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Antilymphocytic globulin (ALG) depressed the antibody responses of mice to SRBC as measured in terms of splenic PFCs of IgM, IgG1, IgG2a and IgG2b specificity. This effect was apparent over a range of antigen does. ALG also reduced the antibody responses of congenitally athymic mice to SRBC and SHI. The possible mode of action of ALG is discussed in the light of these, and other, observations. [ABSTRACT FROM AUTHOR]

Anaphylactic hyposensitization of rat mast cells in vitro was demonstrated by preincubating the cells with specific antigen in suboptimal concentrations. The reduction of the histamine release was more pronounced when the cells were preincubated with increasing concentrations of antigen than with the final concentration. The inhibition remained unchanged for more than 3 hr. Non-specific antigen did not influence the histamine release, whereas a 70% inhibition was demonstrated by specific antigen. In rats sensitized to two different antigens, preincubation with one of these inhibited the subsequent histamine release induced by either of the two antigens. The hyposensitization could not be ascribed to blocking antibodies. [ABSTRACT FROM AUTHOR]

A modification of the leucocyte migration inhibition test, in which cells migrate in an agarose layer, was proposed by Clausen (1971). In the present study, this method was applied to PPD and hepatitis B antigen (HBAg), and its advantages were reassessed. From a technical standpoint, the migration in a monolayer facilitated a study of the morphology of the migrating cells. The proportion of the different kinds of migrating leucocytes was calculated and expressed as a function of the distance of migration. On the whole, 92% of the migrating leucocytes were polymorphonuclears and 8% lymphomonocytes. Three to 10% of the incubated cells were shown to migrate in the agarose layer. The surface area of the controls (migration without inhibition) was sufficiently constant and large (on the average 13 mm²) to allow a clear assessment of inhibition when antigen was added. In addition, the small requirement for antigen facilitated the use of this test in experimentation with purified HBAg. Application of the technique to clinical problems revealed that, with PPD as antigen, a highly significant inhibition was obtained in PPD skin test-positive normal individuals (0.0025

Cellular immune responsiveness, as measured by lymphocyte transformation in one-way mixed leucocyte cultures (MLC) and in phytohaemagglutinin (PHA) stimulated cultures was evaluated in forty patients with systemic lupus erythematosus (SLE) and in seventy-four normal controls. The effect produced by sera from these subjects on in vitro lymphocyte reactivity was tested on autologous cells and on homologous responding cells from a constant panel of ten healthy volunteers. The reactivity of lymphocytes from SLE patients to PHA and to a battery of allogeneic cells was significantly lower than that of normal controls. Sera from some SLE patients inhibited the MLC reactions, while in other cases a distinct stimulatory effect was found. It is suggested that virus-induced modifications of normal histocompatibility antigens cause the appearance of blocking antibody that might bind to the surface of T lymphocytes, impairing their function. [ABSTRACT FROM AUTHOR]

Sera from cases of active chronic hepatitis or acute hepatitis containing smooth muscle antibodies reacted by immunofluorescence with the membrane region of sectioned thyroid cells from thyrotoxic glands. With non-toxic glands the reaction was negative or weak. The prerequisite for a positive reaction was that the complement of the sera had been heat-inactivated. Absorption with smooth muscle antigen abolished the reaction of smooth muscle antibody positive sera with thyroid cells. Somesmooth muscle antibody negative sera from cases with disorders other than liver disease were found to give a similar immunofluorescence staining of the membrane region of sectioned thyroid cells, but these antibodies were not absorbed with smooth muscle antigen. Culture of thyroid cells was found to increase the number of cells reacting with smooth muscle antibody. In contrast, the thyroid cell antigen reacting with smooth muscle antibody negative sera was lost during culture. [ABSTRACT FROM AUTHOR]

Fractions of adult schistosomes (S. mansoni) were tested for their ability to absorb an IgG antibody activity which appears in the sera of rats and Rhesus monkeys infected with the parasite and is lethal to schistosomula cultured in vitro. The same fractions were tested for their immunogenicity in raising lethal antibody when administered to rats in Freund's adjuvant. The results of these experiments indicated that an antigen which both absorbs and induces the formation of lethal antibody is present almost entirely in the water-insoluble portion of adult schistosome homogenates. The antigen appears to be tightly bound to this membrane fraction since the use of strong anionic detergents is necessary in its extraction. Membranes solubilized in sodium dodecyl sulphate were fractionated by gel filtration on Sephadex columns equilibrated with the detergent. The same specific Sephadex fraction was shown to absorb both rat and Rhesus monkey lethal antibodies as well as induce the antibody when administered to rats. Further purification and analysis suggested that the immunogen was one of four proteins with molecular weights estimated between 21,000 and 33,000 Daltons. Rats vaccinated with partially purified antigen developed levels of lethal antibody similar to those found in animals immune to challenge through previous exposure to the parasite, yet were themselves only marginally resistant to infection. These results indicate that while clearly toxic to schistosomula grown in vitro, lethal antibody operating on its own in vivo is incapable of mediating the rejection of schistosome infections. [ABSTRACT FROM AUTHOR]

Lymphokines which were mitogenic for cultured lymphocytes caused germinal centre enlargement within the regional lymph node following their intralymphatic injection. It was found that 17 μg of a lymphokine preparation, produced by 2.5 x 104 peritoneal exudate lymphocytes, resulted in a 14-fold increase in germinal centre area and a 7-fold increase in the labelled cell content of these centres on the 3rd day after injection. Since the daily rate of lymphocyte recirculation through the regional node would supply sufficient numbers of antigen-sensitive lymphocytes to generate this amount of mitogenic lymphokine following antigenic stimulation, it is argued that lymphokine-induced germinal centre enlargement plays a physiological role in immunoregulation. [ABSTRACT FROM AUTHOR]

