RESUMO
Large amounts of immune complexes were present in the serum of infected rats early in infection when parasitemias were low. As the infection progressed and parasitemia increased and then decreased, the amounts of immune complexes in the serum also fell. This result suggests that increased efficiency of complex clearance was an important factor in determining the levels of immune complexes in the serum. In high performance liquid chromatography (HPLC), the complexes in the serum migrated as a peak with material of 350 kDa and greater in mass. They sedimented in a sucrose gradient as a band with a sedimentation coefficient of 22 s, which was calculated to yield a mass of approximately 1100 kDa. Immunoelectrophoresis and radial immunodiffusion showed that IgG was the major immunoglobulin in the complexes. As the IgG content of the complexes increased, the levels of complexes in the serum generally decreased. HPLC analysis of precipitated complexes suggested that they contained loosely bound albumin. Serum proteins were affected by the infection. A depletion of free immunoglobulin was observed during the initial period of immune complex formation.
Assuntos
Complexo Antígeno-Anticorpo/análise , Malária/imunologia , Animais , Anticorpos Antiprotozoários/análise , Antígenos de Protozoários/análise , Centrifugação com Gradiente de Concentração , Cromatografia Líquida de Alta Pressão , Imunodifusão , Imunoglobulina G/análise , Malária/sangue , Plasmodium berghei/imunologia , RatosRESUMO
Acute phase serum (APS) given at the time of challenge with Plasmodium berghei inhibited the generation to immunity to the infecting plasmodia. Administered with a single dose of vaccine, it inhibited induction of immunity by the vaccine. Three weekly doses, the last given two weeks before infection, induced immunity. Administration of vaccine simultaneously with infection neither aggravated nor ameliorated the infection. These results indicate that the effect of administration of APS on immunity generated by immunization or infection is dose- and time-dependent. The depression of immunity induced by this serum is thus temporary, the host finally overcoming the depression and responding to the plasmodial antigen in the serum. The interaction of vaccine and infection observed indicates that the introduction of vaccine is not detrimental to the individual incubating infection; rather, the vaccine is rendered useless, the reducing the aggregate benefit of the immunization to the group.
Assuntos
Malária/imunologia , Plasmodium berghei/imunologia , Vacinas/imunologia , Animais , Imunidade , Camundongos , Ratos , VacinaçãoRESUMO
We investigated the effect of both immune and normal sera on the binding of free Plasmodium berghei by resident and activated macrophages. Resident macrophages bound plasmodia to a greater extent than did activated macrophages, regardless of treatment. Resident macrophages bound free plasmodia, predominantly trophozoites, in the presence of normal serum by a mechanism inhibited by N-acetylglucosamine and N-acetylmannosamine. Macrophages activated through treatment with Propionibacterium acnes ("Corynebacterium parvum"), on the other hand, did not bind free plasmodia in the presence of normal serum through systems inhibited by N-acetylmannosamine or N-acetylglucosamine. The binding of free plasmodia by activated macrophages was greatest in the presence of immune serum and could be inhibited by immune complexes but not by N-acetylmannosamine or N-acetylglucosamine. These results suggest that a receptor for a carbohydrate component of a normal serum opsonin mediates initial adherence of plasmodial antigen onto resident macrophages, triggering both the immunological cascade and macrophage activation. After activation, the macrophages no longer have the carbohydrate-specific receptor but do have functional Fc receptors which mediate the adherence of immune-serum-opsonized plasmodia.
Assuntos
Macrófagos/imunologia , Malária/imunologia , Plasmodium berghei/imunologia , Animais , Complexo Antígeno-Anticorpo , Feminino , Ativação de Macrófagos , Antígeno de Macrófago 1 , Macrófagos/ultraestrutura , Masculino , Microscopia Eletrônica , Microscopia Eletrônica de Varredura , Monossacarídeos/metabolismo , Fagocitose , Plasmodium berghei/citologia , Ratos , Ratos Endogâmicos F344 , Receptores de Complemento/fisiologia , Receptores Fc/fisiologiaRESUMO
Acute-phase serum (APS) collected from Plasmodium berghei-infected rats inhibited phagocytosis of trypsinized rat erythrocytes and of erythrocytes from P. berghei-infected rats. Macrophages (M phi) incubated with APS or heat-aggregated acute-phase serum (HAAPS) for 6 h, followed by 18 h incubation in serum-free medium, exhibited significantly higher levels of phagocytosis than M phi similarly cultured but with normal rat serum. When APS was present at the time of assay, it inhibited erythrophagocytosis by M phi which had been in culture for 0 or 24 h. M phi activation by HAAPS was inhibited by 2-deoxy-D-glucose, which suggests that activation by HAAPS is Fc-receptor mediated. Adsorption of APS with staphylococcal protein A abrogated the ability of APS to inhibit phagocytosis and that of HAAPS to effect M phi activation, suggesting that immune complexes are involved in both processes. Surface-bound immunoglobulins eluted from erythrocytes of P. berghei-infected rats promoted phagocytosis of trypsinized erythrocytes by HAAPS-activated M phi but not by resting M phi. These results indicate that the immunoglobulins which bind to infected or damaged erythrocytes during malarial infections promote erythrophagocytosis by activated M phi and that the immune complexes in serum from rats with acute malaria may inhibit erythrophagocytosis early in the infection but may, over time, induce changes in the M phi which later facilitate erythrophagocytosis.
