bluuebunny/crossref_metadata_2025 huggingface dataset:
Phloem‐derived γ‐aminobutyric acid (GABA) is involved in upregulating nodule N2 fixation efficiency in the model legume Medicago truncatula
Authors: SULIEMAN SAAD , SCHULZE JOACHIM
Published: August 2010

Nitrogen fixation in legumes is downregulated through a whole plant N feedback mechanism, for example, when under stress. This mechanism is probably triggered by the impact of shoot‐borne, phloem‐delivered compounds. However, little is known about any whole‐plant mechanism that might upregulate nitrogen fixation, for example, under N deficiency. We induced emerging N‐deficiency through partial excision of nodules from Medicago truncatula plants. Subsequently, the activity and composition of the remaining nodules and shifts in concentration of free amides/amino acids in the phloem were monitored. Furthermore, we mimicked these shifts through artificial feeding of γ‐aminobutyric acid (GABA) into the phloem of undisturbed plants. As a result of increased specific activity of nodules, N2 fixation per plant recovered almost completely 4–5 d after excision. The concentration of amino acids, sugars and organic acids increased strongly in the upregulated nodules. A concomitant analysis of the phloem revealed a significant increase in GABA concentration. Comparable with the effect of nodule excision, artificial GABA feeding into the phloem resulted in an increased activity and higher concentration of amino acids and organic acids in nodules. It is concluded that GABA might be involved in upregulating nodule activity, possibly because of its constituting part of a putative amino acid cycle between bacteroids and the cytosol.
Use of gaseous 13NH3 administered to intact leaves of Nicotiana tabacum to study changes in nitrogen utilization during defence induction
Authors: HANIK NILS , GÓMEZ SARA , SCHUELLER MICHAEL , ORIANS COLIN M. , FERRIERI RICHARD A.
Published: August 2010

Nitrogen‐13 (t1/2 9.97 m), a radioactive isotope of nitrogen, offers unique opportunities to explore plant nitrogen utilization over short time periods. Here we describe a method for administering 13N as gaseous 13NH3 to intact leaves of Nicotiana tabacum L. (cv Samsun), and measuring the labelled amino acids using radio high‐performance liquid chromatography (HPLC) on tissue extract. We used this method to study the effects of defence induction on plant nitrogen utilization by applying treatments of methyl jasmonate (MeJA), a potent defence elicitor. MeJA caused a significant increase relative to controls in key [13N]amino acids, including serine, glycine and alanine by 4 h post‐treatment, yet had no effect on 13NH3 incorporation, a process that is primarily under the control of the glutamine synthatase/glutamate synthase pathway (GS/GOGAT) in cellular photorespiration. We suggest that the reconfiguration of nitrogen metabolism may reflect induction of non‐photorespiratory sources of nitrogen to better serve the plant's defences.
Affinity Chromatography of β2‐Microglobulin from Human Lymphocytes on Concanavalin‐A‐Sepharose
Authors: PLESNER T.
Published: August 2010

β2 was extracted from human lymphocytes with nononic detergent and separated by affinity chromatography on concanavalin‐A‐Sepharose. The retarded part β2‐micorglobulin is assumed to be associated with HLA antigens. Using a radioimmunoassay for β2‐microglobulin, the average number of presumably free β2‐microglobulin molecules and of presumably HLA‐associated β2‐microglobulin molecules per lymphocyte was estimated to be 3.8 × 105 and 1.4 × 105, respectively.
Interactions Between Ricinus Agglutinin and Human IgA
Authors: SALTVEDT E.
Published: August 2010

By affinity chromatography on Sepharose columns containing insolubilized ricinus agglutinin, all immunoglobulins of the IgA2 subclass and the polymers of IgA1 were retained and could be eluted with lactose. Monomeric IgA1 showed a more heterogeneous reaction with ricinus agglutinin. The major and could part was unretarded, whereas some of it the column and could be eluted with lactose
Evidence for the Release of Mitogenic Factors by Both T and B Lymphocytes in the Human
Authors: BLOMGREN H.
Published: August 2010

Lymphocytes incubated with antigens or nonspecific stimulatory agents may release factors that induce DNA synthesis in other lymphocytes. The aim of this investigation was to determine whether such mitogenic factors (MF) are produced by both T and B cells in the human Both T‐ or B‐cell‐enriched cell preparation from peripheral blood were found to release MF during or after incubation with phytohemagglutinin or pokeweed mitogen Concanavalin A covalently bound to Sepharose triggered B‐cell preparation to higher MF release than either T cells or unfractionated lymphocyte suspensions. These results strongly indicate that both T and B cell in the human are able to produce MF.
Arming of Lymphoid Cells by IgG Antibodies Treated with Protein A from Staphylococcus aureus
Authors: SULICA A. , LAKY M. , GHERMAN M. , GHETIE V. , SJÖQUIST J.
Published: August 2010

Mouse spleen lymphocytes treated with rabbit IgG anti‐sheep erythrocytes (SRBC) complexed with protein A of Stapbylococcus aureus (SpA) form rosettes with SRBC. The attachment of SRBC to lymphocytes was due to the binding of the SpA‐IgG antibody complex to the surface of the lymphocytes and was thus considered ‘arming’ of the cells. Normal mouse spleen cells ‘armed’ with SpA‐rabbit IgG anti‐chicken erythrocytes (CRBC) kill specifically 51Cr‐labeled CRBC ‘in vitro’ in the absence of free antibodies. The killing by these ‘armed’ cells is an effect of the cell‐bound SpA‐IgG antibody complex. Both the SRBC rosette formation and the cell‐mediated CRBC killing was dependent on the concentration of the SpA‐IgG antibody complexes used for ‘arming’ the cell. A 100‐fold increase in rosette formation or in killing of target cells was recorded for lymphocytes treated with SpA‐IgG antibody complexes in comparison with cell treated with noncomplexed IgG antibodies. The specific binding of SpA‐IgG antibody complexes to the Fc receptors of mouse spleen cells was demonstrated by inhibition studies. More than 60% inhibition of the rosette formation and in the killing of target cells was shown for cell treated With normal rabbit IgG or its Fc fragment before addition of the SpA‐IgG antibody complex.
A Method for the Detection and Quantitation of Fc Receptor Sites on Cells
Authors: GHETIE V. , MEDESAN C. , SJÖQUIST J.
Published: August 2010

