bluuebunny/crossref_metadata_2025 huggingface dataset:
PROTECTIVE ANTITOXIC CHOLERA IMMUNITY IN MICE: INFLUENCE OF ROUTE AND NUMBER OF IMMUNIZATIONS AND MODE OF ACTION OF PROTECTIVE ANTIBODIES
Authors: Lange S. , Holmgren J.
Published: August 2010

An adult mouse model has been elaborated for studies of experimental cholera (Vibrio cholerae enterotoxin‐induced intestinal secretion) and protective antitoxic immunity in either ligated small bowel loops or intact small intestine. Mice of different inbred strains varied markedly in intestinal susceptibility to toxin, C57B1 being the most sentive strain tested. Fluid accumulation started within 1 h after the inoculation of toxin and was maximal after 5 h, whereafter recovery gradually ensued. The dose‐response curve was sigmoid, the ED50 of crude toxin being equivalent to about 0.1 μg purified toxin/cm in the loops and 0.3 μg/cm in the nonligated intestine. Two peroral (p.o.) immunizations induced significant protective immunity which increased markedly after two further immunizations by the same route. Additional p.o. immunizations did not appreciably enhance the protective immunity any further. Intravenous (i.v.) vaccination had to be repeated more than 5 times before intestinal immunity could be observed. No correlation between serum antitoxin titers and protective immunity was found. Electron microscopic examination revealed that whereas peroxidase‐coupled cholera toxin bound tightly to intestinal microvilli from unimmunized or 5‐times i.v. immunized mice it did not bind to the microvilli of p.o. immunized animals. The data thus suggest that the protective immunity is mediated exclusively by locally produced antibodies which prevent the binding of toxin to the gut epithelium.
Freezing Of Rat Lymphocytes
Authors: Hem Einar
Published: August 2010

Spleen lymphocyte proliferation, as measured by 3H‐thymidine incorporation induced by phytohae‐magglutinin (PHA), was increased after freeze‐thawing with 10% dimethyl sulphoxide. Depletion or intoxication of macrophages in fresh spleen cell preparations also increased lymphocyte proliferation in response to PHA. On the other hand, freezing of macrophage‐depleted spleen cell suspensions lowered 3H‐thymidine uptake of stimulated cultures. At concentrations above 3%, macrophages added to cultures of fresh purified lymphocytes showed a dose‐dependent inhibitory effect on the PHA response, and fresh macrophages were more inhibitory than frozen‐thawed macrophages. Purified lymphocytes mixed with 10% macrophages showed a higher response after freeze‐thawing. It is concluded that macrophages suppress the lymphoproliferative response to PHA in rat spleen cell cultures, and that these macrophages are more sensitive than lymphocytes to the present freeze‐thaw process.
LOCALIZATION OF Fc RECEPTORS IN HUMAN AND RAT MALIGNANT TISSUES
Authors: TöNder Olav , Krishnan Engikolai C. , Morse Paul A. , Jewell William R. , Humphrey Loren J.
Published: August 2010

Three human malignant tumors and a rat carcinoma (Walker 256), which all showed strong Fc receptor activity in tests with sheep erythrocytes (E) sensitized with rabbit IgG antibodies (A), were used to study the distribution of the receptor in subcellular fractions isolated by differential centrifugation. Smears of the three crude sediments obtained (nuclei, mitochondria rich and membrane rich) all possessed receptor activity. EA incubated in the suspension of cell fractions did not attach to Fc receptor positive tissue sections or to cells in suspension. Cells from which the surface receptors had been eluted still showed activity in sections of cell pellets. Apparently the Fc activity is connected with both external and internal membranes.
IMMUNO‐SUPPRESSIVE EFFECT OF HUMAN MACROPHAGES
Authors: Unsgaard G.
Published: August 2010

Macrophages obtained by culturing blood monocytes in vitro for 8 days showed capacity to inhibit DNA synthesis and cell proliferation in PHA‐stimulated lymphocytes separated from the macrophages by a membrane with pore size 0.2 μm. The DNA synthesis was measured as 3H thymidine incorporation and the proliferation as cell counts. The depression was reduced when the distance between macrophages and lymphocytes was increased concomitant with increased culture volume. Heatkilling of macrophages abolished their lymphocyte depressing capacity. Full inhibitory effect was established within 4 hours when lymphocytes were cultured in the proximity of macrophages. The effect was blocked by molecular filtration membranes with nominal molecular weight limits (nmwl) 1,000 and 10,000 whereas membranes with nmwl 25,000 only partially blocked the effect. No inhibitory effect was registered in the supernatants from macrophage‐depressed lymphocyte cultures. Presumably the lymphocyte depressing effect is mediated via an unstable soluble factor(s).
PRODUCTION OF AN OCHRATOXIN A ANTIGEN WITH HIGH HAPTEN/CARRIER MOLAR RATIO
Authors: Worsaae Helle
Published: August 2010

A mixed anhydride method is described for coupling ochratoxin A to bovine serum albumin. The obtained molar ratio of hapten/carrier in the coupling product was 20. Antibodies against ochratoxin A was obtained from rabbits after immunization with the ochratoxin A‐bovine serum albumin conjugate.
KINETICS OF LOCOMOTION OF HUMAN GRANULOCYTE POPULATIONS
Authors: Schreiner Aksel , Vaula Dag
Published: August 2010

The locomotor kinetics of human granulocytes under gradient and non‐gradient conditions were studied with the Boyden micropore filter method using casein as a stimulant. The in‐filter distribution for 3 different incubation times, and one‐level counts for a wide range of incubation times were determined photographically. Under gradient (chemotactic) conditions, the number of responsing granulocytes depended on the sharpness of the casein gradient, whereas the average migrational velocity of the population appeared to be constant. It was not possible, by comparison with a theoretical model, to demonstrate a directional migration towards increasing casein concentrations. The previously reported Gaussian distribution under non‐gradient (chemokinetic) conditions, was confirmed. In these experiments the number of responding cells appeared to depend on the absolute casein concentration, while their average velocity seemed uninfluenced.
PROTECTION AGAINST EXPERIMENTAL PROTEUS MIRABILIS PYELONEPHRITIS IN RATS AND SIGNIFICANCE OF IMMUNITY
Authors: Larsson Peter , Kaijser Bertil , Olling Sante , Akerlund Arnnica Sohl
Published: August 2010

