tumours and studies are underway to explore this combination in patients with advanced cancers as well. To understand the mechanism of synergy we have studied lymphokine-activated killer (LAK) cell activity in various compartments of mice in response to IFN-α and IL-2 administration. The effects of IFN-γ, TNF-α and IL-4 were also examined. C57BL/6 mice were injected intraperitoneally with HBSS, IL-2 alone, IFN-α alone or both, two times a day for 7 days. On days 4 and 8, LAK activity was tested in a 4-h chromium release in cells obtained from lungs, spleen, and liver using fresh MCA-102 tumour cells as targets. The cells from control mice failed to lyse the MCA-102 target. IL-2 caused the generation of LAK. activity and an increase in total cell yield in all the organs after 3 days of injection. IFN-α failed to generate LAK activity but when administered along with IL-2, caused synergistic enhancement of LAK lysis of MCA-102 target cells. Cell yield in this group was lower as compared with the IL-2-treated group. LAK activity tested after 7 days of IL-2 therapy was significantly decreased compared with that observed after 3 days. However, activity remained at as high a level after 7 days of therapy as after 3 days of therapy in animals treated with IFN-α and IL-2. FACS analysis revealed that asialo GM-1+ (ASGM-1) and NK1.1+ cells were increased in number in IL-2 and IL-2 plus IFN-α-treated spleen; however, the number of these cells was similar in both groups. In the liver, ASGM-1+ cells were higher in the IL-2 plus IFN-α group than in the group treated with IL-2 alone. By in vitro depletion utilizing antibody and Rbc’ experiments, it was clear that both ASGM-1+ and NK1.1+ cells from the spleen mediated most of the cytotoxicity of MCA-102 targets. Pre-treatment irradiation (5 Gy) of mice completely abrogated the capability of IL-2 or IL-2 plus IFN-α to generate LAK activity. IFN-γ also had a stimulatory effect on IL-2 induction of LAK activity. Tumour necrosis factor-alpha (TNF-α) and IL-4 failed to generate LAK activity and, in combination with IL-2, no additional stimulatory effect was observed. These studies indicate that induction of LAK activity may be at least partly responsible in the mediation of synergistic anti-tumour effects of IFN-α and IL-2 or IFN-γ and IL-2.
Interleukin-2 receptor gene expression in kidney transplant recipients treated with cyclosporin A
Authors: YOSHIMURA N , OKA T , AMAGAI T , HORII Y , IMANISHI J
Published: July 2010

We examined the effects of cyclosporin A (CsA) administered in vivo on the capacity of peripheral blood mononuclear cells (PBMC) from kidney transplant recipients to express IL-2 receptor (IL-2R) gene at the level of mRNA after milogen stimulation in vitro. There were no differences in the percentage of IL-2R+ cells among the groups of normal individuals, azathioprine-prednisolone treated, and CsA-prednisolone-treated recipients, using FITC-labelled monoclonal anti-IL-2R antibody (anti-α chain): 40±3±10±1% and 62±8±11±1% of normal PBMC (n= 18), 37±0±9±3% and 61±7±5±8% of PBMC from azathioprine-prednisolone-treated recipients (n = 20), and 37±7±9±6% and 60±7±12±7% of PBMC from CsA-prednisolone-treated recipients (n = 20) expressed IL-2R after 24 h and 48 h of phytohaemagglutinin stimulation, respectively. However, in a study of Northern blotting using cDNA for 1L-2R (anti-a chain specific), both the 3500 and 1400 bp families of IL-2R mRNA were remarkably decreased in PBMC from CsA-prednisolone-treated recipients compared with azathioprine-prcdnisolone-trealed recipients and normal individuals. These studies demonstrated that CsA could inhibit IL-2R gene expression ai the level of mRNA at physiological concentration.
Tissue distribution of mucosal antibody-producing cells specific for respiratory syncytial virus in severe combined immune deficiency (SCID) mice engrafted with human tonsils
Authors: NADAL D , ALBINI B , SCHLÄPFER E , CHEN C , BRODSKY L , OGRA P L
Published: July 2010

Groups of C.B-17 SCID mice were reconstituted intraperitoneally with human tonsillar mononuclear cells (hu-TMC) from children seropositive for antibody to respiratory syncytial virus (RSV) and subsequently challenged intraperitoneally with inactivated RSV or sham-immunized. The synthesis and the distribution characteristics of human antibody to RSV in various murine tissues were studied using an enzyme-linked immunospot assay (ELISPOT). No specific antibody was observed in sham-immunized animals. In contrast, mice engrafted with hu-TMC exhibited the appearance of specific human antibody secreting cells (hu-ASC) after i.p. immunization with inactivated RSV. RSV-specific hu-ASC were detected only in animals engrafted with cells from donors seropositive for antibodies to Epstein-Barr virus. Hu-TMC engrafted mice showed RSV-specific IgM and, in lower numbers, IgG hu-ASC in several tissues including the lungs. Numbers of RSV-specific IgA hu-ASC were low, however, and detected only in the lung. No RSV-specific hu-ASC were detected in the intestine. These data demonstrate for the first time that hu-TMC-SCID chimeras respond to immunization with viral antigen. Furthermore, the results suggest that hu-TMC engraft in lungs but not in the intestinal tissue.
Accelerated diabetes in non-obese diabetic (NOD) mice differing in incidence of spontaneous disease
Authors: BAXTER A G , MANDEL T E
Published: July 2010

