bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunoglobulin Quantitation and Enumeration of Immunoglobulin‐producing Cells: Comparison of Two Indexes of B‐Cell Activation
Authors: MUNOZ J. , PRYJMA J. , FUDENBERG H. H. , VIRELLA G.
Published: June 2006

Human lymphocytes were cultured under different conditions to determine the effects of technical variations on the response to pokeweed mitogen (PWM) and Staphylococcus aureus as measured by (a) quantitation of immunoglobulins (Ig) in the culture supernatants and (b) enumeration of Ig‐secreting cells (ISC) by a reverse plaque technique. The highest numbers of ISC were measured when the cells were cultivated in standing tubes, without glutamine supplementation during the culture, and at a concentration of 1 × 106 cells/ml. The highest Ig concentrations were measured under similar conditions, except that somewhat higher values were obtained with cells cultivated at 1 × 106/ml in 2 ml cultures with PWM stimulation and at 2.5 × 106/ml cultures with S. aureus stimulation. In time‐course studies, peak ISC responses occurred on day 5 with each mitogen, whereas extracellular Ig levels kept rising until day 7, possibly owing to accumulation of secreted Ig. Measurements of the numbers of ISC and of secreted Ig levels in simultaneous cultures of lymphocytes from the same donors showed no correlation: however, co‐stimulation of cultures with PWM and concanavalin A (to stimulate suppressor cells) depressed both Ig levels and ISC numbers. These results suggest heterogeneity in the plaque‐forming cell population with respect to rate of Ig secretion, but indicate that these two techniques both reflect B‐cell activation. They should not, however, be considered interchangeable, and the two probably should be used in conjunction for complete characterization of B‐cell activation.
Dynamics of the Interaction between Entamoeba histolytica and Components of the Immune Response
Authors: AUST‐KETTIS A. , SUNDQVIST K.‐G.
Published: June 2006

Trophozoites of Entamoeba histolytica redistributed specific antibodies reacting to antigens on their surface. The presence of a second layer of anti‐antibodies enhanced the redistribution of anti‐amoeba antibodies but was not a prerequisite for this process. Microfluorometric measurements of the fluorescence intensity in different areas of the cell surface provided objective evidence for the antibody‐induced redistribution. In order to distinguish surface‐bound and internalized anti‐amoeba antibodies, the trophozoites were fixed in PFA. This fixation made the cells impermeable to antibodies. Thus, after fixation in PFA at different times during the redistribution process surface‐bound anti‐amoeba antibodies labelled with one fluorochrome could be detected with anti‐antibodies labelled with another fluorochrome. This approach showed that the amoeba, in addition to the redistribution, also internalized anti‐amoeba antibodies.
Antigen‐dependent Physical Interaction between Human Monocytes and T Lymphocytes
Authors: POWELL L. W. , HART P. , NIELSEN M. H. , WERDELIN O.
Published: June 2006

The physical interaction between human monocytes and autologous T lymphocytes was studied in cultures of cells from tuberculin‐sensitive individuals to which purified protein derivative of tuberculin had been added. Such cultures displayed formation of cell aggregates, each typically consisting of one or two monocytes to which several lymphocytes adhered by means of uropods. The capacity of monocyte‐T‐lymphocyte mixtures to produce this type of aggregate in response to tuberculin correlated with the tuberculin sensitivity of the cell donor as measured by skin test.
Spontaneous Release of Leucocyte Migration‐Inhibitory Factor by Mononuclear Cells Eluted from Rheumatoid Synovial Tissue
Authors: CHATTOPADHYAY H. , CHATTOPADHYAY C. , NATVIG J. B. , MELLBYE O. J. , PAHLE J.
Published: June 2006

Spontaneous leucocyte migration‐inhibitory factor (LIF) was assayed by an indirect system in which mononuclear cells (MNC) eluted from rheumatoid synovial tissues or isolated from peripheral blood of normal donors were cultured without antigen. The supernatants from these cultures and control supernatants, heated to 80°C for 30 min to destroy the LIF activity, were used in the test. In seven out of eleven rheumatoid arthritis patients the eluted synovial MNC produced LIF spontaneously, whereas none of the twelve normal blood donors showed any such production. The ability to show spontaneous LIF production was primarily seen in the joint tissues from sero‐negative patients.
Differential Role of Zn2+ in Antigen‐ and Mitogen‐induced Lymphokine Production
Authors: BENDTZEN K.
Published: June 2006

The effect on human lymphokine production in vitro of phenanthroline, a Zn2+‐chelating agent and an inhibitor of carboxypeptidases A and B, was tested. The elaboration of leucocyte migration inhibitory factor (LIF) by tuberculin‐sensitized mononuclear cells stimulated with the specific antigen was reduced in a dose‐dependent manner, an effect completely restored by addition of excess Zn2+. In contrast, phenanthroline did not affect LIF production by mono‐nuclear cells activated nonspecifically by phytohaemagglutinin. It is hypothesized that the presence of a Zn2+‐dependent molecule, possibly a carboxypeptidase, may be necessary for antigen‐ but not for mitogen‐induced lymphokine production.
Effect of Cyclosporin A on the Generation of Cytotoxic T Lymphocytes in Mouse Mixed Lymphocyte Culture
Authors: NEMLANDER A. , HÄYRY P.
Published: June 2006

Cyclosporin A inhibited at equal concentrations both the proliferate response and the generation of cytotoxic T lymphocytes in one‐way mouse spleen cell mixed lymphocyte culture. The 50% inhibitory concentration was in all experiments 10−2 to 10−1μg/ml. The inhibition was directly proportional to how early the drug was added to the culture: a complete inhibition of both responses was obtained if the drug was added on day.3, and a partial inhibition if added on day 4 of culture. Cytological analysis of the cultured cells demonstrated that resting lymphocytes were not damaged at 100 1000‐fold concentrations of the drug giving complete inhibition of the blastogenic response. The results suggest that cyclosporin A is most effective if present throughout the induction phase of the immune response.
Characteristics of Suppressor Cell Activity Appearing in Cocultures of Two Individuals with Immunodeficiency with Thymoma
Authors: LITWIN S. D.
Published: June 2006