A new rosette technique for identification of Fc‐receptor‐bearing cells is based on the ability of sheep erythrocyte coated with protein A of Staphylococcus aureus (ES) to form rosettes with cell treated with monomeric IgG or aggregated IgG. The IgG is attached to lymphocytes through its Fc region and to a trypsin‐resistant but pronase sensitive receptor (considered an Fc receptor) The ES rosette technique facilitates studies of the interaction of IgG with Fc receptor sites; the binding of any IgG preparation reacting with protein A of Staphylococcus aureus (SpA) can be studied by the technique. Inhibition of rosette formation by SpA was used for quantitation of IgG fixed to the cell surface (that is, the number of Fc reception/cell). The sensitivity of the method permits quantitation of less than 105 IgG molecules bound to the Fc receptors. The relative affinity constant between Fc receptors and IgG ligands was estimated by plotting the percentage of ES rosettes as a function of IgG concentration and calculating the reciprocal of the IgG concentration giving half the maximal number of ES rosettes.
Anti‐Parental‐Strain Lymphocyte Responsiveness of F1 Hybrid‐Strain Lymphocytes
Authors: McCULLAGH P.
Published: August 2010

The interactions between immunocompetent parental‐strain tells and the F1 hybrid rat into which these cells have been injected were studied. This involved examination of the behavior of both parental‐ and host‐strain cells. Host lymphocytes reactive against parental‐strain lymphocytes appeared in the thoracic duct lymph within 12 h of the intravenous injection of parental thoracic duct lymphocytes or thymus cells. The activity of these F1 hybrid‐strain lymphocytes was directed preferentially against parental‐strain lymphocytes with anti‐F1 hybrid potential. This subpopulation of parental cells was absent from the thoracic duct but was well represented in the spleen, this enrichment being masked by a reversible F1 hybrid anti‐parental‐strain lymphocyte response. If the anti‐parental‐strain lymphocyte activity of F1 hybrid cells was interrupted by the destruction of host‐strain cells with antiserum, parental‐strain lymphocytes in the tissues of F1 hybrid rats could be reactivated. By this procedure it was possible to passage graft‐versus‐host reactions initiated by parental‐strain lymphocytes through several F1 hybrid rats, thereby implying that anti‐parental lymphocyte activity is of importance in limiting the serial passage of these reactions. Some of the anti‐parental lymphocyte activity observed on the part of F1 hybrid‐strain cells was probably mediated by anti‐idiotypic antibodies, but other phenomena, especially selective migration of both donor and host lymphocyte subpopulations, are of major importance in the interaction between parental‐strain lymphocytes and an F1 hybrid host.
Certain Polyclonal Antinuclear Antibodies Cross‐React with the Surface Membrane of Human Lymphocytes and Granulocytes
Authors: REKVIG O. P. , HANNESTAD K.
Published: August 2010

Five out of 24 human sera with antinuclear antibody (ANA) titers of 1250 or more contained ANA that bound in vitro to normal viable human mononuclear blood cells and granulocytes, but not to erythrocytes. The antibodies can be eluted off from the cell membranes and shown to possess ANA activity. Anti‐native DNA antibodies and lupus erythematosus factor were not recovered in eluates, indicating that they did not react with the cells. The cells absorbed 75%–87% of the ANA activity from three sera. ANA reacted with both T‐lymphocyte‐depleted and ‐enriched mononuclear cells. No or minimal amounts of ANA bound to mouse spleen cells in suspension; in contrast, the ANA eluted from human cells reacted withnucleiof smeared mouse spleen cells. The cross‐reacting antibodies were predominantly IgG that bound well at 37°C, and F(ab')2 fragments carried both activities. The ANA‐binding plasma membrane antigen was resistant to trypsin and RNAse but was completely inactivated by glutaraldehyde. The data indicate that human leukocyte plasma membranes and cell nuclei from many species contain a cross‐specific antigen. Alternatively, the antigen may be produced in the nucleus and somehow attach to the plasma membrane.
Human Leukocyte Migration Inhibitory Factor (LIF)
Authors: BENDTZEN K.
Published: August 2010

Previous findings suggesting an esterase and protease nature of human leukocyte migration inhibitory factor (LIF) were extended by testing the ability of different protease and esterase inhibitors to reduce LIF activity. The serine‐specific inhibitors phenylmethylsulfonyl fluoride (PMSF) and physostigmine (eserine) markedly reduced LIF activity, whereas the histidine‐specific inhibitors N‐tosyl‐L‐lysine chloromethyl ketone (TLCK) and L‐tosylamide‐2‐phenylethyl chloromethyl ketone (TPCK) were inactive. That LIF might act as an esterase and a protease was further strengthened by the ability of pralidoxime methansulfonate (2‐PAM) to reestablish LIF activity of supernatants previously inactivated by PMSF. Furthermore, the arginine amides N‐benzoyl‐L‐arginine‐p‐nitroanilide (BApNA) and N‐benzoyl‐L‐phenylalanyl‐L‐valyl‐L‐arginine‐p‐nitroanilide (BPVApNA) were shown to satisfy the substrate specificities of the putative LIF enzyme. Indeed, BPVApNA seemed to possess a particularly strong affinity for LIF, indicating its potential role in an enzymatic LIF assay.
Complement‐Dependent Cytotoxicity Against Hepatoma Cells Mediated by IgM Antibodies in Serum from Tumor‐Bearing Rats‐
Authors: LANDO P. , BLOMBERG F. , RAFTELL M. , BERZINS K. , PERLMANN P.
Published: August 2010