The NOD mouse is an established model of autoimmune diabetes mellitus. Various lines of NOD mice differ in their incidence of spontaneous diabetes, e.g. 93% of female NOD/Lt mice compared with 46% of female NOD/Wehi mice develop diabetes by 250 days. These two lines were studied under conditions which greatly accelerate the onset of hyperglycaemia. It was hoped that their responses to these manipulations would reveal characteristic differences which would increase our understanding of diabetes resistance in the low incidence NOD/Wehi line. One dose of 300 mg/kg of cyclophosphamide (CP) produced hyperglycaemia in 50% of NOD mice within 2 weeks in both lines. They were also equally susceptible to diabetes induced by splenocyte transfer at 21 days of age from prediabetic 150-day-old NOD/Lt or NOD/Wehi females. Five daily 40 mg/kg doses of streptozotocin (STZ) resulted in a severity of diabetes in the NOD mice greater than in C57BL or SJL/J mice. While the incidence and severity of diabetes induced in the two NOD lines were similar, this appeared to be principally due to sensitivity to the toxic effects of STZ rather than its ability to exacerbate autoimmune β cell destruction. It has previously been shown that it is possible to prevent diabetes in susceptible NOD mice with simple, relatively benign therapies and here we show that it is possible to induce diabetes in resistant animals at a rate indistinguishable from fully predisposed individuals. It therefore appears that the prediabetic NOD mouse is poised in an immunologically precarious state with the onset of disease being highly dependent on factors which exacerbate or moderate autoimmune destruction.
Corticosteroid can alter antigen expression on alveolar macrophages
Authors: MARIANAYAGAM L , POULTER L W
Published: July 2010

Normal healthy volunteers underwent broncho-alveolar lavage and the cells obtained were cultured for 24 h and 48 h, either alone or in the presence of the corticosteroid, Budesonide. Cell differentials were all normal, the lavages containing > 90% alveolar macrophages. Cytospins of these cells were prepared before and after culture. The cytospins were subjected to immunocytochemical analysis using a panel of MoAbs selected to identify subsets of macrophages and functionally relevant surface antigens. In particular, the expression of RFD1 (antigen presenting cell marker) and RFD7 (mature phagocyte marker) were studied. Before culture, BAL macrophages could be divided into two subsets. Of the cells, 39.3% were RFD1+ and 47.2% were RFD7+. Culture with Budesonide was seen to reduce the proportions of RFD1+ cells to 38% while increasing the RFD7+ population to 69% of total. These changes were relatively specific as Budesonide failed to alter the expression of CD68 or Fc(IgG) receptors. Down-regulation of HLA-DR expression was seen, however, after 24 h contact with Budesonide. As these changes could have functional significance, these data support the hypothesis that steroids may have direct effects on the role of alveolar macrophages in immune responses in the lung.
Monoclonal antibodies VTB-E3, IB5 and HB9 to the leucocyte/epithelial antigen CD24 resemble BA-1 in recognizing sialic acid-dependent epitope(s). Evidence that VIB-E3 recognizes NeuAcα2-6GalNAc and NeuAcα2-6Gal sequences
Authors: LARKIN M , KNAPP W , STOLL M S , MEHMET H , FEIZI T
Published: July 2010

The specificities of seven monoclonal antibodies to the human B cell differentiation marker CD24 have been investigated with respect to sialic acid containing carbohydrates. These are antibodies HB8, HB9, VIB-C5, VIB-E3, ALIa, LC66 and IB5, which are known to bind to polydisperse sialoglycoprotein(s) on Nalm-6 B lymphoblastoid cells. Three of the antibodies, HB9, VIB-E3 and IB5, have been found to resemble the first described antibody in this series, BA-1, in binding also to bovine submaxillary mucin. As with BA-1. the binding of the antibodies is abolished or reduced markedly after desialylation of the epithelial glycoprotein, and the binding to neuraminidase-treated Nalm-6 cells is also reduced. There is evidence for the involvement of non-O-acetylated sialic acid in the determinants recognized by these antibodies, since there is a substantially enhanced binding following mild-alkali treatment of the epithelial mucin which removes O-acetyl groups. One of the antibodies. VIB-E3. is deduced to recognize the oligosaccharide sequences NeuAcα2-6GalNAc and NeuAcα2-6Gal as part of larger antigenic structures. This conclusion has been reached from the results of inhibition-of-binding experiments using a series of structurally defined sialo-oligosaccharides and direct binding experiments using oligosaccharides chemically linked to lipid (neoglycolipids).
A comparison of T cell responses to glycoprotein B (gB-1) of herpes simplex virus type 1 and its non-glycosylated precursor protein, pgB-1
Authors: O’DONNELL C A , CHAN W L
Published: July 2010

