bluuebunny/crossref_metadata_2025 huggingface dataset:
The Immune Response to the Alpha 1–6 Epitope of Dextran is Determined by a Gene Linked to the IgCH Locus
Authors: FERNANDEZ C. , LIEBERMAN R. , MÖLLER G.
Published: June 2006

There is a marked strain variability in the immune response against the alpha 1–6 epitope of dextran B512. Responsiveness was independent of the H‐2 genotype of the strains but completely determined by the IgCH allotypes of the strains. Thus, IgCHb and IgCHj allotypes characteristic of C57BL and CBA strains, respectively, determined high responsiveness, whereas strains possessing all other allotypes studied were low responders. Backcross analysis indicated that a gene(s) determined responsiveness and that this gene was linked to allotypes of the IgCH locus. F1 hybrids between high and low responder strains always gave a response, but the number of plaque‐forming cells was approximately half of that obtained in the high responder strains.
Heating at 56°C does not Eliminate Immune Complex‐bound Clq
Authors: TEPPO A.‐M. , WAGER O.
Published: June 2006

The effect of heating at 56°C on free and complex‐bound human Clq was studied by radioisotope and sucrose gradient centrifugation techniques. The ability of soluble complexes of egg albumin, rabbit anti‐egg albumin, and human Clq to bind extrinsic iodinated Clq did not increase following heating at 56°C. The results of this study did not support the postulated release of complex‐bound Clq on healing at 56°C. They also showed that free Clq was more heat‐labile than Clq in the Clqrs state. The importance of elimination or monitoring of intrinsic Clq in Clq assays for circulating immune complexes is stressed.
Mitogen‐induced Differentiation of Human CLL Lymphocytes to Antibody‐secreting Cells
Authors: ROBÈRT K.‐H. , BIRD A. G. , MÖLLER E.
Published: June 2006

Peripheral blood lymphocytes from nine patients with chronic lymphatic leukaemia (CLL) have been stimulated by photohaemagglutinin, pokeweed mitogen and Epstein‐Barr virus. Our results provide proof that the leukaemic B cells in CLL are capable of responding to mitogenic signals by producing and secreting antibodies, transforming into blast cells, and, probably, increasing their rate of DNA synthesis.
A Thymus‐independent IgG Response against Dextran B512 can be Induced in C57BL but not in CBA Mice, even though both Strains Possess a VHdex Gene
Authors: FERNANDEZ C. , MÖLLER G.
Published: June 2006

CBA and C57BL mice both possess a VHdex gene coding for antibodies against the alpha 1–6 epitope of dextran B512. After immunization with dextran, CBA mice produce IgM plaque‐forming cells (PFC) only, which show regular cyclical fluctuations. The second PFC peak disappeared after injection of dextranase, indicating that it is antigen‐dependent. The anti‐dextran response in C57BL mice is characterized by only one IgM peak, followed I day later by an IgG peak that may exceed the IgM response by a factor of 10. The IgG anti‐dextran response in C57BL mice was thymus–independent. CBA mice gave an IgG response to the hapten fluorescein iso‐thiocyanate (FITC) after immunization with FITC‐dextran, indicating that dextran can function as a carrier for an IgG response in this strain. Attempts to induce IgG PFC against dextran by immunizing CBA mice with thymus‐dependent dextran‐protein conjugates consistently failed, although the conjugates induced IgG PFC in C57BL mice. Spleen cells from CBA mice failed to produce IgG antibodies against dextran after transfer into lethally irradiated C57BL mice. whereas the C57BL spleen cells produced IgG PFC after transfer into CBA mice. The lack of IgG synthesis against the alpha 1–6 epitope of dextran in CBA mice appears to be regulated exclusively at the B cell level and is restricted specifically to the VHdex gene product.
Generation of Cytolytic T Lymphocytes in Vitro
Authors: DEGIOVANNI G. , MARYANSKI J. L. , CEROTTINI J.‐C. , BRUNNER K. T.
Published: June 2006

The memory Cytolytic T lymphocyte (CTL) response of mice was studied in vitro using peripheral blood leucocytes (PBL) of donors previously immunized with allogcneic cells as responding cells, and particulate alloantigen for stimulation. It is shown that alloimmune but not normal PBL respond to paniculate antigen and that srnall numbers of PBL generate high CTL responses provided non‐T accessory spleen cells are added to the cultures. The system provides means to analyse the kinetics of the appearance of memory CTL precursors after primary alloimmunization in individual mice.
Polyclonal Antibody Secretion Induced in Human Mixed Lymphocyte Cultures
Authors: KUNORI T. , RINGDÉN O.
Published: June 2006

In mixed leucocyte culture (MLC), using human spleen cells or thoracic duct lymphocytes, antibody secretion was induced, measured as plaque‐forming cells (PFC) in a haemolysis‐in‐gel assay with fluorescein isothiocyanate (FITC)‐coupled sheep erythrocytes (SRBC) as targets. Peak antibody secretion was seen on day 5. Using protein‐A‐coupled SRBC as targets and developing antisera, antibody secretion in MLC was found to be of IgM, IgG and IgA type. There was no correlation between the number of PFC against FITC‐SRBC in MLC and DNA synthesis. Supernatants from MLC failed to induce antibody secretion.
Macrophage‐Lymphocyte Clusters in the Immune Response to Soluble Protein Antigen in Vitro
Authors: PETRI J. , BRÆNDSTRUP O. , WERDELIN O.
Published: June 2006

