bluuebunny/crossref_metadata_2025 huggingface dataset:
Biochemical and functional characterization of the leucocyte tyrosine phosphatase CD4S (CD4SRO, 180 kD) from human neutrophils.In vivoupregulation of CD45RO plasma membrane expression on patients undergoing haemodialysis
Authors: PULIDO R , ALVAREZ V , MOLLINEDO F , SÁNCHEZ-MADRID F
Published: July 2010

The biochemical and functional characterization, and the regulation of plasma membrane expression of the leucocyte tyrosine phosphatase CD45, have been investigated in neutrophils from healthy donors and patients undergoing haemodialysis. CD45 proteins of 180 kD and 130–150 kD were precipitated from neutrophils from both healthy subjects and haemodialysed patients. Prolonged storing, as well as trypsin treatment of samples containing the 180-kD CD45 protein, generated the 130–150-kD polypeptides. The 130–150-kD CD45 polypeptides carried extracellular CD45 epitopes, including the sialic acid-related UCHLI epitope (CD45RO). Furthermore, these trypsin-generated CD45 polypeptides did not possess phosphatase activity, which could be detected on the 180-kD protein. A remarkable quantitative increase of cell surface expression of the neutrophil CD45 components was detected both after in vitro neutrophil activation and after dialysis treatment with neutropenic membranes. The CD45 biochemical pattern did not qualitatively change upon either in vitro or in vivo dialysis-induced neutrophil activation. The upregulated expression of CD45 on neutrophils from dialysed patients correlated with the neutropenic effect induced by the different dialyser membranes. Maximal upregulation of CD45 expression was observed after 15 min of dialysis with neutropenic membranes, and normal expression levels were restored after I h. By contrast, increase of CD45 plasma membrane expression induced in vitro by treatment of normal neutrophils with the degranulatory agents fMLP or Ca2+ ionophore was maintained. These results demonstrate that neutrophil cell surface expression of the 180-kD CD45 protein is upregulated during the in vivo haemodialysis process, and suggest that a proteolytic activity could regulate the enzymatic activity of CD45 by degranulation of its cytoplasmic phosphatase domains.
Antibodies against sulphatide in sera from patients with autoimmune rheumatic diseases
Authors: AOTSUKA S , OKAWA-TAKATSUJI M , UWATOKO S , YOKOHARI R , IKEDA Y , TODA G
Published: July 2010

We tested sera of patients with various autoimmune rheumatic diseases for the presence of antibodies against sulphatide (an acidic glycosphingolipid), identified as a target antigen for antibodies against the liver cell membrane. Thirty-five percent (7/20) of patients with lupus in the active stage possessed anti-sulphatide antibodies, whereas 10% (2/20) of those in the inactive stage and 20% (4/20) of those in the stationary stage possessed such antibodies. Moreover, 10%. (3/29) of patients with other autoimmune rheumatic diseases also possessed anti-sulphatide antibodies. The level of anti-sulphatide antibodies was significantly correlated with the levels of anti-double-stranded (ds) DNA antibodies (r = 0.634, P <0.001) and dextran sulphate-binding IgG (r = 0.407, P < 0.001). The serum levels of antibodies against sulphatide were correlated with a history of seizures or psychosis in patients with autoimmune rheumatic diseases. Gels coupled with polyanionic dextran sulphate, monoanionic sulphanilic acid and DNA were shown effectively to adsorb anti-sulphatide antibodies in the sera of patients with active systemic lupus erythematosus (SLE) and autoimmune chronic active hepatitis (AI-CAH). These results suggest that the observed reactivity with sulphatide is due to the presence of antibodies capable of reacting with various anionic molecules in the sera of patients with autoimmune rheumatic diseases as well as those with AI-CAH.
Imbalance of CD4+ lymphocyte subsets in patients with mixed connective tissue disease
Authors: BECKER H , LANGROCK A , FEDERLIN K
Published: July 2010

CD4+ (helper/inducer) T lymphocyte subsets were studied in the peripheral blood from patients with mixed connective tissue disease (MCTD) by double-labelling immunofluorescence. The proportion of CD4+CD45RA+ cells was higher (P < 0·01) when compared with controls, whereas CD4+CD29+ cells were markedly diminished (P < 0·001). CD4+CD29+ cells were lower than in patients with progressive systemic sclerosis who were studied in parallel. Upon stimulation with phytohaemagglutinin, CD4+ cells from MCTD patients showed a strong reactivity to acquire the CD29+ phenotype. Expression of high levels of CD29 and other adhesion molecules might lead to facilitated localization of CD4+ cells to inflamed tissue. It is suggested that an increased responsiveness of CD4+ cells to activation signals in vivo and accumulation of CD4+CD29+ cells at tissue sites could result in depletion of this cell subset in the peripheral blood of patients with MCTD.
IL-2 production by intestinal lamina propria cells in normal inflamed and cancer-bearing colons
Authors: PULLMAN W E , DOE W F
Published: July 2010