Assuntos
Eritrócitos/imunologia , Malária/imunologia , Animais , Complexo Antígeno-Anticorpo , Desoxiglucose/farmacologia , Malária/sangue , Fagocitose , Plasmodium berghei , Ratos , Ratos Endogâmicos F344RESUMO
The ELISA test titers and RIPA patterns of sera collected from vaccinated and non-vaccinated rats during P. berghei infection were similar. The sera collected just after clearance of parasitemia from the vaccinated rats, but not that from the non-vaccinated rats protected mice in passive protection tests. After precipitation to remove immune complexes, the sera from the non-vaccinated rats also protected mice. Administration of acute phase serum early in the course of infection aggravated parasitemia and delayed recovery from P. berghei infection in rats. Administration of hyperimmune serum early in the course of infection initially reduced parasitemia but then delayed recovery of rats from P. berghei infection. These results suggest that immune complexes may interfere with antibody mediated immunity to P. berghei and may also retard development of the induced immune response.
Assuntos
Complexo Antígeno-Anticorpo/imunologia , Imunização Passiva , Malária/imunologia , Vacinação , Animais , Formação de Anticorpos , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Malária/prevenção & controle , Camundongos , Plasmodium berghei/imunologia , Ratos , Ratos Endogâmicos , Fatores de Tempo , Vacinas/administração & dosagemRESUMO
In vitro, Plasmodium berghei infected erythrocytes incorporated 35S-methionine into 31 polypeptides with molecular weights from 21 kd to 300 kd. Hemoglobin and additional smaller molecular weight polypeptides were labelled with 35S-methionine by a population of uninfected, reticulocyte-rich rat erythrocytes. 3H-glucosamine was incorporated into at least 3 components by Plasmodium berghei infected erythrocytes. Uninfected, reticulocyte-rich rat erythrocytes did not incorporate 3H-glucosamine. Rabbit antisera against small, free plasmodia formed complexes which contained between 12 and 22 of the 31 labelled polypeptides in the 35S-methionine labelled antigen preparation. Rabbit antisera against soluble antigens washed from small, free plasmodia formed complexes containing many of the same labelled plasmodial polypeptides, however the reactions were particularly strong with those components which yielded polypeptides with molecular weights of 25 kd and 31 kd. Rabbit origin antisera against the 2 preparations did not form detectable complexes with the 3H-glucosamine labelled plasmodial components. Sera from rats undergoing progressive P. berghei infection formed complexes containing an increasing number of 35S-methionine labelled plasmodial polypeptides. Hyperimmune rat serum, the only serum protective upon passive transfer into mice, formed complexes containing 7 polypeptides with molecular weights of 35 kd, 75 kd, 80 kd, 92 kd, 100 kd, 150 kd and 190 kd. Antigens containing 1 or more of these polypeptides may be important in the induction of a protective antibody response against the parasite.
Assuntos
Antígenos de Protozoários/imunologia , Malária/imunologia , Plasmodium berghei/imunologia , Animais , Formação de Anticorpos , Complexo Antígeno-Anticorpo/análise , Eritrócitos/parasitologia , Glucosamina/metabolismo , Imunização , Imunização Passiva , Metionina/metabolismo , Camundongos , Peso Molecular , Biossíntese Peptídica , Peptídeos/imunologia , Plasmodium berghei/metabolismo , Coelhos , Ratos , Ratos Endogâmicos F344 , Ratos Endogâmicos , SolubilidadeRESUMO
The hematologic and immunologic responses to infection with either the Epstein-Barr virus alone or infection with Epstein-Barr virus and Plasmodium knowlesi were studied using common marmosets (Callithrix jacchus). The assays performed included complete blood cell counts, determinations of natural killer cell activity, and determinations of antibody titers to Epstein-Barr virus early antigen, virus capsid antigen and the nuclear antigen. While no animal showed signs of lymphoproliferative disease, it was found that animals infected with Epstein-Barr virus became positive for early antigen, virus capsid antigen and nuclear antigen at low levels. No difference in antibody titers between Epstein-Barr virus infected animals and co-infected animals was observed. An increase also was found in the number of leukocytes in all groups, and an increase in natural killer cells following infection with Epstein-Barr virus. Some depression in natural killer cells was observed in the co-infected animals when compared to Epstein-Barr virus infected animals.