Complement‐dependent cytotoxic antibodies in syngeneic serum from rats carrying transplanted aminoazo dye‐induced hepatomas (D23 and D33) and from rats immunized with irradiated tumor cell or homogenates were studied by a shortterm 51Cr release assay. The tumor‐bearer sera (TBS) were subjected to chromatography on unsolubilized protein A and Sepharose 4B. The cytolytic activity of D23 TBS was recovered in the IgM molecular weight region, whereas no such activity was obtained in the IgG fraction. As judged from immunodiffusion experiments, the IgM molecular weight fraction did not contain any aggregated or complexed IgG. Moreover, in immunofluorescence tests against viable hepatoma cells, using a specific anti‐rat IgG conjugate, the TBS were negative. Cross‐testing of D23 and D33 TBS against the two hepatomas and cross‐absorption of the sera with tumor plasma membranes revealed no tumor specificity in these cytotoxicity reactions. Furthermore, neither fetal nor early postnatal liver cells could absorb out the activity. Absorptions with adult liver plasma membranes, however, abrogated the cytotoxic activity, and even more effective in this respect were homogenates from kidney and small intestine, thus indicating that the cytotoxic antibodies are directed against ‘normal’ adult antigen(s) present also in tissues other than liver.
A Reference System in Crossed Radioimmunoelectrophoresis
Authors: AUKRUST L. , AAS K.
Published: August 2010

A reference system for crossed radioimmunoelectrophoresis (CRIE) has been developed. The system takes into account the variable quality of the 125I‐labeled anti‐IgE and permits a direct comparison of the results from one experiment to another. Simultaneously with the preparation of the CRIE slides, a standard series of small paper discs are prepared containing varying, defined amounts of 125I‐labeled anti‐IgE. These paper discs are laid out for autoradiography together with the CRIE slides. After short periods of exposure, only paper discs with large amounts of isotope are visible on the X‐ray films. Longer exposure times permit paper discs with smaller amounts of isotope to be visible. Each precipitate is assigned to a CRIE class depending on the number of paper discs visible when the precipitate is detected for the first time.
Enhanced Cytotoxicity of Human Lymphocytes Against Rabies‐Infected Cells by Rabies‐Specific Antibodies
Authors: HÄRFAST B. , ANDERSSON T. , GRANDIEN M.
Published: August 2010

Human anti‐rabies immune sera enhanced the in vitro cytotoxicity of human lymphocytes against rabies virus‐infected green‐monkey kidney cells. The immune sera were collected from patients immunized with rabies vaccine produced either in human diploid cells or in nervous tissue. Significant cytotoxicity was observed even with high serum dilutions, indicating that the K‐cell assay might be a sensitive tool for detection of anti‐rabies antibodies.
Generation of Cytotoxic T Lymphocytes Against Ly Alloantigen
Authors: RÖLLINGHOFF M. , PFIZENMAIER K. , STARZINSKI‐POWITZ A. , WAGNER H.
Published: August 2010

Cytotoxic T lymphocytes specific for immune alloantigens controlled by alleles of the Ly system have been induced in vivo. These results were obtained either in a secondary type of response or by treating mice before immunization with a single dose of cyclophosphamide (80 mg/kg).
Increased Sensitivity of Lymphocytes from Atopic Individuals to Histamine‐Induced Suppression
Authors: STRANNEGÅRD I.‐L. , STRANNEGÅRD Ö.
Published: August 2010

Histamine depressed lymphocyte reactivity to phytohemagglutinin and, to a lesser degree, concanavalin A, when administered simultaneously with mitogen to lymphocyte Cultures. Addition of histamine at later times to the cultures appeared to have a slightly enhancing effect on the lymphocyte response. Stimulation of lymphocytes with pokeweed mitogen was in some CMM enhanced, even by high concentrations of Lymphocytes. Lymphocytes from atopic individuals were more sensitive to the inhibitory effect of histamine than Lymphocytes from nonatopic individuals. The sensitivity appeared age‐dependent, but within each age group histamine evoked significantly more suppression on lymphocytes from atopic than from nonatopic individuals. The possibility that the altered reactivity of lymphocytes to histamine, which appears to be associated with atopic allergy, is of pathogenic importance, is discussed, and a hypothesis for the development of atopic disease is proposed.
Suppression of Immunological Activities by Mouse Amniotic Fluid
Authors: ETLINGER H. M. , CHILLER J. M.
Published: August 2010

Mouse amniotic fluid (MAF) was shown to be capable of suppressing those antibody responses observed in euthymic or athymic mouse spleen cell cultures to the T‐independent antigens dinitrophenylated Ficoll (DNP‐Ficoll) and trini‐trophenylated lipopolysaccharide (TNP‐LPS) and to the polyclonal B‐cell activators LPS and purified protein derivative of tuberculin (PPD). Titration experiments demonstrated that the suppressive capacity of MAF for either LPS or DNP‐Ficoll responses was maintained up to a MAF dilution of 1:120. Pre‐incubation of spleen cells obtained from athymic mice with MAF for 8 h significantly suppressed polyclonal B‐cell activation of such cells induced by LPS, although suppression was greater when MAF was present during the entire culture period. In addition, the suppressive activity that MAF demonstrated for antibody production induced by DNP‐Ficoll or LPS was not lost as a result of dialysis. MAF also suppressed the secondary in vitro prolixferative responses of lymph node cells sensitized to the T‐dependent antigen human gamma globulin (HGG). HGG‐induced proliferation of such cells appeared to be more susceptible to suppression effected by MAF than concanavalin‐A‐induced proliferation.
B and T Lymphocytes in Blood and Cerebrospinal Fluid in Acute Aseptic Meningitis
Authors: FRYDÉN A.
Published: August 2010