Biologically significant levels of IL-2 activity were produced by isolated lamina propria mononuclcar cells (LPMC) from normal intestine (n= 12). cancer-bearing colons (n= 35) and inflammatory bowel disease (IBD) affected tissue (n= 12). The levels of IL-2 produced were similar for all three sources of LPMC (normal 252 ± 48 U/ml, I BD-affected mucosa 197 ± 42 U/ml and colon cancer 285 ± 43 U/ml). These levels were significantly greater than those produced by peripheral blood mononuclcar cells (20·5 U/ml, P < 0·01) on a per cell basis. In mucosa from cancer-bearing colons the amount of IL-2 produced by LPMC was unaffected by the invasiveness of the colon cancer. LPMC IL-2 production was markedly suppressed by drugs used in IBD therapy. 5-Aminosalicylic acid (5-ASA) reduced activity in a dose-dependent fashion. At a dose equivalent to the faecal therapeutic level of 0·5 mg/ml activity, IL-2 production by LPMC was suppressed to 3·4% of controls. Similarly, exposure of LPMC to cyclosporin A (CyA) and hydrocortisone (HC) at therapeutic levels reduced I L-2 activity to less than 1% of controls. The major producers of IL-2 activity were shown to be CD3+ T lymphocytes and those bearing the activation markers IL-2R and TFR. Suppression of mucosal IL-2 production represents an important therapeutic mechanism ofdrugs used in the management of IBD including HC, 5-ASA and CyA. These results suggest that mucosal T cells produce appreciable levels of IL-2 activity that may be important in maintaining immune homeostasis in the normal intestine, provide anti-neoplastic cytotoxic activity and contribute to the inflammatory events that characterize the mucosal lesions of IBD.
T cell receptor Vβ variable gene family expression in human peripheral blood lymphocytes at the mRNA and membrane protein level
Authors: HU H Z , DE WEGER R A , BOSBOOM-KALSBEEK K , TILANUS M G J , ROZING J , SCHUURMAN H J
Published: July 2010

A polymerase chain reaction (PCR) was developed, using combinations of an oligonucleotide primer for a T cell receptor Vβ gene family and one for the constant Cβ gene segments, to assess the expression of each of 20 Vβ gene families in RNA after reverse transcription into cDNA. The detection was done after agarosc gel electrophoresis of PCR products and ethidium bromide staining. The positive identification of the PCR products was done by hybridization with a Jβ oligonucleotide probe. For T cell lines, a signal was observed in the Vβ8 combination for Jurkat cells, Vβ 5a in HSB cells, Vβ 2 and Vβ12a in Molt-3 cells and Vβ 2, Vβ 5a and Vβ12a in Molt-4 cells. Using mixtures of RNA from different cell lines, the sensitivity of the method was in the range of 0·1–0·5%. In peripheral blood mononuclear cells from four donors, taken at three different occasions, all Vβ families were detectable. The intensity of the PCR product varied between various Vβ gene families. Flow cytometric analysis of blood mononuclear cells from the same donors with a restricted series of Vβ gene family-specific antibodies also revealed the presence of all families. The approach to assess Vβ gene family expression in heterogeneous populations opens the possibility to study T cell receptor variable gene expression in relation to physiology and pathologic processes.
Immunophenotypical alterations in a subset of patients with common variable immunodeficiency (CVID)
Authors: BAUMERT E , WOLFF-VORBECK G , SCHLESIER M , PETER H H
Published: July 2010

We investigated the expression of surface molecules on lymphocytes from 20 patients with CVID and 40 healthy subjects. Lymphocytes were analysed by dual colour flow cytometry. We identified a subset of patients (8 of 20) characterized by low CD4/CD8 ratio (< 1·1), expansion of T cells co-expressing the activation marker HLA-DR and significant increase in CD8+ T cells co-expressing CD57. Expression of the adhesion molecules LFA-3 (CD58) and ICAM-1 (CD54) was significantly increased in this subgroup. In addition, within the CD4+ T cells the percentage of CD29+ (memory) cells was increased, while the CD45RA and LAM-1 (Leu-8) antigens were depressed. These results indicate that in a subgroup of CVID patients T cells are activated in vivo and the CD57+CD8+ lymphocyte subpopulation, supposed to comprise functional suppressor T cells, is expanded. We suggest a chronic viral infection in these patients, but it is not clear whether this is primary or secondary to the underlying defect.
Kinetics of lymphocyte subsets from peripheral blood during aPlasmodium falciparummalaria attack
Authors: CHOUGNET C , TALLET S , RINGWALD P , DELORON P
Published: July 2010

Variations of lymphocyte subsets were followed longitudinally in 16 patients during an acute falciparum malaria attack. Before treatment, lymphocyte numbers were highly reduced, but the subset distribution was similar to that of healthy individuals. After parasite clearance, lymphocyte counts were normalized and the subset distribution was unchanged. This led to a normalization of all the absolute counts of lymphocyte subsets, except CD8+. The fast normalization of lymphocyte counts suggests that the initial decrease in lymphocyte numbers may reflect sequestration. Magnitude and kinetics of this variation were not related to parasite density or to severity of the attack. Activated T cells (CD3+HLA-DR+) were fewer in African than in European patients, suggesting the importance of the past exposure to malaria parasites in reallocation phenomena. These variations in lymphocyte numbers must be taken into account in the design and the analysis of cellular investigations in patients experiencing a falciparum malaria attack.
Immunoglobulin class and IgG subclass distribution of anticardiolipin antibodies in patients with systemic lupus erythematosus and associated disorders
Authors: LOIZOU S , COFINER C , WEETMAN A P , WALPORT M J
Published: July 2010

