bluuebunny/crossref_metadata_2025 huggingface dataset:
Establishment of Anti‐TNP Antibody‐producing Human Lymphoid Lines by Preselection for Hapten Binding followed by EBV Transformation
Authors: KOZBOR D. , STEINITZ M. , KLEIN G. , KOSKIMIES S. , MÃKELÄ O.
Published: June 2006

Human lymphoblastoid cell lines that produce specific antibody against the haplen trinitrophenyl (TNP) have been established by selecting TNP‐binding human B lymphocytes by TNP‐rosetting and Ficoll‐lsopaque separation, followed by Epstein‐Barr virus (EBV) immortalization. Derived lines secreted polyclonal anti‐TNP antibodies and contained relatively small numbers of specific rosette‐ and plaque‐forming cells against TNP‐RBC. Following rerosetting with TNP‐RBC, the frequency of rosette‐forming cells increased from 2% to 75%. In parallel, the frequency of plaque‐forming cells increased from 0.4% to 30%. The antibody titres in the supernatants increased from 64 to 512 and from 48 to 192, as measured by TNP agglutination and haeniolytic assays, respectively. The antibodies were I9S, IgM. The specificity of the anti‐TNP antibody was confirmed by the hapten inhibition test, in comparison and cross‐reactivity tests with the supernatant of the previously established. EBV‐transformed anti‐4‐hydroxy‐3,5‐dinitrophenacetic acid (NNP) antibody‐producing cell line. Both antibodies were specific: the homologous hapten inhibited them but the heterologous hapten did not.
Human T‐Lymphocyte Subpopulations: Alterations in Systemic Lupus Erythematosus
Authors: MORETTA A. , MINGARI M. C. , SANTOLI D. , PERLMANN P. , MORETTA L.
Published: June 2006

Sharply reduced proportions of T cells with Fc receptors for IgG (TG cells) were observed in blood samples from patients with systemic lupus erythematosus (SLE), mainly with active disease. This T‐cell subset has previously heen shown to be a suppressor in the pokeweed mitogen (PWM)‐dependent B‐cell differentiation. In contrast, the percentages of T cells with Fc receptors for IgM (TM cells), which have been shown to help inimunoglobulin production, were not different from those of normals. TG cells present in the circulation of SLE patients were analysed for their functional capacities in antibody‐dependent cell‐mediated cytotoxicity and in the suppression of a PWM‐induced B‐cell differentiation. In both these assays TG cells from SLE patients had normal effector cell activity. This suggests that the T‐cell defects present in SLE patients are mainly of a quantitative rather than a qualitative type.
Rat Memory T Lymphocytes
Authors: BURCKHARDT J. J.
Published: June 2006

Protein‐coated silica, a macrophage toxin, was used to assess the requirement for accessory cells in the induction of an in vitro proliferative response to (i) antigens from Actinomyces viscosus and (ii) the mitogens conconavalin A (Con A) and phytohaemagglutinin (PHA). T cells were obtained from RIC‐Sprague‐Dawley rats primed in vivo with A. viscosus Nyl by splenectomy and filtering the spleen cell suspensions through Degalan Ig‐anti‐IgG columns. In the presence of 100 mTg silica/ml during 4 days of culture, the proliferative response of T lymphocytes was not diminished. In contrast, when the T cells were precultured with silica for 24 h, washed, and subsequently cultured with the antigen fractions, antigen‐induced proliferation was abolished. This procedure, however, had no influence on mitogen‐induced T‐cell activation. It is therefore concluded that the antigen‐dependent anamnestic in vitro response (but not activation by mitogens) of rat T lymphocytes needs help from silica‐sensitive macrophages.
Lymphocyte Subpopulations in Man: B‐Cell Stimulation Induced by Pokeweed Mitogen and Irradiated T Cells
Authors: JOHNSEN H. E. , MADSEN M.
Published: June 2006

The enhanced stimulasion of human B lymphocytes by pokeweed mitogen in the presence of irradiated T helper lymphocytes has been studied, revealing that the proliferative responses measured hy incorporation of thymidine in cultures of B lymphocytes and irradiated T lymphocytes in a 1:2 or 1:4 ratio is mosily a function of the B cells. Only a minimal number, if any, of the T cells contaminating the B cell suspensions is stimulated to proliferation. This is in contrast to stimulation ofihe B‐cell suspensions without addition of irradiated T cells, where both B cells and T cells proliferate. The irradiated T helper cells have no FcR for antigen‐bound IgG, and as well allogeneic as aucologous T ceils exhibit helper capacity of equal strength. The test system described makes it possible selectively to lest one B‐cell function and the corresponding T helper capacity.
Characterization of IgD
Authors: CORTE G. , VIALE G. , COSULICH E. , BARGELLESI A. , FERRARINI M.
Published: June 2006

Endogenous and surface labelling techniques were used on human lymphoid cells to characterize intracytoplasmic, membrane and secreted IgD. IgD synthesized by lymphocytes and inserted into the cell membrane displayed a single molecular form with the same mobiliiy in sodium dodecyl sulphate polyacrylamide gel eleclrophorcsis (SDS‐PAGE) as the previously described slow migrating serum IgDl. Plasma cells produced and secreted IgD1 and another IgD corresponding to the faster‐moving serum IgD2. Conversion of one molecular form into the other was never observed, thus indicating that neither molecule is a precursor or a degradation product of the other.
Cross‐reactive Cytotoxic T Cells to Alphavirus Infection
Authors: MULLBACHER A. , MARSHALL I. D. , BLANDEN R. V.
Published: June 2006

