Binding of heterologous anti-brain antibodies to mouse B cells.

Antibodies produced in rabbits against mouse brain are able to react with the majority of lymph node and spleen murine lymphocytes, as demonstrated by indirect immunofluorescence. Cells stained by a suitably absorbed rabbit anti-mouse brain serum showed various degrees of fluorescence, from bright fluorescent rings to a few speckles per cell; lymphocytes from the spleen of deprived mice (B mice) predominantly stained in small caps. Double labelling experiments in which B cells were identified by their 'easily detectable' surface immunoglobulin revealed that a great proportion of these cells had two overlapping caps, a large one formed by the anti-immunoglobulin serum and a small one formed by the anti-brain serum. Up to 10 per cent of these spleen B cells, however, appeared as strongly fluorescent as T cells, with the suitably absorbed heterologous anti-brain serum. No anti-immunoglobulin activity was detected in this serum. It is therefore suggested that some B cells may bear on their surfaces a theta or theta-related antigen similar to that on T cells.     

Bone marrow transplantation in the rat. III. Structure of the liver inflammatory lesion in acute graft-versus-host disease.

The liver is a major parenchymal target organ of acute graft-versus-host disease (aGVHD) after bone marrow transplantation in the rat. The authors have analyzed the nature of cellular infiltrates in the liver using monoclonal antibodies against white cell subsets and investigated the anatomic distribution of the inflammatory cell subsets inside the liver parenchyma. Several types of white cells are present in a normal control liver: In the portal area the T-helper (Th) cells predominate, (surface) immunoglobulin-expressing B cells are present in ample numbers, and most of the phagocytes are Ia-positive. In the central vein area the T-suppressor/killer cells (Tsk) dominate, no B cells are present, and most of the phagocytes are Ia-negative. During aGVHD the number of T cells increases rapidly in the portal area; and after an initial strong increase, the Th/Tsk ratio decreases but remains still above 1. In the central vein area there is also an increase in the number of T cells, compared with that in the syngeneic recipient, but the Th/Tsk ratio rapidly decreases and remains uniformly below 1. During aGVHD the B cells entirely disappear from the portal area, whereas a small but distinct number of mature plasma cells with intracellular immunoglobulin appear in the central vein area. Following irradiation the Ia-positive phagocytic cells entirely disappear from the portal area and decrease distinctly in number in the central vein area. During aGVHD the number of Ia-positive phagocytes increases again in both locations. In the central vein area the positive phagocytes are seen over the background level, and, concomitantly, the Ia-negative phagocytes disappear.     

Bone marrow transplantation in the rat. Lytic functions of inflammatory leukocytes during acute graft-versus-host disease

We have isolated inflammatory leukocytes from various lymphoid and parenchymal organs after total body irradiation and bone marrow transplantation from either an allogeneic or syngeneic strain and tested their ability to perform lytic functions in vitro. No direct lytic activity (i.e. cytotoxic T lymphocytes, CTL) to relevant strain-derived target cells in the lymphoid or parenchymal target organs was seen preceding or during acute graft-versus-host disease (aGVHD). Instead, the leukocytes of the spleen and blood and the inflammatory cells of liver and lungs were efficient effector cells against recipient-derived target cells in the presence of relevant antibody (antibody dependent cellular cytotoxicity, ADCC). The NK activity against YAC-1 (natural killer, NK) target cells was first high in the spleen, but when the aGHVD appeared in the allograft marrow recipients the NK activity decreased in the spleen with a concomitant increase in the liver, but not in the other parenchymal target organs. At the same time no NK activity was seen in the syngeneic marrow graft recipients' parenchymal organs. These observations suggest functional differences in the structure of inflammation in the different target organs of aGVHD.     

