RESUMO
A fluorescence assay for the quantitation of tumor cell lysis by activated and non-activated killer (LAK) cells is described. The target cells are labelled with a europium chelate (Eu-diethylenetriaminopentaacetate) and after cytolysis caused by the LAK cells the Eu3+ complex is released into the culture supernatant. The addition of beta-naphthoyltrifluoroacetone to culture supernatant aliquots leads to the formation of a highly fluorescent chelate which can be measured with a time-resolved fluorometer. The influence of various assay parameters has been evaluated including incubation time, effector-to-target cell ratio, the target cell line and different concentrations of interleukin-2 during cell culture. The optimized time-resolved fluorometric assay was found to be as simple and sensitive as the commonly used cytotoxicity assay in which the release of 51Cr from the labelled target cells is measured. In addition the assay is much faster and safer since the label is not radioactive.
Assuntos
Testes Imunológicos de Citotoxicidade , Fluorimunoensaio/métodos , Interleucina-2 , Células Matadoras Naturais/imunologia , Ativação Linfocitária , Comunicação Celular , Quelantes , Radioisótopos de Cromo/metabolismo , Testes Imunológicos de Citotoxicidade/métodos , Európio/metabolismo , Humanos , Células Matadoras Naturais/enzimologia , Células Matadoras Naturais/metabolismo , L-Lactato Desidrogenase/metabolismo , Fatores de TempoRESUMO
When treated with CD3 moAB (aCD3) (1 microgram/ml) and human natural IL-2 (nIL-2) 1000 U/ml, during the exponential phase of growing: 200 U/ml tumor infiltrating lymphocytes (TIL's) from brain tumors could be expanded. Due to the low percentage of lymphocytes in malignant brain tumor tissue (1%) it was not possible to separate the tumor bearing lymphocytes from the tumor cells by a Ficoll Hypaque gradient separation. This resulted in a co-culture of both lymphocytes and tumor cells in which a serum free medium was used. To generate TIL's at all, it was necessary to supplement this culture medium with 20% of the supernatant of cultures of lymphokine--activated killer cells. In 9 out of 12 cases we obtained total growth factors in the range between 10(4) and 10(14) (average 3.6 x 10(7)). These growth were reached after 6 to 13 weeks including up to 3 weeks of an initial lag period during which no cell growth was observed. We noted a decrease in CD8+ cells, whereby the predominant cell type at the end of the cultures was CD3+ and CD4+. The levels of their cytotoxic activities against K-562 and Raji cells were low in contrast to those of CD3 and nIL-2 stimulated cells which were obtained from carcinomas.
Assuntos
Neoplasias Encefálicas/imunologia , Linfócitos do Interstício Tumoral/imunologia , Adulto , Idoso , Neoplasias Encefálicas/terapia , Células Cultivadas , Citotoxicidade Imunológica , Feminino , Humanos , Imunoterapia Adotiva , Masculino , Pessoa de Meia-IdadeRESUMO
To evaluate the Laser-Doppler flowmeter (LD) as a diagnostic tool in microcirculatory alterations of gestosis, cutaneous circulation was measured in the forearm under basal conditions and during reactive hyperaemia by means of an MBF3D (Moor Instrument Ltd., Oxford) in 11 hypertonic and in 14 normotonic pregnant women (+/- proteinuria, +/- pathologic increase in body mass, 16-36 years old, 26-40th week of gestation) with the exception of systolic and diastolic blood pressure, all statistical and clinical criteria were identical in the two groups. There were, however, in the hypertonic patients significant reductions in most of the LD criteria (basal and maximal speed and concentration of erythrocytes, duration of reactive hyperaemia). Regardless of group assignment, there were negative correlations between erythrocytes speed on the one side (basal, maximal), and blood pressure an the other, in some instances also to the increase in body mass and to the APGAR score of newborns. These preliminary findings are considered on an expression of cutaneous vasoconstriction accompanying hypertension in pregnancy. Extensive validation of LD under these conditions seems promising.
Assuntos
Hipertensão/diagnóstico por imagem , Troca Materno-Fetal/fisiologia , Pré-Eclâmpsia/diagnóstico por imagem , Ultrassonografia Pré-Natal/métodos , Adolescente , Adulto , Velocidade do Fluxo Sanguíneo/fisiologia , Feminino , Idade Gestacional , Humanos , Hipertensão/fisiopatologia , Recém-Nascido , Pré-Eclâmpsia/fisiopatologia , Gravidez , Valores de Referência , Vasoconstrição/fisiologiaRESUMO
Pruritic urticarial papules and plaques of pregnancy (PUPPP) is a rare dermatosis of unknown etiology that is most frequently seen in primiparas and twin/multiple pregnancies. The prognosis is favorable. We report a case of PUPPP in a primipara and review the clinical signs, differential diagnosis, possible etiologic factors, diagnosis, and therapy.