Secondary Tc cells immune to alphaviruses (BEB, SIN and SFV) cross‐react between serologically defined subgroups at the level of target lysis and at the level of induction of response. Despite this apparently complete Tc cell cross‐reactivity between BFB, SIN and SFV, amisera raised against BEB and SIN showed virus specificity in their ability to block Tc cell‐mediated lysis of alpha virus‐infected targets. This result suggests that Tc cells recognize the same viral antigen molecule as antibodies, but with less specificity. Other possible interpretations are discussed.
The Effect of Specific Antibody on the Generation of Cytotoxic T Lymphocytes and the Recovery of Mice from Influenza Virus Infection
Authors: YAP K. L. , ADA G. L.
Published: June 2006

A study has been made of the effect of humoral antibody on the generation of specific cytotoxic T cells (Tc) in the spleen or lungs after intravenous injection or intranasal inoculation of infectious influenza virus. Antibody injected before or at the same time as virus inhibited completely the generation of Tc in the spleen. If injected 1 h after virus, the inhibition was reduced by 50% and little inhibition occurred if antibody was injected 6 h after virus. This suggested that antibody failed to influence Tc generation once infection of stimulating cells had occurred. Antibody injected intravenously 24 h after intranasal inoculalion of virus into normal mice did not affect the level of cytotoxic activity present in the lungs, and tracc amounts only (> 1 log10 EID5–0) could be recovered from the lungs at 6 days. As there is a high litre (>6 log10 EID5–0) of infectious virus in the lungs of mice 24 h after infection, this represents a very efficient control mechanism. The same protocol carried out with athymic mice gave only a partial clearance (e. 3 log10 EID50) of virus in the lungs. It was suggested that a major role of humoral antibody was to limit infection by the virus, and in this respect it complemented the action of Tc.
Comparison of the Binding of Radiolabelled Human IgG and Fc Fragments to Murine Spleen Cells
Authors: MORGAN E. L. , SPIEGELBERG H. L. , WEIGLE W. O.
Published: June 2006

The binding properties of an IgG I human myeloma protein, normal IgG, and the Fc and Fab fragments of each were compared in cultures of murine spleen cells. Both 125I‐labelled IgG and Fc fragments bound to splenic lymphocytes, whereas Fab fragments did not bind significantly at the highest concentrations tested. On a molar basis, more Fc bound than intact IgG. According to Scatchard plot analysis, the affinity constant of IgG1 was 1.5± 106± 1±105 L/M and that of the Fc fragments was 7.8 ± 105± 2.6±105 L/M. Approximately 25,000 binding sites/cell were calculated for IgG1 and 102,000 for Fc. Deaggregation of the Fc preparation did not change these values, suggesting that the difference in binding of IgG and Fc did not result from Fc aggregation. Unlabelled IgG inhibited about 25% of the labelled Fc binding, whereas unlabelled Fc inhibited approximately 80% of the labelled Fc binding. IgG antigen‐antibody complexes, however, inhibited 75% of the Fc binding. In the reciprocal experiment both intact IgG and Fc inhibited the binding of labelled IgG by 100%. The major cell population that bound IgG and Fc fragments in the spleen cell preparation were the B lymphocytes. Removal of macrophages did not significantly affect the binding of labelled Fc fragments. In addition, T‐cell‐enriched populations bound an insignificant quantity of Fc fragments.
Dissociation of the Humoral and Cell‐mediated Responses to Alloantigens in Mice by Sublethal Whole‐Body Irradiation
Authors: ENGERS H. D. , LOUIS J.
Published: June 2006

Spleen cells from mice exposed to 500 rad sublethal whole‐body irradiation before immunization with 100 ± 106 allogeneic spleen cells exhibited an enhanced cytolytic T‐lymphocyte (CTL) activity. In contrast, this treatment completely abrogated the humoral alloantibody response, as measured by complement‐mediated lysis. In addition to the observed dissociation between primary humoral and cell‐mediated responses, the character of the in vitro secondary CTL response of spleen cells obtained from mice sublethally irradiated before alloimmunization was also modified. Although these spleen cells were perfectly capable of mounting a secondary response upon restimulation in vitro with intact allogeneic spleen cells, they were no longer capable of responding to subcellular alloantigen preparations. These results suggest that the cellular requirements for the induction of secondary CTL responses in vitro depend upon the nature of the antigen used for restimulation.
Induction of Suppressor Activity on B‐Cell Differentiation in Human T‐Cell Subset without Fc(IgG) Receptors by Levamisole Administration
Authors: MORIYA N. , MIYAWAKI T. , SEKI H. , KUBO M. , NAGAOKI T. , OKUDA N. , TANIGUCHI N.
Published: June 2006

A single oral dose of 150 mg levamisole was administered to five healthy adults. Circulating Fc(IgG) receptor‐bearing T cells (Tγ cells) increased for 5 days after levamisole intake, but total E rosette‐forming cells showed no significant alterations. The generation of immunoglobulin‐producing cells in the peripheral blood lymphocytes (PBL), which was induced in the in vitro pokeweed mitogen (PWM)‐stimulated cultures, was significantly suppressed for 5 days after levamisole administration. Suppressor T‐cell activity on B‐cell differentiation, which was induced by levamisole intake, was evaluated by co‐culturing with allogeneic untreated adult PBL in the PWM system in six other volunteers. A seemingly dose‐dependent suppression on B‐cell differentiation was exerted by T cells isolated on day 3 of levamisole treatment, but not by T cells which were isolated before or on day 14 of the experiment. When T cells were fractionated into two subsets with regard to the presence or absence of Fc(IgG) receptors, suppressor T‐cell activity appeared to be generated by levamisole largely in T cells lacking Fc(IgG) receptors, but not in Tγ cells.
Bacterial Lipopolysaccharides Bind Selectively to Lymphocytes from Lipopolysaccharide High‐responder Mouse Strains
Authors: NYGREN H. , DAHLÉN G. , MÖLLER G.
Published: June 2006