The ability of non-glycosylated precursor glycoprotein B (pgB) to induce T cell responses in herpes simplex virus (HSV) infected mice was compared with fully giycosylated glycoprotein B (gB) and with whole virus. pgB was as effective as gB in priming for virus- and glycoprotein-specific T cells. pgB could also re-stimulate virus or glycoprotein primed cells in vitro as efficiently as gB. In addition, priming with pgB protected mice against a lethal challenge with HSV type 1 (HSV-1) and could induce the early in vivo production of IL-2 and IL-3 in infected mice. In all of these responses, pgB was as effective as gB. Thus, the carbohydrate side chains on gB do not appear to be necessary for T cell recognition of this protein.
Lymphocyte proliferative responses to chlamydial antigens in human chlamydial eye infections
Authors: MABEY D C W , HOLLAND M J , VISWALINGAM N D , GOH B T , ESTREICH S , MACFARLANE A , DOCKRELL H M , TREHARNE J D
Published: July 2010

In order to study the relationship between cell-mediated immune responses to Chlamydia trachomatis and the pathogenesis of human chlamydial eye disease, we have measured the peripheral blood lymphocyte proliferative responses to whole chlamydial elementary bodies in 40 subjects with oculogenital chlamydial infection of varying severity, 13 subjects with genital chlamydial infections and 12 healthy seronegative controls. The mean stimulation index was significantly higher in those with oculogenital infections than in controls. There was a strong correlation between the response to C. trachomatis serotypes B and L1. We studied the relationship between proliferative responses and four clinical parameters: follicular conjunctivitis, papillary hypertrophy, corneal pannus and epithelial punctate keratitis. but were unable to show a significant association with any of these. Nor was there any association between proliferative response and serum antibody titre to C. trachomatis (pooled serotypes D-K), duration of disease or quantitative isolation of chlamydia from the conjunctiva. The depletion of CD8+ cells had no consistent effect on proliferative responses to serotype L1 in 13 subjects.
T cell responses to synthetic thyroid peroxidase peptides in autoimmune thyroid disease
Authors: TANDON N , FREEMAN M , WEETMAN A P
Published: July 2010

Sixteen peptides, representing four different extracellular regions of thyroid peroxidase (TPO) predicted to contain a high proportion of potential T cell epitopes. were synthesized to investigate which parts of this autoantigen may be targets for the T cell response in thyroid autoimmunity. Compared with 25 controls, peripheral blood T cells from 23–37% of 30 patients with Graves’ disease or autoimmune hypothyroidism were stimulated significantly by three peptides, representing amino acids 415–432, 439–457 and 463–481 of the TPO sequence; T cells from individual patients were also stimulated by several other peptides. These results indicate that the T cell response to TPO is directed against several epitopes which may be recognized by different patients.
Qualitative difference of anti-DNA antibody-producing cell precursors in the pre-immune B cell repertoire between normal and lupus-prone mice
Authors: IWAI K , TSUBATA T , KATSURA Y , KUMAGAI S , IMURA H
Published: July 2010

The precursor frequency for anti-DNA antibody-producing cells in the pre-immune B cell repertoire was investigated in young female BALB/c and NZW mice, and in young and aged female NZB ± NZWFl (B/WF1) mice. Spleen cells from these mice were diluted serially and stimulated polyclonally in vitro with lipopolysaccharide (LPS) and IL-4 to induce both IgM and IgG1 production. The results demonstrated that there existed virtually no difference in precursor frequency for IgM anti-DNA antibody-producing cells between normal and lupus mice, confirming previous observations made by other investigators. In contrast, the number of precursors for IgG 1 anti-DNA antibody-producing cells was much higher in young and old B/WF1 mice than in normal mice. These results suggest that the high frequency of precursors for IgG1 anti-DNA antibody-producing cells in the pre-immune B cell repertoire of B/WF1 mice is a crucial factor for the pathogenesis of systemic lupus erythematosus.
Increased expression of class II major histocompatibility complex molecules on B cells in rats susceptible or resistant to HgC12-induced autoimmunity
Authors: DUBEY C , BELLON B , HIRSCH F , KUHN J , VIAL M C , GOLDMAN M , DRUET P
Published: July 2010

Administration of HgCl2 to the susceptible Brown-Norway (BN)rais induces an autoimmune disease characterized by a T-dependent polyclonal activation of B cells responsible for a dramatic increase in serum IgE concentration. The resistant Lewis (LEW) rats injected with HgCi2. do not exhibit such autoimmune manifestations. We show here that, upon HgCl2 injections, major histocompalibility complex (MHC) class II molecule expression is increased very early in lymph nodes and spleen B cells from both strains. So far. it is the earliest marker (day 3)of the effect of HgCI2on the immune system. In both strains this enhancement is transient, but regulatory mechanisms arc much more efficient in the resistant LEW strain than in the susceptible BN strain, In addition, we observed that MHC class II molecule expression on B cells differs according to the organ and the rat strain tested. All these findings are discussed in an attempt to underline the role of MHC class II molecule expression in the occurrence of mercury-induced stutoimmunity.
High serum levels of CD8 antigen in primary biliary cirrhosis: a possible cause of suppressor cell dysfunction?
Authors: NOURI ARIA K T , LOMBARD M , WILLIAMS R
Published: July 2010