Suppressor cell activity in two individuals (S1 and S2) with immunodeficiency with thymoma (ID‐THY) was studied in peripheral blood mononuclear cells in pokeweed mitogen‐stimulated single culture and cocultures. Secreted Ig was measured by radioincorporation and immunoprecipitation after 5‐7 days. Control cocultures of normal/normal cells showed, in most cases, a percentage observed to expected ratio (% O/E) of cpm Ig near 100%. However, augmentation (% of O/E > 150) was often encountered, whereas suppression (% O/E <50) was found only ran In forty‐two cocultures. In ID‐THY/normal cell cocultures the degree of suppression by ID‐THY cells varied widely when the same or different cocultivanls were retested. This finding could be explained in part an inverse correlation between the amount of secreted Ig produced by normal cells in single culture and the degree of suppression of the same normal by ID‐THY cells in cocultures. A panel of normal cells were all suppressed when a range of S1 or S2/normal cell ratios were tested, weighing against genetic differences in suppressibility in the above system. ID‐THY cells failed to block Ig secretion of human lymphoblastoid line cells, suggesting that the mechanism of suppression was related to a block in differentiation rather than interference mith Ig synthesis per se. An experiment using cocultivants separated by a Millipore membrane showed that suppression was mediated by a humoral factor.
d‐Penicillamine in Vivo Enhances Lymphocyte DN A Synthesis: Role of Macrophages
Authors: BINDERUP L. , BRAMM E. , ARRIGONI‐MARTELLI E.
Published: June 2006

Administration of d‐penicillamine (50 mg/kg/day orally) for 4 days increased the uptake of 3H‐thymidine (3H‐TdR) in unstimulated and concanavalin ‐A‐stimulated unseparated lymph node and spleen cells from Lewis rats. Increased 3H‐TdR incorporation was also found in cultures depleted of adherent cells. d‐Penicillamine treatment did not increase the incorporation of 3H‐TdR in lymph node and spleen cells from rats concomitantly treated with the selective macro‐phage toxin silica. in contrast, treatment with d‐penicillamine during the last 4 days of silica treatment sometimes resulted in a marked decrease in 3H‐TdR incorporation. It is suggested that d‐penicillamine treatment in vivo is able to enhance the responsiveness of the lymphocytes, dependent on the presence of functionally intact macrophages. The increased response vanished after 2‐3 weeks, even with continuous administration of d‐penicillamine.
Binding of Murine Myeloma Proteins of Different Ig Classes and Subclasses to Fc‐reactive Surface Structures in Gram‐positive Cocci
Authors: MYHRE E. B. , KRONVALL G.
Published: June 2006

Twelve different murine myeloma proteins were tested for binding to seventy Gram‐positive strains belonging to group A, C and G streptococci and to Staphylococcus aureus. Group A streptococci, known lo hind human IgG. were incapable of binding any of eight murine IgG immunoglobulins tested except for one strain that hound un IgG2b myeloma protein. In contrast, group C and G streptococci interacted with murine immunoglobulins of subclasses IgG2a, IgG2b and IgG3, and G strains also to a lesser extent with IgG1. Bovine and equine group C streptococci did not differ from human group C streptococci in their IgG reactivity. Staphylococcal strains showed a high reactivity with murine myeloma components of IgG subclasses 2a, 2b and 3 and a low but definite binding of an IgG1 myeloma protein. IgA myeloma protein S‐122 interacted with nine of fifteen group A streptococci. This binding could not be inhibited by human IgG and the reactivity is thus different from Fc‐mediated binding of immunoglobulins. One of three IgA myeloma proteins tested, TEPC 15, bound to staphylococci. The Fc specificity of this interaction was confirmed by chromatography on protein A‐Sepharose and by inhibition studies using polyclonal human IgG. The protein A reactivity of this monoclonal protein was detected in IgA aggregates and absent in the monomeric form of IgA.
Detection of Muramidase (Lysozyme)‐secreting Leucocytes at the Single‐Cell Level by a Protein A Plaque Assay
Authors: SMITH C. I. E. , HAMMARSTRÖM L.
Published: June 2006

Using a modification of the protein A plaque assay, muramidase (lysozyme)‐producing leucocytes were detected as plaque‐forming cells. In the presence of anti‐muramidase Ig and complement the secreted lysozyme resulted in lysis of protein‐A‐coated target erythrocytes. By the use of a monolayer technique individual plaque‐forming cells could he identified by staining procedures. Granulocytes as well as monocytes were found to produce muramidase and thus to form plaques. This method could serve as a useful tool when studying lysozyme secretion. Furthermore, by the use of appropriate antisera, this method could be employed for the study of any cell type (any secretion), provided enough molecules in being secreted.
Genetic Specificity of Primary and Secondary Proliferative and Cytotoxic Responses of Human Lymphocytes Grown in Continued Culture
Authors: SCHENDEL D. J. , WANK R. , BONNARD G. D.
Published: June 2006

