[Modified tests basophil degranulation and leukocyte migration inhibition factor in drug allergy. Study 2009-2014]. / Pruebas modificadas de degranulación de basófilos y factor de inhibición de migración de leucocitos en alergia a fármacos. Estudio de 2009 a 2014.

BACKGROUND: Adverse reactions to drugs are increasing and there are few studies for the diagnosis. OBJECTIVE: To determine the utility of modified basophil degranulation (MBD) test and modified leukocyte migration inhibition factor (MLMIF) test to prove drug hypersensitivity. METHODS: 177 patients of both sexes were studied with the diagnosis of drug hypersensitivity, determining MBD, MLMIF, or both, between 2009 and 2014. They were matched with positive and negative controls and the non-allergic population. Applications are issued according to the type of hypersensitivity, considering type I MBD and type IV MLMIF. RESULTS: 170 patients (96.04%) were positive to at least one drug (RR = 4.71). 561 MBD (73.62%) and 201 MLMIF (26.37%) were performed. Female sex was more frequent (64.41%); the average age was 38.5. MBD was positive in 70.23% and MLMIF in 67.16%. The test sensitivity was increased complementarily and with two dilutions. The correlation of MBD and MLMIF was positive and highly significant. CONCLUSIONS: Women have more drug reactions. Modified MBD test is useful at any age. Since medications can activate one or other hypersensitivity mechanism, it is important to request the tests simultaneously.

Antecedentes: Existe incremento de reacciones adversas a medicamentos y pocos estudios para el diagnóstico. Objetivo: Determinar la utilidad de pruebas modificadas de degranulación de basófilos (DB) y del factor inhibidor de la migración de leucocitos (LIF, leukocyte migration inhibition factor) para comprobar la hipersensibilidad a medicamentos. Métodos: Se estudiaron 177 pacientes, de uno y otro sexo, con diagnóstico de hipersensibilidad a medicamentos, en quienes se determinó pruebas modificadas de DB, LIF, o ambas entre 2009 y 2014. Se parearon con controles positivos, negativos y población no alérgica. Las solicitudes se emitieron de acuerdo con el tipo de hipersensibilidad, considerando tipo I a DB y tipo IV a LIF Resultados: 170 pacientes (96.04%) fueron positivos al menos a un medicamento (RR, 4.71). Se realizaron 561 pruebas modificadas de DB (73.62%) y 201 de LIF (26.37%). El sexo femenino fue más frecuente (64.41%); la edad promedio fue de 38.5 años. La prueba modificada de DB resultó positiva en 70.23% y la de LIF en 67.16%. La sensibilidad de las pruebas se incrementó en forma complementaria y a dos diluciones. La correlación de las pruebas fue altamente significativa. Conclusiones: Las mujeres presentan más reacciones a fármacos. La prueba modificada de DB es útil en cualquier edad. Como los medicamentos pueden activar uno u otro mecanismo de hipersensibilidad es importante solicitar las pruebas simultáneamente.

Factor inhibidor de la migración leucocitaria antes del tratamiento con factor de transferencia y después de el / Leukocyte migration inhibitory factor before and after treatment with transfer factor

Se realizaron determinaciones de la actividad del factor inhibidor de la migración leucocitaria "in vitro" en 10 niños con infecciones respiratorias recurrentes antes del tratamiento con factor de transferencia y después de él. Se encontró un incremento significativo (pó0,01) de esta función celular y de la respuesta cutánea a la tuberculina "in vivo", después del tratamiento con este medicamento. Los resultados señalan la importancia del estudio de pacientes con esta entidad y la utilidad de evaluar estas variables en su seguimiento clínico y terapéutico

Comparación de la producción del factor inhibidor de la migración de los leucocitos en recién nacidos y adultos / Comparison of production of leucocyte migration inhibition factor in newborns and adults

Para evaluar la madurez del sistema inmune en recién nacidos que por motivos diagnósticos lo requieren, es imprescindibles conocer los parámetros normales. Conociendo la importancia del factor inhibidor de de la migración de los leucocitos(FIML) y al no tener referencias en cuanto a la producción de esta linfoquina en la etapa neonatal, se decidió comparar la capacidad que tienen los linfocitos T de producir FIML, cuando han sido estimulados con antígenos y con el mitógeno fitohemaglutinina en individuos adultos sanos y niños recién nacidos por parto normal, y en estos últimos se explora la inhibión de la migración de los leucocitos de sangre de cordón umbilical incubados con FIMI, específico para un antígeno. Se concluyó que los recién nacidos no responden a los antígenos de memoria y sí lo hacen a la fitohemaglutinina y al FIML, específico para un antígeno

