bluuebunny/crossref_metadata_2025 huggingface dataset:
In Vitro Generation of Eosinophil Chemotactic Factor from Human and Murine Mononuclear Phagocytes
Authors: CZARNETZKI B. M.
Published: June 2006

Low molecular weight eosinophil chemotactic factor (ECF), which has previously been demonstrated in mast cells, basophils, neutrophils and eosinophils, was shown to be released by several types of mononuclear phagocytes. Highly purified rat peritoneal macrophages and human monocytes produced ECK on stimulation with the calcium ionophore A23187 (Ion) and with phagocytic stimuli in a time‐dependent fashion, whereas lymphocyte‐ or mast cell‐specific stimuli were ineffective. Two murine macrophage lines and a fibroblast cell line (L cells) also generated and secreted ECF with the different stimuli. ECF from macrophages was similar to that from neutrophils in its target cell specificity (eosinophils and neutrophils) and its elution profile on Sephadex G‐Hl columns (300–500 dalton). ECF secretion from monocytes was not affected by mitomycin C or cycloheximide, whereas indomethacin enhanced and a phospholipase A inhibitor decreased its production. These in vitro findings suggest that, through LCF, mononuclear phagocytes may potentially regulate eosinophil and neutrophil influx to sites of inflammatory reactions.
Requirements for Induction of Specific Suppressor T Cells and Detection of their H‐2 Antigen‐binding Receptors by Fractionation on Target Cell Monolayers
Authors: BRONDZ B. D. , KARAULOV A. V. , ABRONINA I. F. , BLANDOVA Z. K.
Published: June 2006

The MC‐resistant specific suppressor T cells that inhibit DNA synthesis and CTL generation in MLC were induced in vitro by γ‐irradiated allogeneic lymphoid cells in a large dose. MLRs were inhibited only slightly when triggered by third‐party cells, even neighbouring with the corresponding sponding stimulators. Unlike the irradiated cells, intact allogeneic lymphoid cells induced a mixture of macrophage‐like and T‐cell suppressors with a pronounced non‐specific component of the action. Syngeneic cells induced low active non‐specific suppressors of macrophage type only. The suppression was not due to a cytotoxic effect, since specific T suppressors differed from CTL by conditions of induction and high sensitivity to γ‐irradiation and from CTL procurers by high sensitivity to CY and HC. The specific T‐suppressors could be selectively removed by adherence to a macrophage monolayer of the corresponding donor. The subsequent elution of adherent lymphocytes with pronase resulted in enrichment of specific T suppressors by a factor of 30 and 2.6, as judged by reduction in the number of lymphocytes required for 50% inhibition of DNA synthesis and 33% inhibition of CTL generation, respectively. The high specificity of this enrichment is shown by using both syngeneic monolayer for fractionation and third‐party stimulators in MLR for testing and by disappearance of slight non‐specific suppression caused by non‐fractionated suppresses. Complete inactivation of the eluted suppressors with anti‐Thy‐1.2 and anti‐T antibodies, their resistance to anti‐Mls antibodies, carrageenin, and carbonyl iron, together with the data of autoradiographic study and total DNA synthesis in the population indicate that the eluted highly specific suppressors represent mainly DNA‐synthesizing large and medium T lymphocytes.
Anti‐Self Receptors
Authors: SIA D. Y. , PARISH C. R.
Published: June 2006

Murine thymocytes and peripheral lymphocytes bind autologous erythrocytes via H‐2L‐region‐restricted receptors. After inhibiting autorosetting with different erythrocyte sonicates the specificity of these anti‐self receptors was examined in F1 hybrid and chimaeric mice. Most F1 lymphocytes simultaneously expressed receptors against both parental haplotypes. Furthermore, analysis of lymphocytes from allogeneic and semi‐allogeneic chimaeras clearly demonstrated that radio‐resistant elements in the recipient thymus did not modify the haplotype specificity of the receptors on donor‐derived lymphocytes.
Analysis of the Expression of I‐Ak‐like Antigens in Murine Fetal and Adult Tissues with the Monoclonal Antibody 10–2.16
Authors: NATALI P. G. , DE MARTINO C. , PELLEGRINO M. A. , FERRONE S.
Published: June 2006

The expression of I‐Ak antigens in normal C3H/FeJ adult and 15‐day embryonic mice has been investigated by indirect immunofluorescence staining of tissue cryostat sections with the anti I‐Ak antigen monoclonal antibody 10–2.16. In adult mice I‐Ak antigens were expressed in Langerhans‐like cells in the skin, epithelium of the gastrointestinal tract, endometrium, thymic reticuloepithelial cells, and several capillary endothelia. On the other hand. these antigens were not detected in Kupffer ceils, alveolar macrophages, brain or mammary gland. In 15‐day‐old embryos the expression of Ia‐like antigens was restricted to thymic reticuloepithelial cells, isolated spleen cells, and capillaries of the gastrointestinal tract.
Histamine Secretion from Human Leucocytes Stimulated by Basophil‐derived Platelet‐activating Factor
Authors: O'DONNELL M. C. , FIEDEL B. A.
Published: June 2006

Crude preparations of platelet‐activating factor (PAF) derived by methanolic extraction of supernatants from antigen‐challenged basophils generally stimulated the secretion of histamine from human peripheral blood leucocytes when co‐incubation with these cells in the absence of an additional stimulator. When these crude preparations were analysed by a preparative thin‐layer chromatography technique established for the isolation of purified native PAF, multiple lipid components were identified. The component that migrated in accordance with native PAF, and which activated platelets to aggregation secretion, was responsible for the induction of histamine release previously seen with the crude PAF preparations and could augment histamine secretion fro m leucocytes undergoing stimulation with either the biologically active peptide F‐Met‐Leu‐Phe or with antigen H. These data suggest that basophil‐derived PAF may serve as an important endogenous feedback mechanism during basophil activation in addition to its modulatory role upon the platelet.
Hybrid Resistance to EL‐4 Lymphoma Cells
Authors: LUEVANO E. , KUMAR V. , BENNETT M.
Published: June 2006