Human T peripheral blood lymphocytes were grown in continued culture using conditioned medium obtained from phytohaemagglutinin‐stimulated, pooled human leucocytes. These cultured T cells (CTC) were tested in mixed lymphocyte culture (MLC) and cell‐mediated lympholysis (CML) assays to determine the genetic specificity of their proliferate and cytotoxic responses. Primary responses were measured after initial in vitro stimulation by allogeneic cells, and secondary responses were measured after a second in vitro stimulation by allogeneic cells. Doth primary and secondary proliferative responses were found to be stimulated by alloantigens controlled by the HLA region and, more specifically, by antigens of the HLA‐D region, in accordance with the responses of normal peripheral blood T lymphocytes, When CTC KW established from unsensitized PBL and then stimulated with allogeneic cells, they could respond by proliferation in MLC, but, in contrast to PBL, they did not show subsequent cytotoxic responses. On the other hand, CTC established from PBL that had been stimulated first with allogeneic cells in either primary or secondary MLC displayed high levels of cytotoxic reactivity in CML. The strongest cytotoxicity was directed against allospecificites controlled by the HLA region and specific for the MLC‐stimulating cells, but lower levels of cross‐reactive cytotoxicity were also observed.
Activation of Human T and B Cells by Rabbit Anti‐Human β2‐Microglobulin
Authors: RINGDÉN O.
Published: June 2006

Rabbit anti‐human β2‐microglobulin (anti‐β2m) increased the highest DNA synthesis in un‐separated lymphocytes or artificially composed mixtures enriched in T and B cells. In enriched T and B cells no or low stimulation was seen. The maximal response by IgG anti‐β2m was seen in proportions of enriched T/B cells, being 3:1 for blood lymphocytes and 1:1 for spleen cells, which are the same as the physiological proportions of T and B cells in these lymphoid organs. Whereas unseparated lymphocytes gave a peat response on day 3, enriched B cells had a peak response on day 6. Fab anti‐β2m did not activate enriched T cells but increased DNA synthesis in enriched B cells to about the same extent as unseparated lymphocytes and mixtures of enriched T and B cells. The proportion of sheep erythrocyte rosette‐forming cells (E‐RFC) decreased after stimulation by anti‐β2m and increased after stimulation by phytohaemagglutinin. However, as revealed by autoradiography, a proportion of lymphocytes activated by anti‐β2m were E‐RFC and this proportion increased with increasing stimulation by anti‐β2m. DNA synthesis induced by anti‐β2m was unchanged for spleen cells and slightly decreased for blood lymphocytes when phagocytic cells were removed by iron treatment. Supernatants from lymphocytes activated by anti‐β2m only induced low DNA synthesis in enriched T or B cells. Experiments with mitomycin treated cells indicate that cooperation and close contact between T and B cells are needed for activation by IgG anti‐β2m. T cells are needed for B cell activation by anti‐β2m and B cells are required for T cell activation to occur.
A New Quantitative Fluorimetric Assay for Phagocytosis of Bacteria
Authors: VRAY B. , HOEBEKE J. , SAINT‐GUILLAIN M. , Leloup R. , STROSBERG A. D.
Published: June 2006

We describe a new quantitative fluorimetric assay for phagocytosis of bacteria. A suspension of fluorescein‐labelled bacteria (Micrococcus lysodeikticus) is mixed and incubated with phagocytes. After fixation with paraformaldehyde, the excess non‐phagocytosed and adsorbed bacteria are lysed with a solution of lysozyme in phosphate‐buffered saline. After washing of the phagocytes, the fluorescence of those that have Ingested the labelled bacteria is. measured with a spectro fluorimeter. We report results obtained with different types of phagocytes which show that this method allows sensitivity, saturation and kinetic studies and the calculation of a phagocytic index.
Mouse/Human T‐Cell Hybrids Rosetting with Sheep Erythrocytes
Authors: SUOMALAINEN H. A. , GOLDSBY R. A. , OSBORNE B. A. , SCHRÖDER J.
Published: June 2006

Hybrid cells have been recovered from selective culture medium after fusion of concanavalin‐A‐activated human lymphocytes with an AKR mouse thymoma (BW 5147). After 6 months of culture twenty‐seven out of forty‐nine clones still contained human chromosomes. Human chromosome 6 was present in 89% of these clones, and human X in 70%. Clones from one hybrid line contained several human chromosomes. In twelve of the clones carrying human chromosomes, the rosetting with sheep Erythrocytes (SRBC) was 3 times as high as In the BW 5147 cell line. All these clones Carried the human chromosome 6, and eight clones contained the human X chromosome as well. In some of these clones (25%) chromosome 6 was the only human one present. In the two clones In which human chromosome 6 was completely missing, the resetting with SRBC was at the level of the BW line. We therefore suggest that genes on human chromosome 6 are responsible for rosetting with SRBC.
Cytochemistry of Human T‐Cell Subpopulations
Authors: BEVAN A. , BURNS G. F. , GRAY L. , CAWLEY J. C.
Published: June 2006

Acid phosphatase and esterase cytochemistry performed on purified normal human T‐cell populations showed that both methods produced distinctive localized dot patterns of reactivity in 60–70% of cells. By examination of rosette preparations formed with ox erythrocytes coated with IgM (EAM), with IgG (EAG), or anti‐human κ and λ light chains, it was shown that this pattern of reactivity was largely restricted to small T lymphocytes possessing receptors for the Fc of IgG (Tμ cells). In addition, both B lymphocytes and T cells with receptors for the Fc of IgG (Tλ cells) were larger lymphocytes with more abundant cytoplasm and usually displayed scattered granular acid phosphatase activity; in esterase preparations both cell types were either negative or possessed similar scattered granular positivity. As compared with Tμ cells, Tγ cells were seen to form loose spontaneous rosettes with sheep erythrocytes. Combined esterase and acid phosphatase staining showed that both enzyme activities in the Tμ. cells are localized in the same area, and ultrastructural acid phosphatase cytochemistry established that this was in distinctive lysosomal structures. Tμ staining by both esterase and acid phosphatase cytochemistry was greatly reduced after rosetting with EAG, but not after rosette formation with EAM or sheep erythrocytes.
Quantitative Study of the Uptake of IgA by Isolated Rat Hepatocytes
Authors: TOLLESHAUG H. , BRANDTZAEG P. , HOLTE K.
Published: June 2006

