bluuebunny/crossref_metadata_2025 huggingface dataset:
Increased Frequencies of the CD29 and CD57 Markers and Decreased Frequency of CD45RA Within CD4+ and CD8+ Subsets after Allogeneic Bone Marrow Transplantation in Man
Authors: MøLLER J. , DICKMEISS E. , RYDER L. P. , JACOBSEN N. , SVEJGAARD A.
Published: June 2006

Two monoclonal antibodies, anti‐CD45RA and anti‐CD29, reciprocally divide the CD4+ and CD8+ lymphocytes into CD4+ CD45RA+, CD4+ CD29+, CD8+ CD45RA+ and CD8+ CD29+ subsets. The CD4+ CD45RA+, CD4+ CD29+ and CD8+ CD45RA+ possess suppressor‐inducer, helper‐inducer and suppressor‐effector functions respectively. Since the role of these subsets has not been established after allogeneic bone marrow transplantation we studied lymphocyte subpopulations in 12 patients 45‐ 227 days after the procedure.

The fraction of CD4+ lymphocytes was significantly (P= 0.0005) decreased to 20±9% versus 43±3% in controls. Within the CD4+ compartment, we found an increase in the fraction of CD4+ cells that co‐expressed CD29 (CD29+/CD4+) to 92±10% versus 48±15% (P=0.008) in controls and a concommittant decrease m CD45RA+/CD4+ to 16±12% versus 56±25% (P=0.008)

Patients were also noted to have an increase in the percentage of CD8+ lymphocytes to 41±5% compared to 23±4% in controls (P=0.0004), Examination of the CD8+ subsets revealed a significant increase in the CD29+/CD8+ fraction to 97±3% versus 64±2% in controls (P= 0.008) and a decrease in the CD45RA+/CD8+ fraction to 36±11% versus 70±21% (P= 0.008).

The number of cells co‐expressing CD57 were also determined within the CD4+ and CD8+ subsets. In patients CD57+/CD4+ were increased to 29±7% versus 1±1% in controls (P=0.04), and CD57+/CD8+ to 49±12% versus 23±9% (P=0.02).

Since CD29+ and CD57+ cells have a poor capability for IL‐2 production and proliferation this shift in subset distribution may account for some of the defects in cellular immunity seen within the first year after allogeneic bone marrow transplantation.
Inhibition of Human Natural Killer Cell Activity by Platelet‐Derived Growth Factor (PDGF)
Authors: GERSUK G. M. , WESTERMARK B. , MOHABEER A. J. , CHALLITA P. M. , PATTAMAKOM S. , PATTENGALE P. K.
Published: June 2006

We have previously reported that platelet‐derived growth factor (PDGF) substantially inhibits human natural killer (NK) cell cytotoxicity, and that NK cells possess high‐affinity surface binding sites for the PDGF‐AB isoform. In this communication, we present direct evidence for the presence of A‐type alpha) PDGF receptors on human NK cells by demonstrating that human NK cells have approximately 150,000 high‐affinity, surface binding sites for recombinant (r)PDGF‐AA and approximately 300,000 high‐affinity, surface binding sites for rPDGF‐BB. This was determined by the competitive binding of 125I‐labelled rPDGF‐AA or 125I‐labelled rPDGF‐BB and homologous unlabelled rPDGF‐AA or rPDGF‐BB to FACS‐sorted, CDl6+ lymphoid (NK)cells, and Scatchard analysis of these data. In addition, we also demonstrated that the various isoforms of PDGF have differential effects on NK‐cell cytotoxicity, Physiological quantities (100 ng/ml) of rPDGF‐BB homodimers, highly purified PDGF‐AB heterodimers from outdated platelets, and rPDGF‐AB heterodimers substantially inhibited NK‐cell cytotoxicity in both a dose‐ and time‐dependent manner. In contrast, pretreatment of NK cells with equivalent nanogram amounts of rPDGF‐AA homodimers resulted in a significantly weaker inhibitory effect on NK‐cell cytotoxicity as compared with the PDGF‐BB und PDGF‐AB isoforms. The implications of these findings are discussed.
Cellular Mechanisms of Graft‐Versus‐Host Disease in a Mouse Model
Authors: DE GIORGI L. , LOWDELL M. W. , MATOSSIAN‐ROGERS A.
Published: June 2006

Female CBA/H (H‐2k, Mlsb) mice alloimmunized prior to and during syngeneic pregnancy with DBA/2 (H‐2d, Mlsa) splenocytes gave rise to offspring which resisted graft‐versus‐host disease (GVHD) following neonatal intraperitoneal inoculation of high doses of DBA/2 spleen cells. Lymphocytes from GVHD‐resistant mice tested after 6 weeks of age were unresponsive to DBA/2 stimulator cells in 72 h mixed lymphocyte cultures. Isotope uptake measured 24 h after culture, however, indicated that a considerable early response was made to DBA/2 which later declined. Proliferative responses to BALB/c were also depressed but no early response to this strain was detected. FACS analysis of T‐lymphocyte profiles of the GVHD‐resistant CBA/H mice revealed a 100% increase in the Lyt‐2+ subpopulation compared to normal CBA/H mice. Significant increases in Lyt‐2+ cells were also noted in in vitro cultures of CBA/H lymphocytes responding to GVHD‐resistant CBA/H stimulators. Lymphocytes from GVHD‐resistant mice suppressed the proliferative responses of normal CBA/H lymphocytes to alloantigenic but not mitogenic stimulation. Suppression of alloantigenic responses were shown to be specific to DBA/2 and did not affect the response to BALB/c stimulator cells, indicating that both anergy and specific suppressor cells were operative in inducing unresponsiveness.
Effect of the In Vitro Ageing on the Restriction of Sheep Erythrocyte Lysis by Horse Serum
Authors: ASSIS A. I. , PEREIRA G. , AZZOLINI A. E. C. S. , BARBOSA J. E.
Published: June 2006

