RESUMEN
1. The effects of experimentally induced heat-stress on the embryonic development of bursa of Fabricius and thymus of the chicken were investigated by means of histological and enzyme histochemical methods. 2. In the experiments, 250 fertile eggs of the Ross 308 broiler strain were divided into two groups. The control eggs were maintained under optimal conditions (378 degrees C and 65 +/- 2% relative humidity, RH) during the whole incubation period. Heat stressed eggs were maintained under normal conditions (378 degrees C and 65 +/- 2% RH) until the 10th d of incubation and then exposed continuously (24 h per d) to high temperature (388 degrees C and 65 +/- 2% RH). Blood and tissue samples were taken from 10 animals of each group at d 13, 15, 18 and 21 of incubation and at d 2, 4 and 7 post-hatch. Tissue samples were processed for enzyme histochemical methods in addition to routine histological techniques. 3. The results revealed that egg temperatures were higher than incubator air temperature. Long-term heat-stress (401-406 degrees C egg temperature) retarded development of thymus and bursa of Fabricius. Peripheral blood ACP-ase and ANAE-positive lymphocyte levels of heat-stressed animals were lower than in the controls. 4. These results give some morphological evidence for immunosuppression induced by high temperature exposure during the embryonic development. Temperature distribution and air circulation in incubator should be questioned in the case of lower broiler flock immunity.
Asunto(s)
Bolsa de Fabricio/embriología , Pollos/inmunología , Trastornos de Estrés por Calor/veterinaria , Timo/embriología , Fosfatasa Ácida/inmunología , Animales , Bolsa de Fabricio/enzimología , Bolsa de Fabricio/inmunología , Embrión de Pollo , Trastornos de Estrés por Calor/embriología , Trastornos de Estrés por Calor/enzimología , Trastornos de Estrés por Calor/inmunología , Inmunohistoquímica/veterinaria , Isoenzimas/inmunología , Naftol AS D Esterasa/inmunología , Fosfatasa Ácida Tartratorresistente , Timo/enzimología , Timo/inmunologíaRESUMEN
Secondary macrophage cell cultures were generated from the primary culture of epiblasts of 8-d-old pig blastocysts. The epiblast-derived macrophagelike (EDM) cells have a morphology and ameboid behavior that is typical of tissue histocytes. The cells reacted positively with monoclonal antibodies specific for pig granulocyte-macrophage lineage cells, and were not reactive with monoclonal antibodies specific for pig B and T lymphocytes. Marked phagocytic behavior and the formation of phagosomes were demonstrated following incubation with FITC-labeled bacteria. The EDM cells stained positively for nonspecific acid esterase that was not inhibited by sodium fluoride. DiI-acetylated-LDL was rapidly taken up by the cells. Transmission electron microscopy of the EDM cells showed phagolysosomes, numerous cytoplasmic vacuoles, large, lobed nuclei, and numerous pseudopods or filopodia at the cell surface. Strong reactivity of the cells with anti-CD14 monoclonal antibody was observed. Further, cytotoxic activity was produced from the EDM cells after exposure to lipopolysaccharide in a concentration and time-dependent manner. The cultures could be maintained and expanded for several months on STO co-culture. Their derivation from the epiblast of the pig demonstrates the possibility of obtaining hemopoietic cell cultures from the preimplantation blastocysts of all mammals.
Asunto(s)
Macrófagos/inmunología , Acetilación , Animales , Linfocitos B/inmunología , Carbocianinas/química , Células Cultivadas , Pruebas Inmunológicas de Citotoxicidad , Antígenos de Histocompatibilidad Clase I/inmunología , Antígenos de Histocompatibilidad Clase II/inmunología , Receptores de Lipopolisacáridos/inmunología , Lipoproteínas LDL/farmacocinética , Macrófagos/citología , Microscopía Electrónica , Naftol AS D Esterasa/inmunología , Fagocitosis/inmunología , Receptores Inmunológicos/inmunología , Porcinos , Linfocitos T/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
We examined the isoenzyme patterns of alpha and beta naphtyl acetate esterase and the IL6 production of two macrophage cell lines, which were cloned from a single fusion of macrophage tumor cells and spleen adherent cells. These clones were phenotypically indistinguishable but differ functionally as line 59 presents antigen to Th 1 lymphocytes while line 63 induces suppressor T lymphocytes. Cell extracts of these lines exhibit different isoenzyme patterns of alpha and beta naphtyl acetate esterase at both pH 7.5 and 5.8 but do not differ significantly in the level of produced IL6. Treatment with nitrogranulogen (NG), a derivative of cyclophosphamide, changes the isoenzyme pattern in line 59 and decreases severalfold IL6 production, while in similarly treated line 63 cells isoenzyme pattern remains unaffected but the production of IL6 is significantly increases. We assume that the observed differences between these two cell lines are due to distinct intracellular translation of the membrane signal delivered by NG. The different behaviour of these two parameters can thus be used as a useful tool to further delineate different macrophage subpopulations. We regard it likely that nonspecific esterases may play a role in intracellular processing or trafficking of antigen.
