The analysis of single haemopoietic colonies grown in methylcellulose and in agar was performed at intervals by a cytocentrifugation method. Correlation was established between morphology of the whole colonies and their cellular content. Three main cell lines predominated: neutrophils, macrophages, eosinophils; a few colonies contained a pure population of basophil-like granulocytes. Development was followed from myeloblasts to polymorphs, both being present in most of the colonies. Cumulated results showed that ( I ) the proliferating compartment remained quite large till day 14, with 58% of cells in S phase, and rapidly decreased after day 16, and that (2) the polymorphs rapidly disappeared from the culture medium.Differentiation proceeded at different rates from one colony to the other, thus suggesting heterogeneity between colony forming cells (CFC). Neutrophil colonies appeared and lysed more rapidly than did eosinophil colonies. Macrophages arose from large immature cells with many promyelocyte features; such cells were present in mixed colonies, containing both neutrophils and macrophages. It is very likely that granulopoiesis results from the development of distinct committed CFC.This work was carried out using normal human bone marrow and may be a useful tool for studying pathological material in the future.
Clusters of cells obtained from a short-term culture of human bone marrow cells in methylcellulose were transplanted in cell-free plates containing conditioned medium. Of 2559 transplants, 439 gave small clusters, 223 gave large clusters, and 70 gave colonies. A better cloning efficiency was achieved from 4-day-old clusters, from aggregates containing 3 cells or more, and from large cells rather than from small cells. Cytocentrifugation could be performed on 215 large clusters or colonies. The population consisted of pure neutrophils in 36.3% and of pure macrophages in 36.8%. A mixture of neutrophils and macrophages was found in 23.2%, thus indicating that these two cell lines originated from the same committed cell (G-CFC). Eosinophils were found in 8 (3.7%) clusters as a pure population, and were never mixed with either neutrophils or macrophages in the other 207 clusters. Despite the small number of eosinophil colonies, one could suggest that using the in vitro technique the committed eosinophil colony-forming cells could be distinguished from the G-CFC.
RésuméLa technique d'immunofluorescence indirecte au cours de la cysticercose a été étudiée à l'aide de quatre antigènes différents : scolex, cysticerques, anneaux de T. saginata et T. solium. La sensibilité de la méthode est bonne, les taux varient dans les cas positifs du 1/40e au 1/640e. Les sérums positifs correspondent aux cysticercoses récentes, s'accompagnant de signes neurologiques notamment, et aux formes calcifiées généralisées encore évolutives. La sensibilité de l'immunofluorescence est supérieure à celle de l'immunoélectrophorèse.La spécificité de la méthode semble bonne, si l'on ne retient comme positifs que les sérums donnant une fluorescence au moins égale au 1/40e. Il n'y a pas de faux positifs, parmi les sérums témoins, si on emploie comme antigène une coupe d'anneau de T. saginata ou T. solium. Les quelques faux positifs ont tous été observés sur coupes de scolex d'E. granulosus, dont l'emploi pris isolément est donc déconseillé.Les réactions croisées ne permettent pas, quel que soit l'anti gène utilisé, de distinguer sérologiquement une cysticercose, un kyste hydatique, une échinococcose alvéolaire ou une cénurose. SummaryHuman cyticercosis studied by the indirect fluorescent techni que. -The authors have studied sera from 35 human cysticercosis cases, with the indirect fluorescent technique, using E. granulosus, C. cellulosae, T. saginata and T. solium as antigens.The sensibility of this method seems good, the titers of the sera varying in the positive cases from 1/40 to 1/640. The 13 positive cases correspond to recent infestations, particularly when neurologic symptoms are noted ; some disseminated calcified cysticercosis give sometimes high titers. Compared with the Immunoelectrophoresis, the indirect IF test is more sensitive.The specificity is quite good when T. solium and T. saginata are employed as antigens, if the minimum positive titer is 1/40 in the controlled sera. The specificity is not so good on slices of E. granulosus with the antihuman polyvalent gammaglobulin fluorescent rabbit serum. Anyway, if it is always possible to recognize sera from parasitoses with the 4 antigens employed, the cross reactions are very important between the Cestode group, and the IF test cannot separate coenurosis, echinococcosis and cysti cercosis. A more precise diagnosis needs then clinical, epidemio logical and histological data.
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