Fresh sheep erythrocytes are resistant to lysis by horse serum but gradually became susceptible to this source of complement after storage (ageing) in the blood at 4°C, This occurs faster when the fresh cells are stored in isotonic buffer. The supernatant of the buffer/red cell suspension stored at 4°C for 10–15 days restrict lysis by horse serum when incubated back at 37°C or 43°C with the ‘aged’ sheep cells or fresh guinea pig red cells. In assays using fresh guinea pig erythrocytes this effect is decreased by reincubation of the cells in buffer, and is specific to horse serum when compared to human serum.
The Presence of Anti‐Protein A Antibodies in Patients with Rheumatoid Arthritis
Authors: TAKEUCHI T. , HOSONO O. , KOIDE J. , AMANO K. , SEKINE H. , ABE T.
Published: June 2006

Sixteen patients with rheumatoid arthritis (RA) were examined for the presence of anti‐protein A antibodies. The F(ab′)2preparations from five RA patients showed significant binding lo IgG‐free protein A on ELISA. The protein A binding was further examined by immunoblotting. The F(ab′)2preparations of high protein A‐binding protein gave A specific reaction with IgG‐free protein A on nitrocellulose paper. This demonstrates the presence of anti‐protein A antibodies in patients with RA. Those RA patients with anti‐protein A antibodies had more active disease as judged by the Lansbury's activity index. The level of serum rheumatoid factor (RAHA) was significantly higher in patients with anti‐protein A antibodies than in those without anti‐protein A antibodies.
Regulation of Human Interleukin‐2 and Interferon‐Gamma Gene Expression by Suppressor T Lymphocytes
Authors: KETZINEL M. , EFRAT S. , SAYAR D. , GEREZ L. , TAL C. , DEUTSCH E. , ARAD G. , KAEMPFER R.
Published: June 2006

Concomitant with induction of inlerleukin‐2 (IL‐2) and interferon‐gamma (IFN‐γ) gene expression in human tonsil cells, milogenic stimulation induces a transient activation of cells able to effectively suppress expression of these genes. Induction of IL‐2 and IFN‐y genes largely precedes appearance of suppressor cell activity, allowing expression of both genes to occur before strong down‐regulation is exerted by activated suppressor cells. Suppressive activity induced in one cell population can inhibit IL‐2 and IFN‐γ gene expression in another population from the same donor. The distinct nature of suppressor cells is supported by the absence of down‐regulation of IL‐2 gene expression in a helper cell line. M LA‐144: yet, in these cells, negative control can be expressed when active suppressor cells are introduced. Our findings support the concept that actual levels of IL‐2 and IFN‐γ gene activity are regulated to a large extent by the differential kinetics of activation of suppressor cells on one hand and of cells expressing the IL‐2 and IFN‐γ genes on the other.
C5 Does Not Play a Major Role in the Immune Response of Mice to SRBC In Vivo
Authors: VAN DEN BERG C. W. , VAN HORSSEN P. J. , HOFHUIS K. M. A. , AERTS P. C. , VAN DIJK H.
Published: June 2006

The role of complement component C 5 in the immune response of mice to sheep red blood cells (SRBC) was investigated, Congenic C5‐suffcieal and C 5‐deficient B10.D2 mice Lind genetically CS‐deficient DBA/2 mice. as such or supplemented with C5‐suficient serum, were used as experimental animals. C5‐substitution of the C5‐deficient mice resulted m measurable CS levels for days. The functional half‐life of C? in C5‐deficienl DBA 2 mice was about 21 h.

No significant differences between the IgM‐responses of C5bearing and naive C5‐delicient animals were observed. This suggests t ha C5 does not play a major role in the primary humoral immune response of mice in vivo, although C5 seems to do so in in vitro experiments, even with the same antigen. Antigen‐induced C5‐production by C5‐defieient mice as one of the explanations of the in vitro, in vivo discrepancy could not be confirmed experimentally.
T‐Cell Receptor Gene Expression by Human γδ T‐Cell Clones from Peripheral Blood and Reproductive Tissues in Relation to Non‐MHC‐Restricted Cytotoxic Function
Authors: CHRISTMAS S. E.
Published: June 2006

T‐cell receptor γ and δ gene expression was determined using V‐region‐specific monoclonal antibodies in conjunction with Southern blot analysis in panels of γδ T‐cell clones from human peripheral blood (n = 77) and reproductive tissue (n = 9). Whereas 53 out of 77 (69%) clones from peripheral blood expressed Vγ9 and Vδ2Jl, only 2 out of 9 (22%) from reproductive tissues expressed Vδ2J1. Two out of eight decidual clones expressed both Vγ9 and VδJ1, while this configuration was rare in clones from peripheral blood. The majority of clones from the peripheral blood of one donor expressed Vγ8 and Vδ3J1. Clones were identified which expressed Vδ1J1 in the disulphlde‐linked Cγ1 form of the receptor and which expressed a gene other than Vδ1 in the non‐disulphide‐linked Cγ2 form, indicating incomplete concordance between expression of Vδ1 and Cγ2. Vδ3 could be expressed in the disulphide‐linked or non‐disulphide‐linked form of the receptor. At least 5 out of 77 peripheral clones were expressing Vδ genes other than Vδ1. Vδ2. or Vδ3 in conjunction with Cγ1 or Cγ2. There was a strong but incomplete correlation between high non‐NHC‐restricted cytotoxic function and Cγ1 expression. Clones from the same donor expressing both Vγ9JPCγ1 and Vδ2J1 showed either high or negligible cytotoxicity. and cytotoxic clones expressing Cγ2 were found. Thus no complete correlation between cytotoxic function and expression of a particular form of the yδ heterodimer was identified. The results also suggest that yδ T cells from reproductive tissues are less likely to express Vδ2J1 than those from peripheral blood.
Human Umbilical Vein Endothelial Cells Synthesize Functional C3, C5, C6, C8 and C9 In Vitro
Authors: JOHNSON E. , HETLAND G.
Published: June 2006