The previous observations of cross idiotypic specificity of IgM cold agglutinins were further investigated in quantitative precipitin assays and attempts were made to relate shared idiotypic specificity to combining specificity for I, i and Pr antigens of red cells. Shared idiotypic determinants were observed among antJ-I and anti-i antibodies which are now known to have combining specificity for determinants present on precursors of the ABH and Lewis blood group substances. There was evidence that one cold agglutinin. Ma, which had previously been shown to have an unusual kind of I specificity was deficient in the idiotypic determinants associated with the other anti-I proteins. Anti-Pr cold agglutinins, which have specificities unrelated to I-i antigens, were found to have distinct cross idiotypic determinants which were not detected on anti-I and anti-i antibodies. IgA and IgM antibodies with Pr specificity isolated from different individuals were found to share idiotypic determinants. Cross specificity antigens were detected at low concentrations in preparations of normal IgG and IgM, but not in any isolated monoclonal macroglobulins lacking cold agglutinin activity. [ABSTRACT FROM AUTHOR]

The interaction in vitro between human lymphocytes and fixed C3 was inhibited by fluid phase C3b and C3c prepared by trypsinization of purified human C3. These C3 fragments suppressed in vitro transformation of human lymphocytes by specific antigens, PPD and Candida albicans somatic antigen, but not by phytohaemagglutinin, a non-specific mitogen. These observations indicate the form of C3 which interacts with complement receptor lymphocytes and suggest that this interaction may play a part in lymphocyte activation by antigen. [ABSTRACT FROM AUTHOR]

Delayed skin hypersensitivity to specific antigen was enhanced when the antigen was given intradermally with Vibrio cholerae neuraminidase in sensitive subjects. No enhancement of skin test response was seen when inactivated enzyme was used. Neuraminidase exhibited no effect on negative skin-test response in non-sensitive recipients and no re!w1ion at the site of this enzyme was observed in any instance, suggesting that the enhancement effect is immunological in nature. [ABSTRACT FROM AUTHOR]

The recognized trend towards the standardization of the leucocyte migration assay prompted the comparison of efficacy of BCG with soluble purified protein derivative (PPD) antigens in this test. Even low BCG concentrations permit high correlations between in vivo and in vitro responses, whilst PPD doses needed to elicit comparable reactions fall within toxic concentrations. BCG-induced inhibition of leucocyte migration is abolished by the protein synthesis inhibitor, cycloheximide. [ABSTRACT FROM AUTHOR]

Rabbit antisera raised against antigens solubilized from the membranes of a transplantable Balb/c adenocarcinoma precipitated an antigen that was not expressed by the normal resting mammary gland or any other normal mouse tissue. The same antisera were used to show the presence of this antigen in other mammary adenocarcinomas, although not in tumours of different histological types. The antigen was, however, transiently expressed by mammary gland cells during the hyperplasia that both precedes and accompanies lactation. A connection is sought between the existence of this antigen and the membrane structural changes hat take place during mitosis. [ABSTRACT FROM AUTHOR]

Delayed hypersensitivity reactions appear to be important in the two main muscular human diseases involving autoimmune anomalies. Using the leucocyte migration test (LMT), a significant abnormality was detected in twelve out of fourteen polymyositis (PM) patients, and in forty out of forty-six myasthenics, in the presence of muscle antigens. No abnormal reactions in the presence of monkey thymus in the LMT were observed amongst the PM patients. as opposed to twenty-one abnormal reactions out of forty-two myasthenics. Secretion of migration inhibition factor (MIF) in the presence of muscular antigens is in accordance with what is known about hypersensitivity reactions during the course of polymyositis. In myasthenia gravis (MG), delayed hypersensitivity to muscle antigens was found to be frequent, and this also applies to thymic antigens, which are considered important in this disease. The role of the T lymphocytes in the neuromuscular junction still remains hypothetical. [ABSTRACT FROM AUTHOR]

Hairy cells from two cases of leukaemic reticuloendotheliosis' were studied for their enzyme content, adherence to nylon wool columns, phagocytosis, and the presence of surface immunoglobulins and lymphocyte-specific surface antigens. The cells reacted negatively for peroxidase, chloroacetate-esterase. alpha-naphthyl-acetate-esterase and naphthol-AS-acetate-esterase. They did not adhere to nylon wool columns nor did they show significant phagocytosis. Most of the hairy cells were found to be positively labelled for surface immunoglobulins of different classes: μ chain-positive hairy cells were predominant in number. In a homogenate of isolated, washed hairy cells from the spleen of one case only IgM could be detected in significant amounts. With peroxidase-coupled anti-thymocyte IgM most of the hairy cells of both cases showed a ring-like labelling very similar to lymphocytes. whereas monocytes and granulocytes showed no labelling. The data suggest that hairy cells are closely related to lymphatic cells of the B-cell type. They exclude with some certains the possibility that hairy cells are directly derived from cells of the myeloid system, particularly monocytes or from classical reticulum cells of the lymphatic tissue. [ABSTRACT FROM AUTHOR]

