肾综合征出血热患者早期骨髓细胞结构异常的研究

目的 观察汉坦病毒(HV)对人体骨髓细胞的作用。方法 应用免疫荧光技术、光镜和电镜细胞学检查对骨髓细胞结构尤其是超微结构进行观察研究，并设对照组。结果 检测了27例肾综合征出血热(HFRS)患者骨髓细胞的病毒抗原，20例呈现阳性。发现粒细胞含HV抗原阳性细胞百分数最高(76％)，其他依次为单核细胞(65％)，淋巴细胞(40％)，巨核细胞(20％)和红细胞(3．7％)。利用光镜、透射电镜和扫描电镜进行细胞学观察发现感染病毒的骨髓细胞、细胞膜和细胞器的损伤明显高于对照组。结论 HV可侵入骨髓细胞中，并对人骨髓细胞具有致病变作用。     

Age-related changes in activation of telomerase in the bone marrow of normal and thymectomized mice

Age-related and thymus-dependent regulation of telomerase activity was studied in the bone marrow of normal (physiological aging) and thymectomized (experimental aging) mice. There was no strong correlation between the age and telomerase activity in bone marrow cells of normal mice. We observed only small individual differences in telomerase activity. Thymectomy 2-fold increased telomerase expression in young (2-month-old) and old (24-month-old) animals. Individual differences in telomerase activity in the bone marrow of thymectomized mice were more pronounced and did not depend on the age. The role of the thymus in cell aging is discussed. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 129, No. 6, pp. 654–656, June, 2000     

Polyclonal antibody synthesis by bone marrow cells of New Zealand Black mice. II. Cellular basis of PAS activity of NZB bone marrow cells

In the preceding report, enhanced polyclonal antibody synthesis (PAS) activity of bone marrow cells (BMC) of NZB mice was shown. Cells involved in its activity among BMC population were explored in this report. Antibody-forming cells (AFC) already existing in the bone marrow were not responsible for this activity since depletion of AFC by passage through a Sephadex G-10 column did not affect the numbers of anti-TNP AFC and immunoglobulin-secreting cells (IgSC) in the spleen of BMC-reconstituted mice when assayed 7 days after the transfer. Thy-1-positive cells among BMC population and host thymus were not relevant to PAS. Suppressor cells seemed to play little role as observed in transfer of mixed BMC population from NZB and BALB/c mice. PAS activity was prominently observed in surface immunoglobulin-positive cells. Significance of enhanced PAS activity was discussed with reference to autoantibody production.     

Cellular composition of rat bone marrow stroma. Antigen-defined subpopulations

Although stromal cells establish the architecture of mammalian bone marrow and organize hemopoiesis, the interrelationships among their macrophage, fibroblastic, endothelial, and adipocyte-like components are not wholly understood. Using murine monoclonal antibodies to cultured adherent cells of rat bone marrow, we observed that the predominant fibroblastoid cells grown from marrow differed from those of non-hemopoietic organs. The marrow type bore a detectable quantity of the ST3 but not ST4 antigen, whereas those from lung, diaphragm, and epididymal fat pad, bore more ST4 than ST3. Those from spleen were an equal mix of both types. Although the tissue distribution of the ST3 antigen was similar to that of Thy-1, it was not identical, and in the brain, the two structures were localized in different areas. While none of the ST3, ST4 (fibroblast directed), or BN(MB)35 (myeloid directed) antibodies recognized fat cells cultured from marrow, the ST10 antibody, selected for binding to marrow derived fat cells, stained peripheral adipose cells, unidentified aglobular cells in areas of fat cell formation, and macrophages, but not fibroblasts. On the basis of these observations, we suggest that the fibroblastoid cells of the marrow are different from those of non-hemopoietic tissues.     

