Comportamiento fractal del repertorio T específico contra el alergeno Poa p9 / Fractal behavior of T specify repertory against Poa p9 alergeno

Antecedentes: basados en una analogía lingüística aplicada al repertorio inmune B y T específicos, el sistema inmune se puede caracterizar por su grado de complejidad como se hace con los lenguajes naturales. Material y métodos: este estudio es la aplicación de una ley matemática al repertorio inmune. Se aplicó la ley de Zipf-Mandelbrot, observada en los lenguajes naturales, al estudio del repertorio T específico contra el alergeno Poa p9. Resultados: se observó un comportamiento a escala de la ley en el repertorio específico para el alergeno obtenido de un paciente alérgico en presencia y ausencia de interferón á, y en los clones de células Th. Los repertorios T en presencia y ausencia de interferón á se comportan fractalmente, con una dimensión fractal de 0.661165 y 0.923895 respectivamente. Conclusión: el grado de complejidad del repertorio T contra el alergeno Poa p9 es una medida matemática objetiva y reproducible del repertorio inmune, la dimensión fractal es un parámetro matemático apropiado para caracterizar la fisiología del sistema inmune. Este comportamiento fractal puede tener implicaciones generales para la inmunología

Comparison of dengue infection in human mononuclear leukocytes with mosquito C6/36 and mammalian Vero cells using flow cytometry to detect virus antigen

Fluorescent activated cell sorter (FACS) analysis is useful for the detection of cellular surface antigens and intracellular proteins. We used this methodology in order to detect and quantify dengue antigens in highly susceptible cells such as clone C6/36 (Aedes albopictus) and Vero cells (green monkey kidney). Additionally, we analyzed the infection in vitro of human peripheral blood mononuclear leukocytes (PBML). FACS analysis turned out to be a reliable technique to quantify virus growth in traditional cell cultures of C6/36 as well as Vero cells. High rates of infection were achieved with a good statistical correlation between the virus amount used in infection and the percentage of dengue antigen containing cells detected in infected cultures. We also showed that human monocytes (CD14+) are preferred target cells for in vitro dengue infection among PBML. Monocytes were much less susceptible to virus infection than cell lines but they displayed dengue antigens detected by FACS five days after infection. In contrast, lymphocytes showed no differences in their profile for dengue specific immunofluorescence. Without an animal model to reproduce dengue disease, alternative assays have been sought to correlate viral virulence with clinical manifestations and disease severity. Study of in vitro interaction of virus and host cells may highlight this relationship.

Producción y análisis de proteínas recombinantes obtenidas de la cepa lisogénica Ro-531

La expresión del cDNA que codifica al antígeno Ro, fue evaluada usando la clona Ro=531 gt11 cuyos productos de traducción fueron inmunorreconocidos por un suero anti-Ro. La producción de proteína Ro recombinante, fue inducida en la cepa lisogénica E. coli Y1089. La antigenicidad de la proteína fue probada por Western blot y por ELISA. En la primera prueba, 15 de los 20 sueros anti-Ro positivos presentaron fuerte reactividad a la proteína de funsión Ro y 4 de los 20 sueros controles, mostraron reactividad débil. Por la técnica de ELISA, se observó una reacción más específica, ya que 15 de los 20 sueros anti-Ro positivos exhibieron afinidad por la proteína Ro recombinante y ninguno de los controles presentó falsos positivos

Kinetics of antigen specific and non-specific polyclonal B-cell responses during lethal Plasmodium yoelii malaria