Intraperitoneal immunization with formalin‐killed bacteria as well as previous hematogenous infection with Proteus O3H1 showed a protective effect against hematogenous pyelonephritis in rats when the homologous strain was used. Transfer of hyperimmune antisera protected against hematogenously induced infection. Neither intravesical or intraperitoneal immunization with formalin‐killed bacteria nor transfer of urines containing antibodies of the IgG class protected against ascending pyelonephritis when the O3H1 strain was used. Data are presented indicating that a rise of pH might decrease the biological effect of antibodies, suggesting that Proteus urease activity is a virulence factor of importance in this context.
IMMUNOCHEMISTRY OF SALMONELLA O‐ANTIGENS:
Authors: Carlsson Hans Erik , Lindberg Alf A.
Published: August 2010

Immunization of rabbits with the synthetic disaccharide‐protein conjugate, tyvelose 1→3 mannose 1→ bovine serum albumin (TM‐BSA) in Freund's complete adjuvant, gave rise to antibodies directed against both the disaccharide hapten and the carrier protein. The hapten antibodies were specific for O‐antigen 9 of Salmonella and were superior in terms of specificity to conventional O factor 9 antibodies, as judged by passive haemagglutination and enzyme‐linked immunosorbent assay (ELISA). A complement dependent bactericidal assay demonstrated that the TM‐BSA antibodies had bacteriolytic activity also directed against the O‐antigen 9 of Salmonella. The high specificity was evident in both the IgG and IgM antibody classes. In the bactericidal assay the IgM antibodies were more active than antibodies of the IgG class. The results suggested that IgG antibodies in hyperimmune serum blocked the complement dependent bactericidal activity of IgM antibodies.
THE PREVALENCE OF IgE ANTINUCLEAR ANTIBODIES IN RHEUMATOID ARTHRITIS AND SYSTEMIC LUPUS ERYTHEMATOSUS
Authors: Permin Henrik , Wiik Allan
Published: August 2010

Antinuclear antibodies (ANA) of the IgE class were studied in sera from patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) and healthy controls. Sixty per cent of 20 RA patients with neutropenia were found to have IgE granulocyte‐specific (GS‐) ANA, whereas only 16% of RA patients without neutropenia had IgE antibodies of similar specificity. About 5 % in each group of RA patients had IgE organ‐nonspecific (ON‐) ANA. Eleven of 15 patients with active SLE and only 4 of 20 with inactive SLE had IgE ON‐ANA. Sera from five patients with lupus nephritis all contained IgE ON‐ANA. None of 100 sera from controls showed presence of IgE ANA. IgE ANA titres in RA and SLE patients correlated to the titres of ANA of the other four immunoglobulin classes. Gel filtration studies at neutral and acid pH of RA sera containing high titres of IgE GS‐ANA indicated the presence of these antibodies in immune complexes. Studies of serum cryoprecipitates supported this conclusion. IgE ANA production may be of pathogenetic importance in RA and SLE by eliciting type‐I reactions.
Fcσ RECEPTORS AND IgG ASSOCIATED WITH HUMAN MALIGNANT TUMOURS
Authors: Wesenberg Finn
Published: August 2010

Thirteen solid tumours were tested for Feσ receptors (FcR) using tissue sections and sheep erythrocytes sensitized with rabbit antibodies as indicator cells. No alteration was found in FcR activity (FcRA) between untreated tissue and tissue homogenized for 2 min or less. More extensive homogenization abolished the FcRA. Tissues homogenized for 1 min were washed at 4° C and eluted at 37, 45 and 56° C. The FcRA was not altered after elution at 37 and 45° C. However, elution at 56° C abolished the activity indicating that the FcR were either eluted or destroyed. Tumours which showed a diffuse pattern of FcRA and no non‐specifically bound IgG displayed an inverse relationship between the FcRA and the eluted IgG. Such relationship was not found with tumours containing non‐specifically bound IgG.
NEUTRALIZING ANTIBODIES AGAINST HUMAN LEUKOCYTE, LYMPHOBLASTOID AND FIBROBLAST INTERFERONS ELICITED BY IMMUNIZATION WITH HUMAN LEUKOCYTE INTERFERON
Authors: PyhäLä Liisa
Published: August 2010

Guinea pigs, rabbits and sheep were immunized with partially purified human leukocyte interferon. The antisera were tested for their neutralizing activities against human leukocyte, lymphoblastoid (Namalva) and fibroblast interferons. Rabbits and sheep developed high levels of neutralizing antibodies against all three interferons. The ratios of the neutralizing activities of leukocyte/lymphoblastoid interferons varied from 1 to 25 and those of leukocyte/fibroblast interferons from 2 to 500. The ratios increased with prolonged immunization. The guinea pigs developed relatively low levels of anti‐leukocyte and anti‐lymphoblastoid antibodies and as a rule failed to produce detectable neutralizing activity against fibroblast interferon. One sheep was given a booster injection of fibroblast interferon after preimmunization with leukocyte interferon. The booster injection stimulated a rise in antibody levels, not only for fibroblast interferon, but also for leukocyte and lymphoblastoid interferons. The antigenic relationships between leukocyte, lymphoblastoid and fibroblast interferons are discussed.
MACROPHAGES AND THE SYNTHESIS OF CONNECTIVE TISSUE COMPONENTS
Authors: KULONEN EINO , POTILA MARITA
Published: August 2010