The cross-reactivity of heterologous erythrocytes was measured at the level of the B and the T cell. B cells produced antibody with restricted cross-reactivity as compared with the cross-reactivity of T cells, assayed by their ability to help B cells in a primary anti-sheep erythrocyte immune response. Helper function was maximally elicited with low antigen doses. The data suggest that the T-cell antigen receptors for erythrocyte antigens differ in specificity from those of B cells and that the sensitivity of T cells to antigen may be very great. [ABSTRACT FROM AUTHOR]

The immunoglobulins secreted into 'antigen-free' isografts of mouse small intestine have been measured by single radial immunodiifusion and immunoelectrophoresis of tissue extracts. IgA was detected in high concentration, and small amounts of IgG I and IgG2 were also present. The IgA content of material within the graft lumen was considerably higher than the IgA content of the graft wall, indicating that IgA had been secreted and stored. Graft IgA increased with time after implantation, and no differences in immunoglobulin contents were found when grafts in thymus-deprived and in normal mice were compared. A group of host mice had been immunized with BSA and had high titres of circulating antibody to BSA; anti-BSA was not detected in the grafts implanted in these mice. However, graft extracts had moderately high titres of bacterial agglutinins when tested against a panel of commensal gut bacteria. These results indicate that secretion of IgA into a segment of small intestine is not dependent upon the presence of antigens within its lumen; however, the immunoglobulins secreted may have antibody activity against antigens present in small intestine elsewhere in the animal. [ABSTRACT FROM AUTHOR]

A membrane antigen on peripheral blood lymphocytes from cases of chronic lymphatic leukaemia (CLL) is described. The antigen was detected by complement-dependent cytotoxicity using serum from a healthy pregnant woman, and appeared to be absent from the normal lymphocyte population in these patients. The serum was also cytotoxic for some acute leukaemia blast cells and for cultured Burkitt lymphoma cells; absorption studies suggested that the CLL antigen is identical to the acute leukaemic antigen. Similar antibody activity was also found in a number of HL-A typing antisera and could be separated from the HL-A specificity by absorption. Although these antibodies are probably directed against foetal antigens, the leukaemic antigen could not be demonstrated on foetal tissues. [ABSTRACT FROM AUTHOR]

Production of antibody against major trasplantation antigens has been studied in sheep receiving osteoarticular allografts at the knee joint. These investigations have shown that this type of allograft leads to the rapid production of serum antibodies in the recipient animal. It was found that a high proportion of the sheep used in these studies possessed antibodies before operation. A large number of the animals which possessed antibody against major transplantation antigens prior to operation developed a wide spectrum of antibodies after operation more rapidly than did those recipients who had no pre-existing antibody. A smaller number showed an increase in antibody spectrum which was comparable with non-sensitized recipients. An analysis of the patters of antibody production was carried out in order to assess the possible benefits of reducing the bony component of the graft in lowering its immunogenicity. It was found that the thickness of bone carried by the graft was largely irrelevant and the immune response was initiated with only small amounts of bone. There is some evidence in this data that the surface area of bone is important in determining the pattern of the immune response. [ABSTRACT FROM AUTHOR]

Continuous lymphoblastoid cell lines (CLC) are more reactive with HL-A antisera in a complement-dependent cytotoxic test than are peripheral blood lymphocytes (PBL). This additional reactivity leads to assignment to a given CLC of more than four HL-A antigens, the maximum allowable under the two locus concept of the genetic control of HL-A. However, absorption of antisera by CLC shows that no more than four HL-A antigens exist on any of the CLC used in this laboratory. The additional reactivity of these cells lines can be explained in three ways. Firstly, it may be due to the presence of sublytic amounts of HL-A antibody in operationally monospecific antisera. Secondly, it may be due to cross-reactivity between HL-A antigens. Both these findings can be explained on the basis of the increased quantity of HL-A antigens on CLC compared to PBL. Thirdly, it may be due to the presence of a heat-labile (56 for 30 min) complement-dependent cytotoxic antibody which is present in 90% of normal human sera, and detects an antigen group tentatively labelled 'D'. [ABSTRACT FROM AUTHOR]

Twenty-eight chimpanzees were divided into six groups according to their history of previous immunization or exposure to hepatitis B antigen (HBAg) and studied for delayed hypersensitivity (DH) to HBAg. Purified HBAg derived from a normal human carrier was used for in vivo skin testing and in vitro leucocyte migration inhibition tests. Of seventeen chimpanzees immunized with HBAg in Freund's complete adjuvant (FCA), nine exhibited positive DH reactions to HBAg with good correlation between the in vivo and in vitro responses. Of the seventeen chimpanzees, fourteen also exhibited positive DH reactions to purified protein derivative of tuberculin (PPD) with marked erythema and induration; the other three exhibited only erythema with no induration. None of the seventeen animals exhibited any immediate reactivity to either HBAg or PPD. Intradermal injection of HBAg-negative human serum failed to elicit DH reactions in four animals who showed positive skin test with purified HBAg; the DH response was thus probably HBAg-specific. Nineteen chimpanzees, including six unimmunized animals, three chronic carriers of HBAg and two which had been injected with HBAg without FCA, failed to show DH response to HBAg. Thus, DH to HBAg was observed only in animals hyperimmunized with HBAg in FCA. [ABSTRACT FROM AUTHOR]

An autoantibody, reacting with a cytoplasmic component of many types of glass-adherent cells in tissue culture has been detected in the serum of patients with infectious mononucleosis. Evidence is presented that the structures with which this antibody reacts are closely related to cytoplasmic microtubules. [ABSTRACT FROM AUTHOR]