Asunto(s)
Esterasas/metabolismo , Interleucina-6/biosíntesis , Isoenzimas/metabolismo , Macrófagos/enzimología , Naftol AS D Esterasa/metabolismo , Animales , Biomarcadores , Esterasas/efectos de los fármacos , Esterasas/inmunología , Hibridomas , Isoenzimas/efectos de los fármacos , Isoenzimas/inmunología , Macrófagos/inmunología , Mecloretamina/farmacología , Ratones , Naftol AS D Esterasa/efectos de los fármacos , Naftol AS D Esterasa/inmunología , Células Tumorales Cultivadas/citología , Células Tumorales Cultivadas/enzimología , Células Tumorales Cultivadas/inmunologíaRESUMEN
The close topographical correlation between naphthol AS-D chloroacetate esterase-positive macrophages and prostaglandin synthase-rich macrophages in the thymus of normal and cyclosporin-treated rats was observed. It seems possible that chloroacetate esterase is an enzyme related to metabolism of arachidonic acid and/or production of its active metabolites.
Asunto(s)
Macrófagos/enzimología , Naftol AS D Esterasa/metabolismo , Prostaglandina-Endoperóxido Sintasas/metabolismo , Animales , Ciclosporina/farmacología , Inmunohistoquímica , Macrófagos/ultraestructura , Masculino , Naftol AS D Esterasa/inmunología , Prostaglandina-Endoperóxido Sintasas/inmunología , Ratas , Ratas Wistar , Timo/citología , Timo/efectos de los fármacos , Timo/enzimologíaRESUMEN
The experiments presented in this paper deal with the question of whether there is an absolute requirement for alpha-naphtylacetate esterase (ANAE)-positive monocytes, HLA-DR+, Leu11a+, and/or Leu7+ cells to stimulate human peripheral blood T lymphocytes by phytohemagglutinin (PHA). Purified (p) T lymphocytes containing less than 0.1% ANAE-positive monocytes were isolated from human peripheral blood mononuclear cells (MNC) by sequential removal of carbonyl-iron phagocytic cells and of low-density cells by density gradient centrifugation and isolation of E-rosette-forming cells (E-RFC). These pT-cells were further depleted of HLA-DR+, Leu11a+, and/or Leu7+ cells using monoclonal antibodies and cell sorting. The T lymphocytes were stimulated by PHA in an ultra-micro culture in glass capillaries at a volume of 1 microliter or 2 microliters, containing 1000 cells per culture. With this method, the accessory cell requirement could be studied under limiting cell number conditions. The results show that pT-cells can be stimulated by PHA in the absence of ANAE-positive monocytes. No ANAE-positive monocytes were found in the culture after stimulation, indicating the lack of differentiation into ANAE-positive monocytes from ANAE-negative precursors. A rabbit antiserum against leukocytic pyrogen (LP, also containing anti-IL 1 activity) only reduced but did not abrogate the stimulation of pT-lymphocytes by PHA. Addition of adherent cells resulted in an enhancement or in an inhibition of the response of pT-lymphocytes to PHA, depending on cell concentration and culture time: The lower the number of cultured T lymphocytes and the shorter the culture time, the higher was the enhancing activity by additional adherent cells, and vice versa. Further purification of the pT-cells using monoclonal antibodies and cell sorting led to the finding that depletion of either HLA-DR+, Leu11a+, or Leu7+ from pT-cells only reduced but did not abrogate the stimulation of the pT-cells by PHA. However, in absence of HLA-DR+ and Leu7+ cells, the pT-lymphocytes totally failed to respond to PHA. This abrogation of the response was not observed when pT-cells were depleted of HLA-DR+ and Leu11a+ cells. In addition, T11+/HLA-DR- T lymphocytes isolated from E-RFC by positive selection in a cell sorter also responded to PHA.(ABSTRACT TRUNCATED AT 400 WORDS)