Three different concentrations of horseradish peroxidase‐labelled lipopolysaccharride (LPS‐HRP) were added in vitro to spleen cells from the LPS high‐responder strain C3H/Tif and to cells from the low‐responder strain C3H/HeJ. After being washed and fixed the cells were exposed to the substrate and prepared for electron microscopy. After addition of 7 and 0.7 μg/ml of labelled LPS only lymphocytes from the high‐responder strain were labelled. About 5–10% of the cells from C3H/Tif bound LPS. which is in accordance with the known frequency of B cells possessing the genetically determined LPS receptor. At the highest dose of labelled LPS (70 μg/ml) a large proportion of lymphocytes from the low‐responder strain also bound LPS. Erythrocytes from both strains bound LPS at all concentrations. It is concluded that LPS‐HRP allows the detection at the cellular level of LPS binding to the genetically controlled membrane receptor for LPS.
The Fc Binding Regions in Protein A are not Responsible for the Polyclonal B Cell Activating Property of Protein A
Authors: SJÖDAHL J. , MÖLLER G.
Published: June 2006

To test whether the polyclonal B cell activating property of protein A is due to its ability to bind to the Fc part of immunoglobulins, experiments were carried out with purified monomeric or dimeric Fc‐binding fragments of protein A. Neither monomeric nor dimeric fragments induced polyclonal antibody synthesis in vitro, whereas intact protein A was a strong activator. It is concluded that the Fc binding part of protein A is not responsible for activation of polyclonal antibody synthesis.
Primary Structure of Pooled, Papain‐solubilized HLA‐A, ‐B, and ‐C Antigens
Authors: TRÄGARDH L. , RASK L. , WIMAN K. , PETERSON P. A.
Published: June 2006

The tentative amino acid sequence of pooled, papain‐solubilized HLA antigen heavy chains has been determined. The amino acid sequence comprises 273 residues. As the structural analyses were performed on HLA antigen heavy chains comprising a mixture of several allelic forms derived from the A, B, and possibly C loci, multiple residues were encountered in several positions. However, a quantitatively dominating residue could always be easily identified. The present data suggest that the amino acid variability of the HLA‐A, ‐B, and ‐C antigens is found in restricted regions of the molecule. The COOH‐terminal third of the HLA antigen heavy chain appears to be less variable than other regions of the molecule. Previous work has shown that the HLA antigen heavy chain contains two immunoglobulin‐like disulphide loops. The COOH‐terminal third of the heavy chain was shown to be similar in primary structure to β2‐microglobulin and the immunoglobulin G constant domains.
A Simple Method to Detect Complement Receptors using Baker's Yeast: YC Rosettes
Authors: RIVERO I. , ABACA H. E. , VALLÉS R. , VANNUCCI J. D. , DIUMENJO M. S. , MORAVENIK M. B.
Published: June 2006

A technique is described to identify complement‐receptor‐bearing cells, using serum‐treated baker's yeast as a ligand. The method consists of incubation of heat‐killed baker's yeasts with fresh AB normal serum, freezing, thawing, and washing of the particles, followed by mixing with the cells. Serum is required to coat the yeasts for the rosette formation. Experiments designed to establish the serum factors responsible for the attachment of the particles to cells show that heat inactivation, chelating agents, or anti‐C3 treatment prevent rosette formation. This is taken as evidence that yeasts (Y) are coated with complement (C) to compose the reagent for the YC rosette technique. The application of this technique to twenty‐five normal individuals demonstrated that a mean of 11.6 per 100 lymphocytes (±4.3) form rosettes; absolute number: 275 (±160) rosette‐forming lymphocytes per mm3. Either AB or autologous fresh serum can be used to coat the yeasts. A combined technique for YC plus E rosettes can be performed allowing the identification and enumeration of four populations of lymphocytes: (a) those having receptors for sheep erythrocytes, (b) complement‐receptor‐bearing lymphocytes, (c) those having both receptors (D lymphocytes), and (d) non‐rosette‐forming non‐phagocytic cells.
Functional Relationship of Macrophages and Basophils to the Thymus Gland
Authors: SCHULTE‐WISSERMANN H. , BORZY M. S. , ALBRECHT R. , HONG R.
Published: June 2006

Human thymus epithelium, depleted of thymocytes and macrophages by means of organ culture, was used in chemotaxis experiments with peripheral blood cells. Such cultured thymus epithelium can attract specifically macrophages and basophils. T‐lymphocytes were attracted only by short‐term (8 day) cultured thymus tissue which still retains some of the original macrophage population. Thymic macrophages formed rosette structures with thymocytes. In other experiments ‘activated’ rabbit macrophages had the capability to destroy thymocytes, whether autochthonous or allogeneic. The possible role of macrophages and basophils in thymus function is discussed.
Characterization of Human Lymphocytes Bearing Fc Receptors for IgE Isolated from Blood and Lymphoid Organs
Authors: HELLSTRÖM U. , SPIEGELBERG H. L.
Published: June 2006

