bluuebunny/crossref_metadata_2025 huggingface dataset:
Avidity of IgA Antibody to Escherichia coli Polysaccharide and Diphtheria Toxin in Breast Milk from Swedish and Pakistani Mothers
Authors: ROBERTON D. M. , CARLSSON B. , COFFMAN K. , HAHN‐ZORIC M. , JALIL F. , JONES C. , HANSON L. Å.
Published: June 2006

The avidity of breast milk IgA antibody was studied with the aid of thiocyanate elution of antibody from solid‐phase bound E. coli polysaccharides and diphtheria toxoid. The relative avidity index for each sample was determined by the molarity of thiocyanate required to elute 50% of the bound IgA antibody under conditions of antigen excess. Milk samples collected from Pakistani mothers during early lactation (2–4 weeks after delivery; n= 12) had a significantly lower median relative avidity index of IgA antibody to E. coli antigens than did early lactation samples from Swedish mothers (n= 11; avidity indices 1.78 m and 2.65 m; P<0.02). Samples collected from Pakistani mothers in mid‐lactation showed a significant rise in the relative avidity index to a median of 2.50 m (P<0.01), with a subsequent fall in late lactation (28–36 weeks after delivery) to 1.75 m (P<0.01). Milk samples from Pakistani mothers in mid‐lactation (n= 12) also had a lower median relative avidity index of IgA antibody to diphtheria toxoid than did samples from Swedish mothers (n= 14;avidity indices 2.35 m and 4.30 m; P<0.002). The lower avidity of breast milk IgA in Pakistani mothers in comparison with Swedish mothers may arise from differences in antigen exposure or nutritional status or could possibly be genetically determined.
Studies on the Role of Lymphocyte Function‐Associated Antigen 1 (LFA‐1) in T Cell Activation
Authors: BERZINS T. , AXELSSON B. , HAMMARSTRÖM M.‐L. , HAMMARSTRÖM S. , PERLMANN P.
Published: June 2006

The effect of polyclonal (anti α and β chain) and monoclonal (anti α chain) antibodies lymphocyte Function‐associated antigen 1 (LFA‐1) on I cell activation was studied. When added at the beginning of activation but not after 24 h or later the antibodies as well as the F(ab')2 or Fab fragments of polyclonal antibodies inhibited concanavalin A (Con A)‐induced proliferation, interleukin 2 (IL‐2) production, and the expression of receptors for IL‐2 and transferrin. The inhibitory effect reached a maximum at the same time as optimal proliferation (72 h) Inhibition of proliferation lasted for 5 days or longer, although IL‐2 production was only inhibited during the first 48 h of culture. Receptors for IL‐2 and transferrin were re‐expressed to the original level after 3 days of activation. Addition of external IL‐2 at the beginning of the anti‐LFA‐1 containing culture prevented the inhibition of IL‐2 receptor expression, while inhibition of transferrin receptor expression was unaffected, supporting the conclusion that the‐expression of these two receptors is regulated by partially independent signals. The polyclonal and monclonal anti‐LFA‐1 antibodies also inhibited phorbol ester (PMA)‐dependent OKT3 activation of highly purified T cells. The results suggest that the LFA‐1 antibodies block an early step in the reactions necessary for IL‐2 production, and that the LFA‐1 molecule participates not only in T cell‐accessory cell interaction but also in T‐T interaction during the early phases of the activation process.
Datura Lectin is Both an Anti‐Mitogen and a Co‐Mitogen Acting Synergistically with Phorbol Ester
Authors: McCURRACH P. M. , KILPATRICK D. C.
Published: June 2006

The lectins from Datura stramonium, Lycopersicon esculentum, and Solanum tuberosum are structurally related and possess a similar carbohydrate specificity, yet the Datura lectin is mitogenic for human lymphocytes while the other two are not. However, the Datura lectin was found to antagonize blast transformation induced by purified protein derivative (PPD), even within the concentration range at which it was optimally mitogenic on its own. The presence of a submitogenic concentration of the phorbol ester 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) enormously enhanced the mitogenic activity of Datura lectin. No such synergy was observed, however, between tomato lectin or S. tuberosum agglutinin (STA) and TPA, nor between Datura lectin and the calcium ionophore A 23187.
Efficient Propagation and Cloning of Human T Cells in the Absence of Antigen by Means of OKT3, Interleukin 2, and Antigen‐Presenting Cells
Authors: LONDEI M. , GRUBECK‐LOEBENSTEIN B. , DE BERARDINIS P. , GREENALL C. , FELDMANN M.
Published: June 2006

The analysis of T lymphocytes infiltrating tissues afflicted by autoimmune diseases may provide major clues towards understanding the pathogenesis of such diseases. Currently the best approach to studying heterogeneous populations such as T lymphocytes involves long‐term culture and cloning. In order to grow and clone T lymphocytes, regular restimulation with the specific antigen is essential, otherwise growth will stop and/or specificity may be lost. In autoimmune diseases the antigens involved in triggering the immunological reaction of T cells are usually unknown. Therefore an alternative way of stimulating T lymphocytes without loss of specificity is clearly needed. Here we describe the cloning and expansion of antigen‐specific T cell clones from the blood of a healthy donor to sizeable numbers of cells (>108) by means of anti‐CD3 monoclonal antibody and recombinant IL‐2 The results obtained showed that this approach can be used to clone and ‘expand’ T lymphocytes that retain antigen specificity over a prolonged period, in this case over 10 weeks. This technique has been used to clone and expand T lymphocytes infiltrating the affected tissues in a variety of autoimmune disorders such as Hashimoto's thyroiditis, Graves' disease, and rheumatoid arthritis, and is an efficient method of propagating T cells, by mimicking the antigenic Stimulus.
Post‐Translational Processing of the Murine Third Component of Complement
Authors: BEDNARCZYK J. L. , CAPRA J. D.
Published: June 2006