(C57BL/6 × DBA/2)F1 hybrid (B6D2F1) mice resist the growth of parental‐strain (B6) EL‐4 lmphoma cells inoculated intraperitoneally; that is, B6D2F1 mice survive longer than B6 mice and do not develop uscites As compared with B6 mice, B6D2F1 mice have higher levels of natural killer (NK) activity against 51Cr‐labelled EL‐4 cells in their lyphiod organs. B6D2F1 mice treated with 89Sr lose NK activity for certain lymphoma cell targets, e.g. YAC‐1, but NK(EL‐4) function is usually intact. However, 89Sr‐treated mice had hybrid resistance to EL‐4 cell in vivo, as determined by survival times and the development of ascites. NK(EL‐4) and NK‐(YAC‐1) activities were stimulated by irrudiated or unirradiated EL‐4 cells, Corynebacterium parvum, or polyinosinic:polycytidylic acid (pl:pC) in spleens of normal B6D2F1 mice, but NK(EL‐4) activity was depressed Within 3 days by such treatment in B6D2F1 mice previously injected with 89Sr. Suppressor cells for NK(EL‐4) but not for NK(YAC‐1) effectors were easily detected in spleens of 89Sr‐treated mice ‘challenged’ with C. parvum. Thus, agents capable of Stimulating NK cell function in normal mice may lead to suppression of that activity in mice depleted of marrow‐dependent cell function by 89Sr. Spleen cells of 89Sr‐treated B6D2F1 mice were also unable to generate anti‐EL‐4 cytotoxic T lymphocytes in a cell‐mediated lympholysis system; this defect appeared also in to be mediated by suppressor cells. Lymphoid cell depleted by 89Sr‐induced marrow aplasia may have two functions in host defences against tumours (especially lymphomas): they may lyse tumour cells directly and they may ‘down‐regulate’ suppressor cells capable of inhibiting other ‘natural’ or ‘induced’ immune functions.
Influence of in Vivo Hydrocortisone on Some Human Blood Lymphocyte Subpopulations
Authors: ONSRUD M. , THORSBY E.
Published: June 2006

The effect of in vivo hydrocortisone (OHC) on natural killer (NK) activity was studied using the K562 cell line as target in a 3‐h 51Cr‐release assay. Peripheral blood lymphocytes obtained from live normal volunteers at 0, 4, 24 and 48 h after Intravenous administration of 300 mg of OHC showed significantly increased NK activity at 4 h, decreased activity at 24 h, with a return toward normal at 48 h. Parallel variation were found in the fraction of lymphocytes bearing receptor for the Fe part of IgG. However, neither the number of these cell nor the NK activity was influenced by the medication when the results were given per millilitre of blood. In vitro pre‐incubation of the effector with OHC for 24 h had no effect on viability, expression of surface markers, or NK activity. It is concluded that under the present conditions NK activity is OHC‐resistant. The variations observed after in vivo administration seem to be due to a reversible redistribution mainly affecting cells other than the NK effectors.
Lupus Erythematosus (LE) Factors Recognize both Nucleosomes and Viable Human Leucocytes
Authors: REKVIG O. P. , HANNESTAD K.
Published: June 2006

The morphological LE test was completely inhibited by mixing LE‐factor‐positive sera with core mononucleosomes, the basic histone‐DNA subunit of chromatin. Free DNA and free histones at physiological ionic strength were not inhibitory. The LE‐factor‐binding antigen was virtually completely regained by mixing DNA and the inner histones H2A, H2B, H3 and H4 in 2mNaCl, followed by step dialysis to 0.15msalt. Trypsin‐digested core mononucleosomes did not inhibit the LE test, indicating that the antigenic site is located on the N‐terminal region of the histones. Twenty‐six out of twenty‐seven LE‐factor‐positive sera contained antinuclear antibodies that cross‐reacted with plasma membranes of viable leucocytes (referred to as X‐ANA). By contrast, weak X‐ANA activity was found in only two out of twenty‐five LE‐factor‐negative sera with high ANA titres. Three LE‐factor‐positive sera were subjected to absorption with viable human leucocytes, which led to the complete disappearance of LE factor activity of all three sera. The data indicate that antibodies with LE factor activity recognize core mononucleosomes and a plasma membrane antigen of viable human leucocytes.
Production of Immune Interferon (Type II) in Cocultures of Mouse Peritoneal Macrophages and Syngeneic Tumour Cells
Authors: OLSTAD R. , DEGRÉ M. , SELJELID R.
Published: June 2006

Mouse peritoneal macrophages were activated by coculture with syngeneic methylcholanthrene‐induced sarcoma cells [7], which grow as an ascites tumour in C3D2 mice [3]. During 4–5 days of cocultivation tumour cells progressively died, leaving highly activated macrophages. Cell‐free supernatant harvested from the cultures contained larger amounts of interferon than either macrophages or tumour cells cultivated alone. Peak activity of interferon occurred on day 2. Non‐adherent cells cultivated together with tumour cells did not produce interferon. Removal of all non‐adherent cells from macrophage cultures and host cells from tumour cell suspension did not abolish interferon production. The macrophages thus seem to be the interferon‐producing cells, but the possibility that the very few remaining lymphocytes may cooperate with the macrophages in interferon production cannot be totally ruled out. The interferon produced could not be inactivated by antibodies against virus‐induced interferon and was destroyed by treatment at pH 2, indicating that the interferon was not of the α or β type.
Microheterogeneity of Alpha‐fetoprotein in Patient Serum as Demonstrated by Lectin Affino‐electrophoresis
Authors: BRĘBOROWICZ J. , MACKIEWICZ A. , BRĘBOROWICZ D.
Published: June 2006