Reduced suppressor cell number and function have been described in a number of autoimmune diseases and this may contribute to pathogenesis. Suppressor cell function depends upon the interaction of the CD8 antigen expressed on suppressor cells with other limbs of the immune system. Recently, soluble membrane antigens including CD8 have been identified in serum and it is possible that the loss of such antigens from viable cells could result in functional deficit. In order to examine whether the decreased suppressor cell function reported in autoimmune type of chronic liver disease is associated with soluble serum CDS levels, sera from 23 patients with primary biliary cirrhosis (PBC). 12 with autoimmune chronic active hepatitis (AI-CAH) and 21 healthy controls were tested using a commercially available enzyme immunoassay. The proportion of cells expressing the CDS antigen and the intensity of its display were also determined using an immunofluorescent technique and an ELISA, respectively, for 12 PBC and 10 healthy controls. The soluble serum CDS levels were significantly higher in PBC (mean U/ml.s.d., 777.331). and AI-CAH (575±291) than controls (322±115) (P < 0.001 and P = 0004, respectively). While the intensity of CDS antigen expression on suppressor/cytotoxic populations was not significantly different in PBC (347±125 per 104 cells) compared with controls (441 ±206). the mean proportion of CDS positive cells was significantly less in PBC (14.1±6.8%) than controls (20.4±7%) (P < 0.05). These data suggest that the apparent reduction in suppressor cell number found for patients with PBC and AI-CAH may be a consequence of the shedding or secretion of CDS antigen from cell membrane of CDS positive lymphocyte. It is also possible that the loss of this antigen is responsible for the reduced suppressor cell function seen in these conditions.
Deficiency of purine nucleoside phosphorylase activity in thymocytes from the immunodeficient diabetic BB rat
Authors: WU G , MARLISS E B
Published: July 2010

The spontaneously diabetic BB (BBd) rat displays marked T lymphopenia. The present study was designed to investigate whether the immunodeficiency in this animal may be associated with deficiency of purine nucleoside phosphorylase (PNP) and possibly adenosine deaminase (ADA). The activities of these two enzymes were measured in lymphoid and non-lymphoid cells from both non-diabetes-prone (BBn) and BBd rats as well as from streptozotocin-induced diabetic (STZ) BBn rats. There were no significant differences between BBn and BBd rats in ADA activities in thymocytes, skeletal muscle or brain. However, ADA activity was increased (P 0.01) by 50% in BBd mesenteric lymph node lymphocytes and splenocytes as compared with BBn cells, but was not altered in cells from STZ-BBn rats. On the other hand, the PNP activity in BBd thymocytes was only 61% (P <0.01) of that observed in BBn cells. This PNP deficiency was not the consequence of diabetes per se, as its activity was normal in thymocytes from STZ-BBn rats. There were no significant differences in PNP activities between BBn and BBd rats in all other cell types examined. The diabetic BB rat may be a novel source of PNP-deficient thymocytes (mainly immature T eells) for studying biochemical mechanisms of immunodeficiency in association with decreased PNP activity. The findings also raise the question of whether a causal relationship exists between PNP deficiency and the recently demonstrated abnormality in T cell maturation in the thymus of the BBd rat.
Low dose streptozotocin causes stimulation of the immune system and of anti-islet cytotoxicity in mice
Authors: KANTWERK-FUNKE G , BURKART V , KOLB H
Published: July 2010

Multiple low doses of streptozotocin arc known to induce immune-mediated insulin deficient diabetes and depression of immune reactivity. We show here that immune depression by streptozotocin is not general but that some parts of the immune system are stimulated. Spleen cells from streptozotocin-treated mice showed enhanced cytotoxicity against syngeneic islet cells and various tumour cells including insulinoma cells. Several cell types served as effector cells, including macrophages, asialo GM1+ and Lyt-2+ lymphocytes. The increased cytotoxic activity towards islet cells was mostly due to macrophages and to non-asialo GM1+ and non-Lyt-2+ lymphocytes. A higher activation state of macrophages in low dose streptozotocin-treated mice was demonstrated by measurements of superoxide anion release. We conclude that multiple low doses of streptozotocin stimulate ‘natural cytotoxicity’, i.e. the non-MHC restricted cytotoxic activity of macrophages, T cells and natural killer lymphocytes.
Actinomycin D upregulates lipopolysaccharide induction of macrophage procoagulant expression and tumour necrosis factor-alpha production
Authors: WHEELER H R , ROCKETT E J , CLARK I , GEGZY C L
Published: July 2010

The antitumour antibiotic actinomycin D (Act D) and the aminosugar D-galactosaminc both enhance the sensitivity of animals to bacterial lipopolysaccharide (LPS), Lipopolysaccharide stimulates macrophage membrane-bound procoagulant activity (MPCA) and tumour necrosis factor-alpha (TNF-a) production in vitro. We investigated the cfTects of LPS combined with either Act D or tJ-galactosaniine on procoagtilant and TNF-x production in vitro. Actinomycin D directly induced procoagulant on the malignant monocytoid cell line WEHI 265. and synergized with LPS to enhance MPCA on both WEHI 265 cells and thioglycollate-induccd peritoneal exudate maero-phages. In the presence of Act D, exudate maerophages expressed procoagulant in response to concentrations of LPS lOOOOO-fold lower than normally required. Pulsing experiments demon-strated that LPS primed these cells within 4 h to respond to Act D. whereas 4 h priming with Aet D inhibited subsequent procoagulant induction by LPS. Although itseflects on TNF-a production were less intense, low levels of Act D more than doubled TNF-a produced by LPS-stimulated exudate maerophages, Procoagulant expression and TNF-a production were not always co-ordinately expressed; interferon-gamtna (IFN-))synergi/ed with LPS to enhance both responses but when IFN-) was combined with Act D only procoagulant was upregulated. t)-galactosaminc failed to alTect these macrophage responses. Results indicate dillerent; vivo mechanisms of enhancement of LPS toxicity by these two agents.
Mitogenic effect of platelet-derived growth factor in human glomerular mesangial cells: modulation and/or suppression by inflammatory cytokines
Authors: FLOEGE J , TOPLEY N , HOPPE I , BARRETT T B , RESCH K
Published: July 2010