To identify monoclonal bone marrow-derived (B) lymphocytes in human myelomatosis specific rabbit antisera were produced against idiotypic specificities on IgG-K myeloma proteins from three patients. The unlisera neither cross-reacted nor reacted with normal immunoglobulins. By indirect immunofluorescence surface Immunoglobulins were demonstrated on 20-47% of peripheral blood lymphocytes from untreated patients after staining with idiotypic antiserum against the patient's own myeloma protein, bul not after staining with other idiotypic antisera. The antisera also stained autologous plasma cells. The monoclonal surface Ig on myeloma lymphocytes was removed by trypsin and regenerated after incubation in serum-free medium. Myeloma protein was not adsorbed onto lymphocytes. It is concluded that monoclonal B lymphocytes belonging to the plasma cell myeloma clone are present In myeloma patients. There were few normal B lymphocytes in untreated patients. During treatment the monoclonal lymphocyte population and the plasma cells content in bone marrow, as well as the concentration of monoclonal immunoglobulin in serum, decreased simultaneously. These findings were associated with other signs of clinical improvement. [ABSTRACT FROM AUTHOR]

Outbred Swiss mice show a stable level of circulating thymic factor (TF) (measured by its action on theta-positive rosette-forming cells (RFC)) until the 6th month of life. Afterwards, this level progressively declines. Swiss mice genetically selected as 'high' and 'low' responders for anti-sheep red blood cell haemagglutinin production show no difference with ordinary Swiss mice with regard to serum TF level. This observation is compatible with previous results showing that the selection mainly induces changes in B-cell reactivity without T-cell modification. Conversely, genetic selection of Swiss mice based on spontaneous antinuclear autoantibody production (tested by immunofluorescence) induces a decrease in TF level, which has already become significant by 2 months of life. The mice thus selected (Swan mice) which present autoimmune manifestations and immune complex disease, like NZB and B/W mice, show the same premature cessation of TF as these other autoimmune mice. Such abnormality is compatible with the T-cell deficiency of NZB, B/W and Swan mice. [ABSTRACT FROM AUTHOR]

Serial sera from nine volunteers, injected 10 years ago with sheep erythrocytes and showing heterophile antibody responses comparable to those seen in infectious mononucleosis, were retrieved from storage and examined for antibodies to Epstein-Barr virus (EBV) related antigens. Eight of the nine volunteers had preexisting antibodies to EBV, as determined by neutralization tests and by indirect immunofluorescence reactions with fluorescein-conjugated antibodies to human IgG, but not with anti-human IgM conjugates. The EBV-specific reactivities remained unchanged after immunization. The results indicate that: (a) EBV- or EBV antigen-containing lymphoblasts from carrier or freshly infected cultures contain no detectable heterophile antigen; (b) immunization with sheep erythrocytes does not evoke antibodies which interact with EBV or EBV-infected cells. These points are supported especially by the results obtained with the heterophile antibody-positive, anti-EBV-negative serum of one of the volunteers who subsequently sustained a primary EBV infection. It is unlikely, therefore, that immunization with heterophile antigen will induce immunity to infectious mononucleosis, apart from the fact that heterophile antibody responses are largely restricted to the IgM class. [ABSTRACT FROM AUTHOR]

Cells binding the antigen Keyhole limpet haemocyanin (KLH) were detected among human peripheral blood leucocytes and antigen-stimulated cultured lymphocytes. They were detected by their formation of rosettes with antigen-coated human O red blood cells. In the peripheral blood 0.13% of the lymphocytes of non-immune and 0.80% of the lymphocytes of immune subjects were antigen binding. In KLH-stimulated cultures of the lymphocytes of non-immune subjects there were 0.1% antigen-binding cells and in those of immune subjects there were 8.5% antigen-binding cells. Binding was specific. Thus, lymphocytes from phytohaemagglutinin- and streptolysin O-stimulated cultures did not bind KLH-coated red blood cells and did not form rosettes with KLH-stimulated lymphocytes. Incubation of KLH-stimulated lymphocytes with heterologous anti-immunoglobulin serum revealed that the KLH receptor was related to IgM. Addition of metabolic blocking agents (puromycin, cycloheximide, actinomycin-D and arabinosyl cytosine) to KLH-stimulated lymphocyte cultures revealed that the generation of antigen-binding cells required both DNA and protein synthesis. After primary immunization of normal human subjects, antigen-binding cells were detected in their appropriate lymphocyte cultures at 7 days and reached their maximum level at 21 days, in five of the six kinetically followed individuals and at 14 days in the other. This data represents the first report of the generation of antigen-binding cells in lymphocyte cultures demonstrated by the rosette method. These methods should be useful in the study of a variety of immunological problems in man. [ABSTRACT FROM AUTHOR]

Purified thyroglobulin was digested with trypsin and papain and was also reduced with dithiothreitol and alkylated with iodoacetamide. The resulting fragments were separated and characterized by immunological techniques. Following enzymatic degradation a small fragment, termed fraction 2, was isolated. It had a sedimentation coefficient of approximately 3S and a mol. wt determined by polyacrylamide gel electrophoresis in presence of SDS was approximately 37,000 daltons. In the Ouchterlony test, it was antigenically deficient as compared with intact thyroglobulin, when rabbit antisera to human thyroglobulin were used. With human autoantisera, fraction 2 did not show any precipitin reaction in the Ouchterlony test. However, it produced weak inhibition in the tanned cell haemagglutination test. The fractions obtained from trypsin and papain digestion appeared to be immunologically identical. However, when these fractions were compared with the similar product obtained from reduced and alkylated thyroglobulin, the latter fraction showed a reaction of non-identity in the Ouchterlony test, and had a weaker inhibiting capacity in tanned cell haemagglutination test. The fraction produced by reduction and alkylation had a sedimentation coefficient of 8S and an approximate mol. wt of 38,500. It can be concluded that the small mol. wt fractions derived from enzymatic breakdown and by reduction and alkylation are different although both possess a number of antigenic determinants recognized by the rabbit antiserum and lack most of the autoantigenic determinants. [ABSTRACT FROM AUTHOR]