Microheterogeneity of alpha‐fetoprotein (AFP) present in the sera of 76 patients was studied by lectin affino‐immunoelectrophoresis. Seventeen patients had benign liver disorders and the remaining 59 patients were treated for primary or secondary liver cancer or yolk sac tumour. By means of Con A, AFP was divided into two variants, while lentil lectin (LCA) made it possible to separate AFP in three variants. In some patients the relative concentrations of Con A and LCA AFP variants were similar; these patients were believed to produce AFP of the same ‘profile’. Fourteen AFP profiles were observed by estimation of the area enclosed by precipitates corresponding to respective AFP variants. It was also possible to estimate the AFP profile on the basis of a simple visual analysis of the electrophoretic plates. The obtained results indicate that the AFP profiles of patients with cancer were variable. In spite of variations of the AFP profile in cancer patients, in most cases it was possible to differentiate primary liver cancer from yolk sac tumour and from liver metastases of cancer. In addition, in two‐thirds of hepatoma patients the AFP profile was different from the profile observed in patients with benign liver disorders.
Differential Binding of IgG Subclasses to Enzyme‐treated Human Lymphocytes
Authors: GILBERT D. , DAVEAU M. , FONTAINE M.
Published: June 2006

Human erythrocytes coaled with each of the four human IgG subclasses were used to detect Fcγ receptors on human peripheral blood lymphocytes (HPBL). Rosette formation was obtained only with erythrocytes coated with IgG3 (EAγ3). Neuraminidase treatment of HPBL induced rosette formation with EAγ1, EAγ2, and EAγ4 complexes. Pronase treatment also induced rosette formation to a lesser extent, but abolished EAγ3 rosetting. Trypsin treatment enhanced EAγ3 rosette formation. These phenomena occurred on both ‘T’ and ‘non‐T’ lymphocytes. Erythrocytes coaled with small quantities of IgG3 did not form rosettes with HPBL. Neuraminidase treatment enhanced their binding, whereas pronase did not. These two phenomena occurred only on non‐T lymphocytes. Rosette formation with EAγ1 was also obtained with lymphocytes stimulated in vitro with mitogens. After 2 days of culture, stimulated lymphocytes expressed receptors that were able to bind both EAγ1 and EAγ3 complexes. These results suggest the existence of cryptic receptors for IgG1, IgG2, and IgG4 that could be disclosed by neuraminidase and pronase treatment and exposed on stimulated lymphocytes. A hypothesis of one or several Fcγ receptors is suggested.
Shared Idiotypes Among Anti‐Albumin Antibodies of Different Species
Authors: JACKSON S. , KULHAVY R. , MESTECKY J.
Published: June 2006

A system designed to detect plasma cells that produce antibodies directed at autologous idiotypic determinants of anti‐human serum albumin (HSA) antibodies in rabbits was used to determine whether anti‐HSA antibodies of horse, goat, swine and chicken origin were cross‐reactive with rabbit antibodies of the same specificity. Fluorochrome‐tagged anti‐HSA preparations of these diverse species were used to stain splenic plasma cells of HSA‐immunized rabbits and a similarly immunized chicken. The degree of idiotypic cross‐reactivity, as detected by binding of anti‐HSA antibodies to anti‐idiotype within plasma cells of HSA‐immunized animals, was sometimes equal to autologous staining. However chicken anti‐Has, the most phylogenetically distant idiotype examined, was demonstrably less cross‐reactive than that obtained from the other species. Likewise, chicken plasma cells usually did not bind mammalian anti‐HSA antibodies to an appreciable degree, as compared with autologous staining. These findings provide evidence for serologic and possibly structural similarities of antibodies of the same specificity from different species.
Regulation of IgG Antibody‐dependent Cellular Cytotoxicity in Vitro by IgM Antibodies
Authors: PERLMANN H. , PERLMANN P. , MORETTA L. , RÖNNHOLM M.
Published: June 2006

IgM antibodies have previously been reported to either inhibit or induce antibody‐dependent lymphocyte cytotoxicity (ADCC). Here we show that human lymphocytes lyse bovine erythrocytes (Eb) in the presence of either IgM or IgG anti‐Eb from rabbits. Seven out of 20 IgM preparations (Sephadex G‐200) were ADCC‐active. IgG‐dependent ADCC was inhibited by human IgG but not by IgM. In contrast, IgM ADCC was inhibited by both IgG and IgM. The effector cells in IgM ADCC were a subpopulation of lymphocytes with distinct Fc receptors for both IgG and IgM. Most of them also had sheep erythrocyte receptors. Extensive purification of the ADCC‐active IgM antibody preparations indicated that very small amounts of contaminating IgG anti‐Eb were responsible for ADCC induction. When purified and ADCC‐inactive IgM antibodies were mixed with suboptimal concentrations of IgG antibodies, a strong enhancement of ADCC was found. To achieve enhancement, the two antibody isotypes had to be present on the surface of the same target cells, and the IgM effect was not due to the release of soluble ADCC‐enhancing factors. Thus, in this system, IgM antibodies are not capable of inducing ADCC on their own. However, they enhance ADCC by improving the contactual interaction between target cells and a special subset of effector cells.
Quantitative Precipitin Analysis of Free and Hidden RF IgM
Authors: UESSON M. , HANSSON U.‐B.
Published: June 2006