Recently dispersed rat hepatocytes showed high capacity for uptake of monomers and polymeric human IgA by simple adsorptive endocytosis. Maximum uptake exceeded 15 million monomer units pit cell. The asialo‐glycoprotein receptor was not involved in this process. J chain‐containing polymers were on the average taken up somewhat better than monomers and substantially better than J chain‐deficient polymers. This result would agree with a partial involvement of the secretory component (SC) in the adsorptive uptake of IgA. Different receptor mechanisms might hence explain the hepatic transport of IgA from blood to bile in the rat and perhaps he involved in the catabolism of IgA. After binding lo the plasma membrane of the rat hepatocyte, most of the human IgA was internalized in endocytic vesicles. A considerable proportion of radiolabelled IgA apparently reached the lysosomes after internalization became degraded.
Isolation and Characterization of Intermediate Complexes and Other Components with Common Antigenic Determinants
Authors: HANSSON U.‐B. , UESSON M. , ALKNER U.
Published: June 2006

Fractions with affinity for heat‐denatured human IgG (HDG) were isolated from four sera that contained intermediate complexes (IC). These IC fractions contained part of the 7S IgG, all IC, and part of the rapidly sedimenting complexes (RC) found in the sera. The IC consisted of IgG1 or IgG3 and the RC of IgG and IgM with kappa and lambda light chains. The IgG in the IC fractions contained an abnormally large amount of neuraminic acid. No correlation between IgG subclass or content of neuraminic acid and complex formation was found. There were indications that the formation of IC was not only the result of a self‐association of IgG molecules with anti‐γ‐globulin activity. Specific rabbit antisera were prepared against two of the IC fractions. Affinity chromatography with immobilized IgG and F(ab')2γ from these antisera confirmed the presence of common antigenic determinants in the sera. These determinants occurred mainly in 7S components in one individual, in IC in one and in RC in another. Only a minor part of the serum components with affinity for HDG contained the determinants. RF activity was found in the components that lacked and in those that contained the common antigenic determinants.
Development of Natural Killer Cell Activity and Genetic Resistance to Bone Marrow Transplantation with Age: Effect of Neonatal Thymectomy
Authors: MILLER S. C. , GALLAGHER M. T. , DATTA S. K. , TRENTIN J. J.
Published: June 2006

The effect of neonatal thymectomy on the development of splenic and bone marrow natural cell‐mediated cytotoxicity and on genetic resistance to bone marrow transplantation was examined in mice. Natural cytotoxicity was measured by a 51Cr release assay; the ability to engraft foreign bone marrow was assayed by the spleen colony method. The natural cytolytic response of spleen cells increased progressively from youth to early adulthood, whereas that of the bone marrow declined during the same age period. Neonatal thymectomy significantly elevated the natural killer cell response of young mice only (4 weeks, spleen; 6 weeks, bone marrow). In other experiments, neonatally thymectomized and sham‐operated mice were lethally irradiated at 4 or 6 weeks of age and injected with 2.5, 5.0 or 10 million rat marrow cells. Six days later spleen colonies were markedly reduced in both 4‐ and 6‐week‐old neonatally thymectomized mice with all rat marrow cell doses tested. Neonatal thymectomy did not alter the percentage of erythroid versus other colonies at either 4 or 6 weeks. In both thymectomized and sham‐operated mice the number of colonies increased with increases in marrow cell dose. The data are suggestive of a production and dissemination to the spleen of cells involved in the natural cytotoxic response from the bone marrow.
Preferential Production of IgA Antibodies by Spleen Fragments Immunized and Maintained in Vitro
Authors: PASANEN V. J. , KARJALAINEN K. , KAARTINEN M. , MÄKELÄ O.
Published: June 2006

Primary immune responses were induced with a hapten conjugate of Ficoll (NIP38‐Ficoll) in cultures of mouse spleen fragments. The production of anti‐NIP antibodies and mouse immunoglobulins IgA and IgG was monitored by solid‐phase radioimmunoassays. Cultures produced IgA and IgG without any stimulation, IgA at a rate of c. 60 ng/mg of wet tissue per day throughout the 10‐day culture period. Background production of IgG gradually declined from almost 1000 ng/mg day at the beginning to 100 ng/mg day at the end. Immunization modestly reduced the decline of the IgG production. It increased the production of IgA fourfold between days 6 and 8. It induced an antibody production that had its peak between days 6 and 8. Up to 50 ng of anti‐NIP antibody was produced during this period per mg of wet tissue per day. Of this antibody 75–90% was IgA. When the same antigen was injected into mice, more than 75% of anti‐NIP antibodies detected in their sera on day 8 were IgG. One cause of the high ratio of IgG/IgA antibodies in vitro is the efficient elimination of oligomeric IgA by the liver. Another possible cause is that the production of IgA is less sensitive than the production of IgG to the adverse conditions of the culture.
Evidence of Aberration of T‐cell Subsets in Aged Individuals
Authors: ABE T. , MORIMOTO C. , TOGUCHI T. , KIYOTAKI M. , HOMMA M.
Published: June 2006

