RESUMEN
An attempt was made at correcting the specific lysosomal enzyme deficiencies in 7 children with Hunter's or Hurler's diseases by transplantation of fetal fibroblasts. In spite of pretreating the young patients with stored blood, following a procedure employed successfully to avoid rejection of kidneys from incompatible donors, the use of serum-free media for culturing the cells before being harvested and incubation of the cells with chorionic gonadotrophin, the transplantation of fetal fibroblasts was not associated with biochemical or clinical changes. None of the seven patients showed immune reactions against the transplanted cells, HLA antigens, or the missing enzymes.
Asunto(s)
Fibroblastos/trasplante , Mucopolisacaridosis II/terapia , Mucopolisacaridosis I/terapia , Animales , Bovinos/sangre , Células Cultivadas , Medios de Cultivo/inmunología , Feto/citología , Fibroblastos/enzimología , Fibroblastos/inmunología , Glicosaminoglicanos/orina , Histocompatibilidad , Humanos , Iduronidasa/deficiencia , Lisosomas/enzimología , Mucopolisacaridosis I/inmunología , Mucopolisacaridosis I/orina , Mucopolisacaridosis II/inmunología , Mucopolisacaridosis II/orina , Oligosacáridos/orinaRESUMEN
A very sensitive (100 pg/ml) solid-phase enzyme immunoassay (ELISA) for the determination of human IgE has been developed. This assay incorporates the avidin-biotin system to increase sensitivity and can detect as little as 100 pg/ml (10 pg/test) of human IgE. The assay is highly specific and allows quantitative determination of human IgE in supernatants of peripheral blood lymphocytes as well as in serum. The very high sensitivity of the assay was accomplished by optimizing concentrations of the following reagents: (1) affinity-purified rabbit anti-human IgE coating antibodies; (2) biotin-conjugated goat anti-human IgE; (3) avidin-horseradish peroxidase (HRP) conjugate. In summary, the assay described is rapid (6 h), reproducible, isotype specific, and has the sensitivity of radioimmunoassays usually employed for the quantification of IgE. This assay may be utilized in establishing concentrations of in vitro IgE levels synthesized by human peripheral blood lymphocytes (PBL).
Asunto(s)
Ensayo de Inmunoadsorción Enzimática/métodos , Inmunoglobulina E/biosíntesis , Animales , Anticuerpos Antiidiotipos/inmunología , Avidina , Biotina , Células Cultivadas , Medios de Cultivo/inmunología , Cabras , Humanos , Inmunoglobulina E/análisis , Linfocitos/inmunología , Linfocitos/metabolismo , Ratones , ConejosRESUMEN
Supernatants from Plasmodium falciparum cultures containing soluble parasite material were mitogenic for normal human peripheral blood mononuclear cells (MNC) in vitro. This was evidenced by blast transformation and significant incorporation of 3H-thymidine and confirms earlier reports of the mitogenic potential of malaria parasites. Lymphocyte activation by these malaria derived products was polyclonal as demonstrated by increased secretion of IgA, IgG and IgM by the stimulated cells. Using rat tissues and Hep-2 cells as substrates, autoantibody activity was found in the IgM fraction of the secreted immunoglobulin. Speckled anti-nuclear (ANA) antibody, anti-globulins (rheumatoid factor) and anti-intermediate filament antibodies were produced by the stimulated lymphocytes. No significant immunoglobulin secretion with autoantibody specificity was found in control cultures in which normal MNC were incubated with supernatants from non-parasitized red cell cultures. The data supports the suggestion that polyclonal lymphocyte activation by parasite derived products occurs in vivo and, in addition, provides an explanation for the presence of autoantibodies in the serum of malaria patients.
Asunto(s)
Autoanticuerpos/biosíntesis , Linfocitos/inmunología , Malaria/inmunología , Animales , Anticuerpos Antiidiotipos/inmunología , Anticuerpos Antinucleares/inmunología , Células Productoras de Anticuerpos/inmunología , Células Cultivadas , Medios de Cultivo/inmunología , Citoesqueleto/inmunología , Humanos , Activación de Linfocitos , Monocitos/inmunología , Plasmodium falciparum/inmunologíaRESUMEN
Mouse spleen cells were treated with concanavalin A (Con A) or aggregated mouse IgG2b for 48 h in culture. When cells thus treated were added to fresh mouse spleen cell cultures immunized with SRBC they depressed the response of B lymphocytes as measured by enumerating plaque forming cells (PFC) on the fourth day of culture. When supernatant from cells cultured with IgG2b was added to immunized cultures this resulted in depression of PFC generation similar to that observed by addition of treated cells. The depression observed was essentially in the same range as that observed by addition of Con A treated cells or their supernatant. These observations extend previous work suggesting that IgG2b-induced PFC depression may result from activation of suppressor T cells with elaboration of soluble suppressor factors. This mechanism of immunomodulation may be important in the pathogenesis of immune complex disorders.
Asunto(s)
Células Productoras de Anticuerpos/efectos de los fármacos , Inmunoglobulina G/inmunología , Animales , Linfocitos B/efectos de los fármacos , Concanavalina A/farmacología , Medios de Cultivo/inmunología , Depresión Química , Técnica de Placa Hemolítica , Ratones , Ratones Endogámicos C3H/inmunología , Ratones Endogámicos C57BL/inmunología , Bazo/citología , Linfocitos T Reguladores/efectos de los fármacosRESUMEN
Many strains of Ureaplasma urealyticum were inactivated, in the presence of complement, by a control antiserum prepared in guinea-pigs against the uninfected culture medium used to grow ureaplasmas. This mycoplasmacidal activity of the control serum, unlike that of the specific antisera prepared against the organisms themselves, was removed by treatment with dithiothreitol or by absorption with the horse-serum component of the medium, suggesting that the activity was due to an antibody of the IgM class acting on horse-serum proteins that had become associated with the surface of the ureaplasmas. The mycoplasmacidal activity in the specific ureaplasma antisera appeared to be due mainly to antibody of the IgG class. A similar complement-dependent mycoplasmacidal antibody to U. urealyticum apparently active against serum components, was present in normal rabbit sera.