Human endothelial cells (EC), cultured serum‐free, synthesize de novo protein which increasingly bind to agarose beads (an alternative pathway activator), until a plateau phase is reached after 24–48 h. EC synthesize functional C3, C5, C6, C8 and C9, which were detected on co‐cultured agarose beads, using relevant polyclonal anti‐complement antibodies. Two monoclonal anti‐C9 neoepitope antibodies (aEl 1, poly C9‐MA) bound to the co‐cultured beads, showing that the terminal complement complex (TCC) (C5b‐9) was assembled on the beads. This also suggests that C7 is synthesized. There seems to be a positive correlation between the amount of agarose‐bound labelled protein and agarose‐bound complement. The results indicate that EC produce and secrete the components for the functional alternative and terminal pathways of complement.
The Effect of Stress In Vivo on the Function of Mouse Macrophages In Vitro
Authors: AARSTAD H. J. , KOLSET S. O. , SEUELID R.
Published: June 2006

Macrophages were harvested from home cage control (HCC) mice, and from mice which had been stressed by repeated brief exposures (3 8 min) to cold water at 10–15 C twice daily for 8 or 14 days. Macrophages obtained from mice stressed 8 or 14 days compared lo macrophages from HCC mice showed in vitro increased amounts of membrane‐bound prothrombinase activity, whereas the thrombin degradation activity was unchanged. Furthermore, macrophages of mice stressed × days showed increased release of coagulation factor X/Xa to supernatant in vitro. These findings suggest an increased amount of prothrombinase complex enzymes on the surface of macrophages from mice stressed 8 days, and increased netivity of the prothrombinase enzyme in macrophages from mice stressed 14 days. The synthesis of proteoglycans(PG)and glycosaminoglycans (GAG) was increased in macrophages from mice stressed 8 days compared to macrophages from HCC mice and mice stressed 14 days. When macrophages from mice stressed 8 days or HCC mice were stimulated in vitro with phorbol myristate acetate (PMA) and IL‐1 or PMA and IL‐2, a changed PG/GAG synthesis was observed only in macrophages from the HCC animals. Furthermore, both the tumour cytotoxicity and the released tumour necrosis factor (TNF) were decreased from macrophages from mice stressed 14 days compared to HCC mice. The results suggest that the macrophages of stressed mice have an altered mode of function more complex than a simple general suppression or activation.
The Adjuvant Action of Cholera Toxin is Associated with an Increased Intestinal Permeability for Luminal Antigens
Authors: LYCKE N. , KARLSSON U. , SJÖLANDER A. , MAGNUSSON K.‐E.
Published: June 2006

This study addresses the question of whether cholera toxin (CT) increases gut permeability for molecules > 3000 Da and whether such an effect is associated with an adjuvant function by CT on the gut immune response. We louiid that CT after oral administration gives rise to strikingly increased gut permeability for Dextran (Mw 3000) eoncomilantly with a strong enhancing effect on the anti‐keyhole limpet heniocyanin (KLII) specific immune response in the lamina propria after oral immunization with KLH plus Dextran and CT. In contrast, the B‐subunit of the hololoxin, which lacks the adenylate cyclase/cAMP‐activating property of CT. failed to increase gut permeability as well as local anti‐KLH immune responses. These results might suggest a causal linkage between the ability of CT to increase gut permeability and its adjuvant property on gut mucosai immune responses. In addition this finding supports the notion that the adenylate cyclase/cAMP system plays a regulatory role in gut permeability and is important in enhancing mucosal immune responses. Based on previous studies and the present data we propose that the mechanism for CT's adjuvant function on mucosal immune responses is by affecting antigen‐presenting cells, T and B cells in the gut to give a net enhancing effect on the stimulation of local immunity, and that the CT‐induced increase in gut permeability might be part of the adjuvant mechanism by facilitating luminal antigens to access the gut mucosal immune system.
The Complex of α2‐Macroglobulin with CD2 in the Plasma of Gastric Carcinoma Patients
Authors: SMORODIN E. P. , KURTENKOV O. A.
Published: June 2006

α macroglobulin (α2M) preparations purified from the plasma of healthy persons and gastric carcinoma patients were further analysed by immunochemical techniques. It was found that only αM from the plasma of gastric carcinoma patients gave u positive reaction with E‐receptor sheep antiserum in rocket immunoelectrophoresis. The αM fraction giving the positive reaction with E‐receptor sheep antiserum was isolated and analysed by the immunobloting technique with monoclonal antibodies to TlI antigen. The presence of the αM‐CD2 complex in the plasma of gastric carcinoma patients was demonstrated. This finding may explain the immunosuppressive effect of the plasma αM in cancer patients.
Granulocyte‐Dependent Extracellular Cytotoxicity is Enhanced by Complement C3bi and Independent of Phagocytosis
Authors: KOJIMA A. , HAZEKI K. , SEYA T.
Published: June 2006

We have evaluated phagocytosis. C3‐binding and cytotoxicity of human leukocytes simultaneously, using IgM‐sensitized C3‐fragment‐bearing sheep erythrocytes (EAC3b. EAC3bi) as targets. In this method, 51Cr‐labelled EAC3b or EAC3bi were added to human peripheral granulocytes anchored by fibronectin onto microtitre plate wells, The degrees of haemolysis, binding and ingestion of the target cells were estimated from the radioactivity released. We found that the granulocytes predominantly lysed EAC3bi but not EAC3b or EAC4b. EAC3bi lysis elicited from granulocytes was as effective as that from lymphocytes under the same assay conditions. However, one difference in the cytolysis of the two effector cells was that EAC3bi bound efficiently to all of the granulocytes similarly to EAC. Tb. whereas it bound Io only 4% of the lymphocytes. The bound cells did not appear to be efficiently phagocylosed by the granulocytes. Blocking studies using antibodies suggested that C3bi receptors. CR3 and CR4. but not other C3‐binding proteins, C3b, C4b receptor (CRI), membrane cofactor protein (MCP) or decay‐accelerating factor (DAF), are involved in gramilocyte‐mediated haemolysis. We speculate that simultaneous stimulation of the C3bi receptors and the fibronectin receptor results in elicitation of cytotoxicity by granulocytes.
Different Activation Signals are Required for the Expression of Interleukin‐1 α and β Genes in Human Monocytes
Authors: HURME M. , SERKKOLA E.
Published: June 2006