1. Macrophage homogenate, added to the incubation medium of granulation tissue slices, decreases the incorporation of labelled proline to collagen and other proteins, to a greater extent in the presence of certain prostaglandins and in oxygen atmosphere, but to a lesser degree if serotonin and cyclic AMP are added. 2. Rheumatoid synovial fluid, connective tissue active peptide (CTAP) of Castor and an extract from CCl4‐damaged liver stimulate collagen synthesis in the presence of macrophages. 3. Preparations from non‐treated macrophages stimulate the incorporation of glucosamine to acid mucopolysaccharides, mainly hyaluronate. 4. In a developing rat granuloma, the formation of collagen was stimulated by homogenized macrophages and by macrophage culture medium, but especially by whole macrophages in the presence of SiO2.
SERUM BACTERICIDAL EFFECT ON CAPSULATED AND NON‐CAPSULATED HAEMOPHILUS INFLUENZAE
Authors: BRANEFORS PAULA , DAHLBERG TERESA
Published: August 2010

In order to test serum bactericidal activity on Haemophilus influenzae, a micromethod was employed. The bactericidal (BC) effect of preimmune and hyperimmune rabbit sera was studied on a type a and a type b strain and their respective non‐capsulated variants. The influence of variation of the experimental conditions on the BC effect was investigated. It was observed that the amount of guinea pig complement added as well as the number of bacteria and reaction time had an influence on the BC effect. The serum dilution showing ≥ 50% killing of the inoculum was chosen as endpoint titre (BC titre). Unheated preimmune rabbit sera, in which no specific antibodies were demonstrable with the complement fixation assay, were bactericidal for the two non‐capsulated strains. Heat treatment to 56°C for 5 min destroyed this effect, which could not be restored by addition of diluted guinea pig serum. Unheated preimmune rabbit sera were not bactericidal for capsulated bacteria. Hyperimmune sera showed heat‐resistant (56°C, 30 min) BC activity on both non‐capsulated and capsulated bacteria, the latter showing quite high BC titres (about 1×105). Serum samples form 30 blood donors had demomstrable, heat‐resistant BC titre against the capsulated bacteria tested, whereas only about one half of the serum samples showed heat‐resistant BC activity against the non‐capsulated bacteria used. Ten non‐capsulated H. influenzae strains isolated from patients with various infections revealed a variable sensitivity to preimmune rabbit sera (normal sera), these strains being more resistant than the two noncapsulated variant strains utilized in the study.
STUDIES ON Cob (COLTON) ANTIGEN AND ANTI‐Cob
Authors: JERNE DONALD , LYLLOFF KIRSTEN
Published: August 2010

Using a reliable and strong reacting antiserum of the rare anti‐Cob specificity, it was shown that the Cob antigen was fully developed at birth. Gene dosage effect could not be demonstrated. The frequency of Co(b+) persons in the Danish population (7.8%) was found not to differ from other Caucasian populations. The antibody belonged to immunoglobulin class IgG without ability of fixing complement in vitro.
INFLUENCE OF AN INTESTINAL MICROFLORA ON THE DEVELOPMENT OF THE IMMUNOGLOBULINS IgG1, IgG2a, IgM and IgA IN GERM‐FREE BALB/c MICE
Authors: NIELSEN EBBE , FRIIS CARL W.
Published: August 2010

The development of immunoglobulins IgG1, IgG2a, IgM and IgA was studied in germ‐free BALB/c mice during the first 14 weeks of life. A transfer of maternal IgG1 and IgG2a was demonstrated during the first 2 weeks of this period. Endogenous synthesis of immunoglobulins was found only for IgM, the level of which was significantly higher at the end of the period than in a conventional group. After oral association of germ‐free BALB/c mice with an intestinal hexaflora, the serum concentrations of IgG1, IgG2a and IgA increased. The levels of IgA, and of IgG1, were higher in the associated group than in the conventional group.
SMOOTH‐MUSCLE ANTIBODIES IN RHEUMATOID ARTHRITIS
Authors: ANDERSEN IRENE , ANDERSEN P. , GRAUDAL H.
Published: August 2010

Smooth‐muscle antibodies (SMA) detected by the indirect immunofluorescence method were found more often in rheumatoid arthritis (RA) patients (15.3%) than in normal controls (7.6%) (0.02 > p > 0.01). The increased frequency was due to IgG‐antibodies which occurred in 12.2% of RA patients. in 4.1% of normal controls and in 7.3% of patients with other arthritic diseases. Eight of 9 IgG‐SMA‐positive RA sera reacted with F‐actin, and one serum contained non‐actin antibodies. In RA, ANA were found in 35.7%, glomerular antibodies in 1.0%, parietal cell antibodies in 4.1% and mitochondrial antibodies in 2.0%. SMA were associated with the occurrence of rheumatoid factors and mitochondrial antibodies (0.02 > p and 2p = 0.04, respectively). In the SMA‐positive group the erythrocyte sedimentation rate was higher and elevated serum alkaline phosphatase values were found more often than in the SMA‐negative group.
ANTIBODY RESPONSE IN PATIENTS WITH PSEUDOMONAS AERUGINOSA INFECTION TO A ‘COMMON ANTIGEN’ FROM P. AERUGINOSA ANALYSED BY MEANS OF QUANTITATIVE IMMUNOELECTROPHORETIC METHODS
Authors: HØIBY N. , HERTZ J. B. , SOMPOLINSKY D.
Published: August 2010

381 sera from 101 patients suffering from Pseudomonas aeruginosa infection have been investigated for antibodies against a ‘common antigen’ (CA) and other antigens from P. aeruginosa by means of crossed immunoelectrophoresis with intermediate gel, using a polyspecific P. aeruginosa antigen/antibody reference system. The earliest antibody response during the P. aeruginosa infections included increased CA antibodies in only 38% of the patients examined. The peak level of the antibody response included increased CA antibodies in 71% of the patients. By means of ‘absorption of antibodies in situ’, it was found that the antibody response to CA was directed against both the common cross‐reactive part of the CA molecule and the Pseudomonas‐specific part, with individual variations. During infections caused by other bacteria containing CA, one‐third of the patients developed antibodies against CA. The diagnostic implications of the results are discussed.
AN INDIRECT HAEMAGGLUTINATION TEST FOR DEMONSTRATION OF GONOCOCCAL ANTIBODIES USING GONOCOCCAL PILI AS ANTIGEN
Authors: REIMANN K. , LIND I. , ANDERSEN K. E.
Published: August 2010