In order to study the kinetics and composition of the polyclonal B-cell activation associated to malaria infection, antigen-specific and non-specific B-cell responses were evaluated in the spleens of mice infected with Plasmodium yoelii 17 XL or injected with lysed erythrocytes or plasma from P. yoelii infected mice or with P. falciparum culture supernatants. Spleen/body weigth ratio, numbers of nucleated spleen cells and Immunoglobulin-containing and Immunoglobulin-secreting cells increased progressively during the course of infection,in parallel to the parasitemia. A different pattern of kinetics was observed when anti-sheep red blood cell and anti-trinitrophenylated-sheep red blood cell plaque forming cells response were studied: maximum values were observed at early stages of infection, whereas the number of total Immunoglobulin-containing and Immunoglobulin-secreting cells were not yet altered. Conversely, at the end of infection, when these latter values reached their maximum, the anti-sheep red blood cell and anti-trinitrophenylated-sheep red blood cell specific responses were normal or even infranormal. In mice injected with Plasmodium-derived material, a higher increase in antigen-specific PFC was observed, as compared to the increase of Immunoglobulin-containing and Immunoglobulin-secreting cell numbers. This suggested a "preferential" (antigen-plus mitogen-induced) stimulation of antigen-specific cells rather than a generalized non-specific (mitogen-induced) triggering of B-lymphocytes. On the basis of these and previous results, it is suggested that polyclonal B-cell activation that takes place during the course of infection appears as a result of successive waves of antigen-specific B-cell activation

Clonal variation within a mucosal isolate derived from a patient with Leishmania (Viannia) braziliensis infection

No transcurso de um periodo de 5 anos foram estudados 3 isolados de um paciente com leishmaniose mucosa recidivante causada pela Leishmania (Viannia) braziliensis e 7 clones de um desses isolados. Este estudo foi feito pela analise dos serodemas e zimodemas. Os resultados indicaram a ocorrencia de variacoes fenotipicas clonais. Oito marcadores isoenzimaticos demonstraram diferencas nos padroes eletroforeticos em Acetato de Celulose (AC), bem como em camada fina de amido. Da mesma forma foram constatadas diferencas em um painel de anticorpos monoclonais especificos e subespecificos. Nossas observacoes indicam ainda que a leishmania (Viannia) braziliensis esta composta por subpopulacoes de parasitas com caracteristicas bioquimicas e antigenicas peculiares.

Anticuerpos monoclonales y clones de linfocitos T contra antigenos HLA: producción y caracterización / Monoclonal antibodies and T-cell clones against HLA antigens: production and characterization

Inmunizando ratones Balb/c con líneas linfoblastoides humanas se produjeron y caracterizaron 21 anticuerpos monoclonales de los cuales 12 parecen estar reconociendo un determinante HLA polimórfo; 4 de ellos reconecen un epítope común a los antígenos Clase I y otro reconece moléculas HLA Clase II. Estos resultados se confirmaron en ensayos de ELISA, citotoxicidad, inmunofluorescencia e inmunopreciptación y la clase y subclase de las inmunoglobulinas se determinó por inmunodifusión de Ouchterlony. Se generaron además 22 clones de células T aloreactivas, por dilución limitante de linfocitos de un donante estimulados en cultivo mixto con linfocitos alogeneicos de sangre periférica; 5 de estos clones se caracterizaron detalladamente. Evaluando su capacidad para proliferar ante un panel de líneas linfoblastoides humanas, se encontró que reconecen las especificidades HLA-A9, A23, A29-31, Bw62 y DR7. Los resultados obtenidos parecen indicar que la generación de clones de células T humanas aloreactivas es mucho más eficiente que la producción de anticuerpos monoclonales múridos en cuanto a la definición de variantes polimórficas de los antígenos HLA

Production and characterization of T-cell clones specific for trinitrophenyl (TNP)-modified syngeneic spleen cells (TNP-self).

Using serial antigenic challenge as the method of selection and stimulation, continuous lines of cytotoxic T-lymphocytes (CTL) directed against TNBS-modified syngeneic spleen cells (TNP-self) have been generated. Spleen cells from C3H/HeJ (H-2k) mice were primed in vitro with autologous spleen cells modified with TNBS, and subsequently cloned by limiting dilution and in soft agar in the presence of IL2. These CTL clones grew continuously in medium supplemented with IL2 and in the presence of antigen. They are antigen specific and H-2 restricted in their target cell recognition. They all express Thyl and Lyt2 surface markers. None of the clones exhibit natural killer (NK) cell activity. All CTL clones tested so far are restricted in their target cell recognition to H-2Kk-TNP and none were found to be restricted to H-2Dk-TNP. These findings demonstrate at the clonal level the H-2K/D restriction of TNP-self specific CTL. These clones provide tools that may facilitate an understanding of the development and regulation of antigen specific CTL. They may also serve as models useful towards an understanding of the mechanism of lysis by CTL.