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The growth of fresh and I issue cultured B-16 malignant melanoma cells in C57BL/6 mice was quantitated, using as parameters the latent time to tumor appearance, tumor incidence, and mortality rate. The tumor was antigenic, as evidenced by the appearance of humoral antibodies to melanoma-associated antigens during the natural growth of this tumor and by the induction of resistance to lethal doses of tumor by immunization with repeated injections of irradiated, or sublethal doses of viable, melanoma cells. This immunity was not entirely specific, since mice immunized to B-16 melanoma were slightly resistant to challenge with BW10232 mammary adenocarcinoma and, conversely, mice immunized to mammary adenocarcinoma were slightly resistant to melanoma. [ABSTRACT FROM AUTHOR]

Three patients with photoallergy to 3,4',5 tribromosalicylanilide (TBS) following the use of a germicidal soap were studies to determine the mechanism of their photoallergy. None of these subjects had undergone previous patch or photopatch testing. Duplicate patches of purified TBS and its photoproducts 4', 5 dibromosalicylanilide (DBS). 4' monobromosalicylanilide (MBS), and salicylanilide were applied to the skin of the back. One set of patches was irradiated using a source of long ultravoilet whereas all ultraviolet radiation was regorously excluded from the second set. In one patient the results were consistent with a plain contact allergy to the photoproducts DBS and MBS. In a second patient irradiation of any of the three brominated salicylanilides resulted in a reaction whereas no reactions were observed in the dark. A free radical mechanism may be responsible for the formation of the complete antigen in this patient. In the third patient both mechanisms appeared to coexist. Reactions to commercial-grade TBS did not differ from those obtained with pure TBS. [ABSTRACT FROM AUTHOR]

The value of HL-A antigens as genetic markers and epidemiologic tools in various disease states is becoming increasingly apparent. The antigens of the HL-A system (the major histocompatibility locus in man) are found on most human cells, especially the leukocytes and cells of the epidermis and the dermis. Associations have recently been found for HL-A antigens in psoriasis. atopic dermatitis, systemic lupus ervthematosus, dermatitis herpetiformis, pemphigus vulgaris,. and malignant melanoma of the skin; in several other diseases, associations with HL-A antigens are also probably present. Two possible mechanisms of association of HL-A with various diseases are (1) a close linkage between a gene governing immunologic responsiveness to a disease and HL-A genes, or (2) antigenic (chemical) cross-reaction between HL-A antigens and the etiologic agent (e.g.. streptococci or viral capsular antigen) associated with the disease. [ABSTRACT FROM AUTHOR]

SV40 is an oncogenic DNA virus capable of both tumorigenesis in vivo and of transformation in vitro. i.e., loss of contact inhibition and subsequent overgrowth of otherwise contact-inhibited cells. Marked increased susceptibility to SV40 transformation of skin fibroblasts is present in Down's syndrome and Fanconi's anemia, both of which are associated with a high incidence of neoplasia. The question arose as to whether skin fibroblasts from patients with xeroderma pigmentosum (XP) which show an otherwise normal incidence of SV40-induced transformation, might, on exposure to UV radiation, show enhanced transformation. Can the biologic defect in relation to carcinogenesis in XP be enhanced susceptibility to the triggering by sun exposure of viral oncogenesis? Both XP and normal skin fibroblasts were exposed to low-dosage UV radiation immediately prior to infection with SV40 and the incidence of T-antigen, an early marker of SV40 infection, and of gross transformation, the morphologic late end point in vitro, were noted. No increase in transformation either absolute or relative to cell survival post irradiation was seen. In no instance was transformation seen in noninfected, but irradiated, control cultures. [ABSTRACT FROM AUTHOR]

Para-toluic acid (1:4 methylbenzoic acid) and ortho-toluic acid were found to be potent allergic sensitizers when applied to human skin. Cross-sensitivity was found between all three isomers, p-toluic, m-toluic, and o-toluic acids. It is suggested that this cross-sensitivity is explicable in terms of carrier protein specificity rather than by the presence of a common chemical sequence in the hapten or by conversion of the cross-reactants to an identical hapten. [ABSTRACT FROM AUTHOR]

