Influence of SHIP on the NK repertoire and allogeneic bone marrow transplantation

Natural killer cell (NK) receptors for major histocompatibility complex (MHC) class I influence engraftment and graft-versus-tumor effects after allogeneic bone marrow transplantation. We find that SH2-containing inositol phosphatase (SHIP) influences the repertoire of NK receptors. In adult SHIP-/- mice, the NK compartment is dominated by cells that express two inhibitory receptors capable of binding either self or allogeneic MHC ligands. This promiscuous repertoire has significant functional consequences, because SHIP-/- mice fail to reject fully mismatched allogeneic marrow grafts and show enhanced survival after such transplants. Thus, SHIP plays an important role in two processes that limit the success of allogeneic marrow transplantation: graft rejection and graft-versus-host disease.     

Male-specific antigen: modification of potency by the H-2 locus in mice

Skin grafts from C57BL/10 (B10) male mice survive significantly longer than do B10.BR male skin grafts on (B10 x X B10.BR)F(1) females. This indicates that the H-2 locus influences the potency of the male-specific antigen in mice.     

Histocompatibility-2 (H-2) polymorphism in wild mice

Red blood cells of 40 wild mice captured at four different locations in the Ann Arbor area were typed for the presence of 15 different H-2 antigens by direct hemagglutination test. Fifteen different phenotypes were found and all were different from those determined by known H-2 alleles of the laboratory mice. Great similarity between phenotypes of mice from the sane location was observed. This is interpreted as further evidence for the deme structure of natural mouse population.     

Homograft rejection delayed by treatment of donor tissue in vitro with antilymphocyte serum

Treatment of rabbit corneal tissue in vitro with pooled rabbit serum delays the onset of corneal homograft rejection in the host. Addition of antilymphocyte serum results in a further significant delay in the onlset of rejection. The mechanisms by which such treatment of donor tissues may modify the antigenic content of the material are discussed.     

Circadian periodicity of bone marrow mitotic activity and reticulocyte counts in rats and mice

Mitotic proliferation in the bone marrow of female rats and mice kept under standardized coniditions with light from 6:00 a.m. to 6:00 p.m. exhibited a significant circadian periodicity with the greatest activity occurring from 6:00 a.m. to 12:00 noon. The reticulocyte levels in peripheral blood were highest at 8:00 a.m.     

海藻多糖对小鼠骨髓放射性急性损伤的保护作用

目的：观察中药海藻多糖对小鼠骨髓放射性急性损伤的保护作用。方法：检测小鼠骨髓有核细胞总数、DNA含量及嗜多色性红细胞微核率。结果：小鼠接受2Gy γ射线照射后，小鼠骨髓有核细胞总数、DNA含量明显降低(P＜0．01)，骨髓嗜多色性红细胞微核率明显升高(P＜0．01)；20、40mg/kg剂量的海藻多糖能有效防止照射小鼠骨髓有核细胞总数、DNA含量降低(P＜0．05)；10、20、40mg/kg剂量的海藻多糖能防止照射小鼠骨髓嗜多色性红细胞微核率升高(P＜0．05)。结论：中药海藻多糖预防性给药对小鼠骨髓放射性急性损伤有一定的保护作用。     

Recognition of H-2 types in relation to the blocking of pregnancy in mice

Inbred BALB/c females were mated and subsequently exposed in a divided cage to "stimulus" males or females whose H-2 type was similar or dissimilar to the stud male's. The incidence of pregnancy blocking was considerably higher when stud and stimulus males differed in H-2 type than when they did not. Similar results were obtained with urine samples of H-2 identical and nonidentical males. Females exposed after mating to other females whose H-2 type differed from the stud male, under the same experimental conditions, also showed an appreciable incidence of pregnancy block. It is therefore concluded that chemosensory recognition of H-2 types affects the reproductive hormonal status of the pregnant female.     