Quantitative precipitin analysis using rabbit IgG anti‐human Fcμ was performed with 16 RF IgM, six macroglobulinaemic IgM and four normal IgM. Abnormal precipitin curves were obtained for all RF, IgM, even when the latter were not readily demonstrated with conventional serological tests for RF, and for macroglobulinaemic IgM with sedimentation rats greater than 19S. These IgM formed significantly more precipitates with IgG anti‐Fcμ in the antigen excess one than did normal IgM, but the precipitin curves for the other zones were similar for all IgM. The underlying mechanisms of some of the reactions were studied and discussed. Because the divergence in the precipitin reaction for normal IgM and RF IgM was so pronounced, a model precipitin curve was constructed. This could be used to detect RF IgM, even when not readily demonstrable with conventional serological tests for RF, by direct analysis of serum. The results obtained for RF IgM suggested that the method might be applied to RF IgG and intermediate complexes comprised of IgG. The mechanisms demonstrated here might be used to develop immunological methods for routine use.
Dimeric IgA in the Rat is Transferred from Serum into Bile but not into Milk
Authors: DAHLGREN U. , AHLSTEDT S. , HEDMAN L. , WADSWORTH C. , HANSON L. Å.
Published: June 2006

Intravenous administration of ducts thoracicus lymph with dimeric IgA antibodies against Escherichia coli 06 to lactating rat dams did not result in transfer of IgA antibodies into the milk, although the antibodies were detectable in serum 1 min after the administration and in bile 60 min later. After intravenous injection of serum from bile‐duct‐occluded (BDO) rats Immunized in the Peyer's patchs into lactating rat dams, IgA antibodies appeared in the serum and remained there up to 230 min. At this time no IgA antibodies were seen in the milk while they were present in bile. IgG and IgM 06 antibodies did not appear in bile or after intravenous administration of lymph or serum from BDO rats.
Clinical Significance of Immunoglobulin‐secreting Cells in the Peripheral Blood in Patients with Multiple Myeloma
Authors: NAGAI K. , TAKATSUKI K. , UCHINO H.
Published: June 2006

Immunoglobulin‐secreting cells were enumerated in the peripheral blood of 31 patients with multiple myeloma, nine patients with so‐called benign monoclonal gammopathy, and ten patients with polyclonal hypergammaglobulinaemia or idiopathic hypogammaglobulinaemia. Immunoglobulin‐secreting cells were detected by a haemolytic plaque assay using protein‐A‐coated erythrocytes in the presence of class‐ or type‐specific rabbit anti‐human immunoglobulin antisera. In about two‐thirds of patients with multiple myeloma, cells secreting the same light‐chain isotype of their serum M‐protein increased in number, whereas this was not the case in the patients with so‐called benign monoclonal gammopathy. Follow‐up studies of immunoglobulin‐secreting cells in multiple myeloma revealed that these cells increased or decreased, correlating with the severity of the disease, and alterations were more rapid than other clinical features. This plaque assay is therefore useful in evaluating the response to chemotherapy in patients with multiple myeloma.
The Influence of Serum on Lymphocyte Cultures
Authors: KRISTENSEN F. , JONCOURT F. , DE WECK A. L.
Published: June 2006

Serum exerts several effects in lymphocyte cultures, one of them being manifested very early. The presence of fetal calf serum (FCS) results in a markedly higher thymidine uptake within a few minutes, as compared with serum‐free cultures. On the other hand, the uptake of uridine and other purine bases seem to be little influenced by serum. Experiments comparing the uptake of thymidine into the cytoplasm or into trichloroacetic acid‐precipitable material suggested that the intracellular thiymidine pool increases in size when FCS is added. Using a Lineweaver Burt plot for thymidine and uridine uptake over a 4‐h period, no changes in uridine uptake were observed in the presence of FCS; on the contrary, serum induced an increased Vmax for thymidine, whereas Km remained constant. Cytofluorographic quantitation of G0 and G1 cells indicated that cells disappear more rapidly from the G1 phase in the presence of FCS. The addition of hydroxyurea to the cultures prevented this disappearance. The results taken together strongly suggest that serum contains a factor promoting the shift of G1 cells into the S phase. When a 4‐h culture period was used, all sera from differed species tested at low concentrations appeared to contain this activity.
Increased Serum IgA Levels in Rats after Portacaval Shunt but not after Portacaval Transposition
Authors: VAERMAN J. P. , LEMAÎTRE‐COELHO I. , McSWEEN R. N. M. , BENJAMIN I. S. , THOMAS H. C.
Published: June 2006

Groups of rats were submitted to end‐to‐side portacaval shunt, portacaval transposition or a control sham operation. There was an 18‐fold increase of the IgA level in the serum of portacavally shunted rats (n=7) at 3–6 weeks after surgery. Gel filtration revealed that this increase was predominantly due to dimeric and polymeric IgA. The serum IgM of these shunted rats, but not the IgG, was also significantly increased (twofold) above that control values. In rats subjected to portacaval transposition (n=6), there was no significant increase of serum IgA, IgM or IgG when compared with their control values. These data confirm the active transfer, by the healthy rat liver, of IgA and, to a lesser extent, of IgM from the blood into bile. Impairment of this function, leading to the accumulation of polymeric IgA and IgM in the serum, in portacavally shunted but not portacavally transposed animals, may be related to impairment of hepatic blood Hot and hepatic atrophy, shown earlier to occur in portacavally shunted but not transposed rats. Such redaction in liver cell mass could also explain the fall in transferrin levels in shunted rats.
Isolation of a Heat‐stable Antigen from Treponema Reiter, Using an Immunoadsorbent with Antibodies from Syphilitic Patients
Authors: PEDERSEN N. STRANDBERG , PETERSEN C. SAND , AXELSEN N. H. , BIRCH‐ANDERSEN A. , HOVIND‐HOUGEN K.
Published: June 2006