The production and mRNA expression of IL‐1α and IL‐1β by human monocytes was examined after two different stimuli, a protein kinase C (PKC) activator phorbol myristate acetate (PMA) and bacterial lipopolysaccharide (LPS). LPS induced production of high levels of both IL‐la and IL‐1β protein (quantitated with type‐specific ELISA assays), while after PMA stimulation only IL‐1/β protein could be detected. The IL‐1α and IL‐1/β mRNA levels quantitated by Northern blotting were in line with the respective protein levels and nuclear run off analysis revealed that PMA did not activate the IL‐la transcription. The production of the IL‐1α and IL‐1/β protein as well as the mRNA expression could be inhibited with protein kinase inhibitor H7, but not with HA 1004, indicating that PKC activation is essential for the activation of these genes. Thus these data indicate that PKC activation alone is sufficient for the induction of the IL‐1/β gene, but some additional signals (provided by LPS) are required for the activation of the IL‐1α gene.
IgG Subclasses in Bronchoalveolar Lavage Fluid from Patients with Asthma
Authors: OUT T. A. , VAN DE GRAAF E. A. , VAN DEN BERG N. J. , JANSEN H. M.
Published: June 2006

We have measured Immunoglobulin G (IgG) subclasses in serum and bronchoalveolar lavage fluid (BALF) from 12 non‐smoking patients with stable asthma and 9 non‐smoking healthy volunteers to obtain information on their possible role in local immunological reactions. The quotients (concentration of IgG subclass in BALF)/(concentration of IgG subclass in serum) were calculated. In controls QIgG3 were lower than QIgGI, QIgG2 and QlgG4.

The IgG subclasses in BALF and epithelial lining fluid (ELF) from patients with asthma were significantly higher than in controls, mainly due to increased leakage from the blood. Again QIgG3 were lower than Q of other subclasses. In the analysis of local production of IgG, albumin or ceruloplasmin was used as reference protein. Several patients showed a local production or a preferential accumulation of one or more IgG subclasses.

We conclude that in healthy persons the IgG subclasses in ELF originate from the systemic circulation by passive permeation. In patients with asthma, the permeability of the respiratory membrane may be increased resulting in increased concentrations of subclasses in lung‐lining fluid. In some patients with asthma, an additional local production of IgG subclasses occurs.
Primary Biliary Cirrhosis (PBC): Characterization of a Monoclonal Antibody (PBC‐MoAb) having Specificity Identical with Disease‐Associated Auto‐antibodies
Authors: BJÖRKLAND A. , MENDEL‐HARTVIG I. , NELSON B. D. , TÖTTERMAN T. H.
Published: June 2006

We have raised a monoclonal antibody (PBC‐MoAb) directed against mitochondria which resembles patent anti‐mitochondrial autoantibodies (AMA) (M2 type) in several respects.

The reaction pattern of PBC‐MoAb was characterized by western blot experiments, immunoaffinity purification and enzyme inhibition studies. PBC‐MoAb reacts specifically with an epitope on the E2 suburil of pyruvate dehydrogenase (dihydrolipoamide acyltransferase) which is essential for enzymatic activity. This was shown as follows: (I) PBC‐MoAb. like PBC‐AMA, completely inhibited PDH enzyme activity and reacted weakly with OGDII; (2) PBC‐MoAb bound strongly to the E2 subunit in western blots, with weaker binding to a doublet of about 56 kDa: and O) in immunosorbent experiments, PBC‐MoAb absorbed most (>95%) 0of the AMA reactive material found in solubilized mitochondria.

The present data together with earlier findings that the majority of PBC patient autoantibodies bind to epitopes defined by the PBC‐MoAb, makes this antibody a valuable tool for characterizing the major PBC‐associated epitope on PDH‐E2 and localizing this epitope in liver tissue.
Mechanisms of Generation of Antibody Diversity as a Cause for Natural Selection of Homoiothermal Animals in the Process of Evolution
Authors: ZAV'YALOV V. P. , TISHCHENKO V. M.
Published: June 2006

The temperature‐dependence of domain interactions in the Fab, Fc. Fb and Fv fragments from humar myeloma immunoglobulins (IgG) samples was investigated by scanning microcalorimetry, NMR and difference spectroscopy. The fragments were found lo be very sensitive to temperature changes. Lowering the temperature below the physiological value 37°C) considerably decreases the energy of interaction of the variable VH and VL domains, resulting at limes in their dissociation. Since the association energies of VH and VL pairs can be affected by the result of somatic recombination and mutation events affecting antibody genes, immunoglobulins can fortuitously acquire The properties of cryoglobulins or cold autoantibodies and induce severe pathological states. It is postulated that this property of immunoglobulins, and by extension, of T‐cell antigen receptors might have been one of the causes for the possible natural selection of homoiothermal animals. In these, the high conformational sensitivity of immunoglobulins to temperature change may be important in the mechanisms of induction of secondary functions In immune responses.
Characterization of High Molecular Weight Amyloid A Proteins
Authors: PRELLI F. , PRAS M. , SHTRASBURG S. , FRANGIONE B.
Published: June 2006

Human amyloid A protein (AA) is usually composed of the NH2‐terminal 76 amino acid residue of serum amyloid A protein (SAA), although lower and higher molecular weight fragments have been reported. We studied the primary structure of six AA proteins with molecular weights of 11 kDA‐15kDA. as determined by SDS‐PAGE. Automated Edman degradation of the intact purified proteins and sequence analysis of enzymatic peptides revealed that the AA proteins were composed of only 74 to 87 residues. Moreover, fragments of apolipoprotein E or histones 2a, 3 and 4 were associated with these AA molecules. Thus, AA heterogeneity may reflect diverse processing of the SAA precursor and a very close association with other proteins.
Production of Heterohybridomas Secreting Autoreactive and Polyreactive Human Monoclonal Antibodies
Authors: SANCHEZ B. , YELAMOS J. , MELERO J. , MAGARIÑO R. , GONZALEZ M. F. , GARCIA R. , RIOS C. , NUÑEZ‐ROLDAN A.
Published: June 2006