The results suggest that the anaemia in mice associated with the injection of C. parvum is mediated by an immune mechanism. Survival studies of 51Cr-labelled red blood cells (RBC) in splenectomized and unsplenectomized mice show that mice immunized with C. parvum three times, clear RBC faster than unimmunized mice. However, hyperimmunized mice stabilized their haemoglobin concentrations above their minimum level and did not clear 51Cr-labelled RBC faster than unimmunized mice. All mice produced antibody against C. parvum but remained direct Coombs'-negative. Serum from immunized animals did not contain opsonins for -51Cr-labelled RBC. The reticulocytosis, normal osmotic fragility tests and normal survival times of 51Cr-labelled RBC from C. parvum-immunized mice all suggest that the main direct influence of the micro-organism is extrinsic of the RBC. Antigens from the micro-organism were adsorbed by erythrocytes in vitro. It is hypothesized that the anaemia is caused by destruction of RBC brought about by antibody reacting with bacteria that have been adsorbed by RBC. [ABSTRACT FROM AUTHOR]

A counterimmunoelectrophoresis (CIEP) technique has been developed for the rapid, simple, specific detection of nucleic acids as antigens, or for the detection of precipitating antibodies to nucleic acids or nuclear antigens. The majority of precipitins could be detected within 1 hr. As little as 0-0015 μg of antigen per ml (e.g. poly A: poly U) could be detected. Specificity of rabbit antisera to nucleic acids was demonstrated by selective reactions using a panel of polynucleotides. 1091 patient sera were examined for precipitins to DNA, single-stranded DNA, nucleoprotein and calf thymus nucleoprotein. Precipitins to DNA were found in 42% of systemic lupus erythematosus sera, 9% of rheumatoid arthritis sera and 4% of juvenile rheumatoid arthritis sera. Results with the CIEP method showed equal sensitivity as results obtained by complement fixation or binding assays, but were more sensitive than double diffusion in agar (Ouchterlony). [ABSTRACT FROM AUTHOR]

T and B lymphocytes in the human foetal thymus and spleen were studied to determine the distribution and degree of development which takes place before exposure to environmental antigens occurs. Tests applied were spontaneous and complement- dependent rosette formation and immunofluorescence to detect surface immunoglobulins. Most thymus lymphocytes were spontaneous rosette-forming cells: the percentage of these cells in the spleen was lower. Complement receptor lymphocytes (CRL) were found in the spleen but not the thymus, suggesting that these tissues contain lymphocytes of different origin. Lymphocytes with surface immunoglobulin (SIg lymphocytes) were more numerous in the spleen than the thymus. Analysis of class-specific heavy chain and light chain determinants suggests that some foetal B cells carry heavy chains of more than one class. A possible model for foetal B-cell development and its relationship to antigen drive is discussed. [ABSTRACT FROM AUTHOR]

The rheumatoid biologically active factor (RBAF) was characterized further with respect to its biological and physical characteristics. The histamine-releasing activity of the RBAF in the guinea-pig lung was influenced in the same manner as soluble immune complexes when the lungs were being perfused at 20°C or 45°C or when the perfusate lacked calcium or magnesium or contained N-ethylmaleimide, phenol, theophylline, adrenaline or succinate. The RBAF was consistently associated with complement-fixing activity and RBAF-positive synovial fluid showed a lower total haemolytic complement level than RBAF-negative fluid. However, RBAF activity was not lost following absorption with anti-human beta IC globulin. There was a higher frequency of free DNA and/or single-stranded DNA in RBAF-positive than negative synovial fluid. RBAF-positive synovial fluid was more active than RBAF-negative fluid in neutrophil chemotaxis when examined at 1:10 but not when undiluted. Mixed IgG-IgM cryoprecipitates failed to show RBAF activity and aggregates seen on analytical ultracentrifugation of rheumatoid synovial fluid did not correspond with the RBAF. The RBAF was stable to freezing and thawing but was labile to acid pH and to heating at 56°C. It was concluded that the RBAF is likely to be a soluble immune complex consisting of IgG and a second constituent which is labile to acid and heat. [ABSTRACT FROM AUTHOR]

The macrophage migration inhibition techique was emplyed to study the debvelopment of human maternal cell-met\dilated immune reactions to plecental antigens during the firs pregnancy. Cell-mediated immune reaction to pooled antigens from five placentas could not be demonstrated during the first trimester, In the th month. peripheral blood leucocyte's from seven out of eight primigravidous women tested were ravtive t placental antigens, The one nonreactor became reactive by the th month. All of the primigravidous women tested during subsequent months were immune reactivity in the course of the first pregancy. [ABSTRACT FROM AUTHOR]