T cells from immune guinea‐pigs produce clusters in vitro with macrophages pulsed with soluble protein antigens. The formation of clusters is antigen‐specific. Time‐lapse cinephotomicrography was used to study the sequence of events leading to cluster formation and the kinetics of already formed clusters. The central lymphocyte attaches through a broad base to the macrophage during the first few hours of incubation. After several hours, during which the central lymphocyte is situated alone on the surface of the macrophage, free lymphocytes begin to interact for shorter or longer periods of time through their uropods with the central lymphocyte, thus acting as peripheral lymphocytes. In spite of a frequent traffic of peripheral lymphocytes in and out, the cluster is a relatively stable structure. Occasionally the entire complex of lymphocytes leaves the macrophage, moves randomly in the culture and attaches to a new macrophage or returns to the same macrophage.
The Humoral Immune Response to Alpha, Beta and Gamma Crystallins of the Human Lens
Authors: SANDBERG H. O. , CLOSS O.
Published: June 2006

By radioimmunoassay the occurrence of antibodies against alpha, beta and gamma crystallins was examined in sera from groups of patients with clear lenses, cataracts or lens injuries. In about half of the sera from each group low anti alpha crystallin activity was found. No significant difference in antibody activity was found between the groups. Antibody activity against beta or gamma crystallins was not detected in any of the sera. Four patients with lens damage were followed for 2 years; two of them developed uveitis and showed increasing anti alpha crystallin activity. It is suggested that in man immunological tolerance to lens crystallins is normally present.
A Rapid Method for the Separation of Functional Lymphoid Cell Populations of Human and Animal Origin on PVP‐Silica (Percoll) Density Gradients
Authors: KURNICK J. T. , ÖSTBERG L. , STEGAGNO M. , KIMURA A. K. , ÖRN A. , SJÖBERG O.
Published: June 2006

Subclasses of lymphocytes can be separated on gradients of non‐toxic polyvinylpyrrolidone‐coated colloidal silica (Percoll) by virtue of differential densities. Such gradients can yield functionally active lymphocyte populations after brief centrifugation. Gradients can he generated in a discontinuous step fashion and centrifuged in standard table‐top laboratory centrifuges or as self‐generating gradients duriny ultracentrifugation. The density medium has low viscosity and can be made isotonic for virtually any use. Gradients have proved useful in both human and experimental animal studies, and high percentage yields allow for separations from small cell numbers. Methods are described for separation of whole blood and lymphotd subpopulations. including blasts stimulated with mitogens or in mixed lymphocyte reactions. The cytotoxic capability of various density fractions was evaluated for mixed lymphocyte culture‐induced allogeneic killing and spontaneous, so‐called ‘natural’ killer cell activity. The lower density associated with blast transformation allows for significant enrichments of stimulated cells from in vitro cultures. Higher thymidine incorporation, restimulation in mixed lymphocyte reactions, and greater cytotoxic capacity are associated with these‘blast’fractions.
Characterization of IgD
Authors: CORTE G. , TONDA P. , COSULICH E. , MILSTEIN CELIA P. , BARGELLESI A. , FERRARINI M.
Published: June 2006

Human IgD present in the serum of normal individuals or of patients with Hodgkin's disease (having high IgD concentrations) was characterized and compared with five IgD myeloma proteins. IgD was isolated using a highly specific anti‐δ insoluble immunoabsorbent from which the bound material was eluted with sodium dodecyl sulphate (SDS) or urea. The latter reagent could be removed by extensive dialysis, thus making possible the renaturation of the eluted molecules. The purity of the IgD thus isolated was confirmed by antigenic analysis. Both K and λ light chain determinants were present on serum IgD, although λ light chain was predominant with a ratio over the K chain of 2:1. SDS‐polyacrylamide slab gel electrophoresis analysis revealed two different molecular forms of serum IgD, one (IgDI) migrating identically to monoclonal IgD, the other (IgD2) having a faster mobility. The difference between the two molecules was entirely, due to the different sizes of their constituent δ chains. Peptide mapping of the two chains (δ1 and δ2 respectively) and of the δ chain of an IgD myeloma protein was carried out using 125I‐labelled material. The three molecules displayed a high degree of homology, the δ2 chain differing by the presence of three characteristic extra peptides. The significance of these extra peptides is discussed in the light of the peptide mapping technique employed.
Demonstration of Electrophoretic Heterogeneity of Serum β2‐Microglobulin in Systemic Lupus Erythematosus and Rheumatoid Arthritis: Evidence against Autoantibodies to β2‐Microglobulin
Authors: PLESNER T. , WIIK A.
Published: June 2006

A sensitive crossed radioimmunoelectrophoretic method (CRIE), originally developed to study lymphocyte‐associated β2‐microglobulin (β2m), was applied in the study of serum β2m in patients with systemic lupus erythemutosus (SLE) and rheumatoid arthritis (RA). In six of seven patients with SLE and nineteen of twenty‐seven patients with RA a considerable elec‐trophoretic heterogeneity of serum β2m was found. In addition to the normally seen symmetric β2m precipitate, a β2 precipitate exhibiting complete immunochemical identity was found in the α‐electrophoretic region. Binding of isolated I25l‐labelled β2m to the abnormal precipitate was demonstrated in crossed immunoelectrophoresis. After gel filtration of sera exhibiting the above‐mentioned β2m binding, all β2m was eluted in low molecular weight fractions corresponding to free β2m. By application of appropriate antisera and a glycoprotein‐binding lectin in intermediate gels in CRIE, it was shown that the possible β2m‐binding ligand is not an antibody, not a major constituent of normal human serum, and not unmodified HLA alloantigen. The abnormality was not restricted to patients with high disease activity but was found more frequently and was more pronounced (mean score 1.6 arbitrary units against 0.57 arbitrary units, P < 0.01) in such patients. Thus our data exelude the possibility that autoantibodies to β2m were present in serum from patients with SLE and RA.
Demonstration of Rheumatoid Factor Idiotypic Antigens on Peripheral Blood B and T Lymphocytes from Patients with Rheumatoid Arthritis
Authors: DOBLOUG J. H. , FØRRE Ø. , NATVIG J. B. , MICHAELSEN T. E.
Published: June 2006