Peripheral blood lymphocytes from two polytransfused renal dialysis patients were transformed by Epstein Barr virus, fused to heteromyeloma and cloned. Eight human monoclonal antibodies from the resulting clones were tested for their binding to variety of antigens by ELISA, indirect immunofluorescence and immunoblotting. Antigens tested included B‐cell lines, T and B lymphocytes, red blood cells, chronic lymphocytic leukaemic B cells, IgG, ssDNA, dsDNA. histones, nucleoprotamine, sperm nuclei, thymus and spleen extracts. MOLT4 cell lysatcs, affinity purified autoantigens, tetanus toxoid, bacterial lipopolysaccharide, insulin, and tissue section screen. These human monoclonal antibodies reacted with more than one antigen to varying degrees and were autoreactive and pulyreactive. One of these heterohybridoma cell lines exhibited cytoplasmic staining with an anti‐CD5 monoclonal. Our findings support the concept that in adult individuals subset of B cells produce heterogeneous IgM antibodies which can bind to variety of different autoantigens and also to foreign antigens. These monoclonals were different from the autoantibodies usually seen in renal dialysis patients in the sense that they were not lymphocytotoxic.
Clustering of IgA‐Producing Immunocytes Related to HLA‐DR‐Positive Ducts in Normal and Inflamed Salivary Glands
Authors: THRANE P. S. , SOLLID L. M. , HAANES H. R. , BRANDTZAEG P.
Published: June 2006

Ig‐producing immunocytes and epithelial expression of secretory component (SC) and HLA‐DR were evaluated by two‐colour immunofluorescence staining in 10 normal and 20 inflamed salivary glands; the latter included specimens from 10 patients with obstructive sialadenitis and 10 with Sjögren's syndrome (SS), Epithelium adjacent to T‐cell infiltrates showed extensive co‐expression of SC and HLA‐DR, suggesting that leucocyte‐derived cytokines were responsible for this concurrent up‐regulation. Clusters (>2 positive cells) of IgA‐producing cells were spatially related to DR‐positive ducts. The possibility is discussed that DR‐expressing epithelium contributes to local terminal differentiation of IgA‐producing plasma cells. A cytokine‐mediated up‐regulation of SC that simultaneously increases the transport capacity for polymeric IgA would constitute an efficient enhancement of secretory immunity in diseased glandular tissue.
Can B‐Cell Signalling be Understood?
Authors: Sinclair N. R. StC.
Published: June 2006

Hie therefore, Robin, overcast the night; The starry welkin cover thou anon With drooping fog as black as Acheron; And lead these testy rivals so astray, As one come not within another's way.

William Shakespeare, A Midsummer Night's Dream Act III, Scene II
In Vitro Activation of Human T Lymphocytes byHaemophilus influenzaeTypebCapsular Polysaccharides
Authors: PEETERS C. C. A. M. , TENBERGEN‐MEEKES A.‐M. , HEIJNEN C. J. , POOLMAN J. T. , ZEGERS B. J. M. , RIJKERS G. T.
Published: June 2006

Polyribosilribitolphosphate (PRP), the capsular polysaccharide fromHaemophilus influenzaetypeb, is a T‐cell‐independent type 2 antigen. In vitro culture of adult peripheral blood T cells with 15 μg/ml PRP leads to induction of interleukin‐2 receptor (IL‐2R) expression on up to 10% of T cells. These cells are CD4+and carry the αβ T‐cell receptor. PRP, at concentrations above 1–5 μg/ml, can also induce in vitro proliferation of both adult and neonatal T cells. We conclude that PRP acts as a human T‐cell mitogen.

The in vitro proliferative response as well as IL‐2R expression was studied in T cells derived from adults after vaccination with native PRP, with PRP conjugated to a carrier protein, or with diphtheria toxoid. Vaccination with conjugated PRP decreased the doses of PRP required for in vitro induction of IL‐2R expression and T‐cell proliferation. This indicates that vaccination with PRP conjugated to a carrier protein improves the in vitro T‐cell response to PRP activation.
Analysis of Monoclonal Rheumatoid Factors obtained from the B‐Cell Repertoire in Rheumatoid Arthritis
Authors: ABDERRAZIK M. , MOYNIER M. , JEFFFRIS R , MAGEED R. A. K. , COMBE B. , SANY J. , BROCHIER J.
Published: June 2006

We have sought to determine whether rheumatoid factors (RF) produced in rheumatoid arthritis (RA) were different from physiological RF produced in normal, healthy adults. RF‐secreting clones were established following Epstein Barr virus (EBV) stimulation of peripheral blood lymphocytes. Ten RF‐secreting clones were established from seven RA patients and 16 from six healthy controls. All monoclonal RF (MRF), except two in each group, were monoreactive and ten of these were shown to have low to medium affinity for IgG, Fc, irrespective of their origin. A majority (74%) of the MRF bound to protein A, indicating that genes of the VHIII family were preferentially used for synthesizing these autoantibodies. The expression of cross‐reactive idiotypes (CRI) by the MRF did not allow distinction between those derived from RA patients and controls. The VHI‐associated CRI G8 and VHIII‐associated CR1 D12 were expressed at low frequency in both panels of RF. These CRI have been shown to be expressed at high frequency in RF paraproteins. However, the idRQ idiotype was expressed within both panels of RF. A possible distinction between polyreactive and monoreactive MRF appeared to be light chain usage since all (four) polyreactive RF used λ chains while the normal κ/λ ratio was observed for monoreactive RF. The frequency of EBV‐activated cells secreting IgM bearing CRI or secreting RF was determined and showed that CRI expression occurred with a higher frequency than did RF, suggesting a dissociation between CRI expression and RF activity.
Characterization of T‐Cell Receptor αβ Repertoire in Synovial Tissue from Different Temporal Phases of Rheumatoid Arthritis
Authors: BUCHT A. , OKSENBERG J. R. , LINDBLAD S. , GRÖNBERG A. , STEINMAN L. , KLARESKOG L.
Published: June 2006