A red cell autoantibody in low titer was demonstrated in one-third of Ugandan sera tested by performing the Combs test. on enzyme (bromelain) treated erythrocytes, The authoantibody was found Keith increased frequency in subjects over 40 years of age and in those with the haemocyte anaemia in pregnancy syndrome. But was not detected in medical students. Two subjects with autoantibody\ie's and normal haemoglobin values has na normal re dell survival The authoantibody was not associated with acute attacks of malaria. The possible relationships of the autoantibody to the he\haemolytic anaemia in pregnancy and the tropical spend meagerly syndromes are discussed. [ABSTRACT FROM AUTHOR]

We have described a new autoantibody present in the serum of diseased NZB mice reactive with autochthonous and syngeneic fibroblasts. allogeneic cells are not affected. This antibody can be specifically absorbed both by fibroblasts and lymphocytes from NZB but not from control mice. The antibody cannot act as a block from diseased NZB mice. This suggests that the precise antigenic specificities of the cell-mediated and antibody responses are different [ABSTRACT FROM AUTHOR]

Cryoglobulin's were detected in the sera of thirteen patients with acute viral hepatitis and of twelve with chronic hepatic diseases tactic chronic hepatitis, primary biliary cirrhosis and cryptogentice cirrhosis). Their nature and antibody activity was studied, In both groups, most of them consisted of mixed cryoimmunoglobulins (Igg and/ire UA) but some were single-class immunoglobulins with one or both types of light chains, Unusual components were also found. fetoprotein was present in four cryoprecipitate's in two as the single constituent and in two associated to immunoglobulins: hepatititsassociated antigen co-existed in one of the latter, some sryoglobulins showed antibody activity against human IgG. smooth mus ele and mitochondrial antigens, In one case. the IgM-Kappa of the cryoprecipitate had antibody against fetoprotein this antigen was also present tin the cryoprecipitate, suggesting immune complex formation. An Autoantibodies were aksi kicked fir bub tee sera if tee tweet fume oatuebts apart frin tee nits cinnib ibesm abtuvidues 10 fetoprotein were found in two patients. [ABSTRACT FROM AUTHOR]

Immune complexes were detected by the platelet aggregation technique in sera of six out of twenty-six patients with sarcoidosis. Five of these patients had acute bilateral hilar lymphoma syndrome, four of them with concomitant erythema nodosum. The size of the immune complexes was 19S or larger. [ABSTRACT FROM AUTHOR]

Immunization of mice with allogeneic tumour antigen in Freund's complete adjuvant (FCA) facilitates the growth of subsequent allografts of R1 and BPS tumours in C57, B1 mice. FCA by itself is less effective. In this particular experimental method, antigen given without FCA is not effective. The extent of facilitation depends on the tumour host combination, on the physical form of the antigen, and on the time interval between pretreatment and tumor allograft. The immunosuppressive effect of FCA appears to be mediated through a direct effect on lymphocytes, largely independent of blocking factors or macrophage effects. [ABSTRACT FROM AUTHOR]

Plaque assays for derepressed autoimmunocompetent B lymphoid cells with specificities for two murine erythrocyte surface autoantigens have been developed. Using purified anti-HB and anti-X anti-erytlirocyte autoantibodies, fluid-phase complement fixation reactions were performed to determine the influence of pH and ionic strength on the efficiency of autoantibody-mediated cytolysis of murine erythrocytes. Anti-HB autoantibody exhibited optimal cytolysis of bromelin-treated erythrocytes at pH 7.0, ionic strength 0.15; haemolysis of intact erythrocytes by anti-X autoantibody was observed over a broad range with maximum sensitivity and specificity at pH 6.6, ionic strength 0.14. NZB spleen ceils secreting anti-HB autoantibody were detected in modified direct haemolysis-in-gel assays, were neutralized by soluble HB autoantigen, and appeared to represent derepressed B lymphoid cells of anti-HB type. Anti-X plaque-forming cells, not demonstrable using conventional assays, were detected under conditions corresponding to the pH and ionic strength optima determined for complement-dependent cytolysis using purified autoantibody. NZB spleen cells secreting anti-X autoantibody were detected in indirect assays employing monolayers of erythrocytes, were neutralized by soluble X autoantigen, and appeared to represent derepressed B lymphoid cells of anti-X type. The distribution of observed plaque diameters in both HB and X assays suggested that more than 95% of all cells secreting autoantibody were detected. [ABSTRACT FROM AUTHOR]

Quantitative measurements of immunoglobulins E, A and G were made in tracheobronchial secretion and serum of thirty-four patients in association with bronchoscopy. Measurable quantities of all three immunoglobulins were found in both secretion and serum in all patients, except in one person, in whom IgA was found neither in the secretion nor the serum, and in another person in whom no measurable IgG was found in the secretion. Calculations indicated that all immunoglobulins were produced locally in the tracheo-bronchial mucosa. This applied especially to IgA and IgE, of which 84 and 73% respectively, were calculated to have been produced locally. Quantitative differences between the three immunoglobulins were also apparent between different groups of patients. The number of patients studied is too small, however, for definite conclusions to be drawn in this respect. [ABSTRACT FROM AUTHOR]

Radioiodine-labelled globulins from antisera directed against mouse immunoglobulin, are fixed by cells of various mouse ascites tumours, thus indicating that the cells are coated in vivo with immunoglobulins. The amounts of tumour-bound immunoglobulin in the tumour-bearing animal increase as the time interval between tumour inoculation and the harvest of tumour cells increases. Tumour cells from mice which had been irradiated prior to the tumour inoculation are associated, in general, with lower amounts of immunoglobulin than tumour cells from unirradiated mice. [ABSTRACT FROM AUTHOR]