In the present study, T‐cell subsets from aged individuals were examined by using anti‐BAT (brain‐associated thymocyte antigen) serum. Anti‐BAT serum was raised against the human fetal brain at 28 weeks of gestation. After absorption with AB erythrocytes, B‐cell lines, and leukaemic cells, anti‐BAT serum was T cell‐specific but unreactive to normal B cells. The ability of anti‐BAT serum‐treated lymphocytes from aged individuals to respond to concanavalin A, phytohaemagglutinin, and pokeweed mitogen (PWM) was unaltered even at a high concentration. In PWM‐stimulated Ig synthesis, T lymphocytes lacking the anti‐BAT serum‐reactive T‐cell subset enhanced the PWM‐stimulated Ig synthesis of autologous B lymphocytes from young individuals. The Con A‐induced suppressor function of lymphocytes from aged individuals was not significantly abolished by treatment with anti‐BAT serum and complement. In the autologous mixed lymphocyte reaction, the decrease in response was minimal when responder cells from aged individuals were treated with anti‐BAT serum even at a high concentration. It is concluded that the T‐cell subset with suppressor function is defective in aged individuals.
A Comparison of EA‐Rosette Formation on Lymphocytes Using Rabbit and Human Sensitizing Antibodies
Authors: DUNNE J. V. , SPIEGELBERG H. L. , VAUGHAN J. H.
Published: June 2006

The IgG receptors of peripheral blood lymphocytes were studied in determine whether those detected by a rabbit anti‐ox erythrocyte antibody (EARa) and a human anti‐human erythrocyte‐antibody (EAHu) were the tame. With cells separated by Ficoll Hypaque and freed of adherent cells by exposure to plastic surface, both EA systems gave similar numbers of rosettes. Mixed rosetting occurred when both types of sensitized cells were added simultaneously. EAHu and EARa rosettes thawed similar kinetics of formation, Depletion of rosette‐forming lymphocytes was possible by using both systems, and depletion by one system showed depletion in the Other, slg‐bearing lymphocytes were also removed, the amount of depletion being dependent on the degree of RBC sensitization With antibody EAHu and ERRa rosettes were equally inhibited by IgG of a given species or subclass. Normal human and rabbit IgG were more inhibitory than bovine or guinea‐pig IgG. All of the human subclasses inhibited, but IgG1 and IgG3 were more inhibitory than IgG4. and IgG4. These results indicate that Fcγ receptors on human lymphocytes react with and IgG configuration that is present on IgG of many species, although it is variably expressed among them. Human IgG1 and rabbit IgG are equivalent in the system and can be used equally well to detect Fc IgG‐receptor‐bearing cells.
Experimentally Induced Transplantation Tolerance
Authors: WATERFIELD J. D. , BICK P. H. , MÖLLER E. , MÖLLER G.
Published: June 2006

Neonatal tolerance was induced in CBA mice to alloantigens expressed by lymphoid cells of the A/Sn strain. 50% of the grafted animals maintained viable skin grafts for greater than 120 days and thus were considered operationally tolerant. However, splenic lymphocytes from these animals were capable of in vitro activation of DNA synthesis in response to the tolerizing alloantigens Likewise, the spleen contained precursors of Cytotoxic effector cells capable, upon activation of similarly recognizing the major histocompatibility complex alloantigens used for tolerance induction. In contrast, in vivo graft‐versus‐host (GVH) effector function was absent, suggesting a possible dissociation of cells involved in the in vitro cell‐mediated lympholysis and the in vivo GVH. Determination of potential serum‐blocking factors as a possible mechanism of graft survival in these tolerant animals proved negative.
Isolation of T Lymphocytes in Chronic Lymphocytic Leukaemia by Depletion of Mouse Rosette‐forming Cells
Authors: KABELITZ D. , FINK U. , REICHERT A.
Published: June 2006

Separation of mouse rosette‐forming cells (MRFC) was used as a technique to isolate T lymphocytes in patients with chronic lymphocytic leukaemia. The resulting interface fraction (non‐MRFC) was highly enriched for sheep rosette‐forming T cells (mean value 71.8% SRFC) as compared with unfractionated peripheral blood lymphoctes (mean value 18% SRFC). A higher degree of T‐cell purity was achieved by this method than by the sheep rosette sedimentation technique in patients with leucocyte counts greater than 50 × 109/I. Functional tests revealed that depletion of MRFC increased not only phytohaemagglutinin and mixed lymphocyte culture (MLC) reactivity but also the stimulatory capacity in the allogeneic MLC. In contrast, separation of MRFC in normal blood donors gave only a poor degree of separation and was not an effective means to isolate lymphocyte subpopulations.
Binding of Fragments of Human IgG to Solid‐phase C3b Measured by Enzyme‐linked Immunosorbent Assay (ELISA)
Authors: HAUTANEN A.
Published: June 2006

The binding of human IgG and different fragment of IgG to C3b adsorbed in polystyrene tubes has been studied by the enzyme‐linked immunosorbent Heat‐denatured polyclonal IgG and F(ab')2 and Fab fragments of IgG bound to solid‐phase C3b Heat‐denatured Fc fragments of IgG also had some reactivity with C3b, but at significantly higher concentrations than F(ab')2 and Fab fragments. The binding of heat‐denatured IgG could not be completely inhibited by the addition of heat‐denatured F(ab')2 fragments in tenfold excess The results suggest that the binding of heat‐denatured IgG to solid‐phase C3b is mediated through the Fab and Fc portions binding of denatured IgG to solid‐phase C3b is mediated through the Fab and Fc portions of IgG molecules.
Immunohistochemical Evaluation of J‐Chain Expression by Intra‐ and Extra‐follicular Immunoglobulin‐producing Human Tonsillar Cells
Authors: KORSRUD F. R. , BRANDTZAEG P.
Published: June 2006