骨髓干细胞移植治疗不同年龄mdx鼠的疗效研究

目的：研究不同年龄的Duchenne型肌营养不良鼠（mdx鼠）与骨髓干细胞移植后缺失蛋白表达的关系。 方法： 获取4-5周C57BL/6小鼠的骨髓干细胞，体外培养3 d，静脉移植到7Gy γ射线预处理的6周龄、8周龄两组各6只mdx鼠。移植12周后，对移植鼠骨骼肌dystrophin蛋白表达情况进行检测。 结果： 6周龄、8周龄两组mdx鼠，静脉移植1.2×107骨髓干细胞，3个月后，分别有16%和7%的骨骼肌纤维表达了dystrophin蛋白。 结论： 静脉移植同种、同系鼠骨髓干细胞的mdx鼠，3个月之后, 不同年龄mdx鼠骨骼肌细胞dystrophin蛋白表达的阳性率不同，幼年鼠骨髓干细胞移植有较高比率的缺失蛋白表达。     

Effect of NK cells on GVHD in H-2 haploidentical bone marrow transplantation in mice

Objective: To study the effect of natural killer (NK) cells on graft-versus-host disease (GVHD) after H-2 haploidentical bone marrow transplantation (BMT) in mice. Methods :Murine model of H-2 haploidentical BMT was established by using Balb/c (H-2d) mouse as recipient, and Balb/c(H-2d)×C57BL/6 (H-2b) (H-2d/b) mouse as donor. Lethally irradiated Balb/c (H-2d) mice were transplanted with the bone marrow cells from Balb/c(H-2d)×C57BL/6(H-2b) (H-2d/b) mice containing donor spleen cells and/or NK cells. GVHD and survival rates were studied by observation of clinical manifestations and pathological changes. Results:In the group of bone marrow +spleen cells, GVHD was induced in 90% mice; but in the group plus with low amount of NK cells,GVHD was induced in 20% mice; and in the group transplanted with high amount of NK cells, GVHD was induced only in 10% mice. Compared to the group transplanted only with BM plus spleen cells, the incidences of GVHD in the latter two groups decreased significantly (P＜0.01) and the survival rates at different periods of 15, 30, 45 and 60 days increased obviously (P＜0.01 ). Conclusion: In mouse H-2 haploidentical BMT, alloreactive NK cells can reduce the incidence of GVHD and increase the survival rate.     

Bone marrow transplantation in the rat. I. Histologic correlations and quantitation of cellular infiltrates in acute graft-versus-host disease.

In an attempt to define which cytologic manifestations of inflammation are characteristic of acute graft-versus-host disease (aGVHD), the authors have analyzed hematologic reconstitution in the bone marrow, spleen, and blood of bone marrow transplant recipients and correlated these events to concomitant cytologic changes in the parenchymal target organs. After bone marrow transplantation from Lewis to BN strain, the strongest inflammatory changes were observed in the liver, a "model" parenchymal target organ for aGVHD in this strain combination. The inflammatory episode of the aGVHD in the liver was characterized by an early lymphoid blastogenesis, the presence of large granular lymphocytes (LGLs), lymphocytosis, and some monocytosis, lacking or significantly less prominent in the liver of syngeneic BN to BN recipients. Concomitantly with the infiltration of the liver with LGLs and lymphocytes, these cells were depleted from blood; and with their disappearance from the liver, they appeared in the recipient spleen. Lack of lymphoid blastogenesis in the bone marrow of allograft recipients and similar though less prominent cytologic changes in the syngeneic graft recipients, make it difficult to differentiate aGVHD-associated changes from normal reconstitution in the lymphoid tissue paper; the minimal changes in the blood make this organ the least suitable site for the monitoring of the aGVHD in the rat.     

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。     

Serum cyclosporine concentration and risk of acute graft-versus-host disease after allogeneic marrow transplantation

To determine the relation between the serum cyclosporine concentration and the risk of acute graft-versus-host disease (GVHD), we studied 179 recipients of bone marrow grafts from HLA-identical sibling donors who received prophylaxis with cyclosporine