With the aim of investigating the distribution of T cells expressing different T‐cell receptors (TCR) in the inflamed synovial tissue of rheumatoid arthritis patients, we have used the polymerase chain reaction to amplify TCR Vα and Vβ transcripts from synovial biopsies obtained by arthroscopy from patients with arthritis of variable duration. From each of nine patients a single biopsy was taken. Southern hybridization analysis of amplified products revealed extensive heterogeneity of TCR Vβ in most patients. On the other hand, restriction in Vα gene expression was seen in several patients. A highly restricted Vα repertoire was observed in all cases with arthritis of short duration. In addition, two of three samples of short duration yielded a more limited number of Vβ transcripts than the others. No conformity was, however, seen in usage of individual Vα and Vβ transcripts among the investigated patients. The present data thus demonstrate variability in synovial TCR expression between rheumatoid arthritis patients, but they also indicate a development towards greater diversity with increasing disease duration, implicating the necessity for careful choice of cases, preferentially selecting for early stages of disease, when further analysing rheumatoid synovial T cells for TCR usage as well as for antigen specificity.
Interleukin‐4 Down‐Regulates Sendai Virus‐Induced Production of Interferon‐α and ‐β in Human Peripheral Blood Monocytes In Vitro
Authors: GOBL A. E. , ALM G. V.
Published: June 2006

Recombinant interleukin‐4 (rIL‐4) caused a 65–70% reduction of the interferon‐α (IFN‐α) and ‐β production induced by Sendai virus (SV) in human peripheral blood monocytes in vitro. Significant inhibition was seen at concentrations as low as 0.1 ng/ml. The rIL‐4 also reduced levels of IFN‐α and ‐β mRNA by 60% and 67%, respectively, as well as the frequency of IFN‐α and ‐β mRNA‐containing cells by 65% and 54%, respectively. The frequency of IFN‐α/β mRNA‐containing cells was inhibited by rIL‐4 throughout the whole course of induction by SV. IL‐4 caused a shift of the grain count distribution towards less heavily labelled cells, suggesting an inhibitory effect of rIL‐4 on most IFN‐α/β mRNA‐containing cells. Antibodies to rIL‐4 did not influence the normal IFN‐α/β response induced by SV, but abolished the inhibitory effect of the rIL‐4.

When rIL‐4 was added to cells 4 h after start of stimulation by SV, at which time much mRNA has accumulated but little IFN‐α/β has been secreted, no inhibition of the IFN‐α/β production by rIL‐4 was seen. IFN‐γ had only a minor reversing effect on the rIL‐4 inhibition, but if cells were precultivated in medium with or without IFN‐γ for 6 h before SV induction, rIL‐4 paradoxically enhanced the IFN‐α/β response.

Our results suggest that rIL‐4 inhibits an early step of IFN‐α/β induction in monocytes, at the level either of transcription of IFN‐α/β genes or of the processing or stability of mRNA. The IL‐4 effects may however depend on the state of activation of the monocytes.
Interferon‐α but not ‐β Genes Require De Novo Protein Synthesis for Efficient Expression in Human Monocytes
Authors: GOBL A. E. , CEDERBLAD B. , SANDBERG K. , ALM G. V.
Published: June 2006

Monocytes produce interferon‐α (IFN‐α) and ‐β when human peripheral blood mononuclear cells (PBMCs) are stimulated in vitro by Sendai virus (SV). We found that about 70% of the IFN‐producing cells (IPCs) expressed both IFN‐α and ‐β mRNA; She rest expressed only IFN‐β mRNA. In the presence of the protein synthesis inhibitor cycloheximide(CHX), the frequency of IFN‐αmRNA containing cells, measured after 6 h, was decreased by 85–90%. Results of nuclear run‐on transcription assays showed that CHX inhibited IFN‐α gene expression. The frequency of IFN‐β mRNA‐containing cells was not reduced by CHX. Actually, a threefold increase was observed at the lower CHX concentrations. Studies on the kinetics of IFN‐α/β mRNA induction showed that CHX accelerated the appearance of IFN‐β mRNA‐containing cells, increased IFN‐β mRNA levels, and delayed the normally occurring post‐inductional decrease of IFN‐β mRNA. Unexpectedly, an initially normal or even accelerated IFN‐α mRNA response was seen in the presence of CHX during the first 3–4 h after SV stimulation. This occurred in a small proportion of the potential IPCs. However, CHX prevented the subsequent marked increase of IFN‐α mRNA levels. Preincubation of PBMCs for 6h in conditioned medium (CM) containing IFN and other cytokines prevented the CHX‐mediated inhibition of IFN‐α mRNA. Without preincubation this was not seen. The preincubation in CM caused an accelerated appearance of IFN‐α mRNA, resembling that of IFN‐β mRNA. The results suggest (that IFN‐α and ‐β genes are differentially regulated in the same monocytes, the former requiring de novo synthesis of intracellular protein(s) for efficient expression.
FcγR‐Dependent Regulation of the Biosynthesis of Complement C3 by Murine Macrophages: the Modulatory Effect of IL‐6
Authors: BAJTAY Zs. , FALUS A , ERDEI A , GERGELY J.
Published: June 2006

The effect of murine IgG isotypes on the gene expression and secretion of the third component of complement (C3) has been studied using the monocytoid cell line P388DI and oil‐elicited mouse peritoneal macrophages. It is demonstrated that the binding of IgG2a and IgG2b but not IgG1 and IgG3 augments the biosynthesis of C3 both in the presence and in the absence of the phorbol ester. phorbol myristate acetate in the case of both cell types. The multifunctional cytokine inlerleukin‐6 (IL‐6) alone reveals no effect on the gene expression of C3, but increases the effectiveness of mouse IgG2a and IgG2b. Confirming the role of FcγRII, a strong up‐regulation of C3 gene expression and C3 secretion was found when macrophages were cultured with the F(ab')2; fragment of the FcγRII‐specific monoclonal antibody 2.4G2.
Effects of Stress on the Growth of a Fibrosarcoma in nu/nu and Conventional Mice
Authors: AARSTAD H. J. , SELJELID R.
Published: June 2006