Antisera were raised against three polyclonal IgM rheumatoid factors (RF). After adequate absorptions, the antisera were rendered idiotype‐specific, as assayed by haemagglutination technique. By using the anti‐idiotype antisera in indirect immunofluorescence on peripheral blood lymphocytes from the patients used as donors for the immunizing RF, it was demonstrated that 3–14% of the lymphocytes were stained, and thus had membrane‐bound structures with idiotypic antigens similar to those of the circulating IgM RF of the same patients. While most of these idiotype‐positive lymphocytes were B lymphocytes, it was demonstrated in one patient that about 7% of the T lymphocytes also had the same idiotypic antigens.
Antigenic Analysis of Mycobacterium leprae
Authors: CLOSS O. , MSHANA R. N. , HARBOE M.
Published: June 2006

About twenty distinct antigenic components have been demonstrated in Mycubacterium leprae (M. leprae) by crossed immunoelectrophoresis against a rabbit antiserum produced by immunization with concentrated M, leprae antigen. This system allows a more detailed analysis of the antigenic relationship between M, leprae and other mycobacteria and a better characterization of the antigenic content of various M. leprae preparations than with previously available antisera which reacted with tar fewer components. The antibody activity in sera of patients with lepro‐matous leprosy was studied by incorporating the sera into the intermediate gel of the M. leprae reference system. Antibodies were found against only seven of the components. Since those compared are all known to be cross‐reacting widely with antigens of other mycobacteria, it is speculated that cross‐immunization may influence the antibody response in lepromatous leprosy.
Quantification of the Pregnancy‐specific β1‐Glycoprotein (SP1) by Immunoprecipitation Techniques: the Influence of a Cross‐reacting High Molecular Weight Ş2‐Protein
Authors: TEISNER B. , FOLKERSEN J. , HINDERSSON P. , JENSENIUS J. C. , WESTERGAARD J. G.
Published: June 2006

Two serum proteins with SPI determinants were separated by means of size chromatography and preparative zone electrophoresis. One of the proteins had β1 electrophoretic mobility and an estimated molecular weight of 90,000, and the other had Ş‐mobility and a molecular weight around 400,000. When available antibody preparations to SPI were used, the high molecular weight protein formed immunoprecipitates only when polyethyleneglycol (PEG) was added to the agarose. It is shown that the high molecular weight protein has a pronounced effect on quantification of SPI by rocket immunoelectrophoresis, whereas the effect was less pronounced but still considerable when quantification was performed by radial immunodiffusion.
Lymphocyte Subpopulations in Man: Characterization of Human Killer Cells against Allogeneic Targets Sensitized with HLA Antibodies
Authors: JOHNSEN H. E. , MADSEN M.
Published: June 2006

Human killer cells mediating antibody‐dependent cytotoxicity against allogeneic lymphoblasts presensitized with HLA antibodies have been studied by rosette fractionation experiments. Enriched and/or depleted cell suspensions have been tested in dose–response studies. Two different populations can act as killer cells. The major cytotoxic capacity is retained among T cells with high‐avidity Fc receptors, whereas a minor cytotoxic capacity was found among non‐T cells with high‐avidity Fc receptors. These two populations have different dose‐response curves, indicating different effector mechanisms.
Con‐A‐activated T Cells Secrete Factors with Polyclonal B‐Cell‐activating Properties
Authors: PRIMI D. , HAMMARSTROM L. , MÖLLER G. , SMITH C. I. E. , UHR JAQUELINE
Published: June 2006

Concanavalin A induced polyclonal antibody synthesis in normal spleen cells in vitro. Optimal responses were obtained by Con A concentrations lower than those optimal for induction of DNA synthesis. T cells, but not macrophages, were necessary for the effect. Spleen cells from nude mice were not activated, whereas cells from the LPS non‐responder stain C3H/HeJ were activated to polyclonal antibody synthesis by Con A. Supernatants from Con A activated spleen cells could by themselves induce polyclonal antibody synthesis in untreated spleen cell cultures, even when Con A had been removed by absorption with Sephadex G‐50 and when alpha‐methyl‐mannoside was present in the secondary cultures. T cells produced the active Supernatants, which were competent to induce polyclonal antibody synthesis, but not DNA synthesis, in both H‐2‐incompatible and compatible strains. When the Supernatants were absorbed with erythrocyte antigens, they specifically induced an enhanced response, in secondary cultures, to the antigen used for absorption. Possible mechanisms of this specific effect are discussed.
Distribution of Ia‐Antigen‐like Molecules on Non‐lymphoid Tissues
Authors: FORSUM U. , KLARESKOG L. , PETERSON P. A.
Published: June 2006

Ia‐antigen‐like molecules are expressed on cells within several different non‐lymphoid tissues of the guinea‐pig. In indirect immunofluorescence analyses anti‐la‐Lintigen antibodies stained epithelial cells lining the intestinal tract, the bile ducts, the respimiory tract and the urinary tract. The rabbit antibodies against la antigens also stained the cells of the parotid and the submandibular glands. Evidence was also obtained suggesting that the reticuloepithelial cells of the thymus, like the Kupffer cells of the liver, express Ia‐antigen‐like molecules. In several cases indirect immunoprecipitation analyses and SDS‐polyacrylamide gel electrophoresis confirmed the immunoprecipitation studies inasmuch as Ia‐anligen‐like subunit with apparent molecular weights of 26,000 and 34,000 could be isolated from ihe non‐lymphoid organs.
Kinetics of the Agglutination of IgG‐coated Latex Particles by Clq: the Influence of Heat‐labile Serum Components
Authors: HÄLLGREN R. , STÅLENHEIM G. , VENGE P.
Published: June 2006