A total of 1223 serum specimens were obtained from 649 consecutive patients attending a dermato‐venereological out‐patient clinic in Copenhagen with a request for venereal disease control. The sera were examined for gonococcal antibodies by both a gonococcal complement fixation test (GCF) and an indirect haemagglutination test using gonococcal pili as antigen (IHA). The diagnosis of current gonococcal infection in 28 per cent of the patients was based on positive culture for Neisseria gonorrhoeae from one or more of the following sites: urethra, rectum and fauces in all patients, and/or cervix in female patients. The specificity, sensitivity and predictive values of positive and negative test results were calculated for the various groups of patients on the assumption that all positive results, both in patients without gonorrhoea but with a previous gonococcal infection and in patients without known current or previous infection, were false positives. The following values were found: Sensitivity: IHA 45–100%, GCF 6–29%. Specificity: IHA 65–89%, GCF 97–100%. Predictive value of positive test result: IHA 43–74%, GCF 69–100%. Predictive value of negative test result: IHA 78–100%, GCF 61–85%.
AN IMMUNOELECTROPHORETIC ANALYSIS OF THE STREP. SANGUIS AND ADULT HUMAN ORAL MUCOSA ANTIGEN EXTRACTS USED FOR IMMUNOLOGICAL INVESTIGATIONS OF RECURRENT APHTHOUS STOMATITIS
Authors: DONATSKY O.
Published: August 2010

Water‐soluble antigens of Strep. sanguis strain 2A, ATCC 10556, and adult human oral mucosa (AHOM) were analysed by four immunoelectrophoretic methods. Crossed immunoelectrophoresis (CIE), using rabbit antibodies raised against the soluble antigens, revealed 26 antigens in the standard Strep. sanguis antigen extract and 16 antigens in the standard adult human oral mucosa extract. All 26 Strep. 2A antigen‐antibody precipitates observed in the CIE were identified as streptococcal components. Eleven of the AHOM antigen‐antibody precipitates in the CIE were derived from human serum proteins and hemoglobin present in the crude AHOM‐Ag extract. Five of the latter precipitates possibly were caused by oral mucosal antigens. The AHOM‐Ag extract contained four bacterial contaminants. None of them influenced the crossed immunoelectrophoretic AHOM antigen‐antibody reference pattern in the present study.
EFFECTS OF POLYCATIONIC COMPOUNDS ON MITOGEN STIMULATION
Authors: HERON I. , LARSEN B. , HOKLAND M.
Published: August 2010

The effects of polycations added to phytomitogen stimulated human lymphocyte cultures have been studied. Within certain dose ranges all polycations tested gave rise to augmented thymidine uptake in mitogen stimulated cultures. The optimum enhancing concentrations of polycations was depending on the serum concentration in the culture medium. This was found to be due to two types of interactions: (a) Interference with mitogen binding serum factors, (b) Reaction with immunosuppressive serum proteins. Suggestive evidence for an effect of polycations also directly on the cells was found by pretreatment and cell density experiments. The direct effect on cells was found not to be due to monocyte bypass or to activation of non T‐cells by the mitogen. It is pointed out that effect of chemicals on in vitro immunresponses have to be considered in relation to the charge properties of the compound.
HAPTEN AND KIDNEY ANTIBODIES IN SERUM FROM CHILDREN WITH URINARY TRACT INFECTION
Authors: AHLSTEDT S. , FASTH A. , JODAL U.
Published: August 2010

The presence of antibodies against the synthetic hapten dinitrophenol (DNP) was investigated in sera from healthy children and adults. Such antibodies were found in all sera. The serum levels were significantly lower in healthy children than in adults. Serum samples from a pilot group of pediatric patients with acute E. coli urinary tract infection (UTI) contained significantly higher DNP antibody levels compared to healthy children and similar to those of the adults. Furthermore. 10 of 17 patients had a significant change in antibody levels in connection with the infection as judged from consecutive serum samples. Relations were found between age and minimum as well as maximum serum DNP antibody values. In a second group of children with E. coli urinary tract infection beside DNP antibodies, antibodies against the Tamm‐Horsfall urinary protein and a pool of O antigens from E. coli common in urinary tract infections were also investigated. In these patients similar findings as for ihe pilot group were made for the levels of the DNP antibodies. An IgG but not IgM antibody response to the Tamm‐Horsfall protein was also found in sera from all the patients over 1 year with acute pyelonephritis. The highest antibody levels against Tamm‐Horsfall protein were noted in patients with vesico‐ureteric reflux. No relation could be recorded between the DNP antibodies and the Tamm‐Horsfall protein antibodies. The antibodies to the pool of O antigens recorded did not correlate to the DNP antibodies. It was concluded that the DNP antibody formation could partly be due to polyclonally stimulatory activities of bacterial substances exposed during infections.
A RADIOIMMUNOASSAY FOR THE DETECTION OF IDIOTYPES IN ALLOANTIGEN REACTIVE MOUSE SERA AND IN NORMAL MOUSE SERA
Authors: NORDFANG OLE
Published: August 2010

A radioimmunoassay was developed for characterization of the binding between rabbit antiserum and idiotypes in B6 anti‐H‐2k antisera. It was a double precipitation assay using iodinated F(ab')2 fragments prepared from B6 anti‐H‐2k antisera as antigen. Suitable absorptions made the non‐specific binding very low, and thus this assay was well suited for detecting idiotypes in preparations, that mainly contained other forms of immunoglobulin than the idiotype. That the rabbit antiserum reacted with idiotypes was demonstrated by the use of F(ab')2 fragments in inhibition assays where B6 anti‐H‐2k antisera could be used as effective inhibitors in contrast to B6 anti‐H‐2q and ABY anti‐H‐2k antisera. Idiotype was also found in the sera of normal B6 mice and as much as 0.05% of the immunoglobulin molecules expressed the idiotype. The inhibition assay was suitable for quantitation of idiotypes, since the reproducibility of the assay through a long period was high.
EFFECT OF PHENYTOIN ON IMMUNOGLOBULIN CONCENTRATIONS IN NASAL SECRETIONS
Authors: GILHUS NILS ERIK , AARLI JOHAN A.
Published: August 2010

Nasal washings from patients receiving phenytoin and from healthy controls were investigated. The concentrations of immunoglobulins and albumin were determined by single radial immunodiffusion. Epileptic patients treated with phenytoin had a higher IgA concentration in nasal washings than controls, 174 × 10−3 g/1 versus 114 × 10−3 g/1. The IgG concentration was also significantly higher in the phenytoin‐treated patients compared to the controls. IgM was detected in nasal washings from 41% of phenytoin‐treated patients and from only 11% of the controls.