BALB/c mice subjected to water stress twice daily, between 3 and 7 min at each exposure 3 days before, 5 days after, or.1 days before and 5 days after intradermal Meth A tumour transplantation, showed an increase of tumour size and tumour growth rate compared with that of home cage control (HCC) mice. However, BALB/c mice subjected to water stress 3 days before and 6 days after tumour transplantation showed a decrease of tumour size and tumour growth rate compared with HCC mice. BALB/c mice subjected to 2 mm of handling stress daily on days 1 through 5, but not BALB/c mice subjected to handling stress on days I through 7 or on days 6 through 7 of tumour growth, showed an increase in tumour size relative to HCC mice. On the other hand. BALB/cBy mice handling stressed on days I through 7, but not BALB/cBy mice handling strewed on days I through 5 or days 6 through 7 after tumour transplantation, showed a decreased tumour size compared with HCC mice. No effect of stress could he observed when nu/nu BALB/c mice were subjected to water stress 3 days before and 6 days after tumour transplantation. However. BALB/c nu/nu mice pre‐injected intraperitoneally with spleen cells from conventional mice had a decreased tumour size und growth rale of the tumour when stressed 3 days before and 6 days after tumour transplantation.
The Complete Amino Acid Sequence of Bovine Serum Amyloid Protein A (SAA) and of Subspecies of the Tissue‐Deposited Amyloid Fibril Protein A
Authors: ROSSEVATN K. , ANDRESEN P. K. , SLETTEN K. , HUSEBEKK A. , HUSBY G. , NORDSTOGA K. , JOHNSON K. H. , WESTERMARK G. T. , WESTERMARK P.
Published: June 2006

Bovine serum amyloid A (SAA) was isolated from the acute phase high density lipoprotein (HDL) fraction of a cow suffering from acute mastitis. The elucidated primary structure revealed a protein consisting of 112 amino acid residues. Compared with SAA proteins from other species, the bovine protein was shown to have an insertion of nine amino acid residues between positions 69 and 70. No microheterogeneity could be observed in the protein.

Amyloid fibrils extracted from the kidneys were found to contain at least three subspecies of protein AA, consisting of 68, 81 and about 110 amino acid residues. The amino acid sequences established for the protein AA subspecies revealed no microheterogeneity, and were identical to that elucidated for protein SAA.
Interferon‐Gamma Production by African Swine Fever Virus‐Specific Lymphocytes
Authors: REVILLA Y. , PENA L. , VIÑUELA E.
Published: June 2006

Peripheral blood mononuclear cells (PBMC) from inbred pigs that were immunized with autologous macrophages infected with the African swine fever (ASF) virus BA71V, a non‐virulent virus isolate, proliferated and produced interleukin‐2 in response to homologous and heterologous isolates of the ASF virus. They produced, however, interferon (IFN) only when challenged in vitro with homologous or attenuated isolates of the ASF virus, but not with heterologous or virulent isolates. The IFN was pH 2 labile and was neutralized by specific serum to porcine recombinant IFN gamma.
Frequency of Chicken CD4+ and CD8+ Cells. Genetic Control and Effect of Rous Sarcoma Virus Infection
Authors: HÁLA K. , BÖCK G. , SGONC R. , SCHULMANNOVÁ J. , TEMPELIS C. H. , VAINIO O. , KEMMLER G.
Published: June 2006

In chickens from congenic inbred lines CB and CC that differ only in the major histocomputibilily complex (MHC), we observed significantly different percentages of CD4+ and CD8+ cells in peripheral blood lymphocytes (PBL) and spleen. Positive cells were detected by indirect immunofluorescence test as analysed by flow cytometry. In both PBL and spleen cell suspensions, the number of CD4+ cells was significantly higher in CB than in CC chickens, whereas in CC birds there was a higher percentage of CD8+ cells than in CB. These statistically significant differences were under the MHC control. We found no statistically significant influence of regressions or progression of Rous sarcoma virus‐induced tumours on the percentage of peripheral T cells and on the interleukin‐2 production in vitro.
TNF Impairs In Vivo and In Vitro Natural Killer (NK) Susceptibility of B16 Melanoma Cells
Authors: PALMIERI G. , MORRONE S. , LOLLINI P.‐L. , DE GIOVANNI C. , NICOLETTI G. , NANNI P. , FRATI L. , SANTONI A.
Published: June 2006

Tumour necrosis factor α (TNF) is a multipotent cytokine which affects many biological properties of both normal and neoplastic cells. Here we show that treatment with TNF reduces B16‐A melanoma cell susceptibility to normal and in vivo‐ and in vitro‐activated NK cell‐mediated killing. This resistance is associated with an enhancement of B16‐A metastatic potential in normal syngeneic mice, but not in anti‐asialo GM1‐treated animals, further supporting the NK dependence of TNF‐induced enhancement of metastatic ability. A significant increase of MHC class I expression on B16‐A murine melanoma cells is observed after TNF treatment. In all these effects TNF interacts positively with interferon γ (IFN γ). Taken together, these results indicate that TNF treatment negatively affects the susceptibility of B16‐A murine melanoma to NK effectors in vivo and in vitro. This decreased susceptibility may be related, at least in part, to enhanced expression of MHC class I antigens on tumour cells.
Comparative Analysis of Syngeneic and Allogeneic Mixed Lymphocyte Reaction of ‘Naturally’ Activated and Resting T Cells
Authors: FUCS R. , DE JESUS S. A. G. , DA NOBREGA A. F.
Published: June 2006

‘Naturally’ activated (NA) or resting T lymphocytes obtained from the spleen of normal BALB/c mice were compared in their capacity to mount a syngeneic mixed lymphocyte reaction (SMLR). Both T‐cell subsets were able to proliferate and secrete IL‐3/GM‐CSF in SMLR cultures. IL‐2 was present in ‘resting’ T‐cell SMLR supernatants, and barely detectable in NA T‐cell SMLR supernatants. Both NA and ‘resting’ T‐cell SMLRs were inhibited with anti‐class II, anti‐CD4, or anti‐IL‐2R MoAbs. NA T cells exhibited a background proliferate and secretory activity in the absence of syngeneic accessory cells. This autonomous activity was susceptible to anti‐CD4, but poorly inhibited with anti‐class II MoAbs. Both NA and ‘resting’ T lymphocytes displayed strong responsiveness to allogeneic stimuli. The analysis of the relative frequency of proliferating cells in the SMLR (BALB/c), or allo‐MLR (B10, B10.A, B10.D2) from NA or ‘resting’ T cells indicated an enrichment for syngeneic reactivity among NA T lymphocytes.