Small amounts of dinitrochlorohenzene (DNCB) in capsule from were fed to 63 adult male volunteers, following which these subjects and 60 control subjects were simultaneously subjected to a sensitizing series of topical applications of the same compound. After sensitization had been achieved, the level of sensitivity was inferred from a quantitative test series. Small amounts of DNCB appeared to have no effect on subsequent sensitization. When more than 20 mgm DNCB were given by mouth, however, a significant decrease in the ultimate level of sensitivity was seen. [ABSTRACT FROM AUTHOR]

Variation in the expression of blood group antigen A in different areas of the human oral mucous membrane was investigated using a double layer immunofluorescence staining technique. The expression of the antigen was semiquantitated by a serial two-fold dilution titration. Comparison between cheek, commissure, lower lip, and hard palate was made. It was not possible to demonstrate any difference in expression of blood group antigen between cheek, commissure, and lower lip. However, in most cases a marked difference in blood group antigen A was found between cheek and hard palate, the latter demonstrating a much weaker expression. [ABSTRACT FROM AUTHOR]

The lymphocyte transformation test is commonly employed as a correlate of delayed hypersensitivity in vitro. As a generalization it is a good correlate, although there are some exceptions. Venous blood is collected in heparin and lymphocytes are collected and placed in a culture with or without antigen. After an appropriate duration of incubation, the precursor of DNA, tritiated thymidine, is added to the cultures. The cells are then harvested and the radioactivity incorporated by the antigen-stimulated cultures and compare it to the counts in the unstimulated cultures. The difference in the counts reflects the increment of DNA synthesis by cells responding to the antigen.

Detergent-soluble antigens from rat liver microsomes and plasma membranes were separated by two-dimensional immunoelectrophoresis. Enzyme activities in the immunoprecipitates were assayed by different staining reactions. Nine different microsomal esterases were detected. Six of these, including one esterase shared with plasma membranes, were inhibited by 0.1 mM difluorophosphate. but not by 1 mM eserine. Plasma membranes and microsomes each contained at least ten nucleoside diphosphatase and triphosphatase-active antigens. Only three precipitates with these activities were detected when plasma membrane extracts were tested against anti-microsomal sera or microsomal extracts against anti-plasma membrane sera. Although the possible immunological identity of these antigens remains to be established these findings suggest that plasma membranes and microsomes have at most three of these nucleoside diphosphatases and triphosphatases in common. Seven microsomal nucleoside diphosphatases and triphosphatases were intimately associated with both acid phosphatase and NADH-neotetrazolium reductase activities. These complexes also contained phospholipids as shown by incorporation experiments either in vivo or in vitro with [14C]-choline or [14C]ethanolamine. Three microsomal UDPase-active antigens did not hydrolyze ATP or ADP and were not associated with acid phosphatase or NADH-neotetrazolium reductase activities. Furthermore, these antigens did not contain any of the labelled phospholipids. Lipid-containing multienzyme complexes of different composition were also detected in the plasma membranes. The nucleoside diphosphatases and triphosphatases in plasma membrane extracts were associated with NADH-neotetrazolium reductase activity and two of them also contained L-leucyl-β-naphthylamidase activity. p-Chloromercuribenzoate (0.1 mM) inhibited all NADH-neotetrazolium reductase activity in the plasma membrane and microsomal precipitates and 10 mM tartrate all acid phosphatase activity in the microsomal precipitates, leaving the other activities intact. This indicated that the various enzyme activities present in the multienzyme complexes depended on separate catalytic sites. The plasma membrane extracts contained three and the microsomal extracts two L-leucyl-β-naphthylamidase-active antigens, respectively. The latter two and one of the plasma membrane antigens were not found to be associated with any other enzyme activities or phospholipids, in contrast to the two plasma membrane antigens mentioned above. [ABSTRACT FROM AUTHOR]