It was attempted to isolate antigens from Treponema Reiter, relevant to syphilis serology, by immunoadsorption with patients' antibodies coupled to CNBr‐Sepharose 4B. One antigen was desorbed by 2 m KSCN in 0.05 m Tris barbital buffer, pH 8.6. The recovery was 3% and 7% in two experiments. A small amount of human antibodies in the isolate was removed on an immunoadsorbant column with insolubilized rabbit antibodies against normal human serum proteins. The antigen thus obtained was immunologically pure when analysed by crossed immunoelectrophoresis. By electron microscopy of immunoprecipitates and by tandem crossed immunoelectrophoresis it was shown that the antigen differed from the flagellar antigen of T. Reiter, but was identical to antigen d previously described in T. Reiter, Antigen d could also be isolated from supernatant of T. Reiter cultures, The d antigen was not denatured at pH 2.8, by 8 m urea or by 3 m KSCN, and it resisted heating to 100°C for 30 min. No protein could be detected in a concentrated preparation, and the antigen might be B polysaccharide. Antigen d is probably present in the sorbent used in the fluorescence treponemal antibody absorption (FTA‐ABS) test and may constitute the active substance of this reagent.
On the Importance of Substrate Adhesion and Cell Aggregation in the Con A‐induced Blastogenesis of Lymphocytes
Authors: RUUD E. , STEEN H. B.
Published: June 2006

The importance of substrate adhesion and cell aggregation for the Con A‐induced blastogenesis of human peripheral lymphocytes was studied by flow cytometric methods. The percentage of lymphocytes responding to the mitogen by growth and DNA synthesis was approximately the same among non‐adherent, single cells as among substrate‐adherent and aggregated cells. However, the response of the former cells was delayed by about 10 h as compared with that of adherent and aggregated cells. This delay increased to about 20 h in monocyte‐depleted cultures. Stimulation with succinyl‐Con A, which caused negligible aggregation, produced a significant response. Blast cells appeared to be largely nonadherent. It is concluded that neither substrate adhesion nor cell aggregation are prerequisites for Con A‐induced blastogenesis, although substrate adhesion seems to have a significant promotive effect that is probably associated with a direct contact of the lymphocytes with monocytes.
Concanavalin A‐induced Suppressor Cell Activity in Lipoid Nephrosis
Authors: OSAKABE K. , MATSUMOTO K.
Published: June 2006

Suppressor cell activity (SCA) was analysed in 28 patients with lipoid nephrosis (LN) and I patients with chronic proliferative glomerulonephritis (CGN). We have assessed the ability of peripheral blood lymphocytes (PBL) stimulated by concanvalin A(Con A) to inhibit the proliferative response of normal allogeneic lymphocytes by both Con A and phytohaemagglutinin (PHA). It was found that the LN patients in the earlier phase of relapse had significantly increased levels of suppression index (SI) when compared with the values obtained with normal controls. In contrast, the mean suppression values in the PBL from LN patients in remission and CGN patients with or without nephrotic syndrome, whether the mitogen used was Con A or PHA, were similar in those of the control subjects. Moreover, when individual patients were followed through their clinical illness. LN patients had high levels of SI, particularly in the beginning of acute exacerbations; the SI levels then decreased sharply in the latter phase of relapse and again increased to relatively normal levels with the onset of clinical remission. These in vitro findings suggest that there exists an alteration in Con A‐induced SCA in a group of patients with LN.
Effect of Tunicamycin on the Assembly and Antigenicity of HLA Antigens: Analysis with Monoclonal Antibodies
Authors: WILSON B. S. , GLASSY M. C. , QUARANTA V. , NG A.‐K. , FERRONE S.
Published: June 2006

The effect of glycosylation on the assembly and antigenicity of HLA antigens was investigated by examining HLA antigens synthesized in the presence of the antibiotic tunicamycin, an inhibitor of asparagine‐linked oligosaccharide addition, with monoclonal antibodies specific for a variety of antigenic determinants. The monoclonal antibody Q5/13 reactive with a determinant expressed on the β chain of human Ia‐like antigens immunoprecipitated α and β subunits with reduced apparent molecular weights from tunicamycin‐treated cells, indicating that glycosylation is not required for association of the Ia‐like antigen α and β subunits. Immunoprecipitation of HLA‐A,B,C antigens from tunicamycin‐treated cells with four monoclonal antibodies specific for the heavy chain and one specific for β2‐microglobulin showed that the heavy‐chain determinant detected by the antibody Q6/64 is absent from the non‐glycosylated molecule. This is the first demonstration that carbohydrate addition during biosynthesis affects the protein conformation of the HLA‐A,B,C heavy chain.
Clones of Human Cytotoxic T Lymphocytes Derived from an Allosensitized Individual: HLA Specificity and Cell Surface Markers
Authors: MALISSEN B. , KRISTENSEN T. , GORIDIS C. , MADSEN M. , MAWAS C.
Published: June 2006

By planned immunization of a volunteer, two stable (≥6 months), specific, alloreactive cytolytic T‐cell clones have been established from his peripheral blood lymphocytes. One clone reacts with all serologically defined HLA‐Cw3 cells from our panel, whereas the other defines a split within the serological HLA‐B40 specificity. The two cytotoxic clones are SmIg‐negative, E‐rosette positive, EA and EAC rosette‐negative. HLA‐A, ‐B and ‐C‐positive, and also HLA‐DR‐ or ‘Ia like‐positive. In addition, they present very similar patterns of iodinated cell surface molecules as analysed by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS‐PAGE), contrasting with that of an EBV cell line derived from the same donor.
Use of Plaque Assays to Study Thyroglobulin Autoantibody Synthesis by Human Peripheral Blood Lymphocytes
Authors: McLACHLAN S. M. , BIRD A. G. , WEETMAN A. P. , SMITH REES , HALL R.
Published: June 2006