The biosynthesis and secretion of the third component of complement (C3) has been studied with the macrophage cell line J774.2. C3 is initially synthesized as a single polypeptide chain precursor termed pro‐C3, of relative molecular weight (Mr) 170,000 that is post‐translationally modified by proteolytic cleavage into two polypeptides linked by disulphide bonds. The larger polypeptide, termed the alpha chain, has an Mr of 110,000–115,000, while the smaller beta chain has an Mr of 55,000–60,000. Pulse‐chase experiments indicate that the proteolytic processing of pro‐C3 occurs intracellularly, just prior to secretion. Unlike human C3, which has carbohydrate on both the alpha and beta chains, only the alpha chain of murine C3 is glycosylated. The carboxylic ionophores monensin and nigericin totally inhibit the proteolytic processing of pro‐C3 at a concentration of approximately in 10−6M. This block on proteolytic processing was shown not to be mediated by changes in intracellular pH induced by the disruption of proton gradients. Rather, data from experiments using carboxylic ionophores and other perturbants of cellular physiology indicated that the enzyme(s) responsible for the proteolytic cleavage of pro‐C3 either reside in a cellular compartment with a neutral pH or are proteinases active over a relatively broad pH range.
Modulation of Sheep Erythrocyte Receptor Expression by Anti‐CD4, Anti‐CD8, and Anti‐HLA Antibodies
Authors: NOGUEIRA‐MACHADO J. A. , CHAVES M. M. , SILVA‐TEIXEIRA D. N.
Published: June 2006

The modulation of E receptors on T cells by anti‐CD4 and/or anti‐CD8 is reported. The percentage of E rosette‐forming cells (RFC) was decreased when sheep erythrocyte and mononuclear cells were incubated in the presence of monoclonal antibodies anti‐CD4 and/or anti‐CD8. This inhibition was not due to shedding of the E receptor and it was reversed by 8‐Br.‐3′,5’cyclic guanosine monophosphate (cGMP). In contrast, anti‐HLA antibodies induced an enhancement of sheep erythrocyte receptor expression evaluated by RFC.
Specific Lymphoproliferation, Gamma Interferon Production, and Serum Immunoglobulin G Directed against a Purified 32 kDa Mycobacterial Protein Antigen (P32) in Patients with Active Tuberculosis
Authors: HUYGEN K. , VAN VOOREN J.‐P. , TURNEER M. , BOSMANS R. , DIERCKX P. , DE BRUYN J.
Published: June 2006

Twenty‐one patients treated for active tuberculosis were examined for immune reactivity to purified protein derivative (PPD) and to a purified 32‐kDa protein antigen (P32) from Mycobacterium bovis, strain BCG. Lymphoproliferation of peripheral blood leucocytes to PPD and P32 was positive in 95% and 71% of the patients respectively. A positive IFN‐γ response was detected in 62% against PPD and in 48% against P32. Low blastogenesis and IFN‐γ production were observed, especially in patients with poor general health and advanced tuberculous lesions. Twelve out of twelve (100%) of the tuberculin‐positive healthy volunteers responded to PPD and P32 with mean lymphoproliferation and IFN‐γ values that were higher than in the patient group. Twelve tuberculin‐negative control subjects were completely unreactive to PPD and P32 antigen. On the other hand, IgG antibodies in the serum were detected in 95% of the patients against PPD, in 77% of the patients against P32 but in none of the tuberculin‐positive or negative healthy volunteers. The highest IgG levels against PPD were found in those patients with the lowest in vitro lymphoproliferation and IFN‐γ production (r=‐0.54; P<0.05). Nonspecific interferon production following induction with Newcastle disease virus, Corynebacterium parvum, or phytohaemagglutinin was comparable in the control and patient groups. Finally, low IFN‐α titres were detected in the serum of about 50% of the patients.
Protein HC‐IgA Complexes Carry Antibody Activities
Authors: TRUEDSSON L. , GRUBB A.
Published: June 2006

Polyclonal protein HC‐IgA complexes (HC‐IgA) were isolated from two different serum pools. Their hydrodynamic volumes were found to be slightly greater than that of monomeric IgA but less than that of dimeric IgA. Sodium dodecyl sulphate (SDS)‐polyacrylamide gel electrophoresis of reduced and carboxymethylated complexes followed by immunoblotting showed that the complexes contained normal light and heavy Ig chains, and one polypeptide chain with Mt= 90,000, which carried both IgA α chain and protein HC epitopes. Enzyme‐linked immunosorbent assays (ELISA) demonstrated that the isolated HC‐IgA carried about 0.1 and 4%, respectively, of the antibody activities against one carbohydrate antigen (Yersinia enterocolitica serotype 0:3 lipopolysaccharide) and one protein antigen (rabbit IgG, i.e. antigen for rheumatoid factors) of the IgA populations of the two serum pools. HC‐IgA with rheumatoid factor activity could also be demonstrated in the unfractionated serum pool. The binding of HC‐IgA in the ELISA was not mediated through its protein HC part. The present observations show that HC‐IgA carries antibody activities and constitutes a unique class of IgA complexes, since it does not dissociate under denaturating conditions alter reduction. It may represent a further biological potential of the humoral immune system.
Correlation of Susceptibility and Cytostatic Factor‐Inducing Activity of Tumour Cells to Peritoneal Macrophages
Authors: HASHIMOTO S. , NAGAOKA M. , YOKOKURA T. , MUTAI M.
Published: June 2006

L929, 3T12‐3, B16, 3LL, and YAC1 cells with cytostatic factor (CF)‐inducing activity from Lactobacillus casei‐elicited murine peritoneal macrophages (LCEPM) were susceptible to the cytostatic activity of LCEPM and to LCEPM‐produced CF, but L1210, P388D1, and Colon 26 cells, which have no CF‐inducing activity, were resistant to that of LCEPM and to the CF. The resistance of P815 cells to that of LCEPM was stronger than that of 3T12‐3 cells, but the CF‐inducing activity of P815 cells was about 5098 weaker than that of 3T12‐3 cells. Release of CF from LCEPM was also caused by heat‐killed (100°C, 10 min) 3T12‐3 or P813 cells, and this release was inhibited by d‐mannose. The CF‐inducing activity of heat‐killed 3T12‐3 or PK15 cells was reduced by mild trypsin digestion (37°C for 10 min). A d‐mannose‐containing glycopeptide or glycoprotein (GP) was separated from 3T12‐3 or P815 cells by concanavalin A (Con A) or wheat germ agglutinini (WGA) affinity chromatography. The CF were released from LCEPM by stimulation with the Con A‐binding GP of the tumour cells, but the WGA‐binding GP had little activity.