B and T cells in blood and cerebrospinal fluid (CSF) were calculated by using the surface membrane immunoglobulin marker for B cells and the capacity to bind sheep erythrocytes to form ‘rosettes’ as a marker for T cells. Patients with acute aseptic meningitis due to mumps virus or other etiologic agents and with various chronic neurological diseases were investigated. Significantly higher T‐cell and lower B‐cell values were observed in CSF than in blood in all three groups. No significant differences were found between the three patient groups. An elevation of B cells and a depression of T cells were observed in blood in patients with aseptic meningitis during the course of the disease.
The Morphology of Echinoid Phagocytes and Mouse Peritoneal Macrophages During Phagocytosis in Vitro
Authors: KAPLAN G. , BERTHEUSSEN K.
Published: August 2010

The morphology of mouse peritoneal macrophages and echinoid phagocytes during phagocytosis in vitro was studied. A striking similarity in the function of the foreign surface receptor is found in the two systems. Glutaraldehydetreated erythrocytes attached randomly over the entire surface of the cells and were internalized without circumferential attachment between the particles and the phagocyte membrane. The particles seemed to sink directly into the cytoplasm of the cells. Tannin‐treated erythrocytes were phagocytosed by the echinoid cells in a similar mode. The complement‐coated erythrocytes were attached only in the perinuclear area of the echinoid phagocyte's membrane, but the morphology of their internalization was similar to that mediated by the foreign surface receptor. A circumferential attachment between the particles and the phagocyte membrane did not seem necessary. This is also the case for mouse peritoneal macrophages.
Labeling Immunoelectrophoresis: A General Method for Increasing the Sensitivity of Rocket Immunoelectrophoresis with 125I‐Labeled Anti‐Antibodies
Authors: RAMLAU J. , BJERRUM O. J.
Published: August 2010

In a new procedure, rocket immunoelectrophoresis is performed at pH 8.7 with rabbit antibodies and 5% polyethyleneglycol 6000 in the agarose gel. After the pH in the gel has been changed to 5 the nonprecipitated immunoglobulins are electrophoresed out of the gel simultaneously with the electrophoresis of 125I‐labeled swine antibodies against rabbit IgG into the gel. The latter antibodies tag the rabbit IgG, which is now present only in the precipitates. The radioactive precipitates are visualized by autoradiography. The method permits quantification of antigens down to an amount of approximately 0.5 ng; well‐defined rockets are not formed below this limit. Compared to conventional protein staining with Coomassie brilliant blue, this represents an increase in sensitivity of up to 20 times.
HLA‐D‐Associated Ia‐Like’ Antigens on Human Macrophages
Authors: ALBRECHTSEN D.
Published: August 2010

Human macrophages procured by culture of adherent peripheral blood mononuclear cells were investigated for reactivity in complement‐dependent cytotoxicity tests with antisera having specificity for HLA‐D‐associated ‘Ia‐like’ B‐cell antigens. The antisera were found to be cytotoxic for 30%–70% of the macrophages from donors whose B cells were also reactive, and absorption with B cells also removed macrophage cytotoxicity. A B‐cell xenoantiserum was also cytotoxic to macrophages. It is concluded that the HLA‐D‐associated ‘Ia‐like’ antigens present on B cells are also expressed by macrophages, at least on a subpopulation.
Influence of an M‐Locus Difference on the Cellular and Humoral Immunological Reactivity Against H‐2‐Determined Antigens of the Mouse
Authors: LILLIEHÖÖK B. , BLOMGREN H.
Published: August 2010

Studies were initiated to determine whether an immune response to the Mls antigen of C3H mice could modify responses of CBA lymphocytes (H‐2‐compatible) to a foreign H‐2 complex. CBA lymphocytes, partially tolerant to the C5H‐determined Mls antigen, generated less effector cell activity against C57Bl cells (H‐2‐incompatible) Chan lymph node cells from normal CBA donors when infused into irradiated C.3H × C57Bl hosts. Effector cell activity was measured as the capacity of the cells in the irradiated spleens to inhibit CBA × C57Bl bone marrow proliferation. In contrast, immunization of CBA mice with C3H × C57Bl cells yielded lower antibody titers against C577Bl cells than immunization with CBA × C57Bl cells. Furthermore, preinjection of CBA mice with C3H × CBA cells strongly reduced the capacity of the animals to produce antibodies against C57Bl cells. Thus, these data support the conclusion that an immune response to a foreign Mls antigen may either enhance or suppress an immune response to H‐2‐incompatible cells.
Enzyme Analysis of Autologous Rosette‐Forming Cells
Authors: PLUM J. , DE SMEDT M. , SABBE L.
Published: August 2010

Autologous rosette‐forming cells (ARFC) have been considered to be post‐thymic precursor cells. Since thymocytes and peripheral T lymphocytes differ considerably in lactate dehydrogenase isoenzyme pattern and in activity of the enzymes of the purine metabolism, we investigated the enzyme profile in the ARFC. The L‐lactate:NAD+ oxi‐reductase analysis showed an isoenzyme pattern that closely resembled the pattern found in peripheral T lymphocytes and was totally different from the thymocytes. The levels of adenosine deaminase and purine nucleoside phosphorylase were identical to those found in the peripheral T lymphocytes and different from thymocytes. In our hands, the ARFC‐enriched suspension contained predominantly OKT4+ and Tμ+ lymphocytes. We propose that ARFC are a heterogeneous population encompassing all known subsets and cannot be considered a separate homogeneous entity.
Mouse B‐ and T‐Cell Colony Formation in Vitro
Authors: CLAESSON M. H. , NISSEN M. HOLST , ROPKE C.
Published: August 2010