Interaction between human Clq and IgG coaled latex particles has been studied by means of a standard aggregometer equipment. A dose‐dependent agglutination was observed and 100 ng of Clq were readily detected. The kinetics of the agglutination was also monitored. Serum, partially puritied Cl, and high molecular weight fractions from Sephadex G‐200 fractionated serum produced agglutination only in the presence of EDTA. In the absence of this chclator these products disintegrated prerormed Clq‐IgG‐latex particle agglutinates. This disagglutin‐ating principle is heat‐sensitive and tentatively macromolecular Cl dependent. The most probable basis of the activity is the competition between Cl, with a high affinity for IgG particles, and Clq. The inability of Cl to induce particle agglutination might be causetl by the Cl subunits Clr and Cls sterically inhibiting the subunit Clq to bridge between the panicles.
Macrophage Requirements of PPD‐induced Blastogenic Factor Production by T and B Lymphocytes in the Guinea‐Pig
Authors: HOJO K. , HIRAMINE C.
Published: June 2006

An attempt has heen made to characterize the cell type which releases blastogenie ractor(s) (BF) on purified protein derivative (PPD) stimulation in complete Freund's atijuvant (CFA)‐sensitized guinea‐pig system and, further, to examine whether macrophages (Mφ) are required for antigen activation of T or B lymphocytes to produce this lyniphokine. Both T and B lymphocyte populations from guinea‐pigs 2–3 weeks after CFA sensitization (early‐stage T and B cells) failed to produce significant amounts of BF in response to PPD, whereas both the populations from 10 to 14 weeks after sensitizalion (late‐siage T and B cells) acquired the ability to produce BF by themselves. It is noteworthy that readded auiologous Mφ not only functioned well to augment BF production by the late‐stage T and B cells but also completely made up for the inability of the early‐stage T and B cells to produce BF. From these data it is reasonable to conclude that in the guinea‐pig sensitized T and B cells can both release BF on exposure to PPD and that Mtp either reconstitute or augment the BF synthetic response of T and B cells.
Heterogeneity and Mechanism of Action of Human Natural Killer Lymphocytes: Differential Distribution of Receptors for Helix pomatia Haemagglutinin (HP Receptors)
Authors: PAPE G. R. , TROYE M. , PERLMANN P.
Published: June 2006

Neuraminidase‐treated lymphocytes from the peripheral blood of normal human donors were fractionated on columns charged with Helix pomatia haemagglutinin (HP) coupled to Sepharose 4B. While lymphocytes lacking HP receptors (HP‐) passed directly through the column (fraction I), lymphocytes with HP receptors (HP+) were subsequently eluted in two distinct tractions with two different concentrations of the competitive hapten N‐acetyl‐D‐galactosamine (fraction II and III. respectively). The natural cytotoxicity of these lymphocytes to various tumour target cells (K562, T24, MANO, HCV29) was tested in a 51Cr release assay. Natural cytotoxicity was found in all three fractions recovered from the HP columns. In general, the cytotoxicity of the lymphocytes in fractions I and II was significantly enhanced over that of the unfractionated lymphocytes. Surface marker analysis and fractionation studies indicated that natural cytotoxicity in these target systems is exerted by both HP+ and HP‐ lymphocytes bearing Fc receptors for IgG. Since the HP receptor is considered to be a marker of T lymphocytes, the findings suggest that a significant fraction of these NK cells may be of T‐cell lineage. The surface marker profiles of these NK cells are very similar to those of antibody‐dependent K cells. Addition of Fab fragments of immunoadsorbent‐purified rabbit antibodies to human immunoglobulin inhibited the natural cytotoxicity of HP‐column‐fractionated lymphocytes to various degrees, indicating that pan but not all of it reflects antibody‐dependent K‐cell reactions. Since cytotoxicity in all three HP fractions was inhibitable in this way, the results suggest that immunoglobulin‐dependent natural cytotoxicity may be displayed by both HP+ and HP‐ effector cells.
Effect of IgM Fractions with or without Rheumatoid Factor Activity on Antigen‐Antibody Reactions
Authors: ALKNER U. , HANSSON U.‐B.
Published: June 2006

The effect of isolated IgM fractions, with or without rheumatoid factor (RF) activity, on reactions between human albumin and rabbit anti‐human albumin, human IgG and rabbit anti‐human IgG, and tetanus toxoid and human anti‐tetanus toxoid was assessed in the antigen excess zone. RF‐active fractions increased and RF‐negative IgM fractions decreased the amount of free antigen. The immune complexes that were affected by RF IgM fractions differed in composition from those affected by normal IgM fractions.
Stimulatory Effect of Human Alpha‐Fetoprotein and its Molecular Variants on in‐Vitro‐induced Lymphocyte Blastogenesis
Authors: SOUBIRAN P. , MUCCHIELLI A. , KERCKAERT J.‐P. , BAYARD B. , MASSEYEFF R.
Published: June 2006