It is suggested that tumour cells with CF‐inducing activity may he susceptible to the cytostatic activity of LCEPM, and those without CF‐inducing activity may be resistant to the cytostatic activity of LCEPM and the release of CF from activated macrophages may be caused by the Con A‐binding GP of the tumour cell surface.
Expression of Platelet‐Derived Growth Factor Receptors is Induced on Connective Tissue Cells During Chronic Synovial Inflammation
Authors: RUBIN K. , TERRACIO L. , RÖNNSTRAND L. , HELDIN C.‐H. , KLARESKOG L.
Published: June 2006

The tissue distribution of the receptor for platelet‐derived growth factor (PDGF) was investigated by immunohistochemistry on frozen sections from normal and inflamed synovial tissue using monoclonal antibodies to the receptor. Non‐inflamed synovial tissue showed no staining, indicating that PDGF receptor expression is low or absent in normal tissue. In contrast, tissue from synovitis with prominent neovascularization showed a strong staining in the tunica media of the proliferating blood vessels as well as on connective tissue cells in the stroma. Tissue from synovitis with prominent proliferation of synovial lining showed intense staining for PDGF receptors in fibroblast‐like cells of the lining and a less intense staining on vascular and connective tissue cells deeper in the stroma. Staining for PDGF receptors was also intense in the pannus tissue close to infiltrated bone and cartilage. In all these forms of synovitis, PDGF receptor staining was associated with increased HLA‐DR staining and infiltration of macrophages and T lymphocytes. The finding that PDGF receptor expression is induced in conjunction with the chronic synovial inflammation associated with rheumatoid arthritis and some other forms of arthritides suggests that PDGF may play a role in the stimulation of mesenchymal cell proliferation that often accompanies chronic inflammatory disease.
Quantification in Enzyme‐Linked Immunosorbent Assay of a C3 Neoepitope Expressed on Activated Human Complement Factor C3
Authors: GARRED P. , MOLLNES T. E. , LEA T.
Published: June 2006

A sensitive double antibody enzyme‐linked immunosorbent assay (ELISA) for quantification of C3 activation products in human plasma, synovial fluid, and cerebrospinal fluid is described. The monoclonal antibody MoAb bH6, which is specific for a C3 neoepitope expressed on C3b, iC3b, and C3c, was used as capture antibody. Detection antibody was a polyclonal rabbit anti‐human C3c followed by development with a peroxidase‐conjugated anti‐rabbit Ig antiserum. The activity in normal human EDTA plasma was 1.5% of that in zymosan‐activated serum (ZAS). The interassay and intra‐assay coefficients of variation were 15% and 3%, respectively. The lower detection limit was 0.00035% of the ZAS standard. Reference range (1.1–2.1% of ZAS) was obtained by measuring EDTA plasma from 40 healthy blood donors. A positive correlation r3=+0.92; P<0.0002) was found between the present assay and an already established C3′g'activation ELISA, when samples from 16 patients were examined in both assays simultaneously. The present assay and an assay detecting the terminal complement complex showed virtually identical activation patterns in consecutively drawn samples from a patient undergoing extracorporal circulation.
The Organ Distribution of F Protein in the Mouse
Authors: GRIFFITHS J. A. , OLIVEIRA D. B. G.
Published: June 2006

F protein is of unknown function and found principally in the liver, with smaller amounts detectable by gel diffusion and immunostaining in kidney and brain. By taking advantage of a newly developed radioimmunoassay, capable of detecting down to 1 ng ml−1 of F protein, the organ distribution of this molecule in the mouse was re‐examined, F protein was not confined to the previously known tissues and indeed was detected at different levels in every organ studied. The highest concentration (6–10 mg g−1 wet tissue) was found in the liver, but significant amounts (2–25 μg g−1) were also found in kidney, heart, and brain. Lower quantities were detectable in all other tissues studied. The presence of F protein in she thymus, and the lack of tolerance to this in bone marrow chimaeras between mice of different F protein types, places certain constraints on the amount and/or location of a protein required for tolerization.
Tonsillar Distribution of IgA and IgG Immunocytes and Production of IgA Subclasses and J Chain in Tonsillitis Vary with the Presence or Absence of IgA Nephropathy
Authors: NAGY J. , BRANDTZAEG P.
Published: June 2006

At the onset or in the course of IgA nephropathy (IgA NP), upper respiratory tract infections and tonsillitis are often followed by periods of gross haematuria. In a search for possible abnormalities in the tonsillar IgA‐ and IgG‐cell system, the palatine tonsils from seven patients with IgA NP and eight controls, all 15 suffering from chronic recurrent tonsillitis, were subjected to an immunohistochemical study. Compared with the controls, tonsils of NP patients contained a significantly (P<0.001) increased proportion of IgA‐producing cells (49.6% versus 35.7%). There was also an increase (P<0.001) in the ratio of IgA polymer‐(J‐chain‐positive) to monomer‐producing cells in NP tonsils compared with controls (35.0% versus 18.8%). Although the tonsillar IgA cells were generally producing mainly IgA1, this subclass was even more predominant in NP tonsils (P<0.03). These results are compatible with the hypothesis that in some patients with IgA NP, the polymeric IgA1 deposited in the mesangium may be of tonsillar origin.
T and B Cell Specific Immune Responses to Purified Protein Derivative in the Cerebrospinal Cavity May be Maintained and Regulated Independently of Systemic Immune Control
Authors: SCHNEIDER E. M. , VON ZITZEWITZ F. , MEIER D. , ZIMMERMANN C. W. , DICHGANS J. , SCHLOTE W. , WERNET P.
Published: June 2006