To exclude the effect of exudation of plasma proteins caused by inflammation, the ratio IgA to albumin was calculated and the median of this ratio estimated both for patients and controls. For patients taking phenytoin as their only drug, the median was 1.89 versus 1.23 in the controls.

There was no relation between the serum concentration of IgA and the ratio between IgA and albumin concentrations in nasal secretions. However, patients with serum IgA deficiency had low concentrations of IgA in the nasal secretion and a reduced IgA to albumin ratio.
HUMAN MONOCYTES INDUCE A REVERSIBLE INHIBITION OF CELL CYCLE PROGRESSION IN THE LEUKEMIC CELL LINE K‐562 DURING CO‐CULTURE IN VITRO
Authors: EGGEN BJØRN MAGNE
Published: August 2010

Human monocytes cultured in vitro exert cytostatic activity against the non‐adherent human leukemic cell line K‐562. K‐562 cells in asynchronous growth were added as target cells to adherent mononuclear blood cells, either freshly isolated or cultured in vitro for 4 and 12 days. After 24 hours of co‐culture, the non‐adherent cells were gently removed and incubated further. Target cell DNA‐synthesis and cellular DNA content were examined by methyl‐3H‐thymidine incorporation and by flow cytofluorometry, respectively. Cell‐proliferation curves were obtained. Monocytes cultured in vitro exert a reversible cytostatic activity, which is a partial cell cycle block in the G0/G1 phase. Target cells exposed to freshly isolated monocytes also showed accumulation of cells in the G0/G1 phase. These target cells did not regain their normal proliferation rate before day 3 after removal from the monocyte cultures. This effect may, however, to some extent be due to loosely adherent mononuclear blood cells removed from the monocyte cultures together with the target cells.
HUMAN ADHERENT MONONUCLEAR BLOOD CELLS: CYTOLYTIC AND CYTOSTATIC ACTIVITY AND CHARACTERIZATION OF EFFECTOR CELLS DURING IN VITRO CULTURE
Authors: HAMMERSTRØM JENS
Published: August 2010

Using a methyl‐3H‐thymidine release assay, the cytolytic activity of human adherent mononuclear blood cells (AC) against three transformed human tumour cell lines of mesenchymal (U‐20S), epithelial (NHIK 3025) and hematogenous (K‐562) origin was compared to the activity against non‐transformed skin fibroblasts (U‐2S) and mesothelial cells (meso). Only transformed target cells were affected by the spontaneous cytolytic activity of normal AC tested after 0, 4 or 8 days of in vitro culture. Lymphokine‐activated 4‐days old AC also preferentially lysed transformed target cells, but low levels of lysis of non‐transformed target cells were observed at high effector cell densities. Freshly isolated AC were spontaneously cytostatic to K‐562, but not to NHIK 3025 cells, while differentiated AC cultured for 8 days were cytostatic to both cell lines. Lymphocyte‐like cells comprised up to 10% of freshly isolated AC, and most of these lymphocytes seemed to be non‐T‐cells as evaluated by α‐naphtyl‐esterase (ANAE) staining. Most of these cells detached during the first 24 h of in vitro culture. and the fraction of AC expressing monocyte markers (diffuse ANAE reactivity, interaction with ox erythrocytes coated with IgG or IgM + complement, interaction with Candida albicans) increased to nearly 100% after 4–8 days of in vitro culture. Thus, the effector cells expressing spontaneous cytolytic activity in this system have monocyte characteristics after 1 day of in vitro culture. However, contribution of adherent non‐monocytic cells to the cytotoxicity observed during the first 24 hours of culture can not be ruled out.
CULTURED HUMAN SYNCYTIOTROPHOBLASTS SYNTHESIZE PREGNANCY‐SPECIFIC BETA‐1‐GLYCOPROTEIN (SP1)
Authors: HEIKINHEIMO M. , SAKSELA O. , LEHTOVIRTA P. , SEPPÄLÄ M. , BOHN H.
Published: August 2010

The synthesis of pregnancy‐specific beta‐1‐glycoprotein (SP1) was studied in human syncytiotrophoblast cultures from first trimester placentas. Incorporation of 35S‐methionine into SP1 was demonstrated by immunoprecipitation and polyacrylamide gel electrophoresis. In double diffusion the precipitation line of neosynthesized SP1 gave a reaction of complete immunological identity with maternal serum SP1. By indirect immunofluorescence intracellular SP1 was localised in the perinuclear region of the cells. The average rate of SP1 release into culture fluid was 600 ng/mg protein in 24 hours. On a molar basis the release of SP1 was 10 times less than that of human chorionic gonadotrophin (hCG). Our results demonstrate, for the first time, that cultured human syncytiotrophoblasts synthesise SP1 immunologically indistinguishable from placental SP1.
T CELL DEPENDENT B CELL DIFFERENTIATION IN THE CHICKEN
Authors: HIROTA YOSHIKAZU , VAINIO OLLI , TOIVANEN PAAVO
Published: August 2010