The class and subclass distribution of an antibody response may give insight into the stimulating mechanism and likely effector functions. IgA, IgG and IgM anlicardiolipin antibodies (aCL) were quantified in a consecutive series of 200 samples sent to an autoimmune serology laboratory to determine the relationships between aCL responses of each of these antibody classes and, in particular, whether there was any utility in the measurement of IgA aCL. Positive results for one of the three aCL isotypes were found in 105 samples (53%). and in 41 samples IgA aCL was detected (21%). However, amongst these unselected samples, little additional information was obtained by measurement of IgA aCL, which was found in conjunction with IgM or IgG aCL in all but five samples, and in these the isolated elevation of IgA aCL was only slight, and showed no disease specificity. The levels of each of the four IgG subclasses of aCL were measured in a subgroup of serum samples from 28 patients with autoimmune disease and from 29 patients with syphilis. Amongst the SLE patients IgGl and lgG3 aCL were the predominant IgG subclasses, consistent with an antigen-driven. T cell-dependent antibody response. However, a subgroup of eight of the autoimmune subjects had predominant elevation of IgG2 aCL, possibly implying a role for T cell-independent antibody production to cardiolipin. Amongst the syphilis patients IgG I and IgG3 aCL were also the predominant subclasses of aCL but lgG4 aCL were also detected in the majority of subjects, consistent with prolonged antigenic stimulation.
Molecular heterogeneity of human IgA antibodies during an immune response
Authors: RUSSELL M W , LUE C , VAN DEN WALL BAKE A W L , MOLDOVEANU Z , MESTECKY J
Published: July 2010

Human IgA occurs in multiple molecular forms (polymeric and monomeric) and two subclasses which show differential distribution between the mucosal and circulatory compartments of the immune system. However, the molecular form and subclass of specific IgA antibodies are influenced, especially during an immune response, by the type of antigen and duration of the response as well as by the route of exposure. These considerations question previously held notions that polymeric IgA and an increased representation of the IgA2 subclass among circulating antibodies or antibody-secreting cells signify their mucosal origin. Although the functional properties of different molecular forms and subclasses of IgA antibodies are incompletely understood, it appears that there is physiological benefit in the diversity of the IgA immune system.
Enhancement of the lytic activity of cloned human CD8 tumour-infiltrating lymphocytes by bispecific monoclonal antibodies
Authors: GORTER A , KRÜSE K M , SCHRIER P I , FLEUREN G J , VAN DE GRIEND R J
Published: July 2010

We have isolated tumour-infiltrating lymphocytes (TIL) clones hum a patient with renal cell cancer. The cloning frequency and the effector function were measured. No difference in cloning frequency (r2= 0·97, frequency =1:13) was observed between TIL expanded with allogencic versus autologous feeder cells. Sixty-four clones expanded with autologous feeder cells and 37 clones expanded with allogeneic feeder cells were assayed for cytolytic activity on an autologous primary renal cell carcinoma (RCC) culture, an allogeneic RCC line, and on the K562 and Daudi cell lines. Most of these clones were also phenotyped. Although TIL clones expressing cytotoxic activity for RCC lines could be generated with both feeder cell preparations, none of the clones tested showed specificity for ceils from autologous primary RCC cultures. However, in the presence of relevant bispecific MoAbs (αOC/TR) all CD8+ TIL clones tested could be induced to lyse autologous RCC cultures. Furthermore, the cytolytic activity of all CD8+ clones tested against allogeneic RCC lines could be induced or further enhanced by αOC/TR or CD3/G250 bispecific MoAbs. In contrast, none of the CD4+ clones tested showed lytic activity. Quantitatively the cytotoxic response in the presence of αOC/TR or CD3/G250 of CD8+ TIL clones against G250+ and MOv18+ cell lines appears to be associated with the level of antigen expression on the target cells. Our results suggest that (i) expansion of TIL with allogeneic or autologous feeder cells does not effect the lytic profile of the clones; (ii) the use of bispecific MoAbs may overcome a lack of specificity of cytotoxic T lymphocytes.
Anti-endothelial cell antibodies in insulin-dependent diabetes mellitus
Authors: WANGEL A G , KONTIAINEN S , SCHEININ T , SCHLENZKA A , WANGEL D , MÄENPÄÄ J
Published: July 2010

The prevalence of IgG class antibodies to endothelial cells (AECA) was studied in 136 young patients with insulin-dependent diabetes mellitus by an enzyme immunoassay using human umbilical cord vein endothelial cells. The patients were divided into four groups according to the time between diagnosis and study and their results were compared with those in control children and blood donors. AECA became progressively more frequent with the duration of diabetes, being 4% in diabetics tested within 2 weeks of diagnosis and reaching 34% after an average disease duration of 11–2 years. They were not more common in patients with neuropathy, retinopathy or nephropathy than in patients without these complications, but were associated with co-existing thyroid disease and IgA deficiency. The results suggest that in insulin-dependent diabetes mellitus AECA are associated with co-existing autoimmune disorders but not with diabetic microvascular disease.
Impaired proliferative capacity and abnormal cytokine profile of naive and memory CD4 T cells from HIV-seropositive patients
Authors: CAYOTA A , VUILLIER F , SCOTT-ALGARA D , FEUILLIE V , DIGHIERO G
Published: July 2010

Purified naive and memory CD4 T cells from healthy donors, HIV+ asymptomatic carriers and AIDS patients were examined for their proliferative activity and their pattern of cytokine secretion (IL-4, IL-6, interferon-gamma (IFN-γ) and tumour necrosis factor-alpha (TNF-α)) upon stimulation with phytohaemagglutinin (PHA), phorbol myristate acetate (PMA) and cross-linked anti-CD3 MoAb, in the presence of recombinant IL-2 (rIL-2). We found a decrease in the proliferative capacity of naive CD4 T cells following stimulation with PHA and PMA, and a sharp decline in this response upon cross-linked anti-CD3 stimulation in both subsets, although it predominated in the naive subpopulation. In AIDS patients, less pronounced impairment of thymidine uptake by the naive subset was found upon PHA and cross-linked anti-CD3 MoAb stimulation. In addition, an altered secretion pattern of the different cytokines was observed, consisting of abnormal secretion of IL-6 by both naive and memory cells, an abnormal pattern of IFN-γ secretion and frequent loss of detectable lL-4 production by HIV patients. These abnormalities were even more pronounced in AIDS patients than in the asymptomatic carriers. Overall, our results extend previous reports indicating functional impairment of memory CD4 subsets in HIV+ subjects by showing that this impairment involves naive CD4 T cells.
Mononuclear phagocytes from human bone marrow progenitor cells; morphology, surface phenotype, and functional properties of resting and activated cells
Authors: KELLER R , KEIST R , JOLLER P , GROSCURTH P
Published: July 2010

