Success of bone marrow transplantation in congenital Diamond-Blackfan anaemia: a case report.

A 6-yr-old girl with congenital corticosteroid-resistant pure red cell aplasia was treated with bone marrow transplant from her HLA-identical, MLC-unreactive sister in November 1984 following conditioning with busulfan and cyclophosphamide. Full engraftment was obtained and the patient at 21 months post-transplant is in excellent clinical condition maintaining normal red cell counts. We conclude that BMT should be considered as a therapy for at least the most severe cases of Diamond-Blackfan anaemia resistant to corticosteroids. Successful outcome of this therapy provides an argument for the stem cell origin of this disorder.

Treatment with antitheta serum affects hemopoietic stem cell HSC proliferation in an in vivo culture system.

Marrow cells from normal WBB6F1 +/+ mice were treated in vitro with either anti-Thy 1.2 serum and complement (C') or normal mouse serum (NMS) and C' and then they were cultured in diffusion chambers implanted into peritoneal cavity of Balb/c mice. Following 1-19 days of culture, chamber contents were injected into W/Wv mice to test for either the capacity to form spleen colonies or to cure the W/Wv anemia. In contrast to NMS-treated marrow, anti-Thy 1.2 serum treated marrow hemopoietic stem cells almost completely failed to proliferate in this system. Number of cells per chamber upon termination of culture was less affected.

Aplastic anaemia with marrow defective in formation of fibroblastoid cell colonies in vitro.

The formation of fibroblastoid colonies by marrow cells in vitro has been used as a putative assay for a stem cell of haemopoietic stroma. Bone marrow from one patient with aplastic anaemic did not form any of these colonies, while its growth in diffusion chambers as an indirect measure of a haemopoietic stem cell was even better than normal. On the other hand, marrow from the other aplastic patient showed only quantitative decrease in the formation of fibroblastoid colonies and simultaneously grew very poorly in diffusion chambers. These patients were indistinguishable by the cytological examination of their marrow, however, the peripheral blood abnormalities were expressed less severely in the first patient. These results suggest the contribution of the defect in marrow cells, which form fibroblastoid colonies in vitro to the development of aplastic anaemia in these patients.

Haematopoiesis in mice heterozygous for the W trait: defective formation of transient endogenous spleen colonies.

It has been determined that W/+ and Wv/+ heterozygous mice, as compared with normal +/+ homozygous littermates, form significantly lower numbers of transient 5-day endogenous spleen colonies in response to X-irradiation. This defect was evident for doses of irradiation between 2-6 Gy (200-600 rad) and was associated with a slightly increased radiosensitivity of the assayed precursor cells (TE-CFU) in W heterozygotic mice. Moreover, the defect was transplantable, i.e., intrinsic to the marrow cells and not to the microenvironment, and was not associated with a similar decrease in cells which form erythropoietic bursts in vitro (BFUe). This study provides a cellular basis for increased radiosensitivity of W/+ and Wv/+ mice and suggests that the 'W' mutation is semi-dominant, both with respect to the white spotting and TE-CFU formation.

Evaluation of the marrow culture on the macrophage layer covering intraperitoneally implanted membrane as an assay for hemopoietic progenitor cells.

When cellulose acetate membranes are implanted into abdominal cavity of mice they turn into a foreign body overgrown with macrophages. Such macrophage layer has been shown by other authors to be able to support the growth of hemopoietic colonies formed by intraperitoneally injected hemopoietic cells. This study confirms and extends this observation by showing that both granulopoietic and erythropoietic colonies may be observed. The number of colonies grown is in linear correlation with that of injected hemopoietic cells. The frequency of erythropoietic colonies was greatly enhanced by blood letting of the host mice. Colony forming cells were most numerous in the bone marrow then in the spleen and peripheral blood and hardly in the thymus. Prior irradiation of the host mice was essential for obtaining colony growth and the optimal dose was determined to be 6.0 Gy. This technique opens the way to studies into hemopoietic progenitor cells for laboratories having no sophisticated tissue culture equipment and where necessary reagents are easily available.

Antitheta immunization affects hemopoietic colony growth in a theta-incompatible mouse system.

Seeking a system with which to study the antitheta-sensitive regulatory cell (TSRC), other than the W/Wv anemic mouse, we tested a model which utilizes mouse strains differing almost exclusively in theta allotype. AKR/FuRd Thy 1.1 mice were immunized against CBA Thy 1.2 mouse thymocytes to produce antibody. The immunized mice were lethally irradiated and grafted with marrow cells from AKR/Cum Thy 1.2 mice. It was reasoned that in this situation antitheta-sensitive cells present in or produced by the graft will be destroyed, while theta-incompatible host cells may not be as capable of collaboration with the graft stem cells as are the graft's theta-compatible cells. Anti-Thy 1.2 immunization did not influence the number of spleen colonies formed by marrow from Thy 1.2 mice; but the colonies formed, especially the erythropoietic colonies, were reduced in size. Also, both the relative and absolute frequencies of erythropoietic colonies were decreased, while those of granulopoietic colonies were increased. Therefore, the destruction of antitheta-sensitive cells in this system resulted in decreased effectiveness of spleen colony growth and altered the differentiation pattern of colony-forming cells. These effects were not observed in control groups immunized with CBA tissues not bearing the Thy 1.2 antigen. This study further stresses the importance of the theta antigen in hemopoietic differentiation and may provide an alternative in vivo model for studying the TSRC.

[Effect of ultrasonic stimulation of the lymphatic system on changes in subpopulations of peripheral blood lymphocytes in multiple sclerosis]. / Wplyw stymulacji ultradzwiekowej ukladu limfatycznego na zachowanie sie subpopulacji limfocytów krwi obwodowej chorych na stwardnienie rozsiane.

In the years 1977-1979 after publication of the experiences of German authors a trial was undertaken of treating 63 patients with multiple sclerosis by means of ultrasonic stimulation of the lymphatic system. It was the purpose of the present work to check the effect of this method on the subpopulations of peripheral blood lymphocytes in 20 patients. These investigations were carried out using rosette tests EAC, AE, E which were performed three times in each case: before the onset of treatment, after the first procedure and after the last procedure. These investigations demonstrated significant differences between the mean values of EAC and EA rosettes before and after the treatment--a rise in the number of lymphocytes B (EAC rosettes) and a fall of lymphocytes T (EA rosettes).