Cytoplasmic J‐chain expression by Ig‐producing cells was characterized immunohistochemically in normal specimens of palatine and nasopharyngeal tonsils (adenoids). Altogether follicular immunocytes, which were mainly of the IgG and IgM isotypes, showed a much higher percentage of J‐chain positivity than the extrafollicular ones, in agreement with the idea that this polypeptide is principally a marker of differentiating or newly differentiated Ig‐producing cells. Thus, in concurrence with the predominating IgG isotype. J‐chain expression was almost 50% in the germinal centres of lymphoid follicles but only about 2% in the extrafollicular compartment. By contrast, a substantial number of J‐chain‐positive IgA immunocytes were found in the Jailer compartment. Since it is believed that extrafollicular Ig‐producing cells are mainly derived from follicular centre cells, this result indicated that significant isotype switching is involved in tonsillar B‐cell differentiation. Most tonsillar IgD immunocytes were J‐chain‐positive, in accordance with the notion that IgD expression is a feature of relatively early clonal development. It is postulated that J‐chain‐positive B‐cell blasts may leave the tonsils and, through isotype switching, contribute to the dimer‐producing IgA‐cell populations normally found in the exocrine glands of the upper aero‐digestive tract.
β‐Microglobulin is Bound to Streptococcal M Protein
Authors: BJÖRCK L. , TYLEWSKA S. K. , WADSTRÖM T. , KRONVALL G.
Published: June 2006

The presence of receptors for three human proteins (fibrinogen. IgG, and β2m‐microglobulin (β2m) was studied in group A streptococcal strains, M type 1, 12 and 14, and in M protein‐negative variants of these strains. Fibrinogen binding was detected in all six strains. IgG did not bind to the two type‐12 strains and uptake was low in both type‐14 strains. The type‐1, M protein‐positive strain was negative in binding experiments with radiolabelled IgG, whereas the M protein‐negative variant was strongly positive. Aggregated β2m showed a high degree of reactivity with the M protein‐positive strains but not with the corresponding M protein‐negative variants. The results indicate that the receptor for aggregated β2m is located on M protein in group A streptococci.
Peroral Immunization of Germfree Piglets: Appearance of Antibody‐forming Cells and Antibodies of Different Isotypes
Authors: TLASKALOVÁ‐HOGENOVÁ H. , ČERNÁ J. , MANDEL L.
Published: June 2006

Precolostral germfree piglets were perorally immunized with sheep erythrocytes daily, beginning on the 5th day of life. At different intervals the numbers of IgM, IgG and IgA antibody‐forming cells were estimated in the spleens, lymph nodes and intestinal mucosa by the haemolytic plaque method. In the beginning of the response IgM antibody‐forming cells were detected in intestinal mucosa and mesenteric lymph nodes. IgA plaque‐forming cells appeared later and were predominant during the whole period in all organs tested. In all lymphatic organs tested the appearance of IgG antibody‐producing cells followed that of IgA plaque‐forming cells. The development of immunoglobulins and antibody of different isotypes in Intestinal contents and sera was compared with the cellular kinetics found in these young animals after local stimulation.
The Induction and Expression of IgA Anti‐DNP Antibodies in Rat Bile and Secretions
Authors: MONTGOMERY P. C. , LEMAÎTRE‐COELHO I. M. , VAERMAN J.‐P.
Published: June 2006

Pregnant rats were immunized with dinitrophenylated type III‐pneumococcal vaccine by the intravenous, gastric, or intramammary routes. Anti‐DNP antibody responses in the IgA. IgG and IgM isotypes were measured in serum, secretions and bile. Gastric intubation was most effective at eliciting IgA antibody responses in bile and secretion, whereas the other routes were more effective at inducing IgG responses in serum and bile. IgM antibody responses were infrequent and were found in fluids most closely associated with the immunization route. Isoelectric focusing studies of IgA antibodies appearing in secretions and bile revealed that the gastric route consistently elicited antibody spectrotypes with shared components. Intravenous and intramammary immunization resulted in IgA spectrotypes possessing less homology, suggesting that these protocols lead to independent antibody responses triggered in spleen, draining lymph nodes, or secretory sites. After gastric stimulation, the appearance of IgA antibodies with identical spectral components in secretions and bile favours the concept that IgA precursor cells with identical clonotype potential migrate from the gastrointestinal area to secretory sites. Antibody expression in bile appears to result from the selective transfer of IgA populations gaining access to serum after synthesis at a secretory site.
Further Structural and Antigenic Studies of Light‐chain Amyloid Proteins
Authors: NATVIG J. B. , WESTERMARK P. , SLETTEN K. , HUSBY G. , MICHAELSEN T.
Published: June 2006

The major subunit protein of amyloid fibrils (758) isolated from a patient with systemic amyloidosis and studied by N‐terminal amino acid sequence analysis was found to be almost identical to the sequence of a VλIV Bence‐Jones protein and a previously described AλIV amyloid protein. The two AλIV amyloid proteins showed strong antigenic cross‐reaction, appearing as antigenic identity in double immunodiffusion tests using anti‐AλlV antiserum raised against one or the other of the two proteins. In addition, another new AλV amyloid fibril protein (R.S.) showed strong amino acid sequence homology and antigenic identity in double immunodiffusions with the prototype of the AλV subgroup (the AR protein). Finally, 20 primary or myeloma‐associated amyloid proteins were characterized using antisera against the AA and several Ig light‐chain‐derived amyloid proteins.
Isolation and Properties of a Complement Inhibitor from Naja haje Venom, Distinct from Known Anticomplementary Factors in Cobra Venom
Authors: VON ZABERN I. , PRZYKLENK H. , DAMERAU B. , ZIMMERMANN B.
Published: June 2006