After 3–4 weeks culture of human bone marrow cells in medium supplemented with IL-3, macrophagc- (M-CSF), and granulocyte-macrophage colony-stimulating factor (GM-CSF), the firmly adherent cells exhibited the morphologic features of mononuclear phagocytes and were strongly esterase-positive. Flow cylometric analysis revealed a rather homogeneous cell population with marked autofluorescence; the large majority of the cells expressed CD14. CD11a,b, and c, Fc receptors for IgG, FcγRI, II, and III, and HLA class II molecules. Interfcron-gamma (IFN-γ). bacteria, and bacterial products modulated expression of some of the surface markers, induced and/ or enhanced respiratory burst, phagoeytic activity, secretion of tumour necrosis factor, and lumouricidal activity; in contrast, these cells were not able to generate reactive nitrogen intermediates.
Cytokine-induced inhibition ofPlasmodium falciparumerythrocytic growthin vitro
Authors: ORAGO A S S , FACER C A
Published: July 2010

The addition of recombinant cytokines to Plasmodium falciparum in vitro cultures retarded the growth of the parasite with the effect of recombinant IL-2 (rIL-2)>interferon-gamma (IFN-γ)>tumour necrosis factor-beta (TNF-β). The process was concentration dependent, being greatest Bl 30000 U/ml and required a 72-h period of continuous exposure for maximum effect. Growth inhibition, as determined morphologically and radiometrically. was a consequence of defective schizont maturation rather than inhibition of merozoite invasion. It was cumulative and detectable within one erythrocytic (48 h) growth cycle.
Evidence for iC3 generation during cardiopulmonary bypass as the result of blood-gas interaction
Authors: PEKNA M , NILSSON L , NILSSON-EKDAHL K , NILSSON U R , NILSSON B
Published: July 2010

Earlier we have shown that iC3 is generated at the blood-gas interface in vitro and that the generation of this molecule is independent of complement activation and the composition of the gas. In order to investigate whether iC3 is also generated during cardiopulmonary bypass where blood comes into contact with oxygen bubbles, two bubble oxygenators were incubated at 37°C with human heparinized blood. A continuous increase in the level of iC3 was shown in the oxygen-perfused bubble oxygenator (up to 100 nmol/l after 180 min) in contrast to the unbubbled control. Similarly, in plasma drawn from patients undergoing cardiopulmonary bypass using either bubble or membrane oxygenators. the levels of iC3 were shown to increase continuously during the operation. Furthermore, this form of C3 was found to be susceptible to cleavage by factor I. The formation of iC3 at the blood-gas interface in vivo could be a mechanism by which gas bubbles induce clinical manifestations associated with complement activation, e.g. during cardiopulmonary bypass, adult respiratory distress syndrome and decompression sickness.
An immunohistochemical analysis of onchocercal nodules: evidence for an interaction between macrophage MRP8/MRP14 and adultOnchocerca volvulus
Authors: EDGEWORTH J D , ABIOSE A , JONES B R
Published: July 2010

We have used a panel of MoAbs to investigate the phenotype of macrophages and other leucocytes infiltrating onchocercal nodules. Nodules were removed from individuals at the end of the second year of a community-based, placebo-controlled trial of annual ivermectin chemotherapy in northern Nigeria. No significant differences were seen in the distribution and phenotype of leucocytes in nodules from ivermectin- and placebo-treated individuals. Live adult worms were only seen in nine of the 21 nodules examined. Three regions were clearly discernible within nodules containing both live and dead worms; an outer fibrovascular capsule (zone A), an inner adult worm bundle with surrounding hyaline extracellular matrix interspersed with solitary cells (zone B), and a dense cellular infiltrate surrounding and in contact with a variable proportion of the worm (zone C). Macrophages were the predominant cell type in all zones of the nodule. Those in zone B were distinguished by their dendritic morphology and strong reactivity with MoAbs directed against class II molecules. FcRl (CD64) and CD68, whereas macrophages in zone C were larger, more heterogeneous in shape, and were distinguished by strong reactivity with MoAbs directed against CR4 (CDlIc, CD18) and MRP8/MRP14, and with MoAb 24. T cells were found primarily in zones A and C, whilst eosinophils were found in only six nodules. A unique staining pattern was seen using MoAbs reacting with the calcium-binding protein MRP8/MRPI4. Most macrophages in zones A and B were negative; however, where the occasional positive macrophage was seen in zone B, MRP8/MRP14 was also found around the cell and on the neighbouring worm surface, giving the impression that MRP8/ MRP14 was being secreted onto the adult worm. Macrophages in zone C were also MRP8/MRP14-positive, and often the whole infiltrate was surrounded with extracellular MRP8/MRP14, with greatest concentration seen adjacent to the worm. M RP8/MRP14 was not identified on the surface of microfilariae (MF) within the same nodules. Since MRP8/MRP14 was seen on the adult worm in the absence of a leucocytic infiltrate, it may have an early role to play in the immune response to Onchocerca volvulus.
Genes on the X chromosome affect development of collagen-induced arthritis in mice
Authors: JANSSON L , HOLMDAHL R
Published: July 2010