Rat spleen coll cultures exposed for 24 h to concanavalin A (Con A‐CM) contain, in addition to inierleukin 2 (IL‐2). factors that promote colony formation in vitro by mouse T cells (TCPA) and B cells (BCPA). TCPA and BCPA are separable on a Sephadex G‐75 column. TCPA has a molecular weight of 15,000 daltons and shows the same elution profile as IL‐2. Absorption studies with Con A‐activated T cells suggested that TCPA and IL‐2 are the same entity. BCPA has an apparent molecular weight uf 45.000 daltons and stimulates colony formation by B lymphocytes seeded at very low cell density (104–5 × 104 cells/ml). In contrast to TCPA, BCPA can only be demonstrated in gel‐filtered material owing to the presence of B colony suppressor activities in crude Con A‐CM. Two B colony inhibitory activities were demonstrated by AcA 34 chromatography of crude Con A—CM with molecular weights of 80,000–130.000 and about 50,000. respectively. Because of the specificity, simplicity and sensitivity of B and T colony formation these assay systems should be valuable tools for in vitro testing of biological activities regulating the immune system.
Mouse Lactate Dchydrogenase (LDH) C4 (Testis) Is Immunochemically Cross‐Reactive with LDH A4 (Muscle) and LDH B4 (Heart)
Authors: WRIGHT L. L. , SWOFFORD J. H.
Published: August 2010

It has been reported that lactale dehydrogenase (LDH) purified from testes (LDH C) of homotherms is not immunochemically cross‐reactive with somatic forms of LDH purified from heart (LDH B) or muscle (LDH A). On the basis of this premise, LDH C has been considered for use as a contraceptive vaccine. Data presented here indicate that mouse anlisera to either mouse or rat LDH C are cross‐reactive with LDH A and B purified from muscle and heart tissues of mice. However, rabbit antisera to mouse LDH C are not cross‐reactive with either mouse LDH A or B. Thus, the degree of cross‐reactivily is dependent on the species from which the immunogen LDH is purified, the antisera are derived, and the LDH used in the assay is purified. The determination that LDH A, B, and C are immunochemically cross‐reactive is of importance in evaluating LDH C as an immunogen in an immunologic approach to contraception.
Autologous Mixed Lymphocyte Reaction in Man
Authors: GUPTA S.
Published: August 2010

The autologous mixed lymphocyte reaction (AMLR) and T‐cell subsets defined with monoclonal antibodies were examined in the peripheral blood of six patients with chronic mucocutaneous candidiasis (CMC). The AMLR was deficient in four of six patients when compared with simultaneously studied healthy controls. These patients had either overt endocrinopathy or circulating autoantibodies. In two of three patients with low AMLR, after the depletion of OKT8+ T cells no enhancement in the AMLR was observed, demonstrating that the deficiency of the AMLR was not due to increased suppressor OKT8+ T‐cell activity. However, the third patient demonstrated almost complete reconstitution of the AMLR response after such depletion, suggesting that OKT8+ suppressor T cells were responsible for decreased AMLR in this patient. Two patients had a low ratio of OKT4/OKT8 phenotype T cells. T cells with Tac antigen (that is, present on activated T cells) were increased in four of six patients. No correlation was observed between the deficiency of the AMLR and the proportions of T‐cell subsets. This study demonstrates a deficiency of the AMLR in some patients with CMC which is associated with increased OKT8+ suppressor T‐cell activity or with the functional deficiency of responder OKT4+ T cells.
Synthesis and Turnover of the Putative T‐Lymphocyte Antigen Receptor
Authors: McKIMM‐BRESCHKIN J. L. , MILLER J. F. A. P.
Published: August 2010

Cells of an antigen‐specific T‐cell clone, A37.4, were treated with the proteolytic enzymes trypsin or pronase to remove the T‐cell antigen receptor. Removal of the receptor, analysed by surface labelling and non‐reduced/reduced gel electrophoresis, stimulated the cells to synthesize new protein rapidly. New membrane protein was readily detectable within 3 h after reculture, and it was able to interact with antigen‐presenting cells, since the T cells could be stimulated to produce interferon. Treatment of cells with metabolic inhibitors demonstrated that there was only a small cytoplasmic pool of protein, and de novo synthesis of mRNA was necessary for quantitative replacement of the membrane protein. The protein also required glycosylation for transport and insertion into the membrane. Despite rapid resynthesis of new receptor, when surface‐labelled cells were recultured either with or without antigen‐presenting cells, there was not a rapid turnover of the receptor. There was also no evidence of any loss of receptor from the membrane of antigen‐stimulated cells or of any shedding of receptor into the culture medium.
Polyclonal Activation of B Cells during Rubella Infection
Authors: HYYPIÄ T. , ESKOLA J. , LAINE M. , SALMI A. , MEURMAN O.
Published: August 2010

Intensive spontaneous production of IgG by peripheral blood lymphocytes in vitro is detected at the onset of rash in patients with rubella virus infection. The IgG synthesized consists not only of antibodies against rvihella virus specific antigens bul also of antibodies to unrelated viruses.
Monoclonal Antibodies Specific for Sendai Virus
Authors: FENNER M. , SIEGMANN K. , BINZ H.
Published: August 2010

