Fourth component of human complement: description of a three polypeptide chain structure.
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The fourth component of human complement (C4) was shown to be composed of three distinct polypeptide chains linked by disulfide bonds and noncovalent forces. The sum of the molecular weights of the chains equalled that of the intact molecule. The mol wt of the alpha-, beta-, and gamma-chains were respectively, 93,000, 78,000, and 33,000 daltons. Action of C1s on C4 affected only the alpha-chain, reducing its mol wt to 87,000 daltons. The size of the activation peptide. C4a, is therefore estimated to be 6,000 and that of the major fragment C4b, 198,000 daltons. Periodic acid-Schiff-stained SDS polyacrylamide gels of reduced C4 revealed carbohydrate to be associated with all three chains. A modification of the original method of isolation of C4 is presented. (+info)
Immune phagocytosis in murine malaria.
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Spleen macrophages from Plasmodium berghei-infected mice are more efficient in the ingestion of parasitized reticulocytes than spleen macrophages obtained from normal animals. Other indications of spleen macrophage activation detected during malarial infection are enhanced macrophage spreading and increased phagocytosis of opsonized and nonopsonized sheep erythrocytes (E). Peritoneal macrophages are not activated to a significant degree. The appearance of antibodies directed against Forssman antigen, but not to other erythrocyte antigens, is also a feature of this infection and explains the ingestion of unsensitized E by spleen macrophages of the diseased animals. The recognition and ingestion of parasitized reticulocytes by infected mice in mediated by cold-agglutinin type immunoglobulins that appear during P. berghei infection and can be blocked by the Fc-binding protein A from Staphylococcus aureus. In advanced stages of the disease, the serum of infected animals inhibits phagocytosis, probably because of the high level of circulating immune complexes. Thus, the clearance of malaria parasites is regulated by several elements of the immune system, in addition to levels of specific antimerozoite antibodies, including the amount of antibodies bound to reticulocytes, the presence of circulating immune complexes, and the degree of macrophage stimulation. (+info)
Forssman-like antibody levels in sera of patients with lung cancer.
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Sera of normal individuals or patients with lung cancer were assayed for Forssman-like antibody by a quantitative and specific method using ethylenediaminetetraacetate-containing buffer to inactivate complement in the test serum. It was shown that although Forssman-like antibody levels were distributed widely, (a) the levels of young (20 to 45 years of age) normal subjects of Blood Groups A and AB were lower than those of Blood Groups O and B, (b) the levels of old (60 to 80 years of age) normal subjects were lower than those of young normal subjects of Blood Groups O and B, and (c) the levels of old lung cancer patients were lower when compared to age-matched normal individuals of their blood group. (+info)
Response of mice to injection of ribosomal fraction from group B Neisseria meningitidis.
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Ribosomes of strain NOR-7 of group B Neisseria meningitidis were isolated by a procedure that included treatment of the cells with sodium dodecyl sulfate, disruption in a French pressure cell, and differential centrifugation. These preparations consisted of 66% ribonucleic acid and 24% protein and sedimented as a single component with a constant of approximately 66S. When used in immunodiffusion tests with homologous rabbit antiserum, untreated ribosomes formed two precipitin lines, when treated with ribonuclease three lines, and when Pronase-digested only one distinct line. Qualitatively indistinguishable reactions were obtained with the same antiserum and ribosomes from group A meningococci, but no precipitation occurred with those of Escherichia coli. When injected into mice, group B ribosomes elicited an increase in the number of antibody-producing spleen cells demonstrable by the hemolytic plaque technique using unsensitized sheep erythrocytes. Sensitization of the erythrocytes with increasing amounts of supernatant fluid of meningococcal cultures progressively reduced the number of demonstrable plaque-forming cells. Neuraminidase treatment of the erythrocytes increased immune hemolysis, whereas Pronase digestion reduced it. Injected mice were protected against homologous and heterologous meningococcal challenge. Both hemolysis and protection-inducing activities of the ribosomes were unimpaired by ribonuclease, but were reduced by Pronase. It is concluded that the immunological response elicited by the meningococcal ribosomes does not involve the group-specific carbohydrate antigen. The immunological mechanism by which the mice are protected against meningococcal challenge remains unknown. (+info)
Heterophile antibodies and tissue injury. 3. A role for platelets in the development of lethal vascular injury during Forssman shock in guinea pigs.