Susceptibility to collagen-induced arthritis (CIA) in mice is associated with a class II gene in MHC (Aq) but also with unknown genes outside MHC. Investigated here is the influence of genes on the X chromosome as well as the role of the X-linked immunodeficiency (xid) mutation. Reciprocal male F1 hybrids, bred to be heterozygous or homozygous for Aq, showed a genetic influence in their susceptibility to develop CIA. Crosses were made between B10.G, B10Q, DBA/I, SWR/J, C3H.Q and CBA/Ca, and all Fi mice were castrated to avoid sex hormone modulation of the susceptibility. A differential timing of arthritis onset and severity were seen in the reciprocal F1 males. An exception was the reciprocal F1 male offspring from SWR/J and DBA/1 crosses which differed only in disease severity late in the course of the disease. The female F1 crosses did not show the same pattern of differential susceptibility to CIA as the F1 males. To exclude the possible influence the Y chromosome, F1 males of reciprocal crosses were back-crossed to the parental strains creating offspring with equal X chromosomes but divergent Y chromosomes. No difference in development of arthritis was observed in these. The influence of the xid mutation was investigated next. The xid loci from the CBA/N mouse was bred into DBA/1 strain which is highly susceptible to CIA. The resulting congenic DBA/l-xid strain was resistant to induction of CIA and did not develop an antibody response to type II collagen. We conclude that polymorphic genes on the X chromosome modulate susceptibility to CIA. The results from the experiments with mice carrying xid mutations confirm that such immune modulating genes exist on the sex chromosomes.
Recognition of Epstein—Barr virus (EBV)-infected cells by T cell colonies from a human chemera: restriction by allogeneic determinants
Authors: PLOTNICKY H , TOURAINE J-L
Published: July 2010

The anti-EBV T cell response was studied in a severe combined immunodeficiency patient (PS) who received two transplants of fetal liver cells. His peripheral blood mononuclear cells (PBMC) were incubated with EBV and cultured during 15 days. Eleven colonies were derived from the T lymphocytes causing the regression of the infected cell foci: nine were constituted with CD3+CD4+CD8− lymphocytes and two with CD3+CD4−CD8+ cells. HLA typing of six colonies showed that two of them derived from the first transplant and four from the second one. The colonies killed the cells of the lymphoblastoid line (LCL) derived from the recipient (PS-LCL), but failed to kill the LCL matched with the transplants. With only one exception, they all lysed also the LCL derived from the mother or from the father, but they were ineffective on the EBV-negative lymphoblasts. Two colonies recognized determinants which did not appear to be HLA antigens, although they were shared by PS and by one of his parents, two (CD4−CD8+) reacted against the LCL which shared HLA-A3 or -A33 with PS-LCL, and four (CD4+CD8−) lysed the LCL sharing HLA-A3, -A33 or -DR5 with PS-LCL, among which only one was demonstrated to interact directly with host HLA-class I determinants. These data indicate that T lymphocytes differentiating in contact with histo-incompatible determinants may express the capability to recognize viral antigens and to lyse virus-infected cells in the context of allogeneic MHC or non-MHC molecules.
Increased soluble CD14 serum levels and altered CD14 expression of peripheral blood monocytes in HIV-infected patients
Authors: NOCKHER W A , BERGMANN L , SCHERBERICH J E
Published: July 2010

Serum levels of soluble CD14 were elevated in HIV-infected asymptomatic patients or those with lymphadenopathy (CDC II/III) 2·9 ± 0·8mg/l compared with normal controls with 2·2 ± 0·47 mg/l P < 0·001. A further rise was seen in patients with ARC (CDCIVA) 3·8±1·1 mg/l, P < 0.01 and patients with AIDS (CDC IVB D) 5·7 ±2·5 mg/l. P < 001. Although absolute numbers of CD14+ cells decrease in the AIDS group, the percentage of CD 14+ monocytes did not change. In contrast, levels of soluble T cell antigens sCD4 and sCD8, which are higher in HIV-infected patients compared with normal subjects, showed no increase with disease progression. Serum levels of sCD14 were correlated positively with β2-microglobulin levels (rs= 0·63, P < 0.0001). Whereas the percentage of CD14+ monocytes did not change, an increase in monocytic CD14 expression in HIV-infected patients was observed (P < 0.01). The percentage of a monocyte subset expressing both CD14 and CD16 increased from 6% in normal healthy persons to 13% in HIV-infected patients (P < 0·001), and did not vary between the HIV patient groups. Incubation of cultured peripheral blood monocytes with azidothymidine had no effect on either normal or EPS-induced or IE-4-inhibited sCDI4 release in vitro. Therefore, an effect of AZT on sCD14 serum values in vivo is considered to be unlikely. Our data further provide evidence that monocytes/macrophages are engaged in HIV infection.
Erythrocytes from young but not elderly donors can bind and degrade immune complex- and antibody-bound C3in vitro
Authors: SHAPIRO S , KOHN D , MILLER B , GERSHON H
Published: July 2010