The monoclonal antibodies A and B specific for the HN molecule of Sendai virus were used to induce polyclonal and monoclonal anti‐idiotypic antibodies. No response was observed in allogeneic Lewis rats, low responses in syngeneic LOU rats, and high responses in allogeneic BN rats and xenogeneic Balb/c mice. The monoclonals A and B share a similar or identical idiotype. since polyclonal anti‐idiotypic antisera to antibody A cross‐reacted completely with antibody B and vice versa. The same was found with the three monoclonal anti‐idiotypes 1, 2, and 3elicited in a BN rat or in Balb/c mice. None of the polyclonal or monoclonal anti‐idiotypes reacted with other monoclonal antibodies specific for Sendai virus, even when these recognized the same epitope as antibodies A and B. The monoclonal anti‐idiotypic antibodies could be used to induce anti‐Sendai antibodies in BN rats.
Altered Natural Killer and Natural Cytotoxic Cellular Activities in lpr Mice
Authors: PAN L.‐Z. , DAUPHINÉE M. J. , AHMED S. ANSAR , TALAL N.
Published: August 2010

Three strains of mice hearing the autosomal recessive lpr gene (MRL. C57BL/6 and C3H)that had spontaneously developed a lupus‐like disease were studied sequentially for functional natural killer (NK) and natural cytotoxic (NC) cell activity. Natural killing was impaired in spleen and bone marrow cells from all the lpr strains, as well as from the congenic strain MRL‐ +/+. which develops a late onset lupus‐like disease. The NK cell activity was found to be depleted as early as 2 months of age in all lpr strains, and decreased further with age. NK activity was augmentable by Poly I: C and interleukin 2 (IL‐2). suggesting that the residual cells can respond to NK modulators. In contrast with NK cell activity. NC activity was not decreased in lpr mice but could be augmented by IL‐3‐rich supernatants. The spontaneous decrease in NK cell activity was associated with an increased autologous plaque‐forming cell (APFC) response to bromelin‐treated mouse red blood cells, which is produced primarily by B cells possessing the Ly‐1 phenotype (Lyt‐1+ B). When NK cell activity was increased by exogenous administration of Poly I:C, the APFC response diminished. Treatment of spleen cells with anti‐asialo GM1 prior to Poly I: C treatment resulted in a decreased NK response but increased both APFC and Lyt‐1+ B cells. The possible regulation of autoreactivity by NK cells is discussed.
Direct Demonstration of Immunoregulatory T‐Cell Defects in Patients with Systemic Lupus Erythematosus
Authors: KOIDE J. , TAKANO M. , TAKEUCHI T. , HOSONO O. , AMANO K. , HOMMA M. , ABE T.
Published: August 2010

The present study was undertaken to determine directly whether immunoregulatory T cells have a defective suppressor function in patients with systemic lupus erythematosus (SLE), and whether anti‐T‐cell antibodies are essential for immunoregulatory T‐cell defects. Peripheral blood T cells and T‐cell subsets were determined in 52 SLE patients. The ratio of T4 to T8 cells was distributed over a wider range in patients with SLE than in the controls. Patients with SLE were divided into three groups (low, normal and high) by the T4/T8 ratio. Lymphocytes from 12 SLE patients (7 with low and 5 with high T4/T8 ratios) were studied extensively. Their disease was inactive or in remission, Anti‐T‐cell antibodies were not detected, and yet the patients had immunological abnormalities characterized by the presence of antinuclear antibodies and hypergammaglobulinaemia. The SLE patients with high T4/T8 ratios had a decreased number of'T8 cells, and defective suppressor‐effector cells. In contrast, patients with low T4/T8 ratios had decreased T4 cells and/or increased T8 cells, and defective suppressor‐inducer cells. Two patients with low T4/T8 ratios had both suppressor‐effector and suppressor‐inducer cell defects. These results indicate that immunuregulaiory circuits in SLE patients are heterogeneous and that immunoregulatory defects exist even when the disease is inactive or in remission. Anti‐T‐cell antibodies were not essential for such immunoregulatory defects. Thus, immunoregulatory T‐cell defects and the development of SLE may be independent conditions due to other unknown causes.
Interleukin 1 Activity in the Acquired Immunodeficiency Syndrome
Authors: ENK C. , GERSTOFT J. , MØLLER S. , REMVIG L.
Published: August 2010

Interleukin 1 (IL‐1) is a monocyte‐derived mediator that participates in the regulation of various T‐lymphocyte activities, among them IL‐2 production. Since IL‐2 deficiency is a central feature in the immunological profile of the acquired immunodeficiency syndrome (AIDS), the production of IL‐1 from peripheral blood monocytes from male homosexuals with AIDS was investigated at the same time as the IL‐1 responsiveness of monocyte‐depleted mononuclear cells (MDC) from the same patients. The IL‐1 was produced by lipopolysaccharide‐stimulated monocytes and assayed by the capacity of monocyte supernatants to amplify the proliferation of phytohaemagglutinin‐stimulated allogeneic MDC from healthy donors as well as murine thymocytes. The IL‐1 responsiveness was measured by measuring the enhancing effect of an IL‐1 standard on the proliferative response of patients' MDC. The IL‐1 production was not reduced compared to the IL‐1 production in a control group, but the IL‐1 responsiveness of the patients' MDC was depressed. The results indicate that depressed IL‐1 production is not one of the immunological disturbances in AIDS, hut that the T‐lympho $$ γ properties $$ IL‐1 are affected.
Enprostil, a Prostaglandin‐E2 Analogue, Inhibits Interleukin‐8 Production of Human Colonic Epithelial Cell Lines
Authors: Toshina K. , Hirata I. , Maemura K. , Sasaki S. , Murano M. , Nitta M. , Yamauchi H. , Nishikawa T. , Hamamoto N. , Katsu K.
Published: August 2010