Assuntos
Anticorpos Antivirais/biossíntese , Infecções por Herpesviridae/imunologia , Herpesvirus Humano 4/imunologia , Malária/imunologia , Animais , Contagem de Células Sanguíneas , Callithrix , Infecções por Herpesviridae/sangue , Infecções por Herpesviridae/complicações , Imunidade Celular , Células Matadoras Naturais/imunologia , Malária/sangue , Malária/complicações , MasculinoRESUMO
Immune complexes produced in vitro by mixing immune serum and soluble Plasmodium berghei antigens and immune complexes precipitated from serum of acutely infected rats blocked macrophage receptor sites for opsonized plasmodia. The immune complexes were precipitated from acute-phase serum, using polyethylene glycol, and their composition was determined by using a Raji cell immunofluorescence assay. The immune complexes contained immunoglobulin G (IgG), IgM, and malarial antigens. These results indicate that immune complexes in the serum of acutely infected animals may protect the plasmodia from the activities of macrophages.
Assuntos
Complexo Antígeno-Anticorpo/imunologia , Macrófagos/imunologia , Malária/imunologia , Plasmodium berghei/imunologia , Animais , Antígenos/imunologia , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Ratos , Ratos Endogâmicos F344Assuntos
Infecções por Anaplasmataceae , Anaplasmataceae/fisiologia , Infecções por Bartonellaceae , Bartonellaceae/fisiologia , Sangue/microbiologia , Infecções por Anaplasmataceae/microbiologia , Infecções por Anaplasmataceae/veterinária , Animais , Antibacterianos/uso terapêutico , Anticorpos Antibacterianos/imunologia , Vetores Artrópodes/microbiologia , Infecções por Bartonellaceae/microbiologia , Infecções por Bartonellaceae/veterinária , Ecologia , Humanos , Imunização , Baço/imunologia , Terminologia como AssuntoAssuntos
Endotoxinas/análise , Plasmodium berghei/análise , Animais , Teste do Limulus , Camundongos , RatosRESUMO
The effect on growth of B. microti of macrophages from normal or B. microti recovered animals was tested in short-term cultures. Normal macrophages did not or only slightly inhibited parasite growth. When immune serum was added to the normal macrophage containing cultures, inhibition greater than that produced by immune serum alone was observed. Macrophages from immune animals had an inhibitory effect on parasite growth even in the absence of immune serum. Inhibition of B. microti growth was greater in cultures containing immune serum and immune macrophages than in cultures containing either immune serum or immune macrophages alone. Supernatants from B. microti cultures which contained immune macrophages or macrophages which had been stimulated by addition of immune serum inhibited parasite growth while cell free supernatant from cultures which contained no macrophages or normal macrophages did not. At 72 and 96 hr of culture erythrophagocytosis by macrophages was more prominent in immune serum containing B. microti cultures than in B. microti cultures not containing immune serum. More macrophages were retained in the immune serum containing cultures than in cultures not containing immune serum. Alternate culture-hamster passage yielded a B. microti strain which grew better in hamster erythrocyte short-term cultures than did the original strain.
Assuntos
Babesia/crescimento & desenvolvimento , Soros Imunes/farmacologia , Macrófagos/imunologia , Animais , Babesia/imunologia , Cricetinae , Eritrócitos/parasitologia , Mesocricetus , Fagocitose , Fatores de TempoRESUMO
Soluble material was obtained from sonically freed plasmodiae by three procedures. Two procedures, cryo-impacting and freeze-thawing, were evaluated for their ability to disrupt the parasites and release soluble material. The soluble materials obtained by these procedures were compared to materials washed from the surfaces of sonically freed parasites. Between 35 and 40% of the total parasite protein was solubilized by freeze-thawing or cryo-impacting. One cycle of freeze-thawing released nearly as much protein as could be released by this method, and additional cycles of freeze-thawing had little additional effect. Cryo-impacting solubilized only a small amount of protein in addition to that which was released by the cycle of freeze-thawing inherent in the procedure. Reductions in the packed cell volume of the material remaining after freeze-thawing or cryo-impacting indicate that the insoluble fragments are broken into smaller pieces as treatment is extended. Electron microscopy of 30-s cryo-impacted and three-times freeze-thawed parasites revealed membrane fragments similar in appearance. Patterns obtained by polyacrylamide gel electrophoresis of the soluble material from freeze-thawed and cryo-impacted parasites were also similar, and approximately 13 protein bands were demonstrated. The material washed from the surfaces of the free parasites, on the other hand, resolved into only two to four major bands on the gel columns. In immunization studies, the soluble and insoluble fractions obtained by freeze-thawing or cryo-impacting and the material washed from the surfaces of the parasites all stimulated a protective immune response. On the basis of the amount of protein required to stimulate roughly comparable immunity, the soluble fraction obtained by freeze-thawing or cryo-impacting free parasites was about twice as potent an immunogen as was the insoluble fraction. The material obtained by gentle washing of the freed parasites was approximately 20 times as potent an immunogen as were the freed parasites and about 7 times as potent as the soluble material obtained by freeze-thawing or cryo-impacting.