Plaque assays have been used to study thyroglobulin autoantibody synthesis and total immunoglobulin production by cultures of peripheral blood lymphocytes from patients with Hashimoto's thyroiditis. Freshly isolated Hashimoto or normal lymphocytes contained small numbers (7–1300) of total IgG plaque‐forming cells (PFC) and total IgM PFC, but specific thyroglobulin antibody PFC was undetectable. After 5–8 days' culture with pokeweed mitogen (PWM), total IgG and IgM PFC were markedly increased to a geometric mean of 10, 140 (8414–12,220) IgG PFC and 3450 (1663–7534) IgM PFC per 106 cultured lymphocytes (95% confidence limits in parentheses). Furthermore, a mean of 63 (22–287) specific IgG thyroglobulin antibody PFC per 106 lymphocytes were detectable in cultures of Hashimoto lymphocytes. The IgG thyroglobulin antibody PFC were stimulated by Epstein‐Barr virus (EBV) infection, suggesting that EBV infection may be useful in obtaining monoclonal thyroid autoantibodies.
Delayed‐Type Hypersensitivity to Ovalbumin in Bone Marrow Chimeric Mice
Authors: LUBET M. T. , KETTMAN J. R.
Published: June 2006

The site of action of the H‐2 linked gene regulating the ability of mice to be sensitized to transfer delayed‐type hypersensitivity was studied using radiation bone marrow chimeras. Lethally irradiated (non‐responder × responder) F1 were reconstituted with bone marrow cells of both parents. Cells of chimeras immunized with limiting doses of ovalbumin responded as did the parental strains. These results suggest that the gene regulating delayed type hypersensitivity is expressed at the level of the delayed‐type hypersensitivity effector cells or that restrictions exits on the collaboration of the helper cells for DTH effectors (which differ at several loci including the major histocompatibility complex).
Serum Inhibitory Factor in Lepromatous Leprosy: Its Effect on the Pre‐S‐Phase Cell‐Cycle Kinetics of Mitogen‐stimulated Normal Human Lymphocytes
Authors: POTTS R. C. , SHERIF M. M. , ROBERTSON A. J. , GIBBS J. H. , BROWN R. A. , BECK J. SWANSON
Published: June 2006

The sera of ten Egyptian men with long‐standing lepromatous leprosy (LL) (mean duration 17.4 years) that had failed to respond in dapsone treatment were shown to inhibit mitogen stimulation responses of normal human lymphocytes. When first tested, the sera partly inhibited the response to phytohaemagglutinin (PHA) and pokeweed mitogen and virtually abolished that to concanavalin A (Con A): after repeated freezing and thawing, the Con A inhibition had disappeared, whereas the PHA response was still partly Inhibited. The inhibitory serum factor(s) had similar actions on lymphocytes from each of six normal donors. Although the sera varied in potency, they showed similar dose response curves when tested against lymphocytes from a single donor. The principal action of the sera was to reduce the number of cells responding to mitogen, without modifying the kinetics of recruitment or rate of volume growth during G1‐phase in those cells that were unaffected by the inhibitory substances(s). Study of PHA dose‐response curves and of the effect of delayed addition of LL serum suggested that the serum factor(s) act by diminishing the responsiveness of the cells, rather than by reducing the concentrations of free mitogen or by blocking cell membrane mitogen receptors The serum from one apparently healthy attendant, who had nursed leprosy patients for 30 years but who did not have leprosy or other chronic infective disease, inhibited completely stimulation by all three mitogens in a manner different from that of LL sera. Serum from the other 13 control patients did not modify the response of normal lymphocytes to stimulation by any of the three mitogens studied. It was concluded that the inhibitory factor(s) in the scrum of patients, with LL were a consequence of the disease and not of the environment in which the patients lived. Microscopy confirmed that the techniques used for recovery of the cultured cells did not introduce bias into the volume spectroscopy measurements.
HLA Restriction of Dinitrophenyl‐specific Cell‐mediated Cytotoxicity in Vitro
Authors: DICKMEISS E. , SØEBERG B. , SVEJGAARD A.
Published: June 2006

Lymphocytes from dinitrochlorobenzene‐sensitized individuals can be stimulated in vitro by autologous dinitrophenyl (DNP)‐conjugated lymphocytes to produce cytotoxic T lymphocytes (CTLs). The activity of these CTLs is specific for DNP‐conjugated target cells, and there is no cross‐reaction with nitrosodimethylaniline or trinitrophenyl‐conjugated target cells. Evidence is presented which makes it improbable that the cytotoxicity is caused by an antibody‐dependent (ADCC‐like) mechanism. Most of the DNP‐specific cytotoxicity is restricted by the HLA‐ABC antigens of the CTL donor, and there is only a low degree of lysis of DNP‐conjugated allogeneic target cells not sharing HLA‐ABC antigens with the donor. The CTLs did not lyse non‐conjugated allogeneic target cells. When CTLs were tested against allogeneic DNP‐conjugated targets sharing only one of the HLA‐ABC antigens of the CTL donor, it was seen that the phenomenon of preferential restriction was pronounced; that is, only some of the antigens of the donor were restricting. A certain pattern has emerged: some antigens (e.g. A2) are good restricting antigens, some (e.g. B12) do not restrict, and some (e.g. B5) function well in one donor but not in another. The sero‐logically cross‐reacting antigens A2 and A28 did not restrict mutually. HLA‐C antigens may in some donors function as restricting antigens.
Alpha‐Fetoprotein on Human Peripheral Blood Lymphocytes Does Not Block Complement‐dependent Lymphocytotoxicity
Authors: REN E. C. , CHAN S. H.
Published: June 2006

Using a direct immunofluorescence assay, we showed that alpha‐fetoprotein (AFP) both in purified form and in hepatocellular carcinoma (HCC) sera was capable of binding onto 10–20% of T lymphocytes and 5–10% of B lymphocytes in human peripheral blood when these lymphocytes were preincubated in AFP‐positive fluids at 4°C in the presence of sodium azide. But when the preincubation temperature was raised to 37°C, most of the membrane‐bound AFP was internalized or shed, and, consequently, less than 3% of the cells showed positive membrane fluorescence. In addition, binding of AFP onto lymphocyte surface membrane and the continuous presence of large amounts of AFP in these lymphocyte cultures did not interfere with the action of cytotoxic antibodies directed against HLA determinants on the lymphocyte surface.
Human Interleukin 2: Production by Both TG Cells and Other T Cells
Authors: SILVA A. G. , ALVAREZ J. M. , BONNARD G. D. , DE LANDAZURI M. O.
Published: June 2006