We previously reported that intracolonic administration of enprostil, a prostaglandin‐E2 (PGE2) analogue, had therapeutic effects on acute colitis induced in rodents by dextran sulfate sodium (DSS). In addition, production of growth‐regulated gene product/cytokine‐induced neutrophil chemoattractant‐1 [GRO/CINC‐1; an interleukin(IL)‐8 like cytokine] was suppressed in the inflamed tissues. In the present study we used a human colon cancer cell line (HT‐29) to investigate enprostil effects on the IL‐8 production of intestinal epithelial cells stimulated by various stimulants. In a MTT assay, concentrations of enprostil >10−5M had cytotoxitic effects on HT‐29 cells. Furthermore, 10−6 M enprostil suppressed IL‐8 production in HT‐29 cells, SW620 and CaCo2 stimulated with interleukin‐1β (IL‐1β) or lipopolysaccharide (LPS), but did not suppress this response when cells were stimulated with tumour necrosis factor (TNF)‐α. These results suggest that enprostil affects a point in the pathway between the IL‐1 receptor or LPS receptor and nuclear factor‐κB(NF‐κB), without affecting the pathway between the TNF receptor and NF‐κB, with the latter factor being required for the IL‐8 gene transcription. The therapeutic effect of exogenous enprostil on DSS colitis may involve the inhibition of IL‐8 production in colonic epithelial cells stimulated by IL‐1β or LPS.
Genomic Organization and Regulation of the Human Interleukin‐18 Gene
Authors: Kalina U. , Ballas K. , Koyama N. , Kauschat D. , Miething C. , Arnemann J. , Martin H. , Hoelzer D. , Ottmann O. G.
Published: August 2010

The human interleukin(IL)‐18 is a key regulator of interferon(IFN)‐γ production and T‐cell differentiation. Here we report the complete genomic structure and characterization of the 5′untranslated promoter region of the human IL‐18 gene. The gene is composed of six exons and five introns, spanning approximately 19.5kb. Promoter activity of the 5′‐flanking region was investigated with a luciferase reporter gene assay. Transient transfection studies demonstrate a constitutive expression of the IL‐18 gene in monocytic U937 and THP‐1 cells. For this constitutive expression at least 92 base pairs of the promoter region are essential as shown by consecutive 5′ promoter deletions in both cell types. DNA protein binding experiments revealed specific binding of activated signal transducer and activator of transcription factor‐5 (STAT5) but not of STAT3 to three consensus sequences upstream in the promoter region. Cotransfection of STAT5 resulted in increased induction of the IL‐18 promoter in the U937 and THP‐1 cells.
Expression of Human TIM‐1 and TIM‐3 on Lymphocytes from Systemic Lupus Erythematosus Patients
Authors: Wang Y. , Meng J. , Wang X. , Liu S. , Shu Q. , Gao L. , Ju Y. , Zhang L. , Sun W. , Ma C.
Published: August 2010

The T‐cell immunoglobulin‐ and mucin‐domain‐containing molecules (TIMs) comprise a new family of cell surface molecules expressed on T cells. TIM‐3 is expressed on T helper type 1 (Th1) cells and implicated in the pathogenesis of Th1‐driven auto‐ and allo‐immune diseases. TIM‐1 is suggested to act as a co‐stimulatory molecule for all T cells, but with potentially stronger effects on Th2 than Th1 cells and is associated with Th2‐related immune diseases. However, the TIM molecules have not been investigated in the systemic lupus erythematosus (SLE). In this study, we examined the expression of TIM‐1 and TIM‐3 on peripheral blood mononuclear cells from SLE patients using quantitative real‐time RT‐PCR. An increased TIM‐1 expression was detected in SLE patients, which correlates with interleukin‐10 expression. We also found that there was a significant increase in the expression of TIM‐1 in SLE patients with quite active disease (SLE disease activity index > 6), indicating that TIM‐1 expression might be related to active clinical phases. In contrast, TIM‐3 expression remained normal in SLE patients with low statistical power (34.89%). However, the expression of TIM‐3 ligand, galectin‐9 increased in SLE patients indicating an enhanced engagement of TIM‐3 with its ligand in SLE, which may result in a decreased regulatory T‐cell function as shown by the decreased expression of FoxP3 and TGF‐β1 in SLE. These data suggest that TIM‐1 and TIM‐3/TIM‐3L are involved in the pathogenesis of SLE.
Circulating Surfactant Protein D is Decreased in Early Rheumatoid Arthritis: A 1‐year Prospective Study
Authors: Hoegh S. V. , Lindegaard H. M. , Sorensen G. L. , Høj A. , Bendixen C. , Junker P. , Holmskov U.
Published: August 2010

Innate immune system abnormalities, e.g., mannan‐binding lectin (MBL) genotype variants, have been demonstrated to modify the disease course of rheumatoid arthritis (RA). Surfactant protein D (SP‐D) shares important structural and functional properties with MBL suggesting that SP‐D may be an additional RA disease modifier. The Met11Thr polymorphism in the N‐terminal part of SP‐D is an important determinant for the SP‐D serum level, but this polymorphism is also essential to the function and assembly into oligomers. We aimed to compare the serum levels of SP‐D in a cohort of newly diagnosed untreated RA patients with healthy matched controls, and to investigate if there was an association to core measures of disease activity within the first year after disease onset. Secondly, we aimed to investigate whether the Met11Thr polymorphism was associated with RA. Serum SP‐D was significantly lower in DMARD naive RA patients compared with healthy controls (P = 0.016). Median SP‐D concentration at inclusion was 878 ng/ml (95% CI: 730–1033) and 1164 ng/ml (95% CI: 1093–1366) in RA patients and matched controls, respectively. SP‐D increased during Methotrexate treatment (P < 0.0001), and at 1‐year follow‐up median SP‐D was 1032 ng/ml (95% CI: 777–1255). SP‐D levels did not correlate with traditional disease activity measures. The Thr11/Thr11 genotype and the Thr11 allele tended to be more frequent in RA patients. In conclusion, the low serum level of SP‐D and the lack of correlation with traditional disease activity measures indicate that SP‐D reflects a distinctive aspect in the RA pathogenesis.
Inhibition of Serine‐Peptidase Activity Enhances the Generation of a Survivin‐Derived HLA‐A2‐Presented CTL Epitope in Colon‐Carcinoma Cells
Authors: Preta G. , Marescotti D. , Fortini C. , Carcoforo P. , Castelli C. , Masucci M. , Gavioli R.
Published: August 2010