A complement inhibitor (CI) has been isolated from cobra (Naja haje) venom which is distinct from the two known anticomplementary factors in cobra venom [1], in functional properties as well as structure. CI is a small (mol. wt 26,000, determined by sodium dodecyl sulphate gel electrophoresis), heat‐labile glycoprotein; the amino acid composition is that of a globular protein. CI interferes at various steps of the complement sequence, including reactions of the classical and the alternative pathway. No effect was observed on C4 fixation and on the assembly of the membrane attack complex from C6–9 (minor inhibiting effects, if present, have not been excluded). Initiation of the alternative pathway is inhibited by CI already at the stage of cleavage of factor B. CI binds to C4, C4b. C3 and C3b: since the major inhibitory action of CI is lost after washing of cell intermediates, complex formation and, as a consequence, steric hindrance may be responsible for the inhibiting effects of CI. CI also interferes with binding of C3b to C3b receptors on human erythrocytes. CI is non‐toxic in mice when given intraperitoneally in doses of 5 μg/g.
Localization of IgGFc and Complement Receptors in Chicken Lymphoid Tissue
Authors: THUNOLD S. , SCHAUENSTEIN K. , WOLF H. , THUNOLD K. S. , WICK G.
Published: June 2006

Chicken lymphoid organs were examined for IgGFc and complement receptors (FcR/CR) by immune adherence on frozen tissue sections. Indicator systems were sheep erythrocytes (E) coated with chicken anti‐E IgG (Each). E coated with rabbit anti‐E IgG (EArab) and normal chicken serum (EAC), and FITC‐labelled zymosan particles coated with chicken serum (ZyC). In the spleen, FcR and CR activity was confined to B‐dependent areas, i.e. the periellipsoidal sheaths and germinal centres and the red pulp. No FcR were found in the thymic or bursal lymphoid tissue, but CR activity was observed in the medulla of bursal follicles. Chicken and turkey IgG, chicken IgGFc, and bovine serum albumin (BSA)‐chicken anti‐BSA complexes inhibited binding of EAch. No inhibition was obtained with chicken IgGF(ab′)2. IgM or albumin, or with BSA‐rabbit anti‐BSA complexes and human or rabbit IgG. E did not adhere to the sections, nor did EArab, EArab incubated with heat‐inactivated chicken serum, or EAC complexes prepared with EArab and guineapig complement. The data suggest that chicken B lymphocytes and macrophages have receptors for avian IgGFc and C which can be demonstrated in tissue sections.
Rabbit Antisera with Specificity for Isolated Mouse T‐Lymphocyte Receptors
Authors: RUBIN B. , BOURGOIS A.
Published: June 2006

Rabbit antibodies obtained after Immunization of mouse Immunoglobulin (Mig)‐tolerant rabbits with B6 anti‐CBA IgG and having specificity for B6 anti‐CBA IgG and T‐cell receptors (antiserum 5936) were used to isolate 5936‐reactive molecules from B6 anti‐CBA mixed lymphocyte culture supernatants. Such 5936‐reactive molecules were produced by the B6 T cells, and they did not react with rabbit anti‐MIg antisera. They had a mol. wt of 50,000–75,000, and were single‐chain polypeptides that did not react with concanavalin A (Con A) Sepharose. These molecules were in turn injected into rabbits, and the antisera thus obtained had the following characteristics: (1) they reacted against B6 anti‐CBA T‐cell receptor material but not against B6 anti‐CBA IgG; (2) they reacted with about 35% of B6 (H‐2b. Ig‐1b) anti‐CBA T cells, 25% of B6 Con A blasts and 0 10%; of normal B6 T cells but not with B6 lipopolysaccharide (LPS) blasts, C3H.B10 (H‐2b, Ig‐I) anti CBA or CBA anti‐B6 T cells, CBA Con A blasts or normal CBA T cells: and (3) they reacted with the same 50,000–75.000 mol. wt, T‐cell‐derived molecules as did antiserum 5936. The implications of these findings are discussed in relation to the nature of T‐cell receptors.
Appearance of Anti‐HLA‐DR‐Reactive Cells in Normal and Rheumatoid Synovial Tissue
Authors: KLARESKOG L. , FORSUM U. , TJERNLUND U. MALMNÄS , KABELITZ D. , WIGREN A.
Published: June 2006

The reactivity of rabbit anti‐HLA‐DR antigen antibodies with cells in normal and rheumatoid synovial tissue was investigated by indirect immunofluorescence on frozen sections of tissue. The antibodies reacted with a significant proportion of the synovial lining cells of both normal and rheumatoid synovial tissue, with endothelial cells, and with a number of, most probably, migratory cells. After dispersion of cells from rheumatoid synovial tissue by digestion with collagenase and DNase, adherent cells of both a macrophage‐like and a dendritic appearance reacted with the anti‐HLA‐DR antigen antibodies. The adherent cells were also found to be potent stimulators in the allogeneic MLR. In addition, it was found that a high percentage of T lymphocytes from both peripheral blood and synovial tissue of rheumatoid patients bound anti‐HLA‐DR antibodies. The present data suggest a role for synovial lining cells in HLA‐D‐locus‐dependent events of importance in the pathogenesis of rheumatoid arthritis and other joint diseases and point to the need for further investigations on T lymphocytes derived from the site of inflammation in the study of rheumatoid arthritis.
T‐Cell Subset Analysis by Monoclonal Antibodies in Primary Immunodeficiences
Authors: PHAN‐DINH‐TUY F. , DURANDY A. , GRISCELLI C. , BACH M. A.
Published: June 2006

T‐cell subsets have been analysed in 23 cases of primary immunodeficiency with monoclonal antibodies. Functional assays investigating T‐cell function—proliferative response to mitogens, antigens, and allogeneic cells, cytotoxicity generated against allogeneic target cells and helper function to pokeweed mitogen‐induced immunoglobulin synthesis—were performed in parallel in the same patients. The results enabled us to delineate four groups of patients. The first group consisted of patients in whom marker and function studies show concordant data, with either normal or strongly decreased T‐cell number and function. The second group consisted of patients in whom various degrees of functional abnormality coexist with subnormal T‐cell number and increased suppressor T‐cell proportion. In the third group, we collected all patients who showed functional deficiencies without marker abnormalities. Finally, there was a small group composed of patients whose T‐cell pattern was strongly suggestive of abnormal differentiation.
Allergenic Synthetic Peptide Corresponding to the Second Calcium‐Binding Loop of Cod Allergen M
Authors: ELSAYED S. , RAGNARSSON U. , APOLD J. , FLORVAAG E. , VIK H.
Published: June 2006