In vivo activated T cells could be isolated from cerebrospinal fluid (CSF) of a patient suffering from chronic meningitis of unclear origin. Although the patient's skin reactivity to purified protein derivative (PPD) was negative, and peripheral T cells did not proliferate to this antigen in vitro, the majority of T cell clones from CSF specifically recognized PPD on either autologous or allogeneic HLA class II compatible macrophages. Remarkably, peripheral blood mononuclear cells potently suppressed the PPD‐specific proliferate responses of healthy donors. The selective enrichment of oligoclonal IgG in the CSF but not in the patient's serum further indicated T and B cell responses lacking systemic feedback control. Analyses of a persisting immune stimulation in the CSF provide a potent diagnostic tool and may explain neurological complications as observed in a number of autoimmune diseases and chronic infections.
Characterization of a Bovine Thymic Differentiation Antigen Analogous to CD1 in the Human
Authors: MACHUGH N. D. , BENSAID A. , DAVIS W. C. , HOWARD C. J. , PARSONS K. R. , JONES B. , KAUSHAL A.
Published: June 2006

Three monoclonal antibodies (MoAb), TH97A, CC13, and CC14, define a thymic differentiation antigen in cattle. The antigen is expressed on 50–60% of bovine thymocytes, located mainly in the cortical areas, but is not expressed on peripheral blood mononuclear cells (PBMC). In cryostat sections of lymph node, the antibodies read with large dendritic‐like cells in the paracortical regions. They also react with a proportion of the large ‘frilly’ cells in afferent lymph and with dendritic‐like cells in the dermis. The antibodies apparently do not react with cells in the epidermis. Biochemical analysis of the antigen recognized by MoAb TH97A reveals two bands of 44 kDa and 12 kDa under reducing conditions. These polypeptides are distinct from bovine class I major histocompatibility complex molecules reactive with the MoAb w6/32. The tissue distribution of positive cells together with results of biochemical analyses indicate that the antigen recognized by these MoAb is the bovine analogue of the human CD1.
Role of Accessory Cells in the Activation of Pure T Cells via the T Cell Receptor‐CD3 Complex or with Phytohaemagglutinin
Authors: HALVORSEN R. , LEIVESTAD T. , GAUDERNACK G. , THORSBY E.
Published: June 2006

The ability of different subpopulations of blood mononuclear cells to serve as accessory cells in the activation of CD4+ and CD8+ T cells via Ti‐CD3 or with phytohaemagglutinin (PHA) was studied. Pure CD4+ or CD8+ T cells did not respond to particle‐bound anii‐CD3 monoclonal antibodies (MoAb) or PHA, whereas responses were seen when non‐T cells served as accessory cells. Removal of class II‐positive cells from peripheral blood mononuclear cells (PBMC) or from non‐T cells diminished, but did not completely abolish, the responses in both T cell subsets, indicating that the accessory cells are mainly found among the class II‐positive cells. However, the class II molecules themselves were not involved, as demonstrated in antibody‐blocking experiments. Removal of monocytes decreased the ability of non‐T cells to serve as accessory cells for both CD4+ and CD8+ cells in PHA activation. In contrast, the removal of monocytes resulted in an enhanced activation by anti‐CD3 MoAb in CD4+ T cells, while the activation of CD8+ T cells was less affected. Positively selected B cells were effective accessory cells in anti‐CD3 and PHA activation. Furthermore, Epstein‐Barr virus (EBV)‐transformed B cell lines were very potent accessory cells both in anti‐CD3 and PHA activation of T cells, and showed the strongest accessory cell function observed in this system on a per cell basis.
Antigenic and Allergenic Determinants of Ovalbumin
Authors: ELSAYED S. , HOLEN E. , HAUGSTAD M. B.
Published: June 2006

The decapeptide 1‐10 of hen's egg ovalbumin (OA), deduced from the known amino acid sequence (Gly‐Ser‐Ile‐Gly‐Ala‐Ala‐Ser‐Met‐Glu‐Phe), was synthesized by Merrifield solid phase peptide synthesis with a yield of >70%. The completeness of the insertion of amino acids during synthesis was monitored by the amino acid compositions of the peptide‐resin, prior to coupling of the preceding residue. The linearity of the synthesis was supported by dansyl Edman degradation and detection of three NH2 terminal residues 5‐dimethylamino‐naphthdine‐1‐sulphonyl (DNS)‐Gly, ‐Ser, ‐Ile, respectively. The peptide was purified by gel filtration chromatography and analytical reversed‐phase high‐performance liquid chromatography (HPLC). The homogeneity of the preparation was calculated both from the amino acid analysis and by integrating the peaks of HPLC to be >83%. The antigenicity of the purified (P2) peptide could be detected by precipitation inhibition with the nephelometric technique. The decapeptide could also specifically react with functional structures on reaginic IgE molecule from the sera of individuals allergic to eggs, inhibiting its further binding to ovalbumin An in‐vivo experiment using direct skin test on two patients allergic to eggs showed no activity, rendering further testing unnecessary. The results suggest that the decapeptide of the NH2 terminal segment of OA encompasses an Ig‐binding haptenic epitope.
Activation of Human T Lymphocytes through CD3 and CD2 (T11) with Anti‐CD3‐Coupled Sheep Erythrocytes
Authors: OCKLIND G.
Published: June 2006