Glomerular mesangial cell proliferation constitutes a frequent pathological alteration in glomerulonephritis. In addition to platelet-derived growth factor (PDGK) inflammatory cytokines such as IL-1. IL-6 or tumour necrosis factor-alpha (TNF-α) have been proposed to have mitogenic activity for mesangial cells. A model was therefore established in which human mesangial cells (HMC) could be reversibly growth-arrested for prolonged times in serum-free medium without suffering irreversible functional or morphological changes. In this model 24 h stimulation with rhPDGF-BB induced an increase of the 3H-thymidine incorporation of 1190.280 (50 ng/ml) %± s.e.m. of medium control. Less growth induction was noted after stimulation with 50 ng ml rhPDGF-AB (925± I26) or rhPDGF-AA (575 ± 24%). Northern analysis confirmed the presence of both α and β-PDGF receptor subunit mRNA in growth-arrested HMCs. rhlL-lα, rhlL-1β, rhTNF-α or rhIL-6 at various doses and times, despite increasing cellular PGE2-release, did not induce significant proliferation in HMCs. Inhibition of PGE2-release did not change the lack ol mitogenicity of lL-l, TNF-α or lL-6. IL-6 did not alter the mitogenic response of the cells towards PDGF. In contrast, both IL-lα and lL-lβ (5 ng/ml) induced a delay but not augmentation of the PDGF growth response. This delay could be reversed by the concomitant addition or recombinant IL-6 or of anti-lL-1 antibody but not by inhibition of prostaglandin synthesis. High doses of TNF-α suppressed PDGF-induced proliferation. These data suggest that in growth-arrested HMCs inflammatory cytokines have a growth-modulating or -suppressive rather than (co-)mitogenic effect while PDGF-BB and-AB and to a lesser degree PDGF-AA are potent mitogens. The findings support the notion that the control of HMC proliferation in pathological situations depends on a complex network of interacting stimuli.
Ontogenesis of the secretory immune system and innate defence factors in human parotid glands
Authors: THRANE P S , ROGNUM T O , BRANDTZAEG P
Published: July 2010

Immunoglobulin-producing cells and epithelial expression of secretory component (SC), amylase, lysozyme (Ly) and lactoferrin (Lf) were studied by immunohistochemistry to obtain information about the development of mucosal immunity. Tissue specimens were obtained from 20 fetal and 40 postnatal parotid glands, (1) Fetal specimens. Occasional IgM- and IgA- But no IgD-, IgG- or IgE-producing cells were seen (ratios, IgM:IgA:IgD:lgG:IgE–4:1:0:0:0). The IgA1 subclass dominated (median 90%, range 50–95%) and these cells were mostly J-chain-positive (median 97%, range 94 98%). Only few lgA2-producing cells were seen (median 10%, range 5–50%) and they were also mostly J-chain-positive (median 99%, range 98–100%). Amylase, Ly and Lf were most prominent in early fetal life, while only small amounts of SC were present. (2) Postnatal specimens. Secretory component increased markedly along with a growing number of IgA- and IgD-producing cells (lgA:IgM:IgD:IgG:lgE–4:2:1 : 1:0). The IgA1 subclass remained predominant (median 65%, range 50-90%) although the proportion of IgA2-positive cells tended to be raised (median 35%, range 10–50%). Most IgA1 (median 97%, range 67–100%) and IgA2 (median 94%, range 75–100%) cells were J-chain-positive. These features probably reflected local activation of the immune system in response to environmental factors. The amount of amylase, Ly and Lf decreased shortly after delivery, perhaps because the cellular stores were emptied by postnatal increase in secretory activity.
COMPLEMENT ACTIVATION AND BIOINCOMPATIBIUTY
Authors: Mollnes T E , Videm V , Riesenfeld J , Garred P , Svennevig J L , Fosse E , Hogasen K , Harboe M
Published: July 2010

The degree of biocompatibility of biomaterials can be evaluated using various assay systems detecting activation of the blood cascade systems, leukocytes or platelets. Activation of complement is one mechanism associated with adverse effects observed when bioincompatible materials are used. We present data showing that the terminal complement compiex, an indicator of terminal pathway activation, is suitable for evaluation of biocompatibility of biomaterials such as cardiopulmonary bypass devices. Furthermore, our results suggest that bioincompatibility is improved when artificial surfaces are modified with end point attached functionally active heparin.
Beneficial effect of human therapeutic intravenous immunoglobulins (IVIg) in mercuric-chloride-induced autoimmune disease of Brown-Norway rats
Authors: ROSSI F , BELLON B , VIAL M C , DRUET P , KAZATCHKINE M D
Published: July 2010