We have investigated the production of interleukin 2 (IL‐2) by human T cells after their stimulation by phytohaemagglutinin (PHA). T cells isolated by rosetting with sheep erythrocytes produced high levels of IL‐2. Further separation of rosette‐forming cells, according to the expression of Fc receptors for IgG, showed that TG and non‐TG cells are equally able to produce IL‐2. The release of IL‐2 by TG cells did not require DNA synthesis or functional Fey receptors, since positively selected TG cells produced IL‐2, even though they lacked lymphoproliferative responses to PHA and antibody‐dependent cell‐mediated cytotoxicity.
The Four Subclasses of IgG Can Be Isolated from Mouse Serum by using Protein A‐Sepharose
Authors: SEPPÄLÄ I. , SARVAS H. , PÉTERFY F. , MÄKELÄ O.
Published: June 2006

We confirmed the findings of Ey and colleagues that mouse IgG is absorbed by protein A‐Sepharose at pH 8.0. Also confirmed was their finding that IgG1 mainly elutes from such a column by means of a buffer with pH 6.0 and that the corresponding pH values for IgG2a and IgG2b were 4.5 and 3.5. We made the new finding that the hulk of IgG2a bearing allotypes a or j eluted already at pH 5, in contrast to IgG2a bearing allotype b. Another newfinding was that IgG3 was mainly eluted at pH 4.5 regardless of the allotype. All four subclasses of IgG could thus be physically separated if the allotype was a or J (the only known exception is allotype b). Separation of IgG2a and IgG3 was achieved even when the allotype was b by using a pH gradient for elution. IgG2a came out al a slightly higher pH than IgG3. Mouse IgG antibodies against group A streptococcal polysaccharide belonged mostly to IgG3 and, to a lesser extent, to IgG2a and IgG2b.
IgA and IgM Antibodies against Mycobacterium leprae in Cord Sera and in Patients with Leprosy: An Indicator of Intrauterine Infection in Leprosy
Authors: MELSOM R. , HARBOE M. , DUNCAN M. E. , BERGSVIK H.
Published: June 2006

A solid‐phase radioimmunoassay was developed for demonstration and quantification of IgA and IgM anti‐M. leprae antibodies. IgA and IgM anti‐M. leprae antibodies were demonstrated in a lepromatous serum pool, in various amounts in individual patients with lepromatous leprosy, and in lower concentration in tuberculoid leprosy and non‐leprosy controls. IgA and IgM anti‐M. leprae antibodies were demonstrated in cord sera from babies of mothers with leprosy. The reliability of fetal IgA and IgM antibody synthesis as an indicator of intrauterine infection in leprosy is discussed.
Degradation of Amyloid Proteins by Different Serine Proteases
Authors: SKOGEN B. , NATVIG J. B.
Published: June 2006

A protein extract was obtained from normal human serum by adsorption to unsubstituted Sepharose 4B. This extract contained one or several enzymes with SAA and AA degrading capacity. The optimal pH for degradation of SAA was about 7.3. On fractionation of the enzyme extract on Sephadex G‐150, the active component Was eluted in the V0peak. The V0 fraction, which on double immunodiffusion analysis was found to contain α2‐macroglobulin, was also active against synthetic substrates used to determine the activity of thrombin and plasma kallikrein. Gel filtration under dissociating conditions and molecular weight estimation further indicated the presence of those enzymes in the preparation. Several serine proteases which are known to be inhibited by α2‐macroglobulin possessed AA and SAA degrating activity. On degradation of SAA, an intermediate split product with molecular weight similar to AA was formed. Kallikrein, plasmin and elastase were also able to degrade intact amyloid fibrils suspended in phosphate‐buffered saline
Expression of the Major Histocompatibility Complex Antigens on Different Liver Cellular Components in Rat and Man
Authors: LAUTENSCHLAGER I. , HÄYRY P
Published: June 2006

We hint analysed the distribution of the major histocompatibility complex (MHC) antigens on different liver cellular components in rat and man. Two types of antisera were used: monospecific rabbit antisera raised against isolated molecules of class I and class II and ordinary hyperimmune alloantisera directed to their allelic specificities. A modified Staphylococcus aureus rosette test, enabling morphological identification of the rosette‐forming cell type, was used. In both species the passenger cells, especially the Kupffer cells, reacted strongly with rabbit anti‐class I and II The passenger erythrocytes reacted with anti‐class I in rat but not in man. Human liver vascular endothelial and bile duct cells displayed a distinct reaction with anti‐class I and II, whereas the hepatocytes reacted very weakly, if at all. In rat liver parenchymal component the expression of these antigens was different and weaker. Rat bile duct cells reacted with anti‐class I and II in approximately 50% of the testings, whereas rat vascular endothelial cells and rat hepatocytes were essential non‐reactive. The alloantiserum reactivity (tested only in the rat) followed closely that of heterologous antisera, and the reactivity of anti‐β2m that of class I alloantisera. Although these differences are probably quantitative rather than qualitative, the results suggest considerable variation in the expression of the MHC antigens on different liver cellular components. These differences partially explain why liver allografts behave differently than, e.g., renal allografts upon transplantation.
The H‐2 Restriction Specificity of Cytotoxic T Cells Derived from Intrathymic Precursors
Authors: HURME M. , SIHVOLA M.
Published: June 2006