An in vitro expression of delayed-type hypersensitivity to tuberculin was tested in Mantoux positive and Mantoux negative persons. Their lymphocytes and monocytes were isolated from venous blood and incubated in vaseline chambers with or without tuberculin. In the presence of tuberculin, a substantially lower percentage of monocytes from Mantoux positive persons spread, than monocytes From Mantoux negative persons. This antigen-induced inhibition of monocyte spreading seems to be a reliable measure of tuberculin hypersensitivity, since it occurs only in Mantoux positive persons and correlates well with the intensity of the tuberculin skin reaction. Reproducibility of the test and the speed with which it is performed could constitute a basis for the use of monocyte spreading inhibition in clinical studies of cell-mediated immune reactions. [ABSTRACT FROM AUTHOR]

Circulating tissue antigens (CTA) were investigated in 143 patients with disorders involving injury of the liver and of other organs and in forty-eight normal subjects by immunodiffusion techniques using rabbit anti-human liver serum containing antibodies to a liver-specific antigen and to tissue antigens of wide organ distribution. Analysis of serum samples by double immunodiffusion showed up to three CTA in the following cases: fifteen out of eighteen, viral hepatitis (VH), two out of thirteen, other infectious diseases, two out of ten, alcoholic cirrhosis, seven out of twenty-one, congestive heart failure (CHF), four out of fourteen, myocardial infarction, ten out of twenty-one, trauma, two out of thirteen, carcinoma and three out of thirty-three, miscellaneous diseases. Forty-eight normal subjects showed no CTA. Immunoelectrophoresis of most of the positive eases showed two to three CTA, while a few cases showed four to six. Absorption tests with organ extracts demonstrated that in most patients, CTA were substances shared by several organs. However, in two cases of VH, in two cases of CHF with liver necrosis and in two cases of trauma to the liver, one of the CTA was shown to be liver specific. The CTA were susceptible to digestion by pronase and were found to be relatively thermolabile. Positive sera showed higher glutamic oxaloacetic transaminase and lactic dehydrogenase activities than the negative sera. These preliminary data suggest that further investigation on CTA in disease involving tissue injury and necrosis may be rewarding. [ABSTRACT FROM AUTHOR]

The serum immunoglobulins IgG, IgA, IgM and IgE were determined together with indirect haemagglutination test in normal subjects and in patients with intestinal and extra-intestinal amoebiasis. The IgG level in intestinal amoebiasis was found to be significantly higher than that of extra-intestinal. Although in intestinal amoebiasis, the concentrations of IgE were comparatively higher than extra-intestinal group, the IgE levels in both groups of amoebiasis were seen to be much higher than that of the normal group. No significant difference was found in the IgA and IgM levels between the three groups of cases. Indirect haemagglutination test was positive in both intestinal and extra-intestinal amoebiasis, but negative in normal subjects. [ABSTRACT FROM AUTHOR]

Patients with breast carcinoma, fibrocystic disease, and fibroadenoma have antibody to an antigen prepared from homologous breast carcinoma. In 50% of patients with breast carcinoma, this antibody, detected by immunodiffusion and complement fixation, appears within 2 weeks of radical mastectomy. This antibody also appears in a lower percentage of the sera of patients with fibrocystic disease and fibroadenoma. Gel permeation chromatography indicates the antigen to be a high molecular weight compound. [ABSTRACT FROM AUTHOR]

Rabbit antisera were prepared against six contractile proteins of smooth muscle- actomyosin, actin, myosin, heavy meromyosin, light meromyosin and tropomyosin -extracted from human uterus by standard procedures. Antibodies were sought by double-diffusion in gel and by immunofluorescence on cryostat sections of rat tissue. All the antisera except anti-actin gave precipitin lines with their respective antigens, and all gave specific fluorescence with smooth muscle fibres and produced staining of liver cells in the region of their cell membranes (polygonal pattern). Anti-actin sera stained in addition renal glomeruli, and also a microfilamentous network in HeLa cells and chick embryo fibroblasts. Anti-heavy meromyosin (and less brightly anti-actin) stained the cytoplasm of bile duct epithelial cells, and the apical and basal regions of intestinal muscosal cells in a linear fashion. These patterns of staining between them reproduce the range of staining patterns seen with human sera positive for smooth muscle antibody by immunofluorescence. [ABSTRACT FROM AUTHOR]

Sera from patients with squamous carcinoma of the cervix uteri and from control patients were examined, by means of indirect immunoflourescence, for antibodies to cell constituents in frozen sections of cervical and other tissues. Cervical carcinoma cells were found to possess cytoplasmie antigens, which reacted with antibodies in autologous and homologous sera. More than half the patients with carcinoma of the cervix had detectable antibodies to cytoplasmic antigens. Comparison of reactions of individual sera and tissues, together with absorption experiments, indicated that either or both of two common cytoplasmic antigens were present in all carcinomas of the cervix and that the same antigens were present in some non-malignant cervices. A high incidence of antinuclear antibodies was found in patients with squamous carcinoma of the cervix, adenocarcinoma of the cervix or body or the uterus, or pre-malignant cervical lesions. [ABSTRACT FROM AUTHOR]

