RESUMO
1. The defects produced on the membrane of the human red blood cell by the action of complement and antibody have been studied by the use of the electron microscope. These are round to slightly ovoid holes and are surrounded by an irregular ring, about 20 A thick. The mean diameter of the holes is about 103 A if human complement is used (regardless of the antibody used for sensitization) and about 88 A if guinea pig complement is used. 2. The holes in normal and PNH red cells appear to be identical, under the same conditions. The membrane defects produced by lysis of PNH cells with acidified normal serum (the Ham's test) are identical to those produced by complement lysis with specific antibody, indicating that complement is undoubtedly the cause of such lysis. 3. Evidence is presented that when human complement acts on human red cells sensitized with anti-I antibody, each complete activation of complement leads to the production of a cluster of holes. This contrasts to the action of guinea pig complement, on sheep cells, each activation of which leads to a single hole. 4. The maximum number of anti-I antibody molecules which can attach to a human red cell (i.e. the minimum number of antigen sites) is about 500,000 for both normal and PNH cells. 5. The number of holes produced during lysis of the PNH cell is the same as that of the normal cell. When all cells are lysed by am excess of C', a mean of about 90,000 holes are present on each membrane. When complement is limited, a larger proportion of PNH cells are lysed due to their peculiar sensitivity to C' but the number of holes on each lysed cell is the same as for normal cells lysed by the same concentration of C'.
Assuntos
Anticorpos , Membrana Celular , Proteínas do Sistema Complemento , Eritrócitos , Hemoglobinúria Paroxística , Hemólise , Humanos , Técnicas In VitroRESUMO
Hapten-antibody complexes prepared at equivalence with the bivalent hapten bis-DNP-octamethylene-diamine and purified rabbit anti-DNP antibody were fractionated by Sepharose gel-filtration and the fractions examined by electron microscopy. Individual fractions were tested for whole-complement fixation and C1 fixation. Dimer forms did not show this type of biological activity, while fractions containing tetramers and larger polymers exhibited both C and C1 fixation, which could be inhibited by prior exposure of the complexes to the univalent hapten epsilon-DNP-caproic acid. The dose-response result indicated that the C-fixation observed was not due to interpolymeric cooperative effects. It was concluded that in the generation of biological activity by soluble antigen-antibody complexes made with complement-fixing antibody, quaternary structural changes following specific combination with antigen may be as important as any tertiary structural alterations that occur in the individual immunoglobulin molecule.
Assuntos
Sítios de Ligação , Testes de Fixação de Complemento , Proteínas do Sistema Complemento/metabolismo , Animais , Anticorpos/análise , Formação de Anticorpos , Soluções Tampão , Cromatografia em Gel , Cromatografia por Troca Iônica , Proteínas do Sistema Complemento/análise , Proteínas do Sistema Complemento/isolamento & purificação , Dinitrofenóis , Proteínas Hemolisinas , Imunoensaio , Microscopia Eletrônica , Coelhos , UltracentrifugaçãoRESUMO
Mitochondrial permeability transition (PT) pore opening and mitochondrial swelling have been reported in association with apoptosis. Conformational alterations of mitochondria induced by tumour necrosis factor-alpha (TNF alpha), and the association with TNF alpha-induced apoptosis, were, therefore, studied in the human acute T-lymphoblastic leukaemia (T-ALL) cell line, CCRF-CEM and its vinblastine-resistant CEM/VLB100 cell line by transmission electron microscopy (TEM). The CEM/VLB100 cell line possessed more condensed (C phase) mitochondria in the resting state compared with its parental cell line, consistent with increased activity of the mitochondrial electron transport chain (ETC). Following exposure to TNF alpha, conformational alterations of mitochondria occurred in both apoptotic and non-apoptotic cells. Orthodox (O phase) mitochondria in non-apoptotic cells underwent C-phase, transitional O-phase and slightly swollen (S-phase) conformational changes. TNF alpha-induced mitochondrial swelling was a late event and was found to a far lesser extent than mitochondrial condensation. No swollen mitochondria were observed in apoptotic cells. Ultracondensed (UC phase) mitochondria were observed in cells undergoing both TNF alpha-induced and spontaneous apoptosis and were seen when TNF alpha-induced apoptosis was inhibited by 3-methyladenine (3MA). The structural integrity of UC phase mitochondria persisted through the apoptotic process. We conclude that TNF alpha-induced mitochondrial swelling and apoptosis are separate events. Mitochondrial ultracondensation is associated with the processes signalling apoptosis and is not a result of TNF alpha-induced apoptotic shrinkage.