The activity of a large number of human erythrocyte antigens was monitored through a procedure which fractionates the erythrocyte membrane proteins. A significant amount of the erythrocyte "ghost" protein could be removed by aqueous extractants without affecting the activity of any antigen. Subsequent extraction of the residual "ghosts" with pyridine solution resulted in a separation of blood group M, N and Pr activities from blood group A, P1, P and I activities. A number of antigens including c, C, D, k, Kpb, Jka and Fya were labile to this treatment. Gel filtration in the presence of detergent showed that the MN and Pr activity was associated with the erythrocyte sialoglycoprotein. In agreement with Whittemore et al. (1969) two types of blood-group-A antigen could be distinguished on the basis of their partition properties on butanol extraction. Type OS-A antigen can be extracted into butanol, while type WS-A antigen is retained in the aqueous phase. The characteristics of blood-group-P antigen were similar to that of type OS-A antigen, while blood-group-I antigen and type WS-A antigen showed similar behaviour.

Salmonella typhimurium 1195 mutant bacteria were allowed to produce clusters of lipopolysaccharide with new, wild-type specificity over the export sites. Ferritin­labeling experiments showed that the lipopolysaccharide diffuses away from the export sites at 37 °C and is found dispersed on the cell surface, whereas at 0 °C it remains immobile. The density of lipopolysaccharide molecules on the bacterial surface, measured as number of lipopolysaccharide molecules per μm2, or as percentage of the bacterial surface covered by lipopolysaccharide was determined in two wild-type S. typhimurium strains and one mutant strain, which was grown under conditions either permissive or nonpermissive for the formation of wild-type lipopolysaccharide. There are 0.7—1.0×105 lipopolysaccharide molecules per μm2 surface area. About 15—25% of the bacterial surface was found to be covered by lipopolysaccharide. The mean intermolecular distance at hypothetical uniform distribution of lipopolysaccharide was calculated to be in the range of 3—4nm. It was of no consequence to the lipopolysaccharide surface density whether the mutant strain produced wild-type or mutant lipopolysaccharide. An estimate of the lateral diffusion constant D (D ≈ 3 × 10-13 cm2/s) of lipopolysaccharide in the outer membrane was made from earlier published data [2]. Possible reasons for the low order of magnitude of D, based on the observed surface density, are discussed. [ABSTRACT FROM AUTHOR]

A purification procedure applied to H-2 antigens (controlled by the H-2 locus) was used for the characterization of HL-A substances (HL-A2 and HL-AT)(controlled by the HL-A locus) solubilized from lymphoid cell membranes by delipidation. Upon Sephadex chromatography the serologiealty active material had an apparent Mr of 32000, and displayed, in relation to H-2 antigens, a faster electrophoretic mobility and higher pI values. One sulfhydryl group and one (intrachain) disulfide bond were present in partially purified HL-A preparations. Reduction and aminoethytation in 4 M urea did not influence the electrophoretic migration of HL-A active substances, suggesting that, as already demonstrated for H-2 alloantigens, in HL-A molecules the genetic determinants are present on single polypeptide chains. Highly purified HL-A alloantigens, like H-2 substances, revealed a high degree of microheterogeneity. These findings further support the concept of the genetic and structural homology of the main histocompatibility systems in mouse and man. [ABSTRACT FROM AUTHOR]

The conformation of rabbit antibody specific for the p-azophenyl-β-lactoside group was studied by small-angle X-ray scattering and the effect of interaction with hapten evaluated. The binding of hapten to the antibody caused a change in the conformation of the protein. The experimental findings indicate that the radius of gyration and the volume of the antibody become smaller by 2 to 3% as a result of interaction with hapten when 50% of the combining sites are occupied. Since the typical shape of the scattering curves remains the same it is inferred that the change of conformation consists mainly of a volume contraction whereas the overall shape, which is best described by T-shaped models, is not essentially modified. [ABSTRACT FROM AUTHOR]