In situ aged erythrocytes (senescent erythrocytes from young donors and both young and old erythrocytes from elderly donors) demonstrate high levels of membrane-bound C3c and C3d and elevated susceptibility to in vitro phagocytosis. In this study we demonstrate that in situ aged erythrocytes are defective in their ability to degrade C3 fragments and clear them from the erythrocyte membrane. Erythrocytes from young donors bind complement-bearing immune complexes via CRI and become susceptible to complement-mediated erythrophagocytosis (“innocent bystander” sequestration). Erythrocytes from elderly donors are defective in their ability to bind such immune complexes, as attested by the lack of an increment in membrane-associated C3 fragments as detected by flow cytometry and lack of an increment in in vitro sequestration. Factor I (serum)-dependent cleavage of C3 fragments and release of immune complexes from the erythrocytes of young donors lead to a drop in erythrocyte-associated C3 fragments and the disappearance of the “innocent bystander” phenomenon. Inhibition of Factor I, and thus inhibition of C3b degradation and immune complex release for the erythrocyte membrane, enhances the levels of ‘innocent bystander’ sequestration of erythrocytes from young donors. Erythrocytes from elderly donors are defective in the dynamic process of CRI binding of complement-bearing immune complexes and Factor I-associated release of membrane-associated C3 fragments, A defect in the ability of all in situ aged erythrocytes to clear their membranes of C3 fragments is also demonstrated for complement bound to the erythrocyte via IgM isoagglutinins. While erythrocytes from both young and elderly donors allow for the IgM-mediated binding of complement to the erythrocyte, only young erythrocytes from young donors demonstrate degradative activity with the release of membrane-associated C3c. Erythrocytes from elderly donors demonstrate lower levels of detectable CRI (CD35). decay accelerating factor (OAF) (CD55) and Protectin (CD59) than do erythrocytes from young donors. Time course studies determine that the defect in handling of immune complexes observed for in situ aged erythrocytes is not due to differences in the kinetics of loading or releasing of immune complexes. These findings on the refractiveness of erythrocytes of elderly donors to uptake of immune complexes and the degradation of C3 fragments may be of importance not only in the understanding of erythrocyte sequestration, but also in the physiology of immune clearance in the elderly.
Some patients with anti-myeloperoxidase autoantibodies have a C-ANCA pattern
Authors: SEGELMARK M , BASLUND B , WIESLANDER J
Published: July 2010

Rapidly progressive glomerulonephritis with or without other signs of systemic vasculitis is often accompanied by antibodies to myeloperoxidase. Such antibodies normally produce a perinuclear pattern on ethanol-fixed neutrophils (perinuclear anti-neutrophil cytoplasm antibodies (PANCA)) at indirect immunofluorescence. We report here sera front three patients that are antimyeloperoxidase-positive in ELISA that instead produce a cytoplasmic pattern (classical anti-neutrophil cytoplasmic antibodies (C-ANCA)), a pattern normally seen in conjunction with antibodies to proteinase 3. These sera did not react with proteinase 3. For two of the sera the specificity of the anti-myeloperoxidase reaction was confirmed with inhibition-ELISA experiments and with immunoblotting. A mouse anti-myeloperoxidase MoAb that produces a cytoplasmic pattern is also described. Competition ELISA experiments show that this antibody and anti-myeloperoxidase sera with cytoplasmic pattern recognize epitopes that are separate from epitopes recognized by another perinuclear pattern producing anti-myeloperoxidase MoAb. Cytoplasmic pattern epitopes as well as ‘perinuclear pattern’ epitopes can be found on all three major myeloperoxidase isoforms. after separation by ion exchange chromatography. Affinity chromatography. using the eytoplasmic pattern producing anti-myeloperoxidase monoclonal antibody, shows that the epitope recognized by this MoAb is present on all myeloperoxidase molecules. This epitope is not confined to any special subpopulation. These findings indicate that all myeloperoxidase do not relocate after ethanol fixation, and that C-ANCA and P-ANCA epitopes exist simultaneously on the same myeloperoxidase molecule. We propose that the two immunofluorescence patterns arise due to different availabilities of the epitopes in the microenvironment where myeloperoxidase Is present.
IgE elevation and IgE anti-malarial antibodies inPlasmodium falciparummalaria; association of high IgE levels with cerebral malaria
Authors: PERLMANN H , HELMBY H , HAGSTEDT M , CARLSON J , LARSSON P H , TROYE-BLOMBERG M , PERLMANN P
Published: July 2010

In the course of studying immunoregulation in human Plasmodium falciparum malaria we have investigated IgE levels and IgE anti-plasmodial antibodies in children and adults from areas of high malaria endemicity in both Africa and Asia. On average, 85% of all donors had significantly elevated levels of total IgE. A fraction of the IgE had anti-plasmodial activity as revealed by ELISA with lysates of infected erythrocytes as antigen. Using synthetic peptides representing antigenic regions of two major plasmodial blood stage antigens, IgE antibody concentrations ranged from 5 to 15 ng/ml serum for each of the peptides. On average, the concentrations of the corresponding IgG antibodies were x500-I000 higher. Immunoblotting of parasite lysates showed that most donors had IgE antibodies against one or several of a restricted number of plasmodial polypeptides, with antibodies against an antigen of mol. wt 45 kD already being present in all donors at an early age. Donors having IgE antibodies to particular antigens also frequently had corresponding IgG4 antibodies, reflecting underlying IL-4-dependent cellular mechanisms controlling formation of these isotypes. As infection with other parasites such as helminths is known lo induce IgE elevation, the results do not prove that plasmodial infections were the primary cause of IgE induction. However, the importance of plasmodial infection for IgE elevation was supported by the finding of significantly higher levels of IgE., but not of IgG, in children with cerebral malaria compared with patients with uncomplicated disease.
Cytokine mRNA expression in endomyocardial biopsies during acute rejection from human heart transplants
Authors: BAAN C C , VAN EMMERIK N E M , BALK A H M M , QUINT W G V , MOCHTAR B , JUTTE N H P M , NIESTERS H G M , WEIMAR W
Published: July 2010