Administration of HgCl2 to the susceptible Brown–Norway (BN) strain of rats induces an autoimmune disease characterized by polyclonal B cell activation, increased serum levels of IgE and the occurrence of anti-glomerular basement membrane antibody-mediated glomerulonephritis. We have observed that the simultaneous administration to BN rats of normal human polyspeciric immunoglobulins for therapeutic use (IVIg) with HgCl2 significantly decreased the occurrence and severity of proteinuria, and reduced serum IgE levels in diseased animals. Hypergammaglobulinaemia was potentiated in animals receiving HgCl2 and IVIg, compared with animals receiving HgCl2 alone. In vitro experiments indicated that F(ab')2 fragments from IVIg inhibited the binding to laminin of pathogenic anti-laminin antibodies from diseased rats, as did antibodies from the resistant Lewis strain of rats but not antibodies from susceptible BN rats. These observations suggest that IVIg may interfere with the immune regulatory mechanisms involved in mercury-induced autoimmune disease in an analogous fashion to the ability of IVIg to suppress the expression of certain pathological autoimmune responses in humans.
Human immune response to rabies nucleocapsid and glycoprotein antigens
Authors: KASEMPIMOLPORN S , HEMACHUDHA T , KHAWPLOD P , MANATSATHIT S
Published: July 2010

Antibodies to two components of rabies virus, nucleocapsid (N) and glycoprotein (G), were compared in 11 rabies patients with those in nine recipients of Vero cell rabies vaccine. All rabies vaccinees had antibodies to N and G components by day 10 after the first vaccine injection. A similar but not identical response was observed in three out of 11 rabies patients. Serum antibodies appeared in rabies patients as early as 3 days after onset of the first symptoms of the disease. In these antibody-positive rabies patients, levels of both antibodies, but particularly of anti-N antibody, were lower than in the vaccinated group. Our results suggest that the process of immune recognition and of antibody development in human rabies is more likely to occur early in the pre-clinical phase, and that reactivity to N protein may be crucial for elicitation of neutralizing antibody.
Killing ofMycobacterium tuberculosiswithin human monocytes: activation by cytokines and calcitriol
Authors: DENIS M
Published: July 2010

Human monocytes were isolated and their ability to harbour growth of virulent tubercle bacilli was assessed, in the presence or absence of various immunomodulators. Calcitriol (1,25 (OH2), vitamin D3) alone, at doses of 10-7 -10-9 m endowed human monocytes with a significant ability to restrict intracellular growth of the tubercle bacilli. Crude immune lymphokines as well as recombinant interferon-gamma (IFN-γ) endowed monocytes with no tuberoulostatic activity. Similarly, other recombinant cytokines tested, notably colony-stimulating factor-1 (CSF-1), interleukin-1 (IL-1), interleukin-3 (IL-3) and interleukin-6 (IL-6) all failed to stimulate anti-tuberculous properties, and even increased growth of the tubercle bacilli in monocytes, in the case of CSF-1. Conversely, incubation of crude lymphokines in combination with calcitriol led to total stasis of the growth of M. tuberculosis. Experiments with recombinant cytokines and immunologically active vitamins showed that a combination of IFN-γ tumour necrosis factor-alpha and calcitriol induced a significant amount of intramonocyte killing of M. tuberculosis. Addition of this cocktail of factors to already infected monocytes fed to substantial killing of tubercle bacilli. These sets of experiments establish clearly that combinations of recombinant cytokines and vitamins may induce substantial intramonocyte killing of M. tuberculosis. The mechanism involved in this killing activity was not clarified.
A 19-year-old man with leucocyte adhesion deficiency.In vitroandin vivostudies of leucocyte function
Authors: DAVIES K A , TOOTHILL V J , SAVILL J , HOTCHIN N , PETERS A M , PEARSON J D , HASLETT C , BURKE M , LAW S K A , MERCER N F G , WALPORT M J , WEBSTER A D B
Published: July 2010

We describe a male patient with leucocyte adhesion molecule deficiency (LAD) or moderate phenotype. Although diagnosis was made only 2 years before his death, the patient survived until 19 yean of age. This enabled us to perform a number of novel investigations, both in vivo and in vitro, relating to his leucocyte biology. Monocytes cultured in vitro matured into morphologically normal, phagocytically capable macrophages, which were able to recognize aged‘apoptotic’neutrophils. By injection of radiolabelled autologous neutrophils we demonstrated a prolonged nculrophil half-life, but normal margination, de-margination on exercise, and splenic pooling. Neutrophil adherence in vitro to vascular cndotnclium was normal. Histological examination of the patient’s lungs at postmortem showed intravascular aggregation of polymorphonuclear leucocytes but a paucity of cells in the interstitium and alveolar spaces. These findings indicate that the peripheral blood leucocytosis commonly observed in these patients may be due to prolonged intravascular neutrophil survival, and suggest that CD11/18 molecules have an important role in facilitating neutrophil emigration from blood vessels at sites of inflammation.
Detection of anti-myeloperoxidase and anti-elastase antibodies in vasculitides and infections
Authors: GALLICCHIO M C , SAVIGE J A
Published: July 2010