Assuntos
Antígenos/imunologia , Malária/imunologia , Plasmodium berghei/imunologia , Animais , Antígenos/análise , Antígenos/isolamento & purificação , Congelamento , Imunização , Masculino , Plasmodium/análise , Ratos , SolubilidadeRESUMO
Short-term in vitro culture of Babesia microti was obtained. Immune hamster serum had an inhibitory effect on the growth of B. microti in vitro. It is suggested that these antibodies act by preventing penetration of erythrocytes by the parasite.
Assuntos
Babesia/crescimento & desenvolvimento , Eritrócitos/microbiologia , Soros Imunes/farmacologia , Animais , Babesia/imunologia , Células Cultivadas , CricetinaeRESUMO
Immune serum delayed the onset of parasitemia in both intact and splenectomized mice, but it neither prevented the development of Babesia rodhaini infection nor protected the mice from death even with further supplementation of immune serum during the infection. The protective antibodies in the serum are more effective in their action on free B. rodhaini parasites than on infected erythrocytes; the parasites (free or inside the red cells) being direct targets for the antibodies. Passive administration of the immune serum seemed to inhibit the active development of the host's own immune response. B. rodhaini, unlike Plasmodium berghei, produces surface changes in the membranes of infected erythrocytes and therefore on incubation of infected erythrocytes and therefore on incubation of infected erythrocytes in immune serum there is either entry and subsequent interaction of antibodies with the parasites within the erythrocytes or there is action of antibody with antigen in the erythrocyte membranes. The effect of immune serum on parasitized erythrocytes is greatest when the parasitized cells incubated in immune serum were administered with additional amounts of immune serum, indicating the ability of the immune serum to extend its action in vivo.
Assuntos
Babesiose/prevenção & controle , Imunização Passiva , Animais , Babesia/imunologia , Babesiose/imunologia , Sangue/parasitologia , Células Cultivadas , Eritrócitos/imunologia , Eritrócitos/parasitologia , Imunidade Materno-Adquirida , Masculino , Camundongos , Baço/imunologiaRESUMO
Evidence is presented to indicate that Plasmodium berghei merozoites, but not trophozotites, have an antiphagocytic capsule. The capsule appears to form around the developing merozoties of the schizont in the parasitophorous vacuole. Serum from animals immune to P. berghei reacts with this capsule. After reaction with immune serum, the antiphagocytic action of the capsule is lost. By the process of binding serum protein, the capsule becomes electron dense and can be readily visuallzed as the surface coat by electron microscopy. At physiological temperatures, phagocytosis by macrophages rapidly follows adhesion of antibody-coated parasites. Both tight and loose phagosomes are formed.
Assuntos
Membrana Celular/imunologia , Macrófagos/imunologia , Fagocitose , Plasmodium berghei/imunologia , Animais , Líquido Ascítico/citologia , Membrana Celular/ultraestrutura , Macrófagos/ultraestrutura , Plasmodium berghei/ultraestrutura , RatosRESUMO
This paper reports the results of a study of the nature of the immune response against Plasmodium berghei parasites by inbred rats. A macrophage-cytophilic antibody specific for malarial antigens was identified and characterized. Detection of the antibody on the macrophage surface was accomplished by the parasite adherence tests and by the indirect fluorescent antibody technique. Isolation and purification of the macrophage-cytophilic and opsonic antibodies from hyperimmune rat serum was accomplished by QAE-Sephadez A-50 elution chromatography, and of the macrophage-cytophilic antibody by adsorption with and elution from syngeneic macrophages as well. Characterization of the cytophilic antibody as immunoglobulin G1 was done by immunoelectrophoresis and by Ouchterlony-type double diffusion in gel. Passive protection tests in weanling inbred rats have demonstrated that the opsonizing antibody conferred some protection against P. berghei. The macrophage-cytophilic antibody, on the other hand, was not protective alone but acted synergistically with the opsonizing antibody.