The immune response to an allograft is regulated by cytokines produced by cells infiltrating the allograft. However, the immunopathogenesis of allograft rejection is not completely understood. To investigate the role of cytokines after clinical heart transplantation, we analysed the expression of cytokine genes in sequentially taken endomyocardial biopsies (EMB) by using the reverse transcriptase-polymerase chain reaction (RT-PCR). We analysed 44 EMB from II recipients: 21 EMB before or during rejection, and 23 EMB without histological evidence of acute rejection. A strong correlation was found between IL-2 gene expression and histologically proved rejection (16/21 versus 1/23 without rejection, P < 0·001; x2 test). Also, expression of IL-4 and IL-6 genes was more often found in EMB during rejection than in EMB without signs of rejection (IL-4, 62% versus 35%; and IL-6, 81% versus 39%. respectively). No relation with rejection or with immunological quiescence was observed for the presence of IL-10 gene transcripts. IL-10, but also IL-6 mRNA were detectable in donor heart tissue before transplantation (9/10), In contrast, IL-2 and IL-4 gene transcripts were absent in these samples. These differences could not be explained by the presence or absence of T ceils, since the gene for the constant region of the β-chain (CG) of the T cell receptor (TCR) not only was expressed in post-transplant EMB but also in pre-transplant donor heart tissue. Our results provide strong evidence that the immunoregulatory cytokines IL-2, IL-4 and IL-6 are important local regulators in the graft during acute rejection. The role of IL-10 in the immunologic response lo the transplanted organ needs further investigation.
Enhanced activation of human T cell clones specific for virus-like particles expressing the HIV V3 loop in the presence of HIV V3 loop-specific polyclonal antibodies
Authors: PEIFANG S , PIRA G L , FENOGLIO D , HARRIS S , COSTA M G , VENTURINO V , DESSI V , LAYTON G , LAMAN J , HUISMAN J G , MANCA F
Published: July 2010

Recombinant virus-like particles (VLP), formed by the yeast Ty p1 protein, carrying the HIV gp120 V3 loop on their surface (V3-VLP) have been tested in vitro for immunogenicity and antigenicity by using VLP pl-specific human CD4+ T cell lines and clones. VLP-specific human T cell lines and clones were generated from normal individuals, indicating that VLP-specific precursor cells present in the peripheral lymphocyte pool can be induced to expand clonally upon antigen challenge in vitro, in the absence of previous immunization. It was also shown that V3-specific polyclonal antibodies enhance V3-VLP-induced activation of VLP-specific T cell clones. Antibody-dependent potentiation has been shown previously in other antigen systems, and it depends on enhanced uptake of complexed antigen by Fe receptor-positive antigen-presenting ceils. Since in this case antigen is internalized by presenting cells as a complex, it can be inferred that a similar event of antibody-mediated antigen uptake can take place with V3-speeific B cells, resulting in presentation by the B cells of T helper epitopes derived from processing of the VLP pi moiety. This suggests that T helper cells specific for the carrier VLP pi protein can be activated to provide help to V3-specific B ceils in the presence of the appropriate antigen construct.
Cross-reactivity studies and differential serodiagnosis of human infections caused by Trypanosoma cruzi and Leishmania spp; use of immunoblotting and ELISA with a purified antigen (Ag163B6)
Authors: MALCHIODI E L , CHIARAMONTE M G , TARANTO N J , ZWIRNER N W , MARGNI R A
Published: July 2010

Results of our studies on the reactivity of chagasic and leishmaniasic sera with the purified T. cruzi- specific antigen 163B6, as assessed by ELISA, and with complex antigenic mixtures from T. cruzi and Leishmania mexicana, by immunoblotting. are presented here. Our objective was to identify the antigens responsible for the exhibited cross-reactivity between trypanosomiasis and leishmaniasis, and to find a specific reactivity pattern corresponding to each parasitosis. In spite of the high cross-reactivity observed with the immunoblotting, the use of 7.5% A-B gels made it possible to identify a characteristic pattern for each parasitosis, that could be distinguished by the naked eye. The characteristic pattern corresponding to chagasic patients was ascribed to reactivity with T, cruzi bands of mol. wts 131, 125, 116, 111, 51-45 and 43 kD, that were not recognized by leishmaniasic sera. Trypanosoma cruzi antigens of mol. wts 85, 81, 70, 65 60, 37 and 32kD were considered as crossing antigens, since they were recognized by leishmaniasis sera. With L. mexicana, most of the chagasic patients presented reaction with antigen of mol. wts 124, 107. 92, 59 and 32kD, while bands of mol. wts 155, 140. 73. 56 and 48kD were recognized only by leishmaniasic sera. In this study we found 12 out of 45 sera of patients with leishmaniasis, from a region endemic for both parasitoses, which exhibited a pattern of bands very similar to those corresponding to chagasic individuals, strongly suggesting a mixed infection. This hypothesis was verified by using a purified specific antigen of T. cruzi, Ag163B6, which would be the major cysteine proteinase of this specie (cruzipain). By HLISA, these 12 sera showed a positive reaction with this purified antigen, as those of chagasic patients, thus leading to the confirmation of the presence of a mixed infection.
Serum antibody responses by male and female C57B1/6 mice infected withGiardia muris
Authors: DANIELS C W , BELOSEVIC M
Published: July 2010