Transferencia in vitro de inmunidad a PPD con extracto dializable de leucocitos de calostro humano / In vitro immunity transference of PPD with human dialysable colostral leukocyte extract

Se pretende demostrar la transferencia de hipersensibilidad a PPD en un modelo in vitro con extracto dializable de leucocitos de calostro de madres PPD- y PPD+ (EDLC PPD- Y PPD-), a través de la medición de la actividad del factor inhibidor de la migración de leucocitos (LIF) de sangre de cordón de recién nacidos de madre PPD+. En los resultados se observa que el EDLC PPD+ incubado con leucocitos de recién nacidos de madres PPD- tuvieron inhibición de la migración de los leucocitos, comprados con la migración de los leucocitos incubados con EDLC PPD-; lo que sugiere que en el modelo in vitro se transfiere hipersensibilidad a PPD con el EDLC.

[In vitro transfer of immunity against PPD with dialyzable extract of leukocytes from human colostrum]. / Transferencia in vitro de inmunidad a PPD con extracto dializable de leucocitos de calostro humano.

The aim of this study is to demonstrate the transference of PPD hypersensibility in an in vitro model, with dialysable colostral leukocyte extract (DCLE) of PPD+ and PPD-mothers, through measurements of leukocyte migration inhibition factor activity (LIF) from blood obtained of the umbilical cord of newborns from PPD+ mothers. The results show that DCLE PPD+ incubated with leukocytes of newborns from PPD- mothers had inhibition of leukocyte migration compared with migration of leukocytes incubated with DCLE PPD-. These results suggest that in this in vitro model, DCLE transfers hypersensibility to PPD.

[Leukocyte migration inhibitory factor and basophil degranulation in drug reactions]. / Factor inhibidor de la migración de leucocitos y degranulación de basófilos en las reacciones a medicamentos.

We performed a prospective study in patients with a medical history of adverse reaction to drugs with the purpose of rule out allergy. We included 31 patients who were attended in the Allergy Service. We compared the sensibility and specificity of the test of inhibition factor of leucocytes migration and degranulation basophil against the exposition. After the statistical analysis, we concluded: the laboratory test, we have already mentioned, have little sensibility and specificity so the exposition test in the quickest, useful, and more simple method to determine drugs allergy, but more dangerous.

Factor inhibidor de la migración de leucocitos y degranulación de basófilos en las reacciones de medicamentos / Leukocyte migration-inhibitory factors and basophile degranulation in drug reactions

Se comunican los resultados de un estudio prospectivo realizado en 31 pacientes con historia de reacción adversa a medicamentos con el propósito de descartar fondo alérgico. Se4 comparo la sensibilidad y especificidad de las pruebas del factor inhibidor de la migración de leucocitos y degranulación de basófilos contra las pruebas de exposición . Los resultados se analizaron estadisticamente y se concluyó que las pruebas de laboratorio antes mencionadas tienen poca sensibilidad y especificidad. La prueba de exposición es el método más rápido, útil y sencillo para determinar alergia a medicamentos, teniendo presente que implica mayor riesgo.

Human leukocyte migration inhibition factor (LIF) increases polymorphonuclear cell endocytosis.

We prepared supernatants of Concanavalin-A activated human lymphocytes containing high titers of leukocyte migration inhibition factor (LIF). A pool of these supernatants was filtered thorough sephadex 6-100 as well as a pool of supernatants from parallel non activated cultures. A migration assay was carried out for each activated fraction, using as control migration the same fraction from non activated supernatants. In this way we found a fraction from activated supernatants with high LIF activity. We assayed the effect of this LIF containing fraction on a yeast endocytosis assay by polymorphonuclear (PMN) cells. We found that the LIF containing fraction increased the number of endocytic PMN in about 80%. This effect was absent from control supernatant and from other fractions from activated supernatant but without LIF activity. The LIF containing fraction did not increase the average number of endocytosed yeast per cell nor the ability to reduce NBT. The endocytosis enhancing effect was blocked by the specific LIF blocker N-acetyl-D-glucosamine. We conclude that LIF can increase the endocytic activity of PMN cells.