Human lymphocytes isolated from adult peripheral blood and cord blood, tonsils, adenoids and spleens were analysed for Fc receptors for IgE (FcE) by rosette assays. The FcE+ cells were also characterized for their class of surface immunoglobulin (sIg), complement receptors, receptors for sheep erythrocytes (E), and Helix pomatia A haemagglutinin (HP), and their abilities to phagocytoze and adhere. The average number of FcE+ cells was in adult peripheral blood 1.2±0.4%, in cord blood 3.0±1.3%, in tonsils 4.2±5.2%, in adenoids 5.8±4.2%, and in spleens varied from 0.8% to 15.8% among individual patients. Overnight culturing of the lymphocytes under conditions that allowed detection of Fc receptors for IgM (Fcμ) usually lowered the number of FcE+ cells. Neuraminidase treatment caused no change. Rosette formation was inhibited by IgE myeloma proteins and their Fc fragments, but not by mildly reduced and alkylated and heated (56°C) IgE, indicating that the receptors are specific for the native configuration of the IgE Fc fragment. Double cell surface marker analyses with fluoresceinated F(ab')2 fragments of purified anti‐μ, δ, and γ antibodies used to label the FcE rosette‐forming lymphocytes from peripheral adult and cord blood showed that 50–80% were sIgM+ but only 0–28% were sIgD+. In contrast, approximately 80% of the FcE+ cells from tonsils, adenoids and spleens were sIgM+ and sIgD+. The FcE+ lymphocytes represented 10–20% of the sIgM+ lymphocytes in both the peripheral adult and cord blood. Depletion and enrichment experiments indicated that most of the FcE+ cells bear complement receptors. Lymphocytes having both E and FcE receptors were not found. Furthermore, the lymphocytes with HP receptors, a marker for T cells and immature B cells, were FcE−. Monocytes and neutrophil granulocytes did not form FcE rosettes. These data indicate that a minor population of human B lymphocytes have FcE receptors. The majority of the FcE+ lymphocytes in the blood differ from those in tonsils, adenoids and spleen in that the majority of the former are sIgM+/sIgD− and the latter sIgM+/sIgD+. The sIgM+/sIgD− FcE+ cells in the blood are probably relatively mature lymphocytes since they lacked HP receptors, which are found on immature B cells
Alternation of Low and High Affinities of Secreted and Cell‐bound Antibodies during the Anamnestic Response of Rabbits to Salmonella senftenberg Microorganisms
Authors: CHABY R. , GFRARD R. , DUFLOT E.
Published: June 2006

We have investigated at the cellular level the variations in affinities of secreted antibodies and of cell receptors during the anamnestic response of rabbits, by using an immunization schedule made up of administrations, over successive intervals of 40 days, of Salmonella senftenberg vaccine. Affinities of free and membrane‐bound antibodies biosynthesized at a given time were measured by inhibition of haemolytic plaque‐forming cells (PFC) and rosette‐forming cells (RFC) with spleen, lymph node, and peripheral blood cells. The immunodominant monosaccharide, methyl‐α‐D‐glucopyranoside (α‐Me‐Glc), was used as inhibitor, and the target cells were coated with the O‐specific polysaccharide of the immunogenie microorganisms. We observed that the monosaccharide concentration causing a 50% inhibition(I50) alternated between low and high values in both PFC and RFC assays. Furthermore, when sheep erythrocytes were coated with polysaccharides from other strains of Salmonella bearing the determinants for serotypes 1 or 3, the percentage ofcross‐reactive PFC obtained exhibited a reverse oscillating pattern of variation. These results demonstrated that affinity of secreted antibodies or of antigen‐receptors of cells, directed against different determinants of the antigenic polysaccharide, alternated between low and high values after each new vaccine administration. Possible mechanisms accounting for this phenomenon are discussed.
Studies on the Cytophilic Properties of Human β2‐Microglobulin
Authors: HESTER R. B. , KUBO R. T. , GREY H. M.
Published: June 2006

The mechanism by which exogenously added β2m binds to lymphoid cells has been explored. In the mouse it has been shown that β2m remains associated with plasma membrane macromolecules following solubilization with NP‐40 and that approximately 25–30% of the binding could be accounted for by binding to H‐2 antigens. No binding to mouse immunoglobulin or Ia antigens could be detected. The sites for binding of the remainder of the cell‐hound β2m were not determined. Whereas normal human lymphocytes showed little or no capacity to bind exogenously added β2m, it was found that phytohaemagglutinin (PHA)‐stimulated cells could bind β2m. This binding occurred optimally 2 days after PHA stimulation. Approximately half of the binding could be accounted for by binding to HLA antigens. The possible significance of these findings with respect to cellular interactions involving major histocompatibility complex gene products in the immune response is discussed.
Antibody Response in Rabbits to Mycobacterium bovis BCG
Authors: MSHANA R. N. , CLOSS O. , HARBOE M.
Published: June 2006

The specificity of the immune response after immunization with Mycobacterimn bovis BCG was studied by crossed immunoelectrophoresis with intermediate gel in a BCG/anti‐BCG system, in which the reaction against thirty distinct components of BCG was recorded. After a single injection of total sonicate of 3 mg (dry weight) bacilli, the antibody response was markedly similar in eight rabbits. The earliest and strongest response was directed against nine components of BCG; all but one of these belonged to the group of thirteen components that cross‐react extensively with other mycobacteria. After repeated immunization with sonicate from about 0.8 μg of BCG bacilli, five components still induced a marked antibody response. All but one of these components are among the most widely cross‐reacting BCG components, and the observations made after subsequent challenge with the higher dose of BCG indicate that low‐zone tolerance was induced against other components of the bacilli. The implication of these findings concerning formation of anti‐mycobacterial antibodies in normal individuals and during mycobacterial infection is discussed.
Characterization of Amyloid of Ageing Obese–Hyperglycaemic Mice and their Lean Littermates
Authors: WESTERMARK P. , SLETTEN K. , NAESER P. , NATVIG J. B.
Published: June 2006

Amyloid fibrits, isolated from 18‐month‐old obese–hyperglycaemic mice and their lean littermates, were characterized immunologically and chemically. The main amyloid fibril subunit protein was protein AA, which cross‐reacted completely with an antiserum against amyloid from mice with experimentally induced amyloidosis and had an amino acid composition and N‐terminal amino acid sequence identical to that protein. These results indicate that the spontaneously occurring amyloidosis in obese–hyperglycaemic mice and their lean littermates corresponds to human, secondary amyloidosis and may serve as a model for that disease.
Effects of Anti‐Guinea‐Pig β2‐Microglobulin Antibodies on Lymphocyte Transformation Induced by Specific Antigens or Mitogens
Authors: LÖGDBERG L. , CIGÉN R. , BERGGÅRD I.
Published: June 2006