Both cortical and medullary thymocytes could give rise to both allospecific and hapten‐specific cytotoxic T lymphocytes (CTLs) if the cultures contained the T‐helper‐cell‐replacing hormone, interleukin 2 (IL‐2). The H‐2 restriction specificity of these CTLs was tested by using three different specificity determinants that are known to be influenced by the H‐2 genotype of the thymus (rather than by the H‐2 of the precursor cell itself): the cross‐killing of trinitrophenyl (TNP) coupled allogeneic targets in anti‐TNP self‐response; the dominance of H‐2k‐restricted C‐TLs over H‐2d‐restricted CTLs in the response of (H‐2K× H‐2d)F1 mice to TNP‐self; and the capacity of alloreactive CTLs to cross‐reactively lyse TNP‐coupled syngeneic cells. The experiments show that, with regard to all of these specificity determinants, CTLs derived from these intrathymic populations are similar to CTLs derived from the adult mouse spleen. These data suggest that the H‐2 restriction specificity is dictated before or at the same time that the cell acquires capacity to respond to allogeneic or haptenated syngeneic cells.
Demonstration of Ia Antigens on Certain Dendritic Cells and on a Novel Elongate Cell Found in Human Synovial Tissue
Authors: WINCHESTER R. J. , BURMESTER G. R.
Published: June 2006

Adherent cells from dissociated human synovial tissue obtained at surgery contain two types of distinctive cells with one or more elongated branching processes that strongly express Ia antigens. One type of cell with Ia antigens is non‐phagocytic and resembles the murine dendritic cell. It is primarily found in patients with rheumatoid arthritis and accounts for a considerable proportion of the identifiable cells with a stellate or dendritic morphology. The expression of Ia antigens progressively diminished in culture. The second type of novel cell with Ia antigens was highly elongate and fibroblastoid. It was readily obtained from patients with osteoarthritis. The cell was frequently characterized by a blunt‐ended filopodium‐like process at one pole of the cell, one or two tapering processes, and zones of microvilli, Evidence was obtained suggesting that this cell, which might otherwise be considered fibroblast‐like, is in the mononuclear phagocyte lineage.
Evidence that Pokeweed‐Mitogen‐reactive B Cells are Pre‐committed in Vivo to the High‐rate Secretion of a Single Immunoglobulin Isotype in Vitro
Authors: STEVENS R. H. , MACY E. , THIELE C. J.
Published: June 2006

Normal human peripheral blood B cells that respond to pokeweed mitogen (PWM)‐activated irradiated T cells with high‐rate immunoglobulin secretion in vitro were analysed with respect to the frequency of the cells stimulated to high‐rate immunoglobulin secretion in vitro and whether the progeny of each cell had the potential to secrete one or multiple immunoglobulin isotypes. In vitro cultures containing limiting numbers of human B cells were initiated in the presence of PWM and excess irradiated T cells, and the quantity of IgM, IgG and IgA secreted was determined after 9 days. The level of immunoglobulin secretion per cell in limiting‐dilution microcultures was shown to be equivalent to that seen in the routinely used macrocultures, indicating that major loss of B‐cell function was not occurring in the microcultures. At limiting B‐cell numbers, individual microcultures were often shown to produce immunoglobulin of a single isotype, either IgM, IgG or IgA. Cultures that did produce multiple immunoglobulin isotypes occurred with a frequency predicted by the random distribution of B cells committed to production of a single isotype. A similar independent distribution of IgM anti‐tetanus toxoid and IgG anti‐tetanus toxoid antibody‐producing precursors was observed when B cells selected on the basis of surface IgM were used in the microcultures. These results suggest that PWM‐reactive B cells are at a stage of maturation in vivo such that they have the potential to secrete a single immunoglobulin isotype when activated in vitro.
Age‐Dependent Allospecific Change in Susceptibility to Local Lymph Node GVHR in Rats
Authors: HERON I.
Published: June 2006

Inoculation of parental strain lymphocytes into the footpads of young F1 hybrid rats induces GVHR, which can be quantitated by estimation of the weight of the popliteal lymph node 7 days later. In a number of strain combinations adult and old F1 animals were also useful for the assay. In some, however, it was observed that aged F1 rats did not support GVHR to the same extent as young did, when one of the parental strains served as donor for the GVHR, but reacted normally if the other parental strain was used. This phenomenon was analysed in serum and cell transfer experiments as well as in vitro in MLC. It was not possible to transfer suppression in vivo, whereas some degree of specific MLC inhibition was observed when serum from old F1 animals undergoing GVHR was assessed. The ‘old’ pattern of response could be reverted partially by splenectomy and completely by low dose irradiation of the recipient before GVHR.
B‐L Antigens (Ia‐like Antigens) of the Chicken Major Histocompatibility Complex
Authors: CRONE M. , JENSENIUS J. CHR. , KOCH C.
Published: June 2006

This paper reviews the present knowledge of B‐L antigens encoded by the chicken B complex as regards to the following aspects: (1) identification and cellular expression, (2) structural studies, (3) evidence for two distinct populations of B‐L antigens, (4) mapping of B‐L loci of the B complex, (5) B‐L and immune response, and (6) the role of the B‐L antigens for the control of mixed lymphocyte reactions (MLR) and graft‐versus‐host (GVH) reactions. It is concluded that B‐L antigens of the chicken exhibit extensive homology with mammalian la antigens. A genetic map of the B complex is presented.
Bone Absorption and the Immune System
Authors: NISBET N. W.
Published: June 2006

The absorption of bone in Grüneberg microphthalmic mice and the op op osteopetrotic rat can be stimulated by an injection of compatible bone marrow, and complete resorption can occur. It is probable that the bone‐absorbing osteoclast responsible is derived from a haematopoietic stem cell resident in the bone marrow (BM). Maintenance of the resorption depends on the survival of donor cells which has an important implication for clinical therapy in man. No evidence was found that the thymus played a leading role in the mechanism of bone absorption in the Grüneberg microphthalmic mouse, or in the op op rat.