Transplantation of bursa cells into surgically thymectomized (STx), cyclophosphamide(CY)‐treated and X‐irradiated (X) newly hatched chicks resulted in a morphological and functional reconstitution of the bursa‐dependent immune system only when the transplanted bursa cells were supplemented with thymus cells. Without supplementation with thymus cells, morphological restoration of the bursa, antibody formation to sheep red blood cells and Brucella abortus, and germinal center formation remained deeply deficient. Bursa cells alone induced a full reconstitution of the bursa‐dependent immunity in non‐thymectomized, CY‐treated and X‐irradiated cell recipients. The incomplete restoration of STx‐CY‐X chickens after transplantation of bursa cells was not due only to lack of T helper cells, as indicated by the incomplete restoration of the bursal morphology and of antibody formation against Brucella abortus, a thymus‐independent antigen. It is concluded that T cells are necessary for the B cell differentiation in the bursa of Fabricius. The mechanism behind this phenomenon remains unclear but our results indicate that a complete T cell function is not a prerequisite for this, since STx‐CY‐X birds reconstituted with bursa and thymus cells together did not have their mitogen (PHA, Con A) responses normalized.
METHODOLOGICAL APPROACHES TO ANTIGEN IDENTIFICATION IN SOLUBLE IMMUNE COMPLEXES; A MODEL STUDY
Authors: HUSBY S. , SVEHAG S. E. , NIELSEN H. , HØIBY N. , SCHIØTZ P. O.
Published: August 2010

The usefulness of different techniques for identification of immune complex‐(IC) bound antigen(s) was investigated in two Ab‐Ag model systems. The following approaches were used: 1) determination of IC‐activity before and after removal of IC by adsorption to antibody‐specific solid phase systems, 2) analysis of PEG‐insoluble IC by reference line‐rocket Immunoelectrophoresis and 3) reprecipitation of PEG‐insoluble IC in the presence of 125I‐labelled antibody to suspected antigens and subsequent analysis of the distribution of the radioactivity. This latter approach, the use of a double PEG precipitation immunoradiometric assay, was found most useful and simple for identification of antigen(s) contained in soluble IC. The technique is most sensitive in demonstrating IC‐bound antigen(s) when the IC are formed in slight to moderate antigen excess.
GENERAL PARALYSIS OF THE INSANE ASSOCIATED WITH HLA‐Aw32
Authors: PEDERSEN N. STRANDBERG , AXELSEN N. H. , SVEJGAARD A.
Published: August 2010

HLA‐A, ‐B, and ‐C locus antigens were determined in 35 unrelated patients with general paralysis of the insane (GPI), and in 13 neurosyphilis patients without dementia. HLA‐Aw32 was found more frequently (corrected p = 4.9 × 10−2) in the GPI patients than in 1009 controls. None of the patients having neurosyphilis without dementia had HLA‐Aw32. These findings indicate that genetically determined mechanisms may play a role in development of GPI. Further studies of such rare patients should be initiated to investigate whether these observations reflects a true association or whether they are due to chance. If possible other well defined groups of tertiary syphilis should also be studied.
DIFFERENT BLOOD PRESSURE RESPONSE TO DOCA‐NaCl TREATMENT IN AUTOIMMUNE AND N,N‘‐ DIACETYLBENZIDINE‐ INDUCED NEPHROPATHIES
Authors: TIKKANEN ILKKA
Published: August 2010

In order to explore the effect of immunologically‐induced nephropathy versus a non‐immunological model on development of hypertension, rats with either Heymann nephritis or N,N'‐diacetylbenzidine (N.N'‐DAB)‐induced nephropathy were subjected to DOCA‐NaCl treatment. Hypertension developed during DOCA‐NaCl treatment in Heymann nephritic rats but not in rats with N,N'‐DAB nephropathy, despite similar degrees of proteinuria and other features of nephrosis. There were no signs of different fluid balance, either. Azotemia did not ensue. Histology and immunohistology showed membranous glomerulonephritis in Heymann nephritic rats whereas glomerular epithelial cell alterations without immune deposits along the glomerular capillary walls were seen in rats with N.N'‐DAB nephropathy. The type of renal damage or factors mediating such damage appear to be important determinants for the hypertensive response of Heymann nephritic rats to DOCA‐NaCl treatment.
ISOLATION AND CHARACTERIZATION OF Fcγ RECEPTORS FROM HUMAN PLACENTA
Authors: MATRE ROALD , KLEPPE GUNNAR , TÖNDER OLAV
Published: August 2010

Placental extract was prepared with Tris‐HCl buffer pH 7.4 containing EDTA and 2‐mercaptoethanol. It agglutinated erythrocytes (E) sensitized with subagglutinating amounts of IgG antibodies from rabbits, guinea pigs and mice, but not E sensitized with IgG from chickens. E or E sensitized with F(ab'h or IgM antibodies were not agglutinated. The agglutination of EA by the extract was inhibited by human, rabbit and guinea pig IgG, but not by bovine and porcine IgG. Aggregated IgG was more inhibitory than monomeric IgG. IgG3 was the most effective subclass. The extract inhibited the formation of EA rosettes with human mononuclear cells, but did not influence the formation of E or EAC rosettes. The significance of the disulfide bonds and the Cγ3 and Cγ2 regions was implied by the finding that the extract neither agglutinated E sensitized with preparations of mildly reduced and alkylated IgG, nor with Facb fragments. These preparations did not inhibit the agglutination. The results strongly indicated that the active component was solubilized placental Fc receptor (FcR). Functionally active FcR was purified by affinity chromatography using aggregated human IgG coupled to Sepharose 4B. SDS‐PAGE of the purified FcR under reducing conditions showed one distinct band corresponding to approx. 47,000 daltons. The band neither consisted of Ig fragments nor CIq. A rabbit antiserum against the FcR inhibited the agglutination of EA by the extract and stained the FcR‐positive areas in placenta.
ANTIBIOTICS AND GRANULOCYTES
Authors: BELSHEIM J. A. , GNARPE G. H.
Published: August 2010

Twelve antibiotics were investigated regarding both their direct in vitro influence on granulocyte chemotaxis, and their indirect effect on the production of chemotactic factors from cultures of Escherichia coli and Pseudomonas aeruginosa. None of the beta‐lactam antibiotics studied caused significant alterations of granulocyte migratory response when incubated with the cells at concentrations of up to 128 μg/ml. The two aminoglycoside preparations, and the two tetracycline preparations caused significant depressions of the migration response. Production of chemotactic factors was stimulated from growing cultures of E. coli by sub‐minimal inhibitory concentrations of beta‐lactam antibiotics and from Ps. aeruginosa by the cephalosporin derivatives only. The differences observed were most probably due to the mode of action at the bacterial cell wall level.
CELLULAR AND HUMORAL IN VITRO CYTOTOXICITY AGAINST AUTOLOGOUS BLADDER TUMOR CELLS IN HUMANS
Authors: JACOBSEN FLEMMING
Published: August 2010