Cytotoxic T lymphocytes eliminate tumor cells expressing antigenic peptides in the context of MHC‐I molecules. Peptides are generated during protein degradation by the proteasome and resulting products, surviving cytosolic amino‐peptidases activity, may be presented by MHC‐I molecules. The MHC‐I processing pathway is altered in a large number of malignancies and modulation of antigen generation is one strategy employed by cells to evade immune control. In this study we analyzed the generation and presentation of a survivin‐derived CTL epitope in HLA‐A2‐positive colon‐carcinoma cells. Although all cell lines expressed the anti‐apoptotic protein survivin, some tumors were poorly recognized by ELTLGEFLKL (ELT)‐specific CTL cultures. The expression of MHC‐I or TAP molecules was similar in all cell lines suggesting that tumors not recognized by CTLs may present defects in the generation of the ELT‐epitope which could be due either to lack of generation or to subsequent degradation of the epitope. The cells were analyzed for the expression and the activity of extra‐proteasomal peptidases. A significant overexpression and higher activity of TPPII was observed in colon‐carcinoma cells which are not killed by ELT‐specific CTLs, suggesting a possible role of TPPII in the degradation of the ELT‐epitope. To confirm the role of TPPII in the degradation of the ELT‐peptide, we showed that treatment of colon‐carcinoma cells with a TPPII inhibitor resulted in a dose‐dependent increased sensitivity to ELT‐specific CTLs. These results suggest that TPPII is involved in degradation of the ELT‐peptide, and its overexpression may contribute to the immune escape of colon‐carcinoma cells.
Retracted: Construction, Expression and Immunoassay Detection of Recombinant Plasmid Encoding Fusion Protein of Roman Chicken Complement C3d and Newcastle Disease Virus F gene
Authors: Liu D. , Niu Z.‐X.
Published: August 2010

The terminal degradation product (C3d) of mammalian complement component C3 plays an important role in modulation of the adaptive immune response through the interaction with complement receptor type 2 (CR2) on B cells. In this study, the gene fragment coding for the complement protein C3d (chC3d) from Roman chicken was cloned and expressed as a fusion protein for its application in the vaccine study of chicken, and for in vitro experiments. The chC3d fragment strengthened B‐cell responses when complexed with antigen. Three potential vaccine construct units were engineered to contain two, four and six copies of chC3d coding gene linked to the F gene of Newcastle disease virus (NDV), an economically important pathogen of chicken that is classified as a list A contagious disease of poultry by the Office International des Epizooties. The cloned chC3d protein and different repeats of C3d proteins in addition to the F gene of NDV were generated separately in Escherichia coli and chicken embryo fibroblast cells with the help of expression vectors. All recombinant proteins were analysed by SDS‐PAGE and Western blotting. Analysis of the immunogenicity of different repeats of C3d revealed that chC3d had an enhancing effect on the immunogenicity of antigens, and that six or more repeats of C3d may be necessary for efficient enhancement of antigen‐specific immune responses. To date, published research into the adjuvant activities of C3d has been limited to experiments in mice, rabbits and cattle. The adjuvant properties of C3d have not been assessed in poultry using homologous C3d in association with antigens relevant to the target species. The Roman chicken C3d fusion proteins described in this study is the first report and will provide a basis for immunization trials in chicken, studies of receptor binding and cell activation of chicken lymphocytes, and investigations of new types of vaccines, including recombinant vaccines and DNA vaccines for future use against other pathogens in chicken.
In Vitro Experiments Demonstrate that Monocytes and Dendritic Cells are Rendered Apoptotic by Extracorporeal Photochemotherapy, but Exhibit Unaffected Surviving and Maturing Capacity after 30 Gy Gamma Irradiation
Authors: Rao V. , Saunes M. , Jørstad S. , Moen T.
Published: August 2010

Extracorporeal photochemotherapy (ECP) has been shown to induce apoptosis in lymphocytes. Until recently the prevailing opinion has been that the monocytes were mainly not affected by this treatment. This study has investigated the effect of ECP and gamma irradiation on monocytes and immature dendritic cells (DC) in vitro and followed the ability of the cells to differentiate and survive post treatment. ECP induced apoptosis in lymphocytes, monocytes and immature DC within 72 h following treatment, in contrast to 30 Gy gamma irradiation, which seemed mainly to affect lymphocytes. The minority of the surviving ECP‐treated monocytes presented a reduced ability to differentiate into immature DC within this time frame. We also demonstrated that immature DC after ECP‐treatment lost their normal ability to mature on stimulation with lipopolysaccharide. As monocytes and immature DC seem to have a reduced ability to differentiate after ECP‐treatment, it is suggested that the therapeutic effect of ECP is caused by in vivo effects of reinfused apoptotic cells, rather than by infusion of monocytes induced to differentiate into immature DC.