雌性小鼠骨髓移植给雄性小鼠后内源性骨髓细胞残存状态的研究

 目的:以雌性小鼠骨髓移植给雄性小鼠的方法,通过检测雄性小鼠血细胞的Y染色体来明确内源性骨髓细胞的残存状态。方法:将雌性或雄性C57BL/6小鼠作为受体,实验组用［137Cs］照射,6 h后每只经尾静脉注射供体小鼠骨髓细胞1×107。统计骨髓移植后14 d动物的存活率,并通过眶静脉采血观察外周血白细胞数量的变化,检测受体雄性小鼠体内Y染色体基因水平的变化以明确骨髓移植的效果。结果:分别用1 000、950和900 rad的照射剂量对受体小鼠进行照射后将供体小鼠的骨髓移植到受体小鼠体内,1 000和950 rad剂量时雌性受体小鼠可迅速恢复造血功能,而雄性受体小鼠则仅有48%的存活率。900 rad照射剂量骨髓移植后,雄性受体小鼠迅速恢复了造血功能,13 d后外周血白细胞计数基本恢复正常。移植后的雄性受体小鼠在5周内已检测不到外周血细胞Y染色体基因,表明雄性受体小鼠的骨髓被完全破坏,雌性供体小鼠的骨髓可完全替代受体雄性小鼠的骨髓并且发挥造血功能。结论: 在照射剂量900 rad照射后,雄性小鼠可以作为骨髓移植受体,为将来应用雄性小鼠作为骨髓移植受体动物开展有关心血管疾病的研究奠定了实验基础。     

Prostaglandin-induced changes on mitogen-activated bone marrow cells

The mitogenic activation of bone marrow cells by dextran sulfate was inhibited by prostaglandin E₂ and enhanced by addition of a prostaglandin synthetase inhibitor such as indomethacin. In contrast, dextran sulfate-induced mitogenesis of spleen cells appeared enhanced by prostaglandin E₂ and inhibited by indomethacin. This dextran sulfate activation of bone marrow cells was partially dependent on phagocytic accessory cells, and the effect of indomethacin on dextran sulfate activation appeared only when macrophages were present in the cultures.When purified bone marrow lymphocytes were used, the dextran sulfate-induced mitogenesis was macrophage-independent, and the presence of indomethacin did not induce any enhancement in the response.Since dextran sulfate acts as a general leukocyte activator, the possibility of a differential modulation of these cells by prostaglandins and prostaglandin synthetase inhibitors is suggested.     

Ovariectomy-associated changes in bone mineral density and bone marrow haematopoiesis in rats

The relationship between the bone mass loss and bone marrow haematopoiesis in osteoporosis remains obscure. We selected 3-month-old female Sprague–Dawley rats and randomly divided them into six groups. Three groups were ovariectomized (OVX), while the other three groups were sham operated (Sham). Four, 8 and 12 weeks after the surgical procedure, the rats were euthanized and sampled. The left femur was used for measurement of bone mineral density (BMD). The right femur distal metaphysic cancellous bone was processed for morphological evaluation. Our results showed that the femur BMD in the 4-week OVX group was not significantly decreased compared with that of the 4-week Sham group, but that the volume of adipose tissue in the bone marrow was markedly increased. The femur BMD in the 8-week OVX group was decreased significantly compared with that of the 8-week Sham group ( P   <   0.05). Meanwhile, the volume of haematopoietic tissue decreased and the volume of adipose tissue increased. The number of megakaryocytes was decreased ( P   <   0.05). Interestingly, the osteoclasts and mast cells were increased in number in the 8-week OVX group ( P   <   0.05). These changes became obvious in the 12-week OVX rats, in contrast to the Sham groups. The volume of trabecular bone and the number of osteoblasts in the 12-week OVX group decreased significantly. Increased reticulin fibres were observed only in the 12-week OVX group. Our studies demonstrated a reciprocal correlation between bone-forming osteoblasts and marrow adipose tissue and suggest that OVX rats may be valuable as an animal model to study hypohaemopoiesis.     

小鼠同种异基因骨髓腔内骨髓移植促进早期造血功能重建

目的探讨同种异基因骨髓腔内骨髓移植(IBM-BMT)对小鼠早期造血功能重建的影响。方法将BALB/c小鼠骨髓单个核细胞(BMNCs)分别用胫骨骨髓腔内注射(IBMI)和尾静脉注射(IV)两种方法移植入经致死量60Coγ射线辐照后的60只C57BL/6小鼠。受鼠随机分为3组:骨髓腔内注射高和低剂量组(IBM1和IBM2组)、尾静脉注射组(IV组),每组20只。在骨髓移植后1、3、6和9d分别计数各组受鼠胫骨骨髓腔内有核细胞总数,并用流式细胞术检测供体植入水平(供体来源有核细胞总数、供体来源髓系细胞数)。结果于移植后6d,IBM1组和IBM2组注射侧胫骨骨髓腔内有核细胞总数、供体来源有核细胞总数、供体来源髓系细胞总数均明显高于IV组(P<0.05或P<0.01?。结论IBM-BMT较IV-BMT更能促进同种异基因骨髓移植后的早期造血功能重建。     

Cellular composition of the spleen after human allogeneic bone marrow transplantation