Recent experiments suggested that alpha‐protein (AFP) may have immunoregulatory properties and play a role in protecting the fetus from rejection by the mother. The present study was performed to determine whether purified human AFP, its molecular variants separated on Lens culinaris agglutinin, and fetal serum albumin showed immunoregulatory activiiy on in vitro transformation of normal human lymphocytes by mitogens or by allogeneic cells. Human AFP and fetal serum albumin were purified from human whole fetuses by immunoabsorption, followed by acid elution. AFP variants (HLI, HL2, HL3, HL4) were separated by lectin affinity chromatography. The proteins were tested in lymphocyte cultures stimulated by concanavalin A, phytohaemagglutinin and pokeweed mitogen and in mixed lymphocyte cultures. Four batches of AFP and one batch of fetal albumin, in the concentration range of 0‐100 μg/ml and in one experiment up to 2 mg/ml, had a stimulatory effect on lymphocytes. One batch of human AFP showed inhibitory effect with low reproducibility. The variants had a stimulatory effect except for HL3, which was found to be slightly inhibitory in phytohaemagglutinin stimulated cultures when compared with cultures without AFP, but there was no significant difference when compared with cultures in which AFP was replaced by its dialysate. Thus, the results reported here are not consistent with the conclusion that human AFP has a significant immunosuppressive effect on man. Hypotheses about causes of discrepancies are analysed.
Restimulation in Secondary MLC by Autologous Non‐T Cells
Authors: ZIER K. S. , HUBER C. , BRAUNSTEINER H.
Published: June 2006

Lymphocytes sensitized in mixed leucocyte culture (MLC) were restimulaied with non‐T or complete cells. Cells primed against autologous non‐T cells and restimulated with autologous cells gave responses with the kinetics of a secondary response. In contrast, cells primed against either type of autologous cell responded to restimulation by allogeneic cells with the kinetics of a primary MLC response. Following priming against alloantigens good secondary responses against both complete and non‐T allogeneic restimulators were seen. Restimulation with auto‐logous non‐T colts, but not autologous complete cells, also led to secondary responses. The observation of” different responses to antigens expressed on non‐T autologous lymphocytes, in comparison to complete cells, implies that the secondary MLC involves more than a response against alloantigen. Allogeneic and autologous MLC may he qualitatively different phenomena. the latter representing the recognition of identifying structure's between interacting populations of cooperating cells.
Characteristics of Individual Polymorphonuclear Leucocyte Motility Obtained with a New Opto‐electronic Method
Authors: DAHLGREN C. , HED J. , MAGNUSSON K.‐E. , SUNDQVIST T.
Published: June 2006

An opto‐electronic device has been used for a quantitative assessment of the motility of individual polymorphonuclear leucocytes (PMNL) adhering to a glass cover slip. One of the oculars in a phase contrast microscope is provided with a mini‐array of 32 × 32 light‐sensitive elements. These are connected lo an electronic unit, capable of recording the number of light‐intensity changes on each element and of visualizing the path of a cell on an oscilloscope screen, as a pattern of dots. The results clearly show that individual PMNL respond differently to environmental conditions; for instance, (ij raising the temperature increased the motility of cells to a maximum at around 39°C and lowering the temperature from 42°C restored their peak motility. (ii) protein was required at attachment depending on the temperature at attachment, (iii) endo‐toxin‐activated normal human serum affected more drastically cells with a low initial motility and cytochalasin B more adversely influenced cells with a high initial motility. (iv) phagocytosis of yeast cells reduced the percentage of motile ceils, which was more pronounced if the PMNL were washed before the mobility measurement. The average motility of the PMNL was also diminished, although individual PMNL retained normal activity after ingestion of one or more yeast cells.
Induction of Autologous Reactivity of Human NK Cells
Authors: TARKKANEN J. , TIMONEN T. , SAKSELA E.
Published: June 2006

‘Mature’ human natural killer cell activity could be enriched by adsorption‐elution with fetal fibroblast as adsorbents against normal allogeneic skin fibroblasts, but not against autologous targets. However, when the natural killer activity was augmented by contact with tumour cells (HeLa), autologous and allogeneic skin fibroblast targets were killed without preference. Adsorption‐elution with augmenting target cells as adsorbents resulted in an efficient enrichment of NK activity showing non‐discriminate cytotoxicity Morphologically, this was associated with an enrichment of large granular lymphocytes previously shown to be responsible for human NK activity. We conclude that the NK activity of human‘mature’NK cells shows a relative autologous exemption, whereas‘pre‐NK’cells augmented to full activity in vitro are non‐discriminative.
Lymphoid Stem Cells in the Intraembryonic Mesenchyme of the Chicken
Authors: LASSILA O. , ESKOLA J. , TOIVANEN P. , DIETERLEN‐LIÈVRE F.
Published: June 2006

Chromosomally marked cells from the 7‐day intraembryonic mesenchyme were transplanted into 14‐day‐old irradiated chick embryos. At the age of 6 weeks donor‐derived T and B lymphocytes were shown to be present in the thymus, spleen and bone marrow, indicating that cells in the 7‐day intraembryonic mesenchyme are capable of developing into functional T and B lymphocytes. In addition to the sex chromosome marker, IgG allotype was used as a marker; the results demonstrate that cells from intraembryonic haemopoietic sites develop into mature IgG‐producing cells. In similar experiments, the 7‐day yolk sac also proved to contain lymphoid stem cells. Since lymphoid cell progenitors are not present in the 2‐day yolk sac, as has been shown previously in the yolk sac‐embryo chimaeras and since circulation is established from day 2 of incubation onwards, lymphoid stem cells present in the 7‐day yolk sac are most likely secondary immigrants originating in the intraembryonic mesenchyme.
Fine Specificity and Cytolytic Activity of Continuously Growing Alloreactive Cytotoxic T Lymphocyte Clones
Authors: SCHAWALLER R. , RÖLLINGHOFF M. , WAGNER H.
Published: June 2006