Migration inhibition factor in acute serum sickness nephritis.

Monocytes have been demonstrated to play an important role in acute serum sickness (AcSS) nephritis. Because accumulation of monocytes within the glomeruli could be the result of local lymphokine production, we studied migration inhibition factor (MIF) activity in supernatants from glomerular cultures, analyzed its temporal relationship with monocyte and lymphocyte accumulation, and tested the effect of anti-T lymphocyte monoclonal antibody on local MIF production. AcSS was induced in 12 rabbits, and one additional rabbit had antigen elimination without proteinuria. Single nephrectomy was performed at the time of antigen elimination in all animals; the remaining kidney was removed four days (4 rabbits) or 14 days afterwards (5 rabbits). In glomerular cross sections (gcs), lymphocytes were identified using monoclonal antibody M108, and monocytes by nonspecific esterase stain (ES). MIF activity was determined in supernatants of cultures of isolated glomeruli by the agarose microdroplet method. Peak of MIF activity (84.3 +/- 2.6%, SEM) was observed the first day of proteinuria in association with peak of lymphocyte infiltration (1.15 +/- 0.1 lymphocytes/gcs) and monocyte infiltration (2.4 +/- 0.3 mean ES score/gcs). MIF activity diminished by day 4 (66.0 +/- 6.3%) and reached control levels by day 14 (12.8 +/- 3.2%). There was a significant correlation between lymphocyte infiltration and MIF activity (r = 0.776, P less than 0.0001) as well as between MIF activity and monocyte accumulation (r = 0.858, P less than 0.0001). In five additional rabbits with AcSS, glomeruli were isolated, treated successively with M108 and normal rabbit serum, and supernatants harvested from 24-hour cultures were tested for MIF activity.(ABSTRACT TRUNCATED AT 250 WORDS)

[Monocyte locomotion inhibiting factor (MLIF) produced by E. histolytica induces an increase of cAMP in human monocytes]. / El factor inhibidor de la locomoción de monocitos (FILM) producido por E. histolytica induce aumento del AMPc en los monocitos humanos.

The supernatant fluid of axenically grown E. histolytica contains a factor (MLIF) which inhibits the locomotion of human monocytes (including chemotaxis) without affecting that of human polymorphonuclear leucocytes. Locomotion, like other cellular functions, is modulated by changes in intracellular cAMP and cGMP. The consensus--with some exceptions--is that while rises in cGMP accompany locomotion, an increase in cAMP (without a concomitant fall in -cGMP) occurs with inhibition of cellular movement. We measured by radioimmunoassay the cAMP concentration of human monocytes exposed to inhibitory concentrations of MLIF. A significant (p less than 0.005) rise in monocyte cAMP was found, comparable to that observed with the use of forskolin, a well known cAMP stimulator. The control studies using plain axenic medium, not only failed to reveal any rise in cAMP but disclosed a small, yet not significant drop in intracellular cAMP. These results suggest that MLIF (like other locomotion inhibitors, i.e. prostaglandins E1, A1 and isoproterenol) produces a significant increase in intracellular monocyte cAMP. This modification in intracellular signals may contribute to the inhibition in monocyte locomotion, an event during which an increase in pericentriolar microtubules has also been observed.

Normalización de una microtécnica para evaluar el factor inhibidor de la migracion leucocitaria in vitro / Standardization of a microtechnique for the evaluation of migration inhibition factor of leukocytes in vitro

Se normalizó la microtécnica directa de la prueba de inhibición de la migraciòn leucocitaria in vitro con la utilización del derivado proteico de la tuberculina (PPD) como antígeno de prueba. Se estableciò el punto de corte de la inhibiciòn de la migraciòn producida por esta linfocina para la concentraciòn del PPD empleado, el cual ajustamos en nuestro laboratorio para el índice de la migraciòn leucocitaria como < 0,65

Production of leukocyte migration inhibition factor (LIF) and migration stimulation factor (MStF) by CD4+ and CD8+ human lymphocytes subsets and T-cell clones.

