Interaction Between Rat Peritoneal-macrophages and Sensitized Erythrocytes - Dependence On Igg Subclass, Antibody Density and the Degree of Hapten Conjugation

The interaction between macrophages (MO) and antibody sensitised target cells was studied by the use of rat peritoneal macrophages, TNP hapten conjugated sheep red blood cells (SRBC) and homologous antibodies of subclasses IgG1 and IgG2a. Under optimal conditions, the great majority of the MO formed rosettes with IgG1-sensitised antibodies while a maximum of 50% was achieved when target cells were sensitised by IgG2a. Using a double rosette technique, the major part of rosette-forming cells was found to bind both of the isotypes. IgG1-mediated rosette formation was observed at very low degrees of sensitisation as opposed to IgG2a-mediated target cell binding. Not only the amount of bound antibody but also the degree of hapten conjugation (epitope density) appear to influence the ratio of rosette-forming cells. IgG1-mediated rosette formation was partially inhibited by monomeric IgG1 and more efficiently by soluble ovalbumin (OVA)-anti-OVA complexes involving IgG1-type antibodies. while IgG2a mediated rosette formation was inhibited by OVA-anti-OVA complexes containing IgG2a type antibodies. and less efficiently by complexes involving IgG1. No inhibition was found by monomeric IgG2a. Based on the present data, we propose that two types of receptors are involved in the interaction of MO and target cells coated by IgG1 and or IgG2a type antibodies. One requires a multiple antibody-receptor interaction, binding both subclasses at overlapping binding sites; the other is able to interact with IgG1 and does not depend on the multiplicity of interactions

Rat Igg Subclasses Mediating Binding and Phagocytosis of Target-cells By Homologous Macrophages

Attachment and ingestion of Cr-51-labelled TNP-SRBC sensitized by rat IgG1, IgG2a or IgG2b-type antibodies by homologous, elicited peritoneal macrophages were studied. IgG1 was found to be the most efficient isotype in mediating these functions. The antibody doses required for a significant attachment were found to differ with the isotype of Ab, while doses needed for a significant phagocytosis and antibody-dependent cellular cytotoxicity (ADCC) varied between 400-700 Ab/SRBC with all the isotypes studied. Both binding and phagocytosis were also influenced by the degree of hapten conjugation when target cells were sensitized by IgG1. Inhibition of these functions by soluble immune complexes and monomeric immunoglobulins suggests the involvement of two Fcgamma in binding of the three isotypes. Based on the present work and on previous results we conclude that IgG2a interacts with a receptor binding complexed IgG only (Fcgamma RII), IgG2b binds to a different receptor which appears to bind monomeric ligand as well (Fcgamma RI), while IgG1 seems to interact with both types of receptor. We propose that phagocytosis can be mediated by both FcgammaRI and FcgammaRII

Antibody Mediated Lysis of Hapten-conjugated Target-cells By Macrophages and By Complement - the Influence of Igg Subclass, Antibody and Hapten Density

Lysis of 51Cr-labeled TNP-SRBC sensitised by rat IgG1 or IgG2a type antibodies by homologous, paraffin oil-elicited peritoneal macrophages (ADCC) or by homologous complement was studied. IgG2a was found to be markedly more efficient in mediating both ADCC and complement dependent lysis compared to IgG1. Inhibition of the ADCC pointed to the involvement of separate but partially overlapping interaction sites for the two isotypes. We suggest that FcRII type receptors play a favoured role in both IgG2a and IgG1 mediated ADCC. The threshold amount of bound antibody required for ADCC was lower than that sufficient for complement dependent lysis regardless on the subclass or on hapten density, The extent of lysis (both ways) was found to depend on hapten density using equal amounts of antibody. The results are interpreted as terms of the possible requirements for association of IgG molecules on the target cell surface

Geographical distribution of hepatitis C virus genotypes in blood donors: An international collaborative survey

The frequency of infection with the six classified major genotypes of hepatitis C virus (HCV) was investigated in 447 infected volunteer blood donors from the following nine countries: Scotland, Finland, The Netherlands, Hungary, Australia, Egypt, Japan, Hong Kong, and Taiwan. Viral sequences in plasma from blood donors infected with HCV were amplified in the 5'-noncoding region and were typed by restriction fragment length polymorphism analysis. Electrophoresis of DNA fragments produced by cleavage with HaeIII-RsaI and ScrFI-HinfI allowed HCV types 1 (or 5), 2, 3, 4, and 6 to be identified. Further analysis with MvaI-HinfI allowed sequences of the type 5 genotype to be distinguished from sequences of type 1 genotype. Types 1, 2, and 3 accounted for almost all infections in donors from Scotland, Finland, The Netherlands, and Australia. Types 2 and 3 were not found in the eastern European country (Hungary), where all but one of the donors were infected with type 1. Donors from Japan and Taiwan were infected only with type 1 or 2, while types 1, 2, and 6 were found in those from Hong Kong. HCV infection among Egyptians was almost always by type 4. Donors infected with HCV type 1 showed broad serological reactivity with all four antigens of the second generation Chiron RIBA-2 assay (Chiron Corporation, Emeryville, Calif.), while infection with divergent HCV genotypes elicited antibodies mainly reactive to c22-3 and c33c. Reactivities with antibodies 5-1-1 and c100-3 were infrequent and were generally weak, irrespective of the geographical origin of the donor. Because the envelope region of HCV is even more variable than the NS-4 region, it is likely that vaccines based on these proteins need to be multivalent and perhaps specifically adapted for different geographical regions.link_to_subscribed_fulltex

Proposition of conception of using micro-additions for High Performance Concrete

The modern, contemporarily used cement composite types make use of various additives. This diploma thesis is focused on designs of mortars that have been enriched with micro and nano additives, which lead to higher mechanical strength through optimized grading of mortar mixes. This thesis also evaluates the influence of material properties on resulting properties of mortars in fresh and hardened state