The effects of a goat anti‐guinea‐pig β2‐microglobulin antiserum (aβ2m) on lymphocyte transformation, induced by specilic antigens or mitogens, were studied, aβ2m was found to exert inhibitory effects on antigen stimulation, with three different antigens (purified protein derivative, hen egg‐white lysozyme, and ovalbumin). and on stimulation induced by the ‘T‐cell mitogens’, concanavalin A and phytohaemagglutinin. and the ‘mixed mitogen’, pokewecd mitogen, Stimulation induced by the ‘B‐cell mitogens’, dextran sulphate and bacterial lipopolysaccharide, did not seem to he inhibited to the same extent by aβ2m. The inhibitory effects seemed specific, as they were not seen with aβ2m that had been absorbed on a column with insolubilized purified guinea‐pig β2m nor with normal goat serum. Time studies indicated that the inhibitions started already during the first day of culture.
Evidence of Similar Idiotypic Determinants on Different Rheumatoid Factor Populations
Authors: FØRRE Ø. , DOBLOUG J. H. , MICHAELSEN T. E. , NATVIG J. B.
Published: June 2006

Strong idiotypic antigens were detected both in monoclonal and polyclonal rheumatoid factors (RF) by applying idiotype‐specific antisera. Shared idiotypic antigens were found between some polyclonal and monoclonal RF as well as between certain IgG and IgM RF.
Peanut Agglutinin
Authors: LONDON J. , PERROT J. Y. , BERRIH S. , LAROCHE L. , NIAUDET P.
Published: June 2006

Peanut agglutinin (PNA) has previously been shown to be a ‘marker’ for early T subpopulations in mice. We have investigaied whether it could also be used in the study of human mononuclear cells. 50–60% of human thymocytes have binding sites for PNA. When separated on a discontinuous Ficoll gradient, the PNA‐positive thymocytes are found preferentially in the layers corresponding to the immunoincompetent cells. In the peripheral blood only 5% of the mononuclear cells are PNA‐positive and we have shown that these cells are monocytes. In tonsils 13% of PNA+ cells are found and they are mostly lymphocytes. Thus, PNA is a marker for some T cell subsets present in the thymus and tonsils.
IgM‐Rheumatoid Factors Cross‐reactive with IgG and a Cell Nuclear Antigen: Apparent ‘Masking’ in Original Serum
Authors: JOHNSO P. M.
Published: June 2006

Fourteen out of fifteen IgM‐rheumatoid factor (IgM‐RF) preparations isolated from high‐titred seropositive rheumatoid sera by affinity chromatography on immobilized human IgG have been shown to cross‐react with a cell nuclear antigen, as manifest by positive immunofluorescent antinuclear antibody (ANA) staining on rat liver and kidney tissue sections. The cross‐reacting ANA activity within isolated IgM‐RF preparations was inhibitabte by both human and rabbit IgG, as well as by isolated rat liver chromatin. and could be isolated from seropositive ANA‐negative sera. It would appear that this cross‐reacting ANA activity can be masked in the original seropositive serum and may often require purification of RF from serum for its manifestation.
Degradation of Thymidine to Thymine by Rheumatoid Arthritis Synovial Tissue Eluate
Authors: NYKÄNEN P. J.
Published: June 2006

A study was made of the effect of rheumatoid synovial tissue eluate on phylohaemagglutinin (PHA) stimulation of peripheral Mood lymphocytes. It was found that rheumatoid synovial tissue eluale caused marked inhibition of 3H‐thymidine incorporation by lymphocytes stimulated with PHA. Synovial tissue from traumatic joints had no effect. The inhibitor was a heat‐sensitive and nondialysable substance. The inhibitory effect on PHA stimulation was diminished by thymine. In thin‐layer chromatogruphy it showed the capacity to convert thymidine to thymine. Because ihymine is not incorporated by DNA‐synthesizing cells, this report explains why negative results arc obtained when lymphocyles are stimulated by rheumatoid synovial tissue using 3H‐thymidine incorporation as an indicator of lymphocyte activation.
Cytotoxic Effects of Activated Human Monocytes and Lymphocytes to Anti‐D‐treated Human Erythrocytes in Vitro
Authors: RANDAZZO B. , HIRSCHBERG T. , HIRSCHBERG H.
Published: June 2006

Incuhation of human monocylcs, derived from peripheral blood, with cell‐free supernalants from mixed lymphocyle cultures resulted in mcirpholngical and funelional changes in the mature macropliages. Activation of monocyle‐derived macrophages by these faciors resulted in a significant increase in their capacity to lyse anti‐D‐trealed human erythrticytcs. The tytic activity of both normal and activated macrophages appeared to be independent of eryihrophagocytosis. T lymphocytes activated by either allogeneic cells or ihe mitogen phytohaemagglutinin were not cytolytic lo treated eryihrocytes even at high effector to target cell ratios.
The Plasma Cell Differentiation Antigen PC.1 is Absent in CH3/Tif and Present in C3H/HeJ
Authors: GAUDERNACK G. , HANNESTAD K.
Published: June 2006

(C3H/Tif × DBA/2)F1 mice, immunized with viable BALB/c plasmacyloma MOPC315 cells, produce antibodies directed against a cell‐surface antigen. The strain and lissue distribution of this antigen was identical to that of the plasma cell differentiation alloantigen PC.1. The antigen is absent in the mouse strain C3H/Tif but is present in the closely relaled substrain C3H/HeJ. This is the third difference between surface structures of the B‐cell lineage of C3H/Tif and C3H/HeJ mice.
Phenotypes of ‘Null’ Lymphoid Cells in Human Blood
Authors: NIAUDBT P , GREAVES M. , HORWITZ D.
Published: June 2006