Proliferative activation of T lymphocytes depends on cell‐cell cooperation, i.e. interactions between specific cell surface receptors and their ligands. My co‐workers and I have recently shown that an activating signal is mediated by interaction between the T cell surface antigen CD2 (T11) and its ligand on sheep erythrocytes (SE), the T11 target structure (T11TS), which is the homologue to the human LFA‐3 antigen. Here I demonstrate that the anti‐CD3 monoclonal antibody (MoAb) UCHT1 coupled to SE (SE‐UCHT1) most efficiently induces accessory cell (AC)‐independent T cell proliferation, and that SE can substitute for AC in stimulation with phytohaemagglutinin (PHA). Inhibition studies with MoAb suggest that (1) concurrent stimulation of CD3 and CD2 is essential for interleukin 2 production and proliferation, since SE‐UCHT1 treated with the anti‐T11TS MoAb L180/1 are not mitogenic; (2) proximity of CD3 and CD2 is required during the stimulation, since a mixture of SE exposing either anti‐CD3 or T11TS is not mitogenic; and (3) that T cell‐AC interactions involving LFA‐1 can be replaced by LFA‐3‐CD2 interactions, since the anti‐LFA‐1 MoAb 60.3 does not inhibit the SE‐UCHT1 response, and only partly the SE+PHA response. These results demonstrate a functional linkage between the CD3 and CD2 structures, making accessory LFA‐1 signals superfluous in proliferative activation of human resting peripheral T cell.
Enhancement of Human Spontaneous Cell‐Mediated Cytotoxicity by a Monoclonal Antibody against the Large Sialoglycoprotein (CD 43) on Peripheral Blood Lymphocytes
Authors: VARGAS‐CORTES M. , AXELSSON B. , LARSSON Å. , BERZINS T. , PERLMANN P.
Published: June 2006

A murine monoclonal antibody, MoAb B1B6 (IgG1χ), which recognizes the large sialoglycoprotein (LSGP)on human peripheral blood lymphocytes (PBL) effectively enhanced the spontaneous cytotoxicity of these cells against She natural killer (NK)‐sensitive target cells K562 and Molt‐4. Whereas preincubation of she lymphocytes with MoAb B1B6 resulted in increased cytotoxicity, preincubation of the target cells had no effect, indicating that the MoAb amplified cytotoxicity at the effector cell level. Kinetic analysis of the data revealed no differences between the control and the MoAb‐treated lymphocytes with regard to Vmax, usually considered to reflect the overall lytic potential of the cells. The slopes of the saturation curves, however, differed significantly for She two cell populations, indicating a substantial increment in the activity of the MoAb‐treated cells. When studied lit the single cell level and with K562 as targets, treatment of PBL with the MoAb resulted in the recruitment of new effector lymphocytes from the pool of non‐binding cells. In contrast, when Molt‐4 cells were employed as targets, no additional effector cells were recruited. These results indicate that the enhanced cytotoxicity induced by MoAb B1B6 is the result of either recruitment of new effector lymphocytes or of an increased recycling capacity of preexisting effector cells. Together with previous observations, these findings support the conclusion that LSGP belongs to the set of surface molecules which regulate human lymphocyte activation.
Differential Induction of Membrane‐Associated Interleukin 1 (IL‐1) Expression and IL‐1α and IL‐1β Secretion by Lipopolysaccharide and Silica in Human Monocytes
Authors: HURME M. , SEPPÄLÄ I. J. T.
Published: June 2006

Bacterial lipopolysaccharide (LPS) and silica dust arc known to be effective inducers of interleukin 1 (IL‐1) in human cultured monocytes. The data reported here show that although the levels of secreted IL‐1 were equally high after in vitro stimulation with an optimal dose of LPS or silica, there were two clear differences: (i) the levels of membrane‐associated IL‐1 (as detected by the comitogenic effect of paraformaldehyde (PFA)‐fixed cells or purified membrane fragments on marine thymocytes) were ca. five times higher after LPS stimulation than after silica stimulation, (ii) the secreted IL‐1 after LPS stimulation was mainly of the pI 7 (IL‐1β) type, while after silica stimulation there were equally high amounts of pI 7 and pI 5 (i e. IL‐1α) forms. In both cases the IL‐1‐active molecules belonged to the 15 kDa class. These data show that the nature of the activating agent has a clear influence on the distribution of the biologically active IL‐1 molecules. Moreover, the finding that after silica stimulation the amount of membrane‐associated IL‐1 (which was recently shown to be of the IL‐1α type) was low, while IL‐1α in the culture Quid was clearly elevated, suggests that the IL‐1α not attached to the cell membrane (or released from it) significantly contributes to the secreted IL‐1 pool.
Major Histocompatibility Complex (MHC) Class II Antigen (HLA‐DR, DQ, and DP) Expression in Human Fetal Endocrine Organs and Gut
Authors: OLIVER A. M. , THOMSON A. W. , SEWELL H. F. , ABRAMOVCH D. R.
Published: June 2006