Lymphocyte‐mediated cytotoxicity (LMC), antibody‐dependent cell‐mediated cytotoxicity (ADCC) and complement‐dependent cytotoxicity (CDC) were measured in a 51Cr assay against autologous tumor cells from 7 patients with non‐invasive and 9 patients with invasive transitional‐cell tumors of the urinary bladder (TCC). Cytotoxicity against the invasive tumor cells were demonstrated in CDC. Heat‐inactivation of sera lead to complete loss of cytotoxicity while addition of allogenic serum restored cytotoxicity. The cytotoxicity of allogenic sera from controls against invasive tumor targets showed no differences when compared with autologous sera. No or very weak cytotoxicity was found against noninvasive tumor targets in autologous or allogenic assays. LMC and ADCC showed weak or no reactivities. Intensive wash or trypsinization of effector cells did not affect the cytotoxicity in LMC. The results indicate the occurrence of complement‐dependent antibodies directed against target cells from invasive tumors of the urinary bladder while no cytotoxic responses were detectable when the target cells originated from non‐invasive tumors.
IgG F(ab‘)2 OF RABBIT ANTI‐M SERA BUT NOT THE UNFRACTIONATED SERA ARE BACTERICIDAL FOR SOME GROUP A STREPTOCOCCI WITH IgG Fc‐RECEPTOR ACTIVITY: Opsonic Effect Ascribable to Anti‐IgG
Authors: SCHALÉN CLAËS , BUROVA LARISSA A. , CHRISTENSEN POUL , GRUBB RUNE , SAMUELSSON GUNILLA , SVENSSON MAJ‐LIS
Published: August 2010

We have earlier reported on a group A streptococcal strain, type M12, which upon serial mouse passage acquired IgG Fc‐receptor activity but lost the M‐antigen. The passaged strain, 12P, was highly virulent for mice and grew well in normal human blood. The present study particularly concerns the opsonic effect on 12P of rabbit anti‐M3, anti‐M 12 and anti‐12P sera, as well as the corresponding IgG F(ab')2‐ Indirect bactericidal tests showed that the homologous anti‐12P serum and IgG F(ab')2 were opsonic. The anti‐M3 and anti‐M 12 had no effect on 12P; surprisingly, however, IgG F(ab')2 isolated from these sera displayed a clearcut opsonic activity. Data are presented which indicate that these »paradoxical« results can be explained by the binding of IgG F(ab')2 with anti‐IgG specificity to human IgG, linked to the streptococcal surface through Fc‐receptors. Only anti‐12P serum, or IgG F(ab')2, were protective for mice on challenge with strain 12P.
SOME CHARACTERISTICS OF IMMUNOFLUORESCENCE TESTS FOR ANTIBODIES AGAINST GLUTEN, USING WHEAT GRAIN SECTIONS OR GLIADIN COATED SEPHAROSE BEADS
Authors: JONSSON JONAS , SCHILLING WALTER
Published: August 2010

Antibodies against gluten and gliadin were determined by the indirect immunofluorescence technique, using frozen wheat grain sections or gliadin‐coated Sepharose beads. The methodological characteristics and diagnostic usefulness of the two techniques were evaluated. The reproducibility of both was improved by introducing a fluorometric reading‐off procedure. Antibody quantitations were preferebly performed as end‐point titrations. The two techniques had different dose‐response relationships. The grain section technique was more discriminative for small variations in antibody concentration than the bead technique. The latter was, however, more reproducible. Besides antibodies against gliadin, a number of patients with gluten enteropathy had antibodies against the main septa of wheat grains. Fluorescence intensity was preferably expressed in multiples of background intensity. A reaction was visually perceived when the fluorometrically‐measured fluorescence intensity reached 2.5 times the background intensity. Using this value as the limit for positive reactions, antibodies were demonstrated in 81 96 of the cases with verified gluten intolerance, compared with 28% in cases with other intestinal allergies and 8% in normals. The diagnostic specificity of both the grain section and Sepharose bead technique for gluten enteropathy increased with increasing antibody concentration and was apparently 100% when the fluorescence intensity produced by a 1/10 serum dilution reached a value 7 to 8 times that of the background. Antibodies against reticulin were demonstrated in 1/4 of the cases having anti‐gluten antibodies but in none of those with non‐gluten‐induced gastro‐intestinal symptoms. Antibodies of the IgG class against cow's milk were demonstrated more often and in higher titre in cases with anti‐gluten antibodies than in those without them.
QUANTITATION OF COMPLEMENT C3b AND C3d SURFACE RECEPTORS ON NEOPLASTIC IgM POSITIVE B LYMPHOCYTES
Authors: JøNSSøN VIGGO , CHRISTENSEN BJARNE EGELUND , VIDEBÆK AAGE
Published: August 2010

C3b and C3d complement receptors on the surface of mononuclear cells from blood, lymph nodes, and spleens were investigated in twenty‐nine CLL patients and in eight patients with non‐Hodgkin malignant lymphoma. Compared with a normal material, especially the lymphoid tissues but also the blood of the patients contained a high number of C3d receptor positive lymphocytes with a strong expression of C3d receptors per cell. The majority of the cells with high C3d receptor expression also had SmlgM. The number of C3b receptor positive cells was decreased in the patients but the C3b receptor expression per cell was normal.
STAPHYLOCOCCAL PROTEIN‐A AGGLUTINATION ASSAY FOR AVIAN VIRUSES
Authors: PANDYA BHARAT V. , THOMPSON KENNETH D. , LINNA T. JUHANI
Published: August 2010