The role of Lyt 1+ T‐cell‐drived secondary CTL inducing factor allowed the cloning of alloreactive cytotoxic T lymphocytes (CTL) by the limiting dilution approach. Several monoclonal cell lines were established in vitro. The lytic activity of some of the cell lines exceeded that of CTL from bulk cultures; that is, 50% of the target cells were lysed at an effector to target cell ratio of 0.04:1. The fine specificity of individual CTL clones is discussed.
Reactivity of Human Lymphoid and Lymphoblastoid Cells with Peanut Agglutinin: Detection of a Blood Cell Subset which Lacks Detectable Membrane HLA
Authors: BALLET J.‐J. , FELLOUS M. , SHARON N. , REISNER Y. , AGRAPART M.
Published: June 2006

The number of cells reacting with flurescein‐labelled peanut agglutinin varies widely among human lymphoid organs. By separation techniques, reactive cell from tonsils were characterized as E‐negative, non‐immunoglobulin‐bearing lymphocytes. In peripheral blood, the lectin reacted with a small percentage of lymphoid and monocytoid cells which lacked HLA membrane determinants but expressed the β2‐mieroglobulin antigens. The tatter phenotype was also found on human lymphoblastoid cell lines. Lectin binding thus provides an effective means of characterizing these cell subsets.
An Evaluation of the Significance of Amino Acid Sequence Homologies in Human Histocompatibility Antigens (HLA‐A and HLA‐B) with Immunoglobulins and Other Proteins, Using Relatively Short Sequences
Authors: STROMINGER J. L. , ORR H. T. , PARHAM P. , PLOEGH H. L. , MANN D. L. , BILOFSKY H. , SAROFF H. A. , WU T. T. , KABAT E. A.
Published: June 2006

A computer search was carried out for homologies between HLA‐A and HLA‐B antigen sequences and the sequences of constant and variable regions of immunoglobulins and of all other sequenced proteins. Searches were made both with relatively short peptide sequences from the HLA antigens and with those longer peptide sequences which were available in 1978. Significant homology of HLA antigen sequences to immunoglobulin constant region sequences was found in two cases: (1) a short decapeptide sequence which includes the fourth cysteine residue of HLA‐B7 and (2) an 89‐amino‐acid residue (Ac‐2) C‐terminal fragment of the papain‐solubilized HLA‐B7 molecule. The difficulty of establishing statistically significant sequence homology with relatively short peptide sequences is emphasized by computer‐based comparisons of the decapeptide sequence with randomly generated peptide sequences. It is concluded that statistically significant homology with short sequences can be assured only when extraordinarily high degrees of homology are present and additional constraints are included in the matches, for example, matches at relatively rare amino acid residues such as Cys, His and Trp. The homology of the 89‐amino‐acid residue sequence to constant region sequences of immunoglobulins is as great as or greater than that of β2‐microglobulin. These findings and the unique domain structure involving a disulphide loop of comparable size strongly favour a common evolutionary origin for this region of HLA‐A and ‐B, β2‐microglobulin and immunoglobulin constant regions.
Decreased Gut‐associated IgA Levels in Patients with Typhoid Fever
Authors: KAUL M. N. , MISRA R. C. , AGARWAL S. K. , SAHA K.
Published: June 2006

Gut and serum immune responses were studied in twenty‐two patients with bacteriologically proved typhoid fever at different stages of the illness and six volunteers after parenteral immunizations with heat‐killed typhoid vaccine. Whereas the former group had some specific antibodies in their intestinal aspirates and significant specific antibody responses in their sera, the latter could only mount a serum antibody response. Moreover, the intestinal IgA levels in the patients remained significantly decreased throughout the whole course of the illness, but a rise of IgG and IgM was observed in their jejunal secretions. In contrast, the levels of all the three major classes of serum immunoglobulins IgG, IgM and IgA were raised in the patients, as opposed to the volunteers, in whom only IgG and IgM levels were increased. It was inferred that the subjects with deficit of intestinal IgA, and thus lacking protective mucosal barrier, are more prone to develop typhoid fever.
Degradation of Amyloid‐related Serum Protein SAA by a Component Present in Rabbit and Human Serum
Authors: SKOGEN B. , NATVIG J. B. , BØRRESEN A. L. , BERG K.
Published: June 2006

After incubation of protein SAA‐containing rabbit serum at 37 C overnight, the strength of the precipitation reaction against antiserum to SAA was decreased. This was at least partly due to enzymatic degradation of protein SAA. The enzymatic activity was not strongly associated with the SAA‐high‐density lipoprotein (HDL) complex, since it segmented in the preparative ultracentrifuge at a density at which the SAA‐HDL complex floats. The degrading component was obtained in concentrated form from rabbit and human sera by adsorption to Sepharose 4B. Degradation of human SAA by the human serum component was inhibited with diisopropyl fluorophosphate, an inhibitor of serine proteases.
Mitogenic Responsiveness of Human T‐Lymphocyte Subpopulations: Regulation by Suppressive Fc‐Receptor‐bearing T Cells and Influence of Fractionation Procedures
Authors: DILLNER‐CENTERLIND M.‐L. , HELLSTRÖM. U. , ROBERTSSON E.‐S. , HAMMARSTRÖM S. , PFRLMANN P.
Published: June 2006

The proliferative response induced by leucoagglutinin (La) in different subpopulations of human I lymphocytes was studied Subpopulations enriched in cells with either high‐ or low‐avidity receptors for sheep erythrocytes (SRBC) (EH.+, Et+) were prepared by sequential E‐rosetting. In addition, T lymphocytes prepared by E‐rosetting under optimal conditions (E+ror) were fractionated on wheat germ agglutin (WGA)‐Sepharose columns, rendering fraction enriched in lymphocytes with either low‐ or high‐avidity receptors for WGA. The T lymphocytes were found to comprise at least three functionally distinct subpopulations, differing with respect to mitogen responsiveness. Cell characterized by high‐avidity receptors for WGA and SRBC were highly responsive to La simulation, regardless of the method used for purification. In contrasts cell with low‐avidity receptors for WGA and probably also for SRBC hut lacking Fe receptors for IgG responded only marginally but were conditioned to respond when subjected to E‐rosetting under optimal conditions. This response was suppressed by lymphocytes with Fc receptors for IgG, which probably also had low‐avidity receptors for WGA and SRBC. The lymphocytes with high‐avidity receptors for WGA and SRBC did not appear to be susceptible to suppression by Fcγ+ cells.
Interaction of Human Polyclonal IgE and IgG from Different Species with Protein A from Staphylococcus aureus: Demonstration of Protein‐A‐reactive Sites Located in the Fab2 Fragment of Human IgG
Authors: INGANÄS M. , JOHANSSON S. G. O. , BENNICH H. H.
Published: June 2006