F(ab′)2 antibody fragments of heteroantisera directed againsi p28,33 (or ‘Ia‐like’ antigen, T‐cell antigen(s) (T), myeloid antigen (M), and immunoglohulin have been used along with rosetting techniques to analyse the antigenic heterogeneity of ‘null’ or ‘unclassified’ lymphoid cells purified ‘negaiively’ from human blood by E rosette depletion of T ceils and B removal on an anti‐immunoglobulin immunoabsorbent. The results confirm that considerable heterogeneity exists wilhin the ‘null’ cell population: the majority of cells are p28,33+ and Fcγ‐receptor‐positive, and a proportion but not all of these p28,33+, Fcγ+ cells are also C3‐receptor‐positive but negative for the other heteroanligens (T and M), 2–6% were ‘pre‐B’ by the criteria of immuno‐fluorescent staining for cytoptasmic IgM in the absence of detectable cell surface Ig. 1–3% stained intensely with anti‐IgM. anti‐IgG or anti‐IgA, or anti‐κ plus anti‐λ and were presumed to be plasma cells or their immediate precursors. No null cells siained wiih antibody to terminal deoxynucleotidyl transferase enzyme, a marker for early T lineage cells, and less than 1% expressed an acute (‘non‐T, non‐B’) lymphoblastic‐leukaemia‐associalcd membrane antigen which is believed to be an ‘early’ lymphoid lineage differentiation marker. A minor subpopulation made sheep ‘E’ rosettes, and another made mouse ‘E’ roseites afier neuraminidase preireatment of the lymphoid cells. A small proportion of ‘lymphoid’, ‘null cells’ were reactive with an anti‐myclomonocytic (M) antibody. It is suggested that ‘null’ lymphoid cells in blood are heterogeneous and that most are not immature T or B precursors but either a relatively mature B lymphocyte population with a very low density of membrane immunoglohulin or a distinct non‐T, non‐B cell type.
In Situ Effector Mechanisms in Rat Kidney Allograft Rejection: III. Kinetics of the Inflammatory Response and Generation of Donor‐directed Killer Cells
Authors: HÄYRY P. , VON WILLEBRAND E. , SOOTS A.
Published: June 2006

We have isolated and identified the infiltrating inflammatory cells from rejecting rat kidney allografts. The first host cells lo appear in the graft, already a few hours after the transplantation are monocytes and lymphocytes. Both T and B lymphocytes contribute to the infiltrate: at early stages of rejection most of the infiltrating lymphocytes have the high electrophoretic mobility of (resting) T cells, whereas later during the rejection most of the infiltrating lymphocytes display the slow mobility of (resting) B cells. The blast response follows 2 days after the influx of lymphocytes. The (B) plasmablast response lakes place somewhat earlier and is higher in magnitude than the (T) lymphoblast response. Macrophages appear 1.5 days after the influx of monocytes. The inflammatory cells proliferate rapidly: after 1 h of pulse‐labelling with 3H‐TdR in vivo up to 24% of the infiltrating leucocytes are labelled. Most labelled cells are blast cells or lymphocytes. although a small but distinct population of labelled monocytes is also detected in situ. The in situ blast and proliferative responses precede the corresponding responses in the host central lymphatic system, i.e. spleen, blood and lymph nodes. The inflammatory leucocytes arc isolated from the allograft parenchymal cells via I e velocity sedimentation. They are strongly and specifically cytotoxic in the 6 h 51Cr release assay to donor‐derived lymphoid target cells in vitro. The peak in situ cytotoxic activity in the graft lakes plate already on day 5 after the transplantation, whereas in the central lymphatic system the cytotoxic cells are detected later and peak values are obtained only after the activity In situ has declined. The findings emphasize the role of the graft as the site of sensitization of kidney transplantation (peripheral sensitization) and the complex nature of the inflammatory response responsible for allograft rejection.
A Live Human B‐Cell Activator Operating in Isolation of Other Cellular Influences
Authors: BIRD A. G. , BRITTON S.
Published: June 2006

Using a reversed haemolytic plaque assay, we demonstrate that live but not inactivated Epstein‐Barr (EB) virus can induce human blood tymphocytes to polyclonal Ig (IgM and IgG) secretion in vitro. Unlike pokeweed mitogen this B‐cell‐activating effect takes place in relative absence of T cells. EB virus thus promises to be a tool for study of B cell function isolated from other cellular influences, although its mechanism of activation appears to be different from that of previously known B cell activators.
Macrophage/T‐Lymphocyte Interaction in the Immune Response to PPD in Humans
Authors: BERGHOLTZ B. O. , THORSBY E.
Published: June 2006

The proliferative response of sensitized human T cells to low concentrations of purified protein derivative of tuberculin (PPD) in vitro has previously been shown lo depend on the presence of autologous or HLA‐D‐compatible macrophages (M). In this paper we present evidence that the M/T cell interaction requires viable M and docs not involve a soluble factor. T cells from neonates do not respond to PPD in our system, while M) from neonates are able 10 present PPD to T cells from HLA‐D matched sensitized adults. It is also demonstrated that the previously reported HLA‐D restriction of T cell/M9 cooperation is not due to T cell mediated killing of allogeneic macrophages in vitro.
Alloantigens Derived from Stimulator Cells and Bound onto M LC‐activated Rat T Lymphoblasts
Authors: FENNER M. , FRISCHKNECHT H. , BINZ H. , LINDENMANN J. , WIGZELL H.
Published: June 2006

Rat T lymphocytes were activated in MLC (mixed leucocyte culture) against allogeneic stimulator cells. A few percent of such T blasts could be shown to carry IgM on their surface when a mixture of T and B lymphocytes was used as responder cells. The immunoglobulin was absent when purified T lymphocytes served as responders, A large proportion of MLC T lymphoblasts could be shown to carry stimulator alloantigens on their surface. These antigens were demonstrated (a) by indirect immunofluorescence using anti‐stimulator alloantibody and FITC (fluorescein isothiocyanate Isomer I)‐labelled F(ab)2 fragments of rabbit anti‐rat Ig; (b) by autoradiography after internal labelling of stimulator cells. which showed the transfer of label from stimulator to responder cells. For biochemical analysis T blasts were restimulated with internally labelled allogeneic or syngeneic stimulator cells, and bound radiolabelled stimulator alloantigens were analysed on SDS‐PAGE (sodium lauryl sulphate polyacrylamide gel electrophoresis). The results suggest that intact SD antigens with a molecular weight of 45,000 daltons and LD determinants with a molecular weight of 34,000 daltons derived from stimulator cells are found on MLC T blasts. No evidence of binding of the 27,000‐dalton la chain to T blasts was found. Possible arrangements of these molecules on the responder blasts is discussed.
Natural Killer Cell Response to Lymphocytic Choriomeningitis Virus in Beige Mice
Authors: WELSH R. M. , KIESSLING R. W.
Published: June 2006