Monoclonal antibodies were used to analyse adrenal, pancreas, thyroid, and gut samples from human fetuses (14–11) weeks estimated gestational age; EGA) for the presence of class II major histocompatibility complex (MHC) antigens (HLA‐DR, DQ, DP) by immunohistochemistry. In the adrenal definitive and fetal cortex, HLA‐DR+, DP−, DQ− cells were clearly demonstrated. These DR+ cells were identified, phentotypically, as predominantly tissue macro‐phages and a small population of CD45R+, IgM+ lymphoid cells. Within the pancreas, numerous cell throughout the tissue were strongly DR+ but DQ−; DP+ cells were not observed until 17 weeks EGA. Using a double‐labelling procedure, minor proportions of these DR+ cells were identified as macrophages or as (CD19+) B cells, while endocrine and endothelial cells were negative. Throughout the thyroid, small numbers of DR+ macrophages and small lmyphoid cells were detected, although the thyroid epithelial cells were DR+, DP+, and DQ−. Large numbers of DR+, DP+, DQ− cells were observed in the stomach wall and mucosa. In the intestine, DR+, DP+, DQ+ cells positive for all MHC class II loci products were abundant throughout the lamina propria and lymphoid aggregates. The class II antigens appeared in the proportion DR>DF>DQ and expression was most prevalent in the mid‐gut region. A small proportion of epithelial cells of the villi along she gut were weakly DR+ but DQ−, DP−. These results show that DR+ cells in fetal endocrine tissue are mainly‘passenger leucocytes' and that, in contrast to recent reports concerning normal adult tissue, the adrenal and pancreatic endothelial cells are DR−.
Efficacy of Influenza Haemagglutinin and Nucleoprotein as Protective Antigens against Influenza Virus Infection in Mice
Authors: ANDREW M. E. , COUPAR B. E. H.
Published: June 2006

Influenza nucleoprotein (NP)‐specific cytotoxic T lymphocytes (CTL) stimulated by immunization of mice with VV‐PR8‐NP6, a recombinant vaccinia virus expressing A/PR/8/34 NP, did not protect mice against challenge with A/PR/8/34 4 days later. Neither were secondary NP‐specific CTL stimulated by reimmunization able to protect mice. These results contrast with the ability of transferred, in vitro‐cultured and stimulated, NP‐specific CTL to protect recipient mice from challenge with A/PR/8/34. Immunization of mice with a recombinant vaccinia virus expressing A/PR/8/34 HA protected mice challenged 4 days later, either via the small amount of antibody already present, or via HA‐specific CTL that would have to be more efficient than NP‐specific CTL in either trafficking to the infected lung or in effector function.
Immune Response in Chronic Schistosomiasis haematobium and mansoni
Authors: FELDMEIER H. , GASTL G. A. , POGGENSEE U. , DAFFALLA A. A. , NOGUEIRA‐QUEIROZ J. A. , CAPRON A. , PETER H. H.
Published: June 2006

Peripheral blood mononuclear cells from Sudanese children heavily infected with Schistosoma haematobium and 5. mansoni were examined for lymphocyte subpopulations, for mitogen and antigen responsiveness, and for natural killer (NK) cell activity before and 5 months after treatment with praziquantel. The humoral immune response was simultaneously investigated by determination of parasite‐specific IgG and IgE antibodies. IgE‐containing circulating immune complexes, and circulating schistosome antigen. A single dose treatment with praziquantel (40 mg/kg) resulted in a normalization of numerical imbalances in lymphocyte sub‐populations, a significant increase in the blastogenic response upon stimulation with adult worm antigen, phytohaemagglutinin (PHA), and concanavalin A (Con A), and restoration of natural killer (NK) cell‐mediated lysis of K562 targets. These findings were paralleled by a remarkable decrease in parasite‐specific IgE antibodies, IgE‐containing circulating immune complexes, and circulating schistosome antigen. The results indicate that the modulation of immune responses in chronic schistosomiasis is associated with active infection and is reversible after successful chemotherapy.
Evidence that the Major Primary Biliary Cirrhosis‐Specific Mitochondrial Autoantigen is a Subunit of Complex I of the Respiratory Chain
Authors: FROSTELL Å. , MENDEL‐HARTVIG I. , NELSON B. D. , TÖTTERMAN T. H. , BJÖRKLAND A. , RAGAN I.
Published: June 2006

Primary biliary cirrhosis (PBC)‐specific antigens were purified from beef heart mitochondria by immunoaffinity chromatography Three major polypeptides (75, 60, and 40 kDa) were detected in the purified antigen fraction both by Coomassie blue staining and by western blot analysis. The 75 kDa antigen was identified as a subunit of Complex I (NADH‐ubiquinone reductase) by the following criteria: (1) antibodies against the purified 75 kDa subunit of beef heart Complex I read with the immunoaffinity‐purified 75 kDa antigen. (2) the 75 kDa subunit present in isolated Complex I, like that in the immunoaffinity‐purified antigen, reacts with PBC sera only after reduction with mercaptoethanol, and (3) the 75 kDa antigen is enriched in isolated Complex I. A relationship between the 75 kDa and the 60 and 40 kDa antigens is suggested, since optimal binding of anti‐mitochondrial autoantibodies (AMA) to the latter antigens also requires prior reduction with mercaptoethanol. A fourth major antigen (70 kDa) was also detected by western blot analysis, but only in samples that had not been boiled prior to electrophoresis. This antigen, which is also present in isolated Complex I, resembles the 75, 60, and 40 kDa antigens in its response to mercaptoethanol and its reaction with antibodies against the 75 kDa subunit of Complex I. A scheme is presented which relates all of the PBC antigens to the parent 75 kDa subunit of Complex I, probably as proteolytic products of the latter.
Blood Transit and Recirculation Kinetics of Lymphocyte Subsets in Normal Rats
Authors: WESTERMANN J. , PUSKAS Z. , PABST R.
Published: June 2006

Thoracic dud lymphocytes (TDL) were labelled with fluorescein isothiocyanate (FTTC) and injected into normal, unanaesthetized rats with either a central venous catheter or a thoracic duct cannula. The blood transit time and the appearance in the lymph was calculated and then the percentages of B, T, T helper, and T suppressor lymphocytes were determined with monoclonal antibodies (Ox12, Ox19, W3/25, and Ox8, respectively). The blood transit time of all subsets was about 30 min. However, the percentage of H lymphocytes from 5 min after injection onwards is reduced (14.5±1.9%) compared to the injected TDL (32.7±1.7%), These cells are not in the lung vascular pool. The recovery of FITC‐labelled TDL in the thoracic duel within 48 h is much higher (56.8±5.3% of the injected lymphocytes) than in previous studies using radioactive markers. B lymphocytes appear later and in a reduced number in the thoracic duel. The mean transit time is 26 h and the recovery 31.5±4.2% in contrast to T lymphocytes (18 h and 66.3±6.5%, respectively). Tile technique of combining FITC in vitro labelling with surface staining after completion of migration does not interfere with lymphocyte migration. It can therefore be used to study the migration of lymphocyte subsets, in normal, untreated animals.
Presence of Mycobacterium leprae‐Reactive Lymphocytes in Lymph Nodes of Lepromatous Leprosy Patients
Authors: DESAI S. D. , BIRDI T. J. , ANTIA N. H.
Published: June 2006