The meaning of these results for NA T‐cell function and repertoire is discussed.
Ia Expression in Keratinocytes Following Ultraviolet Radiation
Authors: GILHAR A. , ETZIONI A. , EIDELMAN S.
Published: June 2006

Injections of murine gamma interferon (IFN‐γ) into BALB/c nude mice induced Ia expression by keratinocytes The aim of the present study was to use this murine model to determine the effect of ultraviolet radiation (UV) or cyclosporine A (CyA) on Ia expression by keratinocytes. Two sets of experiments were performed. In the first, mice were injected intraperitoneally with IFN‐γ for 6 days. The mice were divided into three groups. One group, with one ear protected by electrical tape, was exposed to UVB radiation for 15 days starting 4 days before the injection. The second group received subcutaneous injections of CyA simultaneously with the IFN‐γ and during the 10 days following the IFN‐γ injection. The third group received only IFN‐γ injections. Fifteen days alter the IFN‐γ injection all mice were killed and evaluations of la positive cells were performed. In the second set of experiments the nude mice were treated with CyA or UVB only 10 days after the last IFN‐γ injections. In both experiments UVB inhibited and down‐regulated la expression by keratinocytes. This effect on keratinocytes was not observed in the protected ears. Thus it appears that the effect of UVB on keratinocytes is local and not systemic. CyA failed to inhibit or down‐regulate Ia expression. This study may shed some light on understanding the mechanism effect of UV radiation in a variety of skin diseases.
Effects of Immunomodulators on Ecto‐5′‐Nucleotidase Activity on Blood Mononuclear Cells In Vitro
Authors: CHRISTENSEN L. D. , ANDERSEN V. , NYGAARD P. , BENDTZEN K.
Published: June 2006

In this study the effects of immunomodulators on the ecto‐5′‐nucleotidase (ecto‐5′‐NT) activity on blood mononuclear cells (BMC) were examined in vitro. Interleukin‐4 (IL‐4) and interferon‐γ (IFN‐γ) decreased the level of ecto‐5′‐NT activity on BMC whereas prostaglandin E2 (PGE2) increased the ecto‐5′‐NT level. All three immunomodulators influenced the ecto‐5′‐NT activity of isolated monocytes whereas only IL‐4 and PGE2 had an effect on the enzyme level on isolated lymphocytes. The effect was dependent upon protein synthesis. The effect was dose dependent: IL‐4 was effective at concentrations down to 0.5 U/ml, IFN‐γ down to 40 U/ml and PGE2 at nanomolar concentrations. These data indicate that immunomodulators may also take part in the regulation of ecto‐5′‐NT activity on BMC in vivo.

BMC from 7 patients with different immunodeficiency syndromes showed decreased ecto‐5‐NT activity on freshly isolated cells. However, following culture ecto‐5′‐NT activity was increased above the level found on freshly isolated BMC from healthy persons. On BMC from 3 patients with hypogammaglobulinaemia, the effect of IL‐4 on the level of ecto‐5′‐NT activity was identical to that found on BMC from healthy donors, whereas PGE2 increased ecto‐5′‐NT activity on BMC from only 1 of the 3 patients investigated. The decreased ecto‐5′‐NT activity of BMC from patients with immunodeficiency may thus be due to a defective regulation of ecto‐5′‐NT activity
T‐Cell‐Dependent Production of IgG by Human Cord Blood B Cells in Reconstituted SCID Mice
Authors: UENO Y. , ICHIHARA T. , HASUI M. , MARUYAMA H. , MIYAWAKI T. , TANIGUCHI N. , KOMIYAMA A.
Published: June 2006

Reconstitution of severe combined immunodeficient (SCID) mice with human lymphocytes has recently allowed the elucidation of abnormalities of immune responses in various immunological disorders. In the present study, mononuclear cells (MNC) from neonatal cord blood and adult peripheral blood were intraperitoneally injected into SCID mice to examine induction of human Ig in respective mice recipients. Human IgG was consistently detected in the serum of SCID transferred with adult MNC, but only a few SCID recipients of cord blood MNC showed detectable but low levels of IgG in the serum. The combination experiments of isolated B and T cells disclosed that some interactions between B and T cells might be necessary for IgG production in transferred SCID mice. Notably, transfer of cord blood B cells with adult but not cord blood T cells resulted in efficient induction of IgG, associated with a change in subclass distribution. The results suggest that inability of neonatal B cells to produce IgG can be overcome by transfer with adult mature T cells into SCID mice.
Immune Responses During Human Schistosomiasis Mansoni. XVIII. Immunologic Status of Pregnant Women and Their Neonates
Authors: NOVATO‐SILVA E. , GAZZINELLI G. , COLLEY D. G.
Published: June 2006

The immune status of peripheral blood mononuclear cells (PBMC)and sera of pregnant woman infected with the helminth Schistosoma mansoni was studied during pregnancy, and the cord blood mononuclear cells (CBMC) and sera from their offspring were studied at parturition. PBMC pokeweed mitogen‐induced responses were maintained in gravid women, but the responses to both schistosome and non‐schistosome antigenic preparations declined progressively during pregnancy. Schistosomal antigens simulated proliferative responses by the CBMC of many neonates born of infected mothers, but not those of uninfected mothers. These specific responses by CBMC of only neonates born of infected mothers are indicative of in utero, cell‐mediated sensitization of the neonates, which could be due either to circulating schistosomal antigens or to anti‐idiotypic antibodies which cross the placenta during gestation. Sera from infected mothers and the cord blood sera from their babies showed the same levels of specific IgG anti‐schistosomal activity. Anti‐schistosomal IgM levels were maintained during pregnancy to some antigens and not to others, while such antibodies were rarely found in cord blood sera, and then only at very low levels.