Lymphocytic choriomeningitis virus (LCMV) induced low levels of natural killer (NK) tell activity in C57BL6 mutant beige (bg/bg) mice, which had previously been reported to have no cytotoxic NK cells. NK cell‐mediated lysis by bg/bg splenocytes was observed against a cell line (YAC‐I) very sensitive to NK cell cytotoxicity, but not appreciably against a less scum live cell line (L‐929), The bg/bg mutant mice with this very low NK cell activity and control stains of mice (bg/+, C57BL6) with high NK cell activity synthesized comparable amounts of virus and interferon in the spleen, suggesting that NK cells may not play a significant role in curtailing viral synthesis before the advent of the specific immune response mechanisms
Presence of Receptors for IgD on Human T and Non‐T Lymphocytes
Authors: SJÖBERG O.
Published: June 2006

Ig‐coated latex particles were used to study the presence in human blood of lymphocytes with receptors for various Ig classes. A significant proportion of cells bound to particles coated with IgG, IgA and IgM. In addition, 0–6.5% (mean 2.2%) peripheral blood lymphocytes from normal blood donors were able to form rosettes with IgD‐coated latex particles. Inhibition studies showed that the latter receptors were distinct from those directed against IgG, IgA and IgM. IgD receptor‐bearing cells seemed to exist both among T cells and non‐T cells but were 3–10 times more frequent in the non‐T‐cell population.
The Role of Viral Glycoproteins in Mumps‐Virus‐dependent Lymphocyte‐mediated Cytotoxicity in Vitro
Authors: HÄRFAST B. , ÖRVELL C. , ALSHEIKHLY A. , ANDERSSON T. , PERLMANN P. , NORRBY E.
Published: June 2006

Human peripheral blood lymphocytes (PBL) from healthy donors express enhanced natural cytotoxicity to target cells after a brief exposure 10 mumps virus in vitro, We describe here experiments aiming at elucidating the mechanism of this virus‐dependant cytotoxicity. Treatment with proteolytic enzymes resulted in virus particles depleted of one or both kinds of their glycoprotein spikes. Removal of both of these components from the virion abrogated their ability to enhance cytotoxicity. This virus‐dependent cytotoxicity was significantly but not completed reduced when one of the spike glycoproteins (gp 75, HANA) was removed selectively. Similarly, nucleic‐acid‐free preparations of the spikes, obtained by detergent treatment of mumps virions, also dialed enhanced cytotoxicity. However, the activity of these preparations was lower than that of untreated virions Further evidence for the importance of II AN A was provided by the use of F(ab′)2 fragments of anti‐HANA‐specific rabbit antibodies. When these fragments were allowed to react with virus before addition of the virus to PBL. no augmentation of cytolysis was observed. Antibody fragments specific for the other spike protein (gp 61, F) failed to inhibit the virus‐dependent enhancement of PBL‐mediated cytotoxicity. However, anti‐HANA and anti‐F blocked this reaction when added directly to the mixture of virus‐treated PBL and target cells. The results are compatible with the hypothesis that virus‐dependent cytotoxicity requires HANA for anchoring the virus to PBL receptors (and perhaps to bring effector and target cells into closer contact), whereas F may be involved in subsequent events increasing effector cell function.
Phospholipase A2 and Arachidonic Acid: a Common Link in the Generation of the Eosinophil Chemotactic Factor (ECF) from Human PMN by Various Stimuli
Authors: TESCH H. , KÖNIG W.
Published: June 2006

An eosinophil chemotactic factor (ECF) of low molecular weight can be generated and released from human polymorphonuclear neutrophils by the calcium ionophore, phagocytosis of zymosan particles, arachidonic acid, and phospholipase A2. Since the activation of cells by the ionophore and during the phagocytic event leads to phospholipid turnover, with the subsequent generation of arachidonic acid, it is reasonable that phospholipase A2 represents the common link for ECF production. The kinetics of ECF release by phospholipase A2 is similar to the pattern observed with the various stimuli. After a rapid rise in activity a decline occurred at later times of secretion, suggesting a mechanism of inactivation. During subcellular fractionation of cells an ECF‐generating component was enriched in the 200,000 g supernatant fraction, which represents the cytosol. Addition of arachidonic acid or phospholipase A2 induced ECF generation. On gel filtration analysis the ECF‐generating component revealed a molecular weight of about 80,000 daltons. It is suggested that this component represents a lipoxygenase.
Lymphocyte Subpopulations and IgG Concentrations in Cerebrospinal Fluid and Blood from Patients with Myasthenia Gravis
Authors: NÆSS A. , GILHUS N. E. , AARLI J. A.
Published: June 2006

Twelve patients with myasthenia gravis (MG) were examined for cerebrospinal fluid (CSF) T lymphocytes and for peripheral blood lymphocyte subpopulations (E‐, EAET‐, EA‐ and EAC‐rosette‐forming cells). The patients had a significantly increased percentage of CSF T cells (88.2±8.7%) when compared with controls (78.0±9.1%). Absolute concentrations of CSF T cells were not significantly increased. In peripheral blood no significant change in lymphocyte subpopulations was observed, but there was a slight leucocytosis in the patient group. The patients had increased CSF IgG concentrations, CSF IgG to protein ratio, and CSF to serum IgG ratio, indicating an intrathecal production of IgG. No oligoclonal bands could be demonstrated in the agarose gel electrophoresis. The two patients with thymoma had anti‐muscle antibodies in CSF and serum, with ratios of 1:256 and 1:1024, respectively. Serum IgG levels were increased, but not significantly, in the patient group. These results suggest an involvement also of the central nervous system in some patients with MG.
Neutrophil Chemotaxis Defect in IgA Deficiency Evaluated by Migration Agarose Method
Authors: D'AMELIO R. , LeMOLI S. , ROSSI P. , AIUTI F.
Published: June 2006