A critical problem in leprosy is the relative deficiency of antigen‐specific T cell‐mediated immunity. We were successful in detecting a significant response to viable M. leprae in mononuclear cells isolated from the lymph nodes of lepromatous leprosy patients in contrast to the apparent M. leparae‐specific energy seen in the peripheral blood. This observation suggests that antigen‐reactive lymphocytes are generated in the lymph nodes of lepromatous patients but the inability to detect them in the circulation may be due either to a different processing and presentation of mycobacterial antigens within the peripheral blood and lymph node compartments or to a selective sequestration of lymphocytes within the lymph node.
The Leucocyte L1 Protein: Identity with the Cystic Fibrosis Antigen and the Calcium‐Binding MRP‐8 and MRP‐14 Macrophage Components
Authors: ANDERSSON K. B. , SLETTEN K. , BERNTZEN H. B. , DALE I. , BRANDTZAEG P. , JELLUM E. , FAGERHOL M. K.
Published: June 2006

The partial amino acid sequence of L1 protein light and heavy chains reveals an overall structure identical to the two macrophage proteins, MRP‐8 and MRP‐14, deduced from the sequence of the cDNA encoding the polypeptides. The light chain of L1 protein (L1‐L) was shown to contain two modified amino acid residues.
IgG in Murine Intestinal Secretions Aging Effect and Possible Physiological Role
Authors: SENDA S. , CHENG E. , KAWANISHI H.
Published: June 2006

To assess the role of, and age‐associated alterations in, intraluminal IgG in the intestine, total IgG and subclass distribution in small intestinal perfusates and pilocarpine‐stimulated intestinal secretions were measured under non‐fasting conditions in three groups of BALB/c mice: 2–3, 12–14, and over 24 months old. Total intraluminal IgG and its subclasses (i.e. IgG1, IgG2a, and IgG2b) increased with age. However, the binding capacities of natural intraluminal IgG antibody (Ab) for outer membrane antigen (Ag) from normal habitant bacteria (Escherichia coli and Enterobacter agglomerans) were markedly reduced in the aged lavage samples as compared with such binding in samples from the other two age groups. The striking similarity to serum in the magnitude of the increase of intraluminal IgG and also in the distribution patterns of its subclasses in the small intestinal perfusates from also age groups suggests that the serum is a major source of the IgG. The intraluminal IgG may play some role in humoral immune protection under physiologic conditions of the gut lumen.
Recombinant Tumour Necrosis Factor (TNF) Fixed to Cell Monolayers Retains Its Cytotoxic and Growth‐Stimulatory Activity
Authors: HOFSLI E. , NISSEN‐MEYER J.
Published: June 2006

Recombinant human tumour necrosis factor (rTNF) retained its cytotoxic activity after being fixed with paraformaldehyde to adherent cell monolayers. The cytotoxicity appeared to be mainly due to fixed rTNF and not to any free soluble rTNF that could have leaked out from the lived rTNF cell preparations. The fixed rTNF cell preparations also stimulated the growth of human diploid fibroblasts, under conditions where little growth‐Stimulatory activity was found in suspension. These results indicate that TNF may exert its effect on target cells without internalization, perhaps through a receptor‐mediated process that may alter the levels of a second messenger within the target cells. This signal transduction does not appear to involve cAMP or cGMP, since we were unable to detect significant changes in the levels of these two second messengers in TNF‐exposed WEHI 164 clone 13 cells.
Purification of a Major Mycobacterium bovis Antigen for the Diagnosis of Bovine Tuberculosis
Authors: FIFIS T. , PLACKETT P. , CORNER L. A. , WOOD P. R.
Published: June 2006

A Mycobacterium bovis antigen has been purified from culture filtrate by chromatofocusing. This antigen is a major component of culture filtrate and cell extracts and shows a considerable degree of micro‐heterogeneity in electric charge and molecular weight. Studies with monoclonal and polyclonal antibodies raised against the purified antigen show that some of its antigenic determinants also occur in higher molecular weight species in culture filtrate and particularly in whole cell preparations. Immunoblotting and ELISA studies, using sera from M. bovis‐infected animals, showed that this antigen is one of the most immunoreactive components of M. bovis, recognized by the majority of animals with detectable antibody response to M. bovis. The specificity of the purified antigen is far superior to that of the crude culture filtrate, with very few false positive results. The purified antigen also elicits strong in vivo and in vitro cell‐mediated responses The amino acid compositions of two variants of this antigen have been determined and found to be similar to that of MPB‐70.
Distribution of κ and λ Light Chain Isotypes among Human Blood Immunoglobulin‐Secreting Cells after Vaccination with Pneumococcal Polysaccharides
Authors: HEILMANN C. , BARINGTON T.
Published: June 2006