The glycopeptides containing either the disaccharide or the heteropolysaccharide chains were purified, after enzymatic digestion of glomerular basement-membrane antigens, by gel filtration and ion-exchange chromatography. Their antigenicity was investigated in seven patients with anti-glomerular-basement-membrane antibody-mediated glomerulonephritis, by the leukocyte migration test and radioimmunoassay. The leukocyte migration of the seven patients is inhibited in presence of membrane antigens with both types of oligosaccharide chains. The disaccharide-containing glycopeptide inhibits the leukocyte migration of these patients using 10-times less antigenic material; the heteropolysaccharide-containing glycopeptide does not modify the leukocyte migration of the same patients. Similarly, only the disaccharide-containing glycopeptide decreases the binding of labelled membrane antigens with anti-glomerular-basement-membrane antibodies, using the radioimmunoassay. This glycopeptide carries therefore the major antigenic determinants of humoral and cellular anti-basement-membrane auto-immunization. The amino acid sequence of the glycopeptide is Hyl-Gly-Glu-Asp-Gly the disaccharide prosthetic group being linked to the hydrolysine residue of the peptidic chain; its immunodominant antigenic site is formed by the Galp-Hyl unit. [ABSTRACT FROM AUTHOR]

Mouse histocompatibility antigens were solubilized from lymphocyte membranes by limited papain degradation. Spleens from Balb/c mice (H-2d), enlarged by administration of lymphoma cells, were used for the membrane preparations. The solubilized protein components were purified by ion-exchange chromatography, gel filtration and discontinuous polyacrylamide-gel electrophoresis. A further characterization was achieved in a two-dimensional protein mapping system using disc-electrophoresis and isotachophoresis in the first and second dimension, respectively. The purification was monitored for alloantigens H-2.4(D) and H-3.21(K) (H-2D and H-2K represent two regions of the H-2 system). On Sephadex G-100 partially purified H-2-alloantigen chromatographed in a region of Mr = 35500. The pI of these substances showed a maximum at pH 5.5 and 5.3 for H-2.4 and at 5.2 for H-2.31. Free SH- and dithio-groups were determined with 5,5′-dithiobis(2-nitrobenzoic acid). 1.56 μmol SH/μmol alloantigenic material was found, but no SS-bond could be detected after alkaline cleavage. This suggests the absence of covalently linked protein subunits in papain-solubilized molecules bearing single private specificites. This was confirmed after reduction and alkylation in 4 M urea: 30-50% of the alloantigenic activity was retained and shown, in polyacrylamide-gel electrophoresis, to migrate in the original region of activity (RBPB 0.29 and 0.33 for H-2.4; about 0.33 for H-2.31, BPB = bromophenol blue). After reductive alkylation gel filtration resulted in a further purification of the alloantigenic material. Highly purified substances showed a diffuse staining pattern and a broad serological profile after electrophoresis. We assume this heterogenity to be a genuine property of H-2 antigens. Structural and genetic implications of these findings are discussed. [ABSTRACT FROM AUTHOR]

By applying analytical gel chromatography technique the association behaviour of human erythrocyte phosphofructokinase has been studied. The molecular weight distribution of the polymerized forms of this enzyme covers a range of 380000 to about six millions. The degree of polymerization has been found to depend on the protein concentration, on the presence of various substrates and effectors and on the pH-value. ATP favours the aggregation of the enzyme with increasing pH, whereas in presence of fructose 6-phosphate or fructose 1,6-bisphosphate respectively the degree of association appears to be independent of the pH value between pH 6 and 9. In the microgram range the specific activity of the enzyme decreases with an increase of the enzyme concentration. [ABSTRACT FROM AUTHOR]

A three-step mild procedure is described, that permits isolation of an adenovirus type-2 soluble hexon, penton and fiber antigens in an extensively purified form. This procedure comprises ammonium sulfate precipitation. Sephadex ion-exchange chromatography and adsorption chromatography on an hydroxyapatite column. The final antigen preparations were homogeneous with regard to immunological, electrophoretic and electron microscopic criteria. [ABSTRACT FROM AUTHOR]

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]