The peptic fragments F(ab′)2 and Fc″ of human polyclonal IgE were tested by the SpA‐IgE radioimmunoassay for interaction with protein A from Staphylococcus aureus. The Fab″2ɛ fragment but nut the Fc″ɛ fragment retained the ability to react with protein A. IgG preparations from different species were tested for their capacity to inhibit the reaction between 125I‐IgE and protein A. IgG preparations from man and the dog, pig, guinea‐pig and rhesus monkey were all potent inhibitors whereas those from the rabbit, rat, sheep, horse, cow, goat and mouse were very poor inhibitors. The inhibition of the binding of 125I‐IgE to protein A by Fab′2γ fragments, but not by Feγ fragments, suggests a novel type of interaction of protein A with immunoglobulins.
Study of the Mechanism for in Vitro Activation of Mouse NK Cells by Interferon
Authors: SENIK A. , KOLB J. P. , ÖRN A. , GIDLUND M.
Published: June 2006

The enhancement of the lytic capacity of mouse splenic ‘natural killer’ (NK) cells by interferon has been studied in vitro as a model for NK cell differentiation from inactive immediate precursors. We show that the increased cytotoxicity is a function of interferon concentration, and that two stages in the NK cell differentiation pathway can be distinguished. The first, very brief, can he performed at 0°C and without protein synthesis and probably corresponds to the fixation of interferon on its cell surface receptors. The second, resulting from the inductive signal given by interferon, proceeds for several hours and requires both RNA and protein synthesis. Our results also indicate that target cells for interferon‐induced cytotoxicity are cells for interferon‐induced cytotoxicity arc cells with ‘null’ characteristics, similar to the mature NK cells. Finally, they suggest that no soluble Intermediary factor other than interferon is involved in the enhancing of NK cell cytotoxicity.
A Gene of the Immunoglobulin H‐Chain Cluster Controls the Murine Antibody Response to Pneumococcal Polysaccharide Type 14
Authors: MÄKELÄ O. , PASANEN V. J. , SARVAS H. , LEHTONEN M.
Published: June 2006

Mice of various strains were immunized with pneumococcal polysaccharide type 14 (pneumo‐14), and their anti‐pneumo‐14 antibodies were measured by the Farr test. Mice of strains BALB/c, ST/b, NZB and CBA (Ig allotypes a, e or J) had 300–1700 ng of antibody nitrogen per millilitre of serum on day 7. The corresponding values for C57BL/Ka, RF or AKR mice (allotypes b, c or d) were 40–300 ng/ml. Two families of congenic strains were tested, one with the C57BL and the other with the BALB/c background genome. Their response was either high or low depending on the VH genes, and other gene loci had little effect on the concentration of anti‐pneumo‐14 antibodies.
Activation of Human Peripheral Blood Lymphocytes: Effect of Concanavalin A and Lipopolysaccharide on in Vitro Synthesis of DNA and Immunoglobulins
Authors: NESPOLI L. , VITIHLLO A. , MACCARIO R. , LANZAVECCHIA A. , UGAZIO A. G.
Published: June 2006

We studied the Interaction of lipopolysaccharide (LPS) and concanavalin A (Con A) with regard to IgM and IgG production in in vitro cultures of human peripheral blood lymphocytes (PBL) In our system LPS alone over a wide range Of concentration did not stimulate detectable IgM or IgG production, while Con A at optimal (6 μg ml) and suboptimal (0.6 μg/ml) mitogen concentration induced synthesis of small amounts of Ig. A marked enhancing effect was present when both Con A and LPS were added to the cultures. The different doses of LPS had similar effect on both classes of Ig, and typical dose‐response curves were obtained. To evaluate the cellular basis of this synergism, the effect on cell proliferation was studied undo identical experimental conditions in normal subject and patients with X‐linked agammaglobulinaemia (X‐LA). Parallel cultures were set up after monocyte depletion by adherence on petri dishes. On day 3, increasing doses of LPS were associated with progressive decreases in 3H‐thymidine (3H‐TdR) Incorporation. Similar result were obtained with normal lymphocytes and those from X‐LA patients. Monocyte depletion did not substantially after the lymphocyte response pattern. The preferential Induction of helper activities, either directly by helper stimulation or indirectly by suppressor minimum, is suggested us a possible mechanism of the Interaction observed.
A Synthetic Hexadecapeptide Derived from Allergen M Imposing Allergenic and Antigenic Reactivity
Authors: ELSAYED S. , TITLESTAD K. , APOLD J. , AAS K.
Published: June 2006