Donor-derived cells in the central nervous system of twitcher mice after bone marrow transplantation

The twitcher mouse is an animal model of galactosylceramidase deficiency, comparable to Krabbe's disease, a lysosomal storage disease in humans. As in most lysosomal storage diseases, neurological deterioration is a prominent feature of the disease in these mice. Transplantation of enzymatically normal congenic bone marrow was earlier found to result in prolonged survival and increased levels of galactosylceramidase in the visceral organs of twitcher mice. It is now reported that bone marrow transplantation results in increased galactosylceramidase levels in the central nervous system (CNS). Concomitantly, the levels of psychosine, a highly toxic lipid that progressively accumulates in the CNS of untreated twitcher mice, stabilized at much lower levels in the CNS of treated twitcher mice. Histologically, a gradual disappearance of globoid cells, the histological hallmark of Krabbe's disease, and the appearance of foamy macrophages capable of metabolizing the storage product were seen in the CNS. By immunohistochemical labeling it was demonstrated that these foamy macrophages were of donor origin. The infiltration of enzymatically competent, donor-derived macrophages was accompanied by extensive remyelination in the CNS. It is concluded that after bone marrow transplantation, donor-derived macrophages infiltrate the affected brain tissue and are capable of inducing a partial reversal of the enzyme deficiency.     

Drug-induced tolerance for skin allografts across the H-2 barrier in adult mice

Mice treated with methylhydrazine derivatives for 3 to 4 weeks before the transplantation showed markedly prolonged survival times for allogeneic skin grafts differing at the H-2 histocompatibility locus. Presumably permanent tolerance was induced in about 20 to 30 percent of the mice when the drug treatment was combined with a single injection of additional donor antigen. The tolerance persists without further drug treatment and is specific for donor tissue.     

Prevention of organ allograft rejection by a specific Janus kinase 3 inhibitor

Because of its requirement for signaling by multiple cytokines, Janus kinase 3 (JAK3) is an excellent target for clinical immunosuppression. We report the development of a specific, orally active inhibitor of JAK3, CP-690,550, that significantly prolonged survival in a murine model of heart transplantation and in cynomolgus monkeys receiving kidney transplants. CP-690,550 treatment was not associated with hypertension, hyperlipidemia, or lymphoproliferative disease. On the basis of these preclinical results, we believe JAK3 blockade by CP-690,550 has potential for therapeutically desirable immunosuppression in human organ transplantation and in other clinical settings.     

Engraftment and development of human T and B cells in mice after bone marrow transplantation.

A model for human lymphocyte ontogeny has been developed in a normal mouse. Human bone marrow, depleted of mature T and B lymphocytes, and bone marrow from mice with severe combined immunodeficiency were transplanted into lethally irradiated BALB/c mice. Human B and T cells were first detected 2 to 4 months after transplantation and persisted for at least 6 months. Most human thymocytes (30 to 50 percent of total thymocytes) were CD3+CD4+CD8+. Human immunoglobulin was detected in some chimeras, and a human antibody response to dinitrophenol could be generated after primary and secondary immunization.     

A substance in liver which inhibits thymine biosynthesis by bone marrow

A substance was extracted from rabbit liver which inhibited the incorporation of formate into bone-marrow thymine in vitro. In view of the important role of thymine biosynthesis in cell division, it is suggested that the inhibitor present in liver is a naturally occurring mitosis inhibitor.     

BMP4在诱导骨髓间充质干细胞向生殖细胞分化中的作用

为了探讨骨形态发生蛋白4(bone morphogenetic protein 4,BMP4)在诱导骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)向生殖细胞分化中的作用,将全骨髓培养法获取小鼠BMSCs,在含10%胎牛血清(fetal bovine serum,FBS)的DMEM培养液中培养传代;取生长状态良好的第三代(P3)BMSCs,分别添加不同浓度(5、10、20、40和80ng·mL~(-1))的BMP4作为实验组,未添加BMP4为对照组。4d后,MTT法和台盼蓝染色检测细胞的增殖率和存活率,实时定量PCR(RT-qPCR)检测生殖细胞阶段特异性基因(Oct4、Dazl、Fragilis、Mvh、Nobox、Stella、Stra8和Gdf9)的表达。结果发现,BMP4浓度为5~20 ng·mL~(-1)时细胞存活率和增殖率最高;BMP4浓度为20 ng·mL~(-1)组Mvh、Fragilis、Oct-4、Dazl及Stella表达水平最高,均显著高于对照组(P0.05);Nobox表达水平则在BMP4浓度40ng·mL~(-1)组最高,且各浓度组均显著高于对照组(P0.05);各浓度BMP4组Stra8和Gdf9的表达水平均与对照组间无显著性差异(P0.05)。此结果提示,BMP4浓度在20 ng·mL~(-1)时,BMSCs的存活率、增殖率及原始生殖细胞特异性基因的表达最高;BMP4在诱导间充质干细胞向生殖细胞分化过程中起重要作用。     