The light chain isotype of immunoglobulin‐secreting blood cells was investigated by means of monolayer plaque‐forming cell assays allowing direct immunofluorescence staining for cytoplasmic δ and λ light chains in center cells. The study revealed that cultured, polyclonally activated pokeweed mitogen (PWM) and Epstein‐Barr virus (EBV), IgM‐, IgG‐ and IgA‐secreting cells expressed the δ light chain isotype in approximately 65% of the cells. IgM and IgG‐secreting cells induced by vaccination with pneumococcal polysaccharides had a similar percentage of δ light chain‐containing cells. In contrast, IgA‐secreting cells induced by vaccination with pneumococcal polysaccharides showed a different (bimodal) distribution as regards expression of δ light chain. The majority (56%) of the Investigated individuals expressed δ light chain in approximately 50% of the cells and the rest expressed δ light chains in approximately 80% of the cells. The percentage of cells containing δ light chains among spontaneous IgA‐secreting cells in unvaccinated individuals was approximately 50% and thus also differed from the general pattern for mitogen‐activated B cells. The light chain pattern of IgA‐secreting cells from individuals vaccinated with pneumococcal polysaccharides and from unvaccinated individuals probably indicates that these cells are being derived from B‐cell clones with a limited idiotypic heterogeneity, which have been selected and clonally expanded by naturally occurring antigens at the mucosal membranes.
IgG Subclasses of Pneumococcal Antibodies —Effect of Allotype G2m(n)
Authors: SARVAS H. , RAUTONEN N. , SIPINEN S. , MÄKELÄ O.
Published: June 2006

Antibody responses to three pneumococcal polysaccharides (types 3, 14, and 18C) were analysed after vaccination with a 23‐valent polysaccharide vaccine. Antibodies to all three polysaccharides could be detected before immunization. Clear cut IgG, IgA, and/or IgM antibody responses to the polysaccharides were seen in three‐quarters of the vaccines IgG2 was the predominant and IgG1 the second most abundant subclass of anti‐pneumococcal IgG antibodies both before and after the vaccination. The relative proportions of IgG2 and IgG1 antibodies exhibited a continuous variation from 1:0 to approximately 0:2. After vaccination. G2m(n)‐positive homozygotes had about four times more IgG2 antibodies (anti‐14 und anti‐18C than G2m(n)‐negative vaccinees. Heterozygotes occupied an intermediate position. The same pattern was seen less clearly in type 3 antibodies after vaccination, and in all three antibodies before vaccination. The G2m(n) allotypes had no detectable effect on the levels of IgG1, IgG4, or IgM antibodies, and possibly a weak effect on IgG3 and IgA antibodies (G2m(n)‐positive homozygotes responded strongly).
Stimulation of Interleukin 6‐Like B‐Cell Differentiation Factor Production in Human Adherent Synovial Cells by Recombinant Interleukin 1
Authors: HARIGAI M. , HARA M. , NORIOKA K. , KITANI A. , HIROSE T. , SUZUKI K. , KAWAKAMI M. , MASUDA K , SHINMEI M. , KAWAGOE M , NAKAMURA H.
Published: June 2006

Abnormal production of immunoglobulin in the joint space is frequently observed in patients with rheumatoid arthritis (RA). We have previously demonstrated that adherent synovial Cells (ASC) from patients with RA are involved in B‐cell differentiation by their spontaneous production of B‐cell differentiation factor (BCDF). The regulation of the production of this factor, however, has not yet been described. We investigated the effects of recombinant interleukin 1α and β (rIL‐1α and rIL‐1β) on the production of BCDF in ASC Increased production of BCDF was observed with increased rIL‐1 concentration. Production of BCDF was detected 3 h after exposure of ASC to rIL‐1 and increased throughout a 48‐h culture. This BCDF, assayed on SKW6‐CL4 cells, was found to share a common active site with interleukin 6. The effect of rIL‐1 was almost neutralized by anti‐IL‐1 antibody and the addition of polymyxin B did not diminish the effect of rIL‐1, indicating that rIL‐1 itself stimulates ASC in vitro. These results suggest that IL‐1 may play a regulatory role in the production of BCDF in synovial tissue.
Translocation of Protein Kinase C to Subcellular Fractions of Human Neutrophils
Authors: CHRISTIANSEN N. O. , BORREGAARD N.
Published: June 2006

The subcellular localization of protein kinase C in unstimulated human neutrophils and neutrophils stimulated by phorbol‐myristate‐acetate (PMA), 1‐oleoyl‐2‐acetyl‐rac‐glycerol (OAG), and ionomycin was investigated in subcellular tractions obtained by nitrogen cavitation and Percoll density gradient centrifugation. Protein kinase C was found to he localized mainly in the cytosol in unstimulated cells, whereas significant translocation to fractions containing the plasma membrane was observed after stimulation by PMA, OAG, and ionomycin. At the same time, phospholipid‐insensitive protein kinase activity appeared in the cytosol and the plasma membrane fractions. To determine whether binding of protein kinase C occurred to the plasma membrane or to intracellular membranes that had translocated to the plasma membrane, we investigated the ability of isolated azurophil, specific and secretory granules, and plasma membrane vesicles to bind protein kinase C in response to addition of PMA and OAG. Only fractions containing plasma membranes and secretory granules were able to bind protein kinase C. The observation explains the selective activation of plasma membrane structures by protein kinase C.
Effects of 12‐O‐Tetradecanoyl Phorbol‐13‐Acetate (TPA) on Rat Thymocytes
Authors: SASAKI R. , TAKAKU F. , MIURA Y.
Published: June 2006

The short‐term incubation of rat thymocytes with 12‐O‐tetradecanoylphorbol‐13‐acelaic (TPA) resulted in a significant increase in sialyltransferase (S‐T) activity and a decrease in terminal deoxyribonucleotidyl transferase (TdT) activity. The ratio of peanut agglutinin (PNA)‐positive cells and of TdT‐positive cells in TPA‐treated cells also decreased. However, TPA had no significant effects on the viability, morphology, DNA synthesis, and DNA polymerase α activity of the cells. More marked changes were observed by incubating a non‐T, non‐B human lymphoid leukaemia cell line with TPA. Similar findings were also noted in TPA‐treated mouse thymocytes. These changes may represent an aspect of TPA‐induced differentiation of murine thymocytes.