Autoantibodies that produce a perinuclear pattern on indirect immunofluorescent examination of ethanol-fixed neutrophils (pANCA) are found in about half of all cases of microscopic polyarteritis. These antibodies are often directed against myeloperoxidase or elastase and we have developed sensitive reproducible ELISAs for their detection and study. Seven sera from 19 patients with microscopic polyarteritis or segmental necrotizing glomerulonephritis contained anti-myeloperoxidase or anti-elastase antibodies or both. In contrast, only one of 18 sera from patients with Wegener’s granulomatosis, where the pattern of immunofluorescence is predominantly cytoplasmic, had anti-myeloperoxidase antibodies and no anti-elastase antibodies were detected. Using sera from patients with microscopic polyarteritis, both anti-myeloperoxidase and anti-elastase antibodies were demonstrated to be of high affinity. There was no immunoglobulin class, subclass or light chain restriction noted. Anti-myeloperoxidase and anti-elastase antibodies were also found occasionally in anti-glomerular basement membrane disease, mixed connective tissue disease, systemic lupus erythematosus, post-streptococcal glomerulonephritis and atypical pneumonia. In further studies these antibodies were not associated with other lung infections, although anti-elastase antibodies were noted in one of 14 sera positive for ASOT that were tested. Anti-myeloperoxidase antibodies were found more frequently than anti-elastase antibodies and these antibodies were occasionally present together. In addition some sera with pANCA had neither anti-myeloperoxidase nor anti-elastase antibodies. The target molecules in these cases remain unclear.
UCHL1+ (CD45RO+)‘memory’T cells predominate in the CD4+ cellular infiltrate associated with allergen-induced late-phase skin reactions in atopic subjects
Authors: FREW A J , KAY A B
Published: July 2010

We have previously shown that CD4+ T lymphocytes accumulated at the site of allergen-induced late-phase reactions (LPR) in the skin of atopic subjects. In order to determine whether these were predominantly‘memory’or‘naive’cells, monoclonal antibodies recognizing isoforms of the CD45 common leucocyte antigen and immunocytochemical methods were used to study the composition of the T cell infiltrate. Allergen-induced late-phase skin reactions were biopsied6, 24 or 48 h after allergen challenge. Memory (CD45RO+/UCH L1+) T cells predominated and few naive (CD45RA/Leu 18+) cells were identified. Double immunofluorescence was used lo confirm that the UCHLI+ and Leu 18+ cells were CD4+ T lymphocytes. The selective recruitment of memory T cells to LPR sites is consistent with the active involvement of T lymphocytes in alopic allergic inflammation. A possible alternative explanation for apparently selective recruitment is the differential expression of endothelial adhesion molecules on memory and naive T lymphocytes.
Changes in leucocyte populations following murine bifunctional antibody infusion in colon cancer patients
Authors: GRIDLEV D S , STICKNEY D R
Published: July 2010

This study was undertaken to determine whether infusion of a unique ZCE/CHA bifunctional antibody (BFA, 5–40 mg) could alter the composition and functions of peripheral blood leucocytes in 18 patients with colon cancer. The BFA is made by combining chemically the Fab’fragments of two murine monoclonal antibodies. One fragment (ZCE 025) binds to the carcino-embryonic antigen (CEA) and the other (CHA 255) to an epitope, present on an In -benzyl EDTA analog of bleomycin (BLEDTA IV) and on 111In-hydroxy-ethyl-thiourca benzyl EDTA (EOTUBE). The radiolabelled epitope (111ln-BLEDTA IV or 111In-EOTUBE) was given 4 days after pre-localization with BFA. Peripheral blood samples were tested before BFA infusion, at the end of infusion (1 h later), and at 4 and 7 days post-infusion. A 50% or greater suppression in lymphocyte responsiveness to phytohaemagglutinin (PHA) and concanavalin A (Con A) was seen in 13 out of 18 and 12 out of 18 subjects, respectively, at some time after BFA infusion; this was especially evident in those patients with prc-infusion stimulation indices of > 50 (PHA) and/or > 10 (Con A). In contrast, natural killer (NIC) cell cytotoxicity and oxygen radical production increased in five out of 15 and in seven out of 18 subjects, respectively. Little or no change was observed in CD3, CD4, CDS. CD16, and CD19 markers on lymphocyte subpopulations as determined by flow cytometry. These data suggest that significant changes in mitogen-induced lymphoproliferation. NK cell cytotoxicity. and oxygen radical production can occur in a substantial proportion of cancer patients after infusion of the ZCE/ CH A bifunctional antibody system. The immunomodulation was unrelated to initial BFA dose, dose of BFA as a carrier, or to subsequent infusion of either form of the 111In cpilope. The clinical significance of these phenomena, if any, remains to be determined.
Studies in vivo and in vitro of serum amyloid P component in normals and in a patient with AA amyloidosis
Authors: HAWKINS P N , TENNENT G A , WOO P , PEPYS M B
Published: July 2010