骨髓间充质干细胞移植在卵巢早衰小鼠卵巢功能重建中应用

探讨骨髓间充质干细胞(BMSCs)移植在卵巢早衰小鼠卵巢功能重建中的应用价值。通过一次性腹腔注射环磷酰胺和白消安建立小鼠卵巢早衰(POF)模型,并随机分为建模组、BMSCs移植组和空白对照组。移植组于建模14 d经尾静脉注射绿色荧光蛋白(GFP)转基因鼠BMSCs悬液(5×10~7个·mL~(-1)),每4 d重复1次,共5次;建模组和空白对照组注射等量生理盐水。观察建模后及BMSCs移植后小鼠的一般情况、血清FSH和E2水平、卵巢组织学变化以及GFP荧光信号在卵巢内的分布。结果发现,建模组小鼠食欲明显减退,活动减少,与空白对照组相比体重显著减轻(P0.05),FSH水平显著增高(P0.05),E2水平显著降低(P0.05),卵巢体积减小,间质纤维化严重,始基卵泡、生长卵泡和成熟卵泡数均显著减少(P0.05)。BMSCs移植后,移植组小鼠体重增加,食欲增强,活动增多;28 d后FSH浓度显著低于建模组(P0.05),E2浓度、始基卵泡、生长卵泡数及成熟卵泡数均显著高于建模组(P0.05),但各项指标仍与空白对照组间存在显著性差异;荧光显微镜下可见绿色荧光信号散在分布于卵泡周围。此结果提示,BMSCs可在小鼠损伤卵巢组织中定位存活,对卵巢组织结构及内分泌功能有修复作用。     

Prevention of type I diabetes in nonobese diabetic mice by virus infection

The nonobese diabetic (NOD) mouse is an animal model of type I diabetes and develops a characteristic autoimmune lesion in the islets of Langerhans with lymphocytic infiltration and destruction of pancreatic beta cells. The result is hypoinsulinemia, hyperglycemia, ketoacidosis, and death. Diabetes usually begins by the sixth month of age but can occur earlier when young NOD mice are infused with lymphocytes from older NOD donors. When newborn or adult NOD mice were infected with a lymphotropic virus they did not become diabetic. The interaction between viruses and lymphocytes is pivotal in aborting diabetes, as established by experiments in which lymphocytes from virus-infected donors failed to transfer diabetes. In contrast, lymphocytes from age- and sex-matched uninfected donors caused disease. Therefore, viruses and, presumably, their products can be developed to be beneficial and may have potential as a component for treatment of human diseases. Further, these results point to the utility of viruses as probes for dissecting the pathogenesis of a nonviral disease.     

Transplantation of allogenic bone marrow in canine cyclic neutropenia

Transplantation of normal bone marrow cells to a gray collie dog with cyclic neutropenia resulted in normal granulocytopoiesis. The finding suggests that cyclic neutropenia occurs because the hematopoietic stem cells are defective. Because of the similarity of human and canine cyclic neutropenia, it also suggests that the human disease may be curable by marrow transplantation.     

山羊BST-2基因的表达及其亚细胞定位

将山羊的BST-2基因 (Capra hircus bone marrow stromal antigen 2 gene)分别克隆至原核表达载体pGEX-4T-1和真核表达载体p3×Flag-CMV-14中,获得了重组表达质粒pGEX-BST-2和p3×Flag-BST-2。将重组质粒pGEX-BST-2转化至E.coli BL21(DE3)感受态细胞中,用IPTG进行诱导表达。SDS-PAGE及Western blot结果显示,诱导表达的重组蛋白分子量约为42 ku,与预期蛋白一致。利用脂质体介导转染法将重组质粒p3×Flag-BST-2转染Vero细胞,经激光共聚焦显微镜检测,结果显示,BST-2蛋白在细胞中可以良好表达,并主要定位于细胞膜上。