We compared the levels of serum antibodies in male and female C57BI/6 mice during the primary and after challenge infection with Giardia muris. Male mice began passing cysts in their faeces earlier than females, and were shedding cysts for over 60 days, while females stopped shedding cysts by day 20 after infection. In both males and females there were significant increases in parasite-specific IgM 10 and 20 days after infection. No differences in parasite-specific serum IgA were observed until 40 days after infection. Parasite-specific IgG (whole) levels were elevated on days 20 and 40 in females, while males showed no significant increases. In addition, females had a much stronger IgG2b and IgG3 response than males. After challenge with either cysts or soluble parasite protein only the females had significant increases in specific anti-parasite IgG2b. Our data show differential ability of males and females to control the infection with G. muris is paralleled by a difference in the anti–parasite serum IgG response of the mice.
Immune complex glomerulonephritis is induced in rats immunized with heterologous myeloperoxidase
Authors: YANG J J , JENNETTE J C , FALK R J
Published: July 2010

Anti-neutrophil cytoplasmic antibodies (ANCA), including anti-myeloperoxidase (MPO) antibodies, are associated with pauci-immune necrotizing small vessel vasculitis or glomerulonephritis, 1n order to substantiate a pathogenic role for ANCA, an animal model of pauci-immune ANCA-induced glomerulonephritis or vasculitis is required. Brouwer et al. reported pauci-immune glomerulonephritis in rats immunized with human MPO followed by perfusion of kidneys with lysosomal enzyme extract combined with H2O2, and suggested that this could serve as a model of ANCA-induced disease. We repeated these studies in spontaneously hypertensive rats (SHR) and Brown Norway rats (BNR). We immunized rats with human MPO, When circulating anti-MPO antibodies were detectable by indirect immunofluorescence microscopy and ELISA, blood pressure was measured, then perfusion of the left kidney of each rat was done via the renal artery in a closed, blood-free circuit with either MPO + H2O;, MPO, H2O2 alone or MPO + H2O2 neutral protease. Rats were killed on day 4 or day 10 after perfusion, and specimens were examined by light and immunofluorescence microscopy. Pathological lesions and deposits of IgG. C3, and MPO were found in immunized rats perfused with MPO + H2O2 with or without neutral protease, or MPO alone, in both rat strains and on both day 4 and day 10, The degree of histologic injury was proportional in intensity to the amount of IgG immune deposits. Spontaneously hypertensive rats sustained more damage and higher blood pressure than Brown Norway rats. No lesion was observed in immunized rats perfused with H2O2 or in the non-perfused right kidneys. Some of the non-immunized rats perfused with MPO + H2O2 developed pathological lesions. In conclusion, these rat models are examples of immune complex-mediated glomerulonephritis, and therefore are not similar to human ANCA-associated disease.
Quantification and localization of HLA-DR and intercellular adhesion molecule-1 (ICAM-1) molecules on bronchial epithelial cells of asthmatics using confocal microscopy
Authors: VIGNOLA A M , CHANEZ P , CAMPBELL A M , PINELJ A M , BOUSQUET J , MICHEL F-B , GODARD P H
Published: July 2010

An increased expression of HLA-DR and lCAM-1 molecules on bronchial epithelial cells has been observed in asthmatic patients. The aim of this study was to evaluate the localization and to quantify the spontaneous expression of HLA-DR and ICAM-1 on bronchial epithelial cells recovered by bronchial brushing of nine asthmatics and nine controls. Epithelial cells constituted over 95% of cells recovered as shown using an anticytokeralin MoAb. Expression of HLA-DR and ICAM-1 was studied using indirect immunofluorescence and confocal microscopy. The intensity of fluorescence of epithelial cells expressing HLA-DR and lCAM-1 was significantly (P< 0·003) increased in asthmatics. In asthmatics, but not in controls, the expression of both molecules was localized in the cytoplasm on the apicolateral portions of the cells. This study shows an up-regulation in the expression of HLA-DR and ICAM-1 molecules by bronchial epithelial cells from asthmatics and a localization of these molecules within the cell.
Intestinal and circulating antibody-forming cells in IgA-deficient individuals after oral cholera vaccination
Authors: FRIMAN V , QUIDING M , CZERKINSKY C , NORDSTRÖM L , LARSSON L , ERICSON D , BJÖRKANDER J , THEMAN K , KILANDER A , HOLMGREN J , HANSON L Å
Published: July 2010

In search for a possible explanation for the different susceptibility to mucosal infections in IgA-deficient (IgAd) individuals, the frequency of total immunoglobulin-secreting-ells (ISC) and vaccine-specific antibody-secreting cells (ASC) in intestinal mucosa and peripheral blood was determined by the enzyme-linked immunospot (ELISPOT) assay before and after peroral vaccination with a B subunit-whole cell cholera vaccine. Two groups of IgAd individuals, frequently infected and non-infected respectively, and normal controls were studied. Before cholera vaccination there were significantly higher frequencies of total IgM and IgG ISC in the gut, but not in the blood, in the IgAd individuals than in the controls. However, there were no significant differences between healthy and infection-prone IgAd individuals in this respect. In response to oral cholera vaccination, intestinal cholera toxin (CT)-specific IgG and IgM ASC were significantly more abundant among the IgAd individuals with a history of frequent infections than among ihc healthy IgAd individuals and controls. A similar difference in IgG and IgM ASC, although not significant, was also noted in blood. In IgAd individuals with frequent infections the vaccine induced variable anti-CT IgM ASC responses in the gut, ranging from no increase to a few strikingly high responses. In the controls, the CT-specific responses were dominated by IgA ASC. The data show that oral cholera vaccination evoked strong CT-specific IgG ASC responses, and in some cases also strong IgM ASC responses in the intestinal mucosa of IgAd patients with a history of frequent infections. The healthy IgAd individuals unexpectedly responded with lower numbers of CT-specific IgG ASC and did not show any increase of CT-specific IgM ASC in the intestinal mucosa. Thus, inability to mount a mucosal immune response to an oral antigen cannot in itself explain recurrent infections among many IgAd individuals.