Pure serum amyloid P component (SAP) was isolated from a normal donor pool, from individuals with the different genotypes of an MspI restriction fragment length polymorphism (RFLP) linked to the SAP gene, and from a patient with AA amyloidosis. The SAP preparations were all identical and all behaved as a single homogeneous species in polyacrylamide gel electrophoresis, isoelectric focussing, reverse-phase chromatography, binding in vitro to phosphoethanolamine-Sepharose (binding constant 2.4× 107l/mol) and AL amyloid fibrils (1.6 × 108l/mol), and binding to amyloid deposits in vivo in mice with casein-induced amyloidosis. The in vivo metabolism of 125I-SAP from a single donor was normal and identical in three healthy individuals representing the three different MspI RFLP genotypes. There is thus no frequent polymorphism of SAP in normal subjects, and SAP altered with respect to the characteristics studied here is not a necessary condition for pathogenesis of systemic AA amyloidosis.
Expression of the fibronectin receptor VLA-5 is regulated during human B cell differentiation and activation
Authors: BALLARD L L , BROWN E J , HOLERS V M
Published: July 2010

We examined the expression of VLA-5, a libronectin receptor, during human B cell development and activation. VLA-5 is a member of the integrin supergene family; VLAs arc heterodimers of at least six unique a chains sharing a common (i chain; most arc involved in cell attachment to extracellular matrix (ECM). A hypothesis of haematopoietic development is lhat maturing cells leave the bone marrow because of the loss of VLA-5 during differentiation. However, mature B ceils are not primarily circulating cells, and the role of ECM receptors in homing to peripheral lymphoid tissue and inflammatory sites is unknown. To examine the expression of VLA-5 during B cell development, cell lines blocked at specific stages of differentiation were evaluated for their synthesis and surface expression of VLA-5 using VLA-5-specific antibody and cDNA probes. VLA-5 mRNA and surface expression were found in the pre-B cell lines, REH and Nall 1, but not in more differentiated Raji cells or in several EBV-transformed peripheral B cell lines. Circulating peripheral B lymphocytes and resting tonsillar and splenic B lymphocytes expressed no VLA-5 by FACS analysis. Interestingly, mRNA and surface expression of VLA-5 were found in SKW. a highly differentiated, IgM-secreting line. In addition, low levels of staining for VLA-5 expression could be demonstrated when tonsillar or peripheral blood B lymphocytes were stimulated by Staphylococcus aureus Cowan (SAC). All cell lines expressed VLA-3 and VLA-4. two other receptors reported to mediate fibronectin binding in some cell types. Thus, our studies provided no evidence for developmental or inflammatory regulation of these receptors. Binding studies, however, demonstrated that adherence of both pre-B REH cells and SKW cells to fibronectin was almost completely inhibited by a monoclonal antibody to VLA-5o. In addition, Raji cells, which lack VLA-5 but express VLA-3 and VLA-4. showed very low level binding to Hbronectin. This demonstrates that for some B lymphocytes VLA-5, rather than other possible fibronectin receptors, primarily mediates attachment to fibronectin. These data also suggest that human VLA-5 expression is regulated during B cell development, with expression at a very early stage and then again after activation. This pattern of loss and reacquisition of an ECM receptor may be relevant to normal B cell maturation and to function during immunologic injury.
Identification of a myeloperoxidase inhibitor from normal human serum
Authors: YEA C M , DULARAY B , ELSON C J
Published: July 2010

An inhibitor of myeloperoxidase (MPO) has been identified in normal human serum. Initial experiments confirmed that high levels of MPO inhibitory activity are present in sera and that the inhibitor did not act by interfering with the assay. Purification of the inhibitor activity by salt precipitation followed by ton exchange and affinity chromatography revealed the presence of a protein of 150 kD. The purified inhibitory activity displayed dose and lime dependency and was not associated with IgG or IgA. It is considered that human serum contains an inhibitor of extracellular MPO capable of protecting against hypohalous acid release in host tissues and that if inhibitor levels are reduced such protection may fail.
Cord blood contains cells secreting antibodies to nervous system components
Authors: FREDRIKSON S , SUN J , XIAO B-G , LINK H
Published: July 2010

Umbilical cord blood of newborns and peripheral blood or healthy adults were investigated by an immunospot assay for cells secreting IgG, IgA and IgM antibodies against myelin basic protein (MBP), proteolipid protein (PLP), myelin-associated glycoprotein (MAG) and myelin oligodendrocyte glycoprotein (MOG) which represent putative antigens for an autoimmune attack in multiple sclerosis (MS) and against acetylcholine receptor (AChR) which is considered an important autoantigen in myasthenia gravis. Cells secreting antibodies against one or more of these autoantigens were detected in 18 out of 24 newborns, and in eight out of 20 adults. Eight of the cord blood samples contained cells secreting antibodies of IgG, IgA and/or IgM isotypes to one antigen, five to two antigens, two to three antigens, two to four antigens, and one to five antigens. Most prominent were anti-MBP IgG antibody secreting cells which were detected in 13 newborns at a mean number of 1/20000 cord blood cells, and in six adults at a mean number of l/105 peripheral blood cells. Anti-AChR IgG antibody secreting cells were detected in four out of 12 newborns versus four out of 14 peripheral blood specimens, at mean values of 1/105 cells in both instances. Cells secreting autoantibodies of IgA and IgM isotypes were less frequent both in cord blood and peripheral blood. The occurrence of nervous tissue autoantibody secreting cells in newborns must be related to a possible primary role of such autoantibodies in MS and myasthenia gravis.