Assuntos
Apoptose/efeitos dos fármacos , Leucemia-Linfoma de Células T do Adulto/patologia , Mitocôndrias/ultraestrutura , Dilatação Mitocondrial , Fator de Necrose Tumoral alfa/farmacologia , Adenina/análogos & derivados , Adenina/farmacologia , Fragmentação do DNA , Resistencia a Medicamentos Antineoplásicos , Humanos , Proteínas Recombinantes/farmacologia , Células Tumorais CultivadasRESUMO
Electron and light microscopic studies of the intestinal epithelium in Crohn's disease demonstrated localized areas of damage to the superficial epithelium, occurring without accompanying acute inflammation. In a blind study of rectal biopsy specimens from seven patients with Crohn's disease, four with ulcerative colitis, and four normal controls, this finding of patchy necrosis without acute inflammation was observed exclusively in four of the seven cases of Crohn's disease.
Assuntos
Doença de Crohn/patologia , Reto/patologia , Biópsia , Colite Ulcerativa/patologia , Epitélio/patologia , Feminino , Técnicas Histológicas , Humanos , Masculino , Necrose/patologiaRESUMO
Anti-haemagglutinin-labelled antibodies have been used to search for influenza entry into cells by fusion of viral and plasma membranes. The plasma membranes of infected cells were stained by immunoperoxidase but not by immunoferritin reagents. It is suggested that the staining obtained with the peroxidase conjugate was due to diffusion of the diaminobenzidine reaction product away from the enzymic site. Immunoferritin labelling provided no evidence for entry of influenza by fusion of viral and plasma membranes under conditions of physiological pH.
Assuntos
Membrana Celular/microbiologia , Vírus da Influenza A/fisiologia , Fusão de Membrana , 3,3'-Diaminobenzidina , Animais , Linhagem Celular , Células Cultivadas , Galinhas , Chlorocebus aethiops , Fibroblastos , Hemaglutininas Virais/imunologia , Técnicas Imunoenzimáticas , Microscopia Eletrônica , Coloração e RotulagemRESUMO
The electron microscopic appearances of the corneoscleral and iris tissue removed at operation from a child with Hurler disease and glaucoma showed distinctive swollen cells with intracellular inclusions similar to those which are observed in other tissues in these patients and which are due to abnormal lysosomal storages of mucopolysaccharides. Some recent observations on the possible relationship between mucopolysaccharides and the drainage of fluid from the anterior chamber are briefly reviewed and correlated with the present observations. The development of glaucoma in this patient is thought to be associated with the presence of the mucopolysaccharide-containing cells in the region of the aqueous drainage channels.
Assuntos
Glaucoma/etiologia , Mucopolissacaridose I/complicações , Criança , Córnea/ultraestrutura , Feminino , Glaucoma/patologia , Humanos , Corpos de Inclusão/ultraestrutura , Iris/ultraestrutura , Microscopia Eletrônica , Esclera/ultraestrutura , Pele/ultraestruturaRESUMO
Electron microscopy of differentially centrifuged synovial fluid from patients with differing types of arthritis revealed a few membranes bearing complement lesions. It was shown that this is due to activation of the alternate pathway of complement by leukocytic enzymes. It is not clear where the site of action is in inflammatory exudates; it may be on internal membranes of the cells, as no detectable cytotoxic effect could be found using Rb(86) as a marker for specific release. In addition, lysosomal enzymes have the net effect of inhibiting the hemolysis of indicator cells caused by activation of the alternate pathway by inulin.