The chemotactic and random mobility functions of twelve selectively IgA‐deficient patients were evaluated by a method using agarose gel. A severe polymorphonuclear cellular chemotactic defect was found in ten out of twelve patients, but only five of them also showed a marked associated impairment of random locomotory function. Furthermore, in one subject, levamisole therapy resulted in a dramatic improvement of both chemotactic and random mobility functions. These results are discussed in the paper with respect to the possible pathogenetic implications.
Mechanisms of Cell‐mediated Cytotoxicity in Mice Rejecting Xenogeneic Human Lymphoblastoid Cells
Authors: CARNAUD C. , ALTMAN J. , ERRASTI P. , VAN DER GAAG R.
Published: June 2006

The in vivo immunization of mice with human lymphoblastoid cell line LHN13 generates direct cell‐mediated cytotoxicity by spleen cells. The lytic activity appears as early as day 3 after the intraperitoneal inoculation of 7.5 × 105 cells and persists at least until day 11. The killer cells do not adhere to plastic and are not retained on nylon wool columns or on Degalan heads coaled with mouse Ig plus rabbit‐anti‐mouse Ig The effector cells are partly inhibited by treatment with anti‐Thy‐1.2 serum plus complement, but this inhibition appears to be non‐specific since antiserum alone or normal mouse serum plus complement have the same effects. Heat‐aggregated IgG strongly inhibit cytotoxicity, indicating that the effector cells are Fc‐positive and that such receptors are implicated in lysis. Altogether, these features strongly argue for an ADCC phenomenon. The involvement of antibodies is demonstrated by the fact that eluates (56°C, 30 min) from immune cells alone induce lysis in the presence of normal spleen cells as effectors The lytic activity of these eluates can be removed by specific adsorption on protein A coupled to Sepharose heads and on the human lymphoid target cells. Positive complementation between immune and non‐immune spleen cells suggests that the arming process may occur in vitro during the assay, when antibodies are released by plasmacytes
HLA‐D/DR Restriction of Macrophage‐dependent Antigen Activation of Immune T Lymphocytes: Cross‐reacting Allogeneic HLA‐D/DR May Partly Substitute for Self HLA‐D/DR
Authors: BERGHOLTZ B. O. , THORESEN A. B. , THORSBY E.
Published: June 2006

Optimal proliferative response of T lymphocytes to purified protein derivative of tuberculin (PPD) III vitro requires that antigen be presented by autologous macrophages or allogeneic macrophages sharing HLA‐D/DR determinants with the T cell donor. In some cases, however, T cells may respond to a limited extent to PPD in association with macrophages expressing different HLA‐D/DR determinants. In this paper experiments are presented where various combinations of T cells and HLA‐D/DR disparate macrophages were stimulated with PPD. We often found a stronger PPD‐specific response in HLA‐D/DR‐incompatible combinations in which the macrophages carried HLA‐DR antigens known from serological studies to be cross‐reactive with those of the T cell donor than in combinations in which this was not the case. Thus, the cross‐reactions detected by serology may sometimes be reflected on a functional level in T lymphocyte/macrophage cooperation.
A γl Heavy‐Chain Disease Protein (EST) Lacking the Entire VH and CH1 Domains
Authors: BIEWENGA J. , FRANGIONE B. , FRANKLIN E. C. , VAN LOGHEM E.
Published: June 2006

Aγl heavy‐chain disease protein (EST) is described which lacks the entire VH and CH1 domains and starts with the normal sequence of γ1 H‐chains corresponding to the beginning of the hinge region (position 216). Although degradation cannot be excluded with certainty, it is probable that this protein is synthesized as an internally deleted γ heavy‐chain disease protein. Presumably a DNA recombination has occurred resulting in the deletion of the genes coding for the VH and CH1 domains with splicing of the precursor RNA to the sequence coding for the hinge region.
A Two‐Signal Mechanism for the Induction of Cytotoxic T Lymphocytes
Authors: SYMINGTON F. W. , TEH H.‐S.
Published: June 2006

Depletion of adherent cells from stimulator and responder lymphocyte by a single filtration through nylon wool columns, led to complete abrogation of the cytotoxic response to the stimulating alloantigen. Cytotoxic responses were restored by adding anti‐Thy‐l‐complement‐treated normal peritoneal exudate cells (PECl)syngeneic or allogeneic to the responding population. Alternatively, the response could be reconstituted With coslimulator, a lymphokine obtained by stimulation of spleen cells with concanavalin A. Costimulator was not itself cytotoxic and induced few or no cytotoxic T lymphocytes (CL) in the absence of stimulator cells. Costimuiator was also more efficient than allogeneie PEC, which in turn were more efficient than syngeneic PEC, in reconstituting the cytotoxic response. The number of CL produced to the activating alloantigen was shown to increase with increasing concentration of dissimulator. More Interestingly, in the presence of a relatively high concentration of cosnmulator. CL were also activated to target cells that differ in H‐2 haplotype from the stimulating alloantigen. Lysis of the third‐party target cells could not lie inhibited by cold targets syngeneic to the activating alloantigen. A clonal assay for cytotoxic precursors was used to confirm that CL. for the activating alloantigen and CL for the third‐party H‐2 antigens were derived from different progenitors. Only about 37% of the cytotoxic clones produced were specific for the activating alloantigen. These observations are explained in terms of a two‐signal model for CL activation.