A peptidewith the sequence of the second calcium‐binding loop (EF loop) of cod Allergen M was synthesized by automatic solid‐phase technique. The synthetic peptide corresponded to residues 88–103 of the known primary structure of Allergen M. The immunochemical reactivity of this loop, previously demonstrated for the overlapping enzymic fragments, was confirmed by using the synthetic preparation. The purified hexadecapeptide was shown to bind specifically to reaginic IgE from sera of cod‐allergic individuals, in both in vivo and in vitro test systems. It could also bind rabbit anti‐Allergen M, as shown by rocket line immunoelectrophoresis and quantitative precipitation inhibition techniques. The findings emphasized that the immunological reactivity of the synthetic peptide (88–103) was compatible with a monovalent haptenic function: blocking and not eliciting allergic reactions.
Characterization of Human Null Cells Isolated from Peripheral Lymphocytes by a Simultaneous Double‐Rosetting Procedure
Authors: NG A.‐K. , INDIVERI F. , RUSSO C. , QUARANTA V. , PELLEGRINO M. A. , FERRONE S.
Published: June 2006

Human null cells were isolated from peripheral blood lymphocytes (PBL) by differential centrifugation on a Ficoll‐Hypaque gradient of the PBL following simultaneous rosetting with erythrocyte indicators specific for B and T lymphocytes. Specifically, the T lymphocytes were rosetted with 2‐aminoethylisothiuroniuum bromido‐treated sheep erythrocytes (ShE), whereas the B lymphocytes were either rosetted with ShE coated with xenoantibodies against human gamma globulin or first sensitized with monoclonal antibodies to human Ia‐like antigens and then rosetted with ShE coated with xenoantibodies against mouse gamma globulin. Approximately 90% of the lymphocytes isolated were null cells that did not bear detectable B‐cell markers—that is, surface immunoglobulin and/or Ia‐like antigens—or T‐cell markers—that is, ShE receptors. The large majority of the null cells expressed receptors for the IgG Fc fragment (53–92%), C3 component (65–92%), and monkey erythrocytes (60–91%) but lacked receptors for the IgM Fc fragment and murine erythrocytes. The null cells exhibited high natural killer cell activity and antibody‐dependent cellular cytotoxicity and were two‐ to four‐fold more active than the total PBL and the T‐enriched cell fractions. The null cells, however, did not respond to stimulation with phytohaemagglutinin and failed to function as either stimulation or responders in an unidirectional mixed lymphocyte reaction.
Properties of Cell Subpopulations of Starfish Axial Organ: In Vitro Effect of Pokeweed Mitogen and Evidence of Lymphokine‐like Substances
Authors: LECLERC M. , BRILLOUET C. , LUQUET G. , AGOGUE P. , BINAGHI R. A.
Published: June 2006

The in vitro effects of pokeweed mitogen (PWM) on axial organ (AO) cells of the echinoderm starfish Asteria rubens have been studied. PWM stimulates 3H‐thymidine incorporation by the whole population of AO cells (index of stimulation 4–5). On the basis of surface adherence to nylon wool the AO cell population can be fractionated into adherent and non‐adherent subpopulations. PWM strongly stimulates the non‐adherent cells (index 14) but not the adherent cells (index 1.2–1.8). Specific soluble mediators obtained from stimulated non‐adherent cells were able to stimulate the whole AO cell population but not the fractionated adherent or non‐adherent cells.
Differences in Specific Antibody Responses of Human Tonsillar Cells to an Oral and a Parenteral Antigen
Authors: PAGANELLI R. , LEVINSKY R. J.
Published: June 2006

Tonsillar lymphocytes stimulated in vitro with either β‐lactoglobulin or with tetanus toxoid were shown to produce specific antibodies by a direct plaque assay and by radioimmuno‐precipitation of the culture supernatants. There was a sixfold increase in the number of IgA‐secreting cells in response to β‐lactoglobulin; no such effect was seen in response to tetanus toxoid, where a fivefold rise in IgG‐secreting cells occurred. These differences in antibody response are probably due to the route of initial antigen presentation. Those antigens priming the mucosa‐associated lymphoid system stimulate mainly IgA‐producing cells, in contrast to parenteral antigens, which elicit a predominantly IgM and IgG response.
ADCC of Cultured Human Melanoma Cells: Analysis with Monoclonal Antibodies to Human Melanoma‐associated Antigens
Authors: IMAI K. , NG A.‐K. , GLASSY M. C. , FERRONE S.
Published: June 2006

Four monoclonal antibodies to human melanoma‐associated antigens (MAA) were found to be able to mediate specific cell‐dependent lysis (ADCC) of cultured human melanoma cells. The extent of specific lysis of melanoma cells was influenced by the effector to target cell ratio, the amount of antibody added to the reaction mixture, and the incubation time. Cultured melanoma cells treated for 16 h with puromycin or cycloheximide (final concentration, 1.0 μg/ml) displayed increased Susceptibility to ADCC even though the cell surface expression of MAA was not changed. On the other hand, treatment of melanoma cells with tunicamycin (final concentration, 1.0 μg/ml) reduced the expression of MAA but did not affect susceptibility to ADCC. These findings suggest that other properties of the target cell membrane besides antigen density play a role in the outcome of ADCC reaction.