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Platelets appear to be pathogenetic determinants in the development of lethal Forssman shock, which was provoked in guinea pigs by an intravenous injection of rabbit antiserum to sheep erythrocyte stromata. Within moments, circulating platelets (prelabeled with (14)C-serotonin) were removed from the blood stream and impacted in the lungs, where they liberated (14)C into the tissues. When animals were depleted of platelets prior to the production of shock, they survived for prolonged periods of time or were protected against death. Pretreatment with antiinflammatory compounds capable of inhibiting platelet aggregation and release phenomena had a similar protective influence. It would appear, therefore, that Forssman shock is a convenient and accessible model for investigating the mechanisms whereby platelets mediate immune vascular damage. (+info)
Cell mitotic cycle synthesis of NIL hamster glycolipids including the Forssman antigen.
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The synthesis of phospholipids and glycolipids during the cell mitotic cycle of an established hamster line, NIL, has been studied. Cells were synchronized with excess thymidine and mitotically harvested by shaking. Cells were radioactively labeled for 4 h with palmitate, glucosamine, or galactose. Lipids were analyzed by thin-layer chromatography. As cells progressed through the mitotic cycle, incorporation into phospholipids increased but the fraction represented by each remained constant. Similarly, ceramide monohexoside, dihexoside, and hematoside were labeled equally in all phases. Ceramide trihexoside and tetrahexoside were labeled only during G(1) and S. Ceramide pentahexoside (the Forssman antigen) shows density-dependent synthesis, accumulation, and reactivity. Ceramide pentahexoside was labeled during all phases of the mitotic cycle but the rate of incorporation decreased in S and G(2). The total amount of lipid assayed immunologically in cell extracts gradually increased. Exposure of the Forssman antigen in untreated or trypsin-treated cells was studied using binding of chemically labeled antiForssman antiserum. The amount of antigen detected in trypsinized cells increased during G(1) and early S but then remained constant. Mitotic cells exposed all detectable antigen. As cells progressed through the mitotic cycle, a large fraction of the Forssman antigen became cryptic. (+info)
Inhibition of anaphylactic histamine release by Forssman antiserum. I. Characteristics of the reaction and inhibitor.
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Forssman antiserum produced in rabbits immunized with sheep erythrocyte stromata was found to contain an IgG antibody which inhibited both passive anaphylactic sensitization of guinea-pig lung and also histamine-releasing activity of soluble immune complexes. This Forssman antibody did not itself cause histamine release or depletion of lung histamine stores. The IgM haemolysin component of the Forssman antiserum was not associated with inhibitory activity. The inhibition by the IgG Forssman antibody differed from that of normal rabbit gammaglobulin both in its irreversible character and in being absorbed by sheep erythrocytes. The inhibitory antibody had no effect on the histamine-releasing activity of compound 48/80, anaphylatoxin or reversed anaphylaxis. It was concluded that IgG Forssman antibody probably blocks the tissue receptor(s) for anaphylactic antibody in guinea-pig lung. (+info)
Haptenic activity of galactosyl ceramide and its topographical distribution on liposomal membranes. Effects of temperature and phospholipid composition.
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The relation between the immune reactions of phosphatidylcholine liposomes containing galactosyl ceramide and the physical properties of the glycolipid in membranes was studied. The immune-agglutination of dipalmitoyl phosphatidylcholine liposomes was affected both by reaction temperature and by cholesterol content. Fatty acyl chain length of phosphatidylcholine also influenced the immune-agglutination. The electron spin resonance and calorimetric studies indicated that the fatty acyl chain length of phosphatidylcholine and cholesterol content, as well as temperature, affect the physical properties of galactosyl ceramide in liposomal membranes. In the absence of cholesterol, most galactosyl ceramide molecules were clustered on the phosphatidylcholine liposomes below the chain-melting transition temperature of the phospholipid, whereas they were randomly distributed in the membrane above the transition temperature. Upon addition of cholesterol to the membranes below the chain-melting transition temperature, the number of glycolipid molecules in the cluster phase decreased. Cholesterol increased the ordering of galactosyl ceramide molecules in the phase of random distribution on membranes above the transition temperature. The change in topographical distribution of galactosyl ceramide in membranes was parallel with that of immune-reactivity. (+info)