The cellular composition of the spleen has been assessed in 18 patients who died 15-326 days after receiving allogeneic marrow for leukaemia. The white pulp showed marked lymphocyte depletion with no germinal centres, very few B cells, and rare plasma cells. The marginal zone was unrecognizable but there were moderate numbers of T cells in the periarteriolar lymphatic sheaths (PALS), showing great variation in CD4/CD8 ratio. The percentage of CD4+ cells decreased with time post transplant. CD8+ cells were reduced in patients with graft-versus-host disease (GvHD) who also showed no increase in cells staining for activation markers. No T cells were detected expressing immature phenotypes and no differences were detected between patients who received marrow purged or unpurged of T cells. Macrophage numbers appeared normal. Extramedullary haemopoiesis (EMH) was predominantly in the red pulp greater than 30 days after transplantation but more commonly in the white pulp before then. Pyknotic cells were common in seven cases and appeared to be associated with EMH rather than GvHD. Chimaeric studies demonstrated small numbers of donor cells in the PALS at 26 days and larger numbers at 56 days.     

Stromal changes in leukaemic and related bone marrow proliferations

The stromal structure of the bone marrow was studied in 96 cases of leukaemia and related disorders.     

Ultrastructural changes in guinea-pig bone marrow basophils during anaphylaxis

A sequence of ultrastructural changes related to the degrees of severity of the anaphylactic reaction was observed in the bone marrow basophils of guinea-pigs. This sequence of changes might represent the sequential stages of a process of degranulation which consisted of (1) formation of haloes of low electron-density around the granules, (2) development of vacuoles by intercommunicating the adjacent haloes and of confluent vacuoles by joining the small vacuoles, and (3) disruption of cell membrane with concomitant expulsion of granules into the intercellular space or sinusoid. The vacuoles might communicate with the cisternae of the endoplasmic reticulum. Degranulation was found to be a result of an antigen—antibody reaction in which the reaginic antibodies produced had a selective affinity for the cell membrane of the basophils. The mechanism of degranulation might involve two factors: a physicochemical factor of changing the nature of the cell membrane resulting from the antigen—antibody interaction, and a mechanical factor of pressure exerted on the cell membrane by the confluent vacuoles. The alterations in the granules themselves, such as relatively larger size, lighter electron-opacity and looser texture, appeared to be associated with the histamine release from the granules. The structural changes of the granule contents encountered in anaphylaxis suggest that the ultrastructure of the basophil granules may be composed of an orderly interlacing framework of microfilaments onto which other constituent substances are bound.     

Heterotopically induced bone marrow. I. Cellular composition of bone marrow derived from the heterotopic ossicles induced in mice by xenogeneic epithelia of human amnion and dog's transitional epithelium.

Following heterotopic osteogenesis by implantation of xenogeneic epithelia (FL and WISH cell line, transitional epithelium of dog) in mice a biogenesis of hemopoietic tissue among induced ossicles is observed. Precursors and mature forms of all types of blood cells are found in the induced bone marrow. The concentration of lymphocytes in the induced bone marrow is higher, and that of erythropoietic cells lower as compared with orthotopic femur bone marrow. The yield of myeloid cells varied from 0.14 to 3.61 x 10(6) cells per induced bone-containing nodule.     

Cytokines transduced bone marrow stromal cell lines promote immunohematopoietic reconstitution in mice after allogeneic bone marrow transplantation

Impaired immune reconstitution following allogeneic T-cell depleted bone marrow transplantation (allo-TCD-BMT) is a major obstacle to its clinical application. Stromal cell line QXMSC1, established from bone marrow cells of BALB/c(H-2d), was transfected with murine IL-3 and/ or IL-2 gene, and injected into lethally irradiated C57BL/6(H2b) mice. We evaluated its effects on immunologic and hematopoietic reconstitution after allo-TCD-BMT. The results showed that QXMSC1-IL-3 + IL-2 could significantly increase the numbers of hematopoietic primitive progenitors (CFU-S), committed progenitors (CFU-GM, and BFU-E), and lymphocytes (CD8+ cells, CD4+ cells, and B cells). Similarly, immune functions of recipient mice were significantly enhanced in the QXMSC1-IL-3 + IL-2 group. In addition, QXMSC1-IL-3 or QXMSC1-IL-2 also exerted apparent effects on accelerating immune reconstitution, but these effects were far less than that of QXMSC1-IL-3 + IL-2. Our results demonstrated that stromal cell-mediated IL-3 and IL-2 gene therapy may be a potent approach in promoting immunologic and hematopoietic reconstitution after allo-TCD-BMT.