A macrolipoprotein of human, rabbit and rat livers could be purified and characterized. The proteins were located on the membranes of isolated liver cells by indirect immunofluorescence technique using monospecific heterologous antisera. Antibody assays and immunofluorescence studies demonstrate a complete organspecificity, but incomplete species specificity of the membrane protein. The technique of isolating liver cells without the use of proteases is an improvement which will facilitate immunological in vitro studies on living hepatocytes. [ABSTRACT FROM AUTHOR]

Human spermatozoal heads could be made to swell by treatment with trypsin or a combination of dithiothreitol and trypsin. On these swollen spermheads a new auto-antigen was detected. Sera containing antibodies against swollen spermheads gave a homogeneous fluorescence covering the swollen head when tested in the indirect immunofluorescence test. Sera from men, women and children were tested for antibodies against these swollen spermatozoa in the indirect immunofluorescence test. In sera from men the frequency of a positive reaction correlated with a high serum spermagglutinating anlibody titre. Sera from women and children were negative. Antibodies directed against this antigen were primarily IgG. The intensity of the immunofluorescent staining for one auto-antiserum on swollen spermheads was measured with a Leilz microscope photometer and expressed in standard fluorescence units. Spermheads were solubilized by treatment of swollen spermatozoa with deoxy ribonuclease (DNase). The solubilized spermhead preparation was analysed for the presence of auto-antigens by means of quantitative immunofluorescence inhibition and spermagglutination inhibition tests. Antibodies against swollen spermheads, the front part of the acrosome and the postnuclear region could be absorbed with solubilized spermhead preparation, whereas spermagglutinating antibodies could not be absorbed. [ABSTRACT FROM AUTHOR]

A cytomegalovirus (CMV) induced early antigen (EA) was demonstrated in fibroblasts infected in the presence of the DNA inhibitor cytosine arabinoside (Ara C). In the dose used (20 /(g/ml) Ara C blocked DNA synthesis completely, as judged by thymidine incorporation and autoradiography. The percentage of EA positive cells was 0.6 8 hr post-infection, increasing to 50 and 65% on the two following days. EA differed from late antigens (LA), present in CMV-infected but otherwise untreated fibroblasts, with regard to distribution and antigenic specificity. Sera differed in their reactivity to EA at comparably high anti-LA titres. Direct immunofluorescence staining of EA-positive cells was only blocked with high titred anti-EA sera, but not with high titred anti-LA sera with a low or negative anti-EA-titre. Anti-CMV-EA did not cross react with antibodies against EBV, Herpes simplex or varicella/zoster. A collection of 115 serial sera from forty-five acutely CMV-infected patients was tested for anti-EA and anti-LA antibodies. The diagnosis was confirmed by the occurrence of a significant rise in CF antibodies and/or virus isolation. Sera taken before illness (five patients) had neither anti-LA nor anti-EA antibodies when tested al a 1 :40 dilution. All patients in the acute phase of this disease had anti-LA titres between 1:640 and ≧ 1 : 10240. 43/45 of these patients developed anti-EA at titres between 1:40 and ≧ 1:2560. As a rule EA-antibodies were detected 10-19 days after the onset of symptoms. Anti-EA tended to peak later than anti-LA and declined again within a few months. Anti-LA was maintained at nearly constant levels. This applied also for antibodies detected in the CMV complement fixation (CF) test. Sera from healthy seropositive donors or from patients with unrelated bacterial or viral infections contained anti-LA-antibodies in titres ranging from 1:40 to 1 : 1280 as a rule, but had no anti-EA-antibodies. Diagnostically significant increments or declines in the anti-HA titres were observed more frequently than corresponding changes in the levels of anti-LA and CF antibodies. In view of the transitory nature of the anti-EA response, the presence of this antibody in an acute phase serum at a titre of ≧ 1: 80 can be taken as suggestive of a current primary infection. [ABSTRACT FROM AUTHOR]

Rabbits were immunized with peripheral cells, thoracic duct cells and various lymphoblast cell lines. Bleedings were taken approximately 4 weeks after each injection corresponding to the primary and secondary immune response. The immunofluorescence (IMF) titre of each serum was estimated using the specific antigen cell and the cells used to raise the other sera. In general the titres were higher when the specific antigen cells were used. In absorption experiments it was found that anti-peripheral cell sera and anti-thoracic duct cell sera were readily absorbed by the specific ornon-specie fie cells, but the anti-lymphoblast sera, although readily absorbed by the specific or other lymphoblast cells, could not be completely absorbed with peripheral and thoracic duct cells. The IMF test was, by minor modifications, adapted to allow preliminary studies to be made of the desorption of anti-human lymphocyte globulin from the lymphocytes and it is concluded from these investigations that, while peripheral cells, thoracic duct cells and cultured human lympho blasts may contain similar and/or identical antigens, they also express similar but not identical antigens, while cultured lympho blasts in addition possess antigens absent from peripheral and thoracic duct lymphocytes. [ABSTRACT FROM AUTHOR]

The frequency of cryoglobulinaemia was statistically significant in acute proliferative endocapillary glomerulonephritis, in membrana-proliferative glomerulonephritis and in lupus glomerulonephritis. They all have in common glomerular cellular proliferation and glomerular deposits of C3. The cryoglobulin's in most of these cases contain IgG and IgM. Except for acute proliferative endocapillary glomerulonephritis, a rather good correlation was observed between the cryoglobulin constituents and the immunoglobulins present in the glomerular tuft. Rheumatoid factor was very rarely found in the serum of these patients. A cryoglobulin characterized by the presence of the three immunoglobulins was also found in cirrhosis of the liver with or without proven glomerulonephritis. [ABSTRACT FROM AUTHOR]