A solid phase synthesis (SPPS) hexadecapeptide corresponding to residues 49–64 of allergen M from cod was found to bind specifically IgE antibodies from sera of cod‐allergic individual. The SPPS peptide was similarly reactive in IgG‐mediated reactions; it could interfere with Allergen M line precipitate in rocket‐line immunoelectrophoresis, giving a distinct deflection. It is the only reported synthetic polypeptide imposing allergenic and antigenic reactivity. The hexadecapeptide is composed of two homologous tetrapeptides located at its termini, tentatively concluded to be mutually critical for the immunological specificity. The data obtained confirmed the predicted reactivity of the Ca2+‐binding CD loop of the intact Allergen M.
The Induction of Suppressor T Cells by Concanavalin A is Independent of Cellular Proliferation and Protein Synthesis
Authors: SHAND F. L. , ORME I. M. , IVANYI J.
Published: June 2006

Factors that govern the induction of suppressor T cells after stimulation with concanavalin A (Con A) were investigated in a two‐stage culture system. Normal mouse spleen cells were incubated with Con A in the presence of a variety of drugs and then assayed for suppressive activity by means of a secondary anti‐sheep erythrocyte response in vitro. The inclusion of inhibitors of mitosis (vinblastine sulphate or mytomycin C) or protein synthesis (cycloheximide or pactamycin) into normal spleen cell cultures containing Con A tailed to inhibit the subsequent development of suppressor cells. Furthermore, spleen cells from mice previously irradiated with 900 rad or injected with cyclophosphamide expressed a level of suppressor activity after Con A stimulation which was equivalent to that of normal spleen cells. However, the inclusion of drugs that inhibit microtubule or microfilament function (colchicine or cytocholasin B) did prevent suppressor cell induction. Kinetic studies also revealed that significant suppressor activity was detectable in normal spleen cells after only 3 h exposure to Con A. These results indicate that the induction of suppressor T cells in this system is a maturation event involving changes in the cell membrane and is entirely independent of protein synthesis and cellular proliferation.
The Antigens of Mycobacterium bovis, Strain BCG, Studied by Crossed Immunoelectrophoresis: a Reference System
Authors: CLOSS O. , HARBOE M. , AXELSEN N. H. , BUNCH‐CHRISTENSEN K. , MAGNUSSON M.
Published: June 2006

A reference system for the soluble antigen of Mycobacterium bovis, strain BCG is described. The system is based on crossed immunoelectrophoresis with intermediate gel. A Commercially available immunoglobulin preparation made from rabbit anti‐BCG hyperimmune serum was used as reference antibodies, while a concentrated BCG culture filtrate was used as reference antigen. The pattern obtained was highly reproducible, and most of the components were stable when the fiftyfold‐concentrated culture filtrate was stored in –20°C. About thirty different antigenic components were selected as reference antigens and numbered. The majority of the reference antigens was present in extracts prepared from BCG by ultrasonication or bacterial press extraction. Use of the system for studies of antigenic relationship between mycobacteria, identification and quantification of antigens, and characterization of antimycobacterial antibodies are illustrated by examples. The antigens of two preparations of tuberculin purified protein derivative (PPD) were identified. The antigen designated BCG60 was found to be a major constituent of tuberculin PPD. Evidence is presented that this antigen is cell wall associated.
Method to Prove Ingestion of Particles by Macrophages with Light Microscopy
Authors: VAN FURTH R. , DIESSELHOFF‐DEN DULK M. M. C.
Published: June 2006

Research on phagocytosis is often hampered by the inability to distinguish whether (opsonized) particles have been ingested by phagocytes or are only attached to the surface of these cells. Treatment of the cells after phagocytosis to remove all extracellular particles makes it possible to evaluate phagocytosis with certainty by light microscopy. Opsonized erythrocytes attached to the macrophage surface are usually removed by hypotonic lysis. The present report describes the advantages of the use of lysostaphin to lyse Staphylococcus aureus and of xylene, chloroform or dioxane to dissolve polystyrene latex beads on the surface of peritoneal macrophages and embryonic fibroblasts. This procedure facilitates differentiation between professional and facultative phagocytes.
Cross‐reacting Xenoantibodies to the Heavy Chain of H‐2 and HLA‐A, B Antigens: Serologic and Immunochemical Characterization
Authors: CALLAHAN G. N. , PELLEGRINO M. A. , FERRONE S.
Published: June 2006

H‐2 cross‐reactive antibodies present in HLA xenoantisera were purified by absorption on and elution from murine cells. Antibodies isolated from one of these antisera. 78‐E48, were found to mediate complement‐dependent lysis of both human and murine lymphoid cells but not of Daudi cells or of human lymphoid cells coaled with Fab2 fragments from a cow anti‐human β2‐microglobulin (β2m) antiserum. In indirect immunoprecipitation analyses 78‐E48 reacted only with proteins of approximately 45,000 and 12,000 MW present in NP‐40 extracts of both human and murine lymphoid cells. Sequential precipitation experiments with rabbit anti‐human β2m and allo‐anti‐H‐2Kk, Iak sera established that these proteins were in fact H‐2 and HLA‐A, B antigens. It was also found that 78‐E48 reacted only with the heavy chain of HLA‐A, B and H‐2 antigens, since this eluate was unreactive with β2m in a radioimmunoassay, and its capacity to immunoprecipitate the 45,000 and 12,000 MW proteins from human cell extracts was unaffected by prior reaction with purified human β2m. These data show for the first time that H‐2 and HLA‐A, B antigens share properties that probably depend upon their tertiary structure.
Separation and Functional Analysis of Subpopulations of T Cells in the Bone Marrow of the Mouse
Authors: RÖPKE C.
Published: June 2006

Functional capacities of T‐cell precursors have been evaluated by cell culture experiments after fractionation of bone marrow cells from normal and nude mice by 1 g velocity sedimentation. The results indicate that the T‐cell precursors in the bone marrow are cycling cells, that T‐cell precursors in the bone marrow of nude mice have more limited potentials than those of normal mice, and that both Thy‐1.2‐positive and Thy‐1.2‐negative precursors are stimulated by T‐cell activators.