The production of the lymphokines leukocyte migration inhibition factor (LIF) and migration stimulation factor (MStF) at the level of CD4+ and CD8+ human lymphocyte subsets was investigated. In a first series of experiments, anti-CD4 and anti-CD8 monoclonal antibodies capable of inhibiting the activation by concanavalin-A (Con-A) of the respective T-cell subset were used. It was observed that when CD8+ cell activation was blocked, LIF was always produced after Con-A activation. When CD4+ cell activation was blocked, MStF was produced in five out of nine experiments (no activity in the other four). The addition of N-acetyl-D-glucosamine to block LIF in supernatants of anti-CD8 treated cells was unable to show evidence of masked MStF activity. In a second series of experiments, T-cell clones were established from continuous growing T-lymphocyte cell lines developed from cultures of Con-A activated normal human leukocyte cultures. The phenotype of 22 clones was determined and their ability to produce LIF or MStF investigated. Four clones produced MStF after Con-A activation and all of them were CD3+, CD4-, CD8+. Three clones produced LIF after Con-A activation and all of them were CD3+, CD4+, CD8-. We conclude that LIF is produced by CD4+ cells and MStF by CD8+ cells.

Neutrophil migration induced by inflammatory stimuli is reduced by macrophage depletion.

Previous experiments of our group have shown that neutrophil migration induced by inflammatory stimuli is reduced by agents which block the release from macrophages of a specific factor for neutrophil migration (MNCF, [1, 2]). The present paper evaluated the influence of macrophage depletion induced by lavage of the peritoneal cavity on neutrophil migration. In both normal and thioglycollate-stimulated peritoneal cavities, lavage with saline reduced the resident macrophage population by about 80% and significantly blocked neutrophil migration induced by inflammatory stimuli such as carrageenin, zymosan and E. coli endotoxin. Peritoneal lavage, however, did not affect neutrophil migration induced by MNCF. Thus, these results support the suggestion that macrophages participate in the control of neutrophil migration induced by acute inflammatory stimuli.

Human lymphocytes produce both leukocyte migration inhibition (LIF) and simulation (MStF) factors after mitogen, alloantigens or antigenb activation

Utilizando un sistema desarrollado para producir factor inhibidor de la migración de leucocitos (LIF) en altos títulos, hemos encontrado que cuando activamos linfocitos humanos con concanavalina-A, obtenemos LIF en 55% de los experimentos y factor estimulador de la migración en 38%. Cuando los linfocitos fueron activados por medio de cultivos mixtos de linfocitos encontramos resultados muy parecidos. Se seleccionaron sobrenadantes de linfocitos humanos activados con concanavalina-A que tuvieron una alta actividad de LIF. Al bloquear el LIF con N-acetil-D-glucosamina se pudo observar que estos sobrenadantes contenían también factor estimulador de la migración. En pruebas de LIF directo usando como antígeno a la estreptocinasa-estreptodornasa, encontramos que cuando el ensayo se realiza en presencia de N-acetil-D-glucosamina el valor de índice de migración se incrementa de manera importante. Concluimos que cuando los linfocitos humanos son estimulados ya sea por mitógenos, aloantígenos o antígenos, ellos producen tanto factor inhibidor como estimulador de la migración de leucocitos. Finalmente se discute la importancia de este hallazgo en la determinación de immunidad celular in vitro en humanos por medio de la prueba de LIF

Mitogen induced LIF-production in T-lymphocyte subpopulations

El método de rosetas con eritrocitos de pollo recubiertos de IgG se ha utilizado junto con separación por flotación en ficoll-hypaque para obtener subpoblaciones de células enriquecidas o disminuidas de linfocitos T con receptor para Fc de IgG (Tg). Estas poblaciones celulars se han utilizado para determinar producción de LIF a diferentes concentraciones de los mitógenos, utilizando un método en microgota de agarosa. De esta manera, hemos podido observar una respuesta semejante en las diferentes poblaciones celulares hacia concanavalina A y una respuesta diferencial hacia fitohemaglitinina donde por lo general se observa una respuesta negativa de la población enriquecida con células Tg mientras que las poblaciones de linfocitos totales o aquellas en las que se han eliminado las Tg dan una respuesta positiva al segundo mitógeno