Protein‐A rich staphylococci were used to develop an agglutination assay for the detection and quantitation of avian reticuloendotheliosis virus (REV) and Herpes virus of turkeys (HVT). Rabbit anti‐REV or anti‐HVT were used to coat the staphylococci and these reagents were mixed with REV or various other viral antigen preparations. An agglutination titer was determined for antigen preparations of varying purity and the results indicated that in addition to being rapid, this assay allows the detection of as little as 0.3 to 0.5 μg of partially purified viral antigens. No crossreactivity was observed with the two antisera and the various antigen preparations. This study also confirmed the reports in the literature that chicken immunoglobulins do not bind to protein‐A of the Cowan I strain of Staph, aureus.
DEMONSTRATION OF BOVINE VIRAL DIARRHEA VIRUS ANTIGEN IN CRYOSTAT‐ AND PARAFFIN‐SECTIONS OF BOVINE TISSUES BY THE IMMUNOPEROXIDASE TECHNIQUE
Authors: OHMANN H. BIELEFELDT , JENSEN M. HOLM , SøRENSEN K. J. , DALSGAARD K.
Published: August 2010

An immunoperoxidase technique (IPT) for the demonstration of bovine viral diarrhea virus (BVDV) antigen in tissue sections is presented. The method was applied to cryostat sections and to alcohol fixed, paraffin embedded specimens. The specificity as well as the sensitivity of the IPT corresponded to the immunofluorescence technique (IFT), and the specimen processing methods used represented an improvement of the histomorphological preservation of tissue structures.
LYMPHOCYTE SUBPOPULATIONS IN MAN: T‐CELLS WITH FcR FOR IgG IN THE ALLOGENEIC RESPONSE IN VITRO
Authors: JOHNSEN H. E.
Published: August 2010

T‐cell suspensions enriched and depleted for the subpopulation with FcR for antigenbound IgG (EA‐RFC) have been tested for the responding capacity in MLC/CML. The results reveal that the EA‐RFC depleted T cell suspensions (enriched for FcR‐ve T cells) have a greater proliferative capacity but unchanged cytotoxicity compared to the unfractionated T cell suspensions. On the contrary, the EA‐RFC enriched T cell suspensions (enriched for FcR + ve T cells) have only weak proliferative capaticy and cytotoxicity. These results indicate that the major part of proliferating T‐amptifiers as well as cytotoxic T‐precursor cells are FcR‐ve. Fractionation of cytotoxic effector cells day six of culture reveal that cytotoxic cells are found among EA‐RFC depleted as well as enriched suspensions. The cytotoxic capacity of effector cells was found to be substantially reduced after EA rosette formation due to a blocking and sterical inhibitory influence of the EA complexes on the close effector‐target contact necessary for cytotoxicity. These results indicate predominance of FcR + ve cytotoxic T cells. The rosette indicator system used here for identification and definition of FcR cannot separate cytotoxic T‐precursor cells and strongly proliferating T‐amplifiers found to be FcR‐ve. On the contrary, cytotoxic T‐effector cells are predominantly FcR + ve.
STUDIES ON THE THYMUS‐DEPENDENT IMMUNITY IN THE RAT MUTANT »NUDE«
Authors: Svendsen Ulrik Gerner , Koch Claus , Werdelin Ole , Christiansen Peter
Published: August 2010

Different T‐cell capacities in nude, athymic rats were tested. Spleen cells from nude rats were unable to give significant responses to Con A as could spleen cells from normal rats. No precipitating antibodies after immunization with chicken serum were observed in nude rats, although such antibodies were found in the haired rats. The experiments confirmed that the used stock of nude rats had a similar T‐cell defect as the original Rowett strain of nude rats.
COMPLEMENT‐DEPENDENT IN VITRO CYTOTOXICITY AGAINST AUTOLOGOUS INVASIVE BLADDER TUMOR CELLS IN HUMANS
Authors: JACOBSEN FLEMMING
Published: August 2010

Complement‐dependent serum‐mediated cytotoxicity (CDC) was measured in a 51‐chromium release assay against autologous tumor cells from 7 non‐invasive and 9 invasive transitional‐cell tumors of the urinary bladder. CDC was demonstrated against tumor cells from invasive tumors. Heat‐inactivation of autologous sera lead to complete loss of cytotoxicity. There were no differences in CDC of autologous sera from patients and allogenic sera from controls. The cytotoxic response seems to be strongly dependent on the target cell. CDC was significantly reduced by use of an allogenic C 2 deficient serum. Direct immunofluorescence did not reveal any tumor cell associated immunoglobulins of IgG or IgM classes. Indirect immunofluorescence with autologous heat‐inactivated sera demonstrated in most cases both IgG and IgM attachment to the tumor cells, but there were no obvious relations between indirect immunofluorescence and CDC. Absorption of allogenic sera to trypsin‐ or neuraminidase‐treated erythrocytes did noi affect CDC of these sera against invasive tumor targets. The results indicate a complement‐dependent cytotoxicity against target cells from invasive bladder tumors. Complement seems to be activated through the classical pathway, but the possible role of naturally‐occurring antibodies against invasive tumor targets is not clarified.
LOCALIZATION OF THE C3b RECEPTORS IN HUMAN RENAL GLOMERULI
Authors: MATRE ROALD , NILSEN RLNE , IVERSEN BJARNE M.
Published: August 2010

Cryostat sections of human kidney were incubated with C3b containing immune complexes. The complexes were horseradish peroxidase (HRP) and Flab'b fragments of rabbit IgG anii‐HRP incubated in C4‐deficient guinea pig serum. The peroxidase activity was revealed histochemically. The sections were studied using light and electron microscopy. The bound complexes were localized to the foot processes of the visceral epithelial cells and to the surface of the parietal epithelial cells facing Bowman's space. No receptors could be demonstrated on the endothelial or mesangial cells.
RED BLOOD CELLS COATED WITH TUMOR CELL EXTRACTS AS TARGETS IN 51Cr RELEASE ASSAYS
Authors: KARJALAINEN H. E. , MÄNTYJÄRVI R. A.
Published: August 2010

Sheep red‐blood cells were coated with tumor cell antigens prepared by extractions with 3M KCI. The coated cells were labelled with 51Cr and used as targets in immuno‐assays. The technique was found suitable for complement‐mediated as well as antibody‐dependent cell‐mediated reactions, and it offers several applications for use in tumor immunology.