Assuntos
Membrana Celular , Proteínas do Sistema Complemento , Eritrócitos/imunologia , Animais , Reações Antígeno-Anticorpo , Parede Celular , Escherichia coli/imunologia , Cobaias , Proteínas Hemolisinas/farmacologia , Humanos , Lipopolissacarídeos , Membranas Artificiais , Microscopia Eletrônica , Modelos Biológicos , CoelhosRESUMO
Electron microscopic study of the events occurring at the cell membrane during reactive lysis by complement, showed that a foliaceous particle was formed at the C5b-7 stage, that enlarged to a particle with a variable number of arms at the C5b-8 stage. Up to this point, no typical complement lesions were found. At the C5b-9 stages, the particles were completely converted to typical complement lesions, i.e. hollow cylinders projecting from the cell membrane and partly penetrating it. C5b-9 complexes assembled in the fluid phase did not show the typical structure of the lesions, but were amorphous masses of fibres.
Assuntos
Proteínas do Sistema Complemento/imunologia , Membrana Eritrocítica/ultraestrutura , Eritrócitos/ultraestrutura , Hemólise , Complemento C5/imunologia , Complemento C6/imunologia , Complemento C7/imunologia , Complemento C8/imunologia , Complemento C9/imunologia , Membrana Eritrocítica/imunologia , Humanos , Microscopia EletrônicaRESUMO
Inhibitors of glycolysis, oxidative phosphorylation, protein synthesis, membrane Na&-K& transport and microfilament and microtubule function have been employed to elucidate the mechanism of influenza virus uptake by CAM and CEF cells. Electron microscopy demonstrated uptake of virus by viropexis in the presence of all these inhibitors. Utilizing a pulse labelling technique, virus entering CEF cells in the presence of inhibitors was shown to initiate specific virus polypeptide synthesis after neutralization of remaining extracellular virus and removal of the inhibitors. As a consequence of these findings an energy independent mechanism of viropexis has been proposed.
Assuntos
Vírus da Influenza A/crescimento & desenvolvimento , Receptores Virais , Animais , Azidas/farmacologia , Membrana Celular/microbiologia , Embrião de Galinha , Colchicina/farmacologia , Técnicas de Cultura , Cicloeximida/farmacologia , Citocalasina B/farmacologia , Desoxiglucose/farmacologia , Corpos de Inclusão Viral , Ouabaína/farmacologia , FagocitoseRESUMO
Organ cultures of 10 Fallopian tubes were inoculated with a genital strain of Chlamydia trachomatis and seven were infected. Infection was enhanced by centrifuging the organisms on to the tissues, larger numbers of organisms being reisolated from the tissues after this procedure. There was evidence of chlamydial multiplication because the number of organisms which were recovered from the tissues three to five days after inoculation had increased. Recovery was rare, however, after the sixth day, thus suggesting a self-limiting infection. Organ cultures of two bovine oviducts were infected with the bovine abortion strain of Chlamydia psittaci, but in these experiments centrifugation of the inocula did not enhance infection. The organisms were found in both the tissue and medium of cultures up to 18 days after inoculation and in much greater numbers than in the C. trachomatis-infected Fallopian cultures. Chlamydial infection was not entirely host-tissue specific, because C. trachomatis organisms were isolated from bovine oviduct cultures. Inclusions, however, were not detected histologically or electron microscopically in the epithelium of C. trachomatis-infected cultures, but they were detected by these means in C. psittaci-infected bovine cultures. All the elements of the chlamydial growth cycle were seen by electron microscopy, organisms being found in ciliated and possibly non-ciliated cells, and shedding of some infected epithelial cells was observed. No evidence of extensive epithelial cell damage was observed, however, and no loss of ciliary activity was detected in cultures infected with either C. trachomatis or C. psittaci when compared with uninoculated cultures. Thus acute salpingitis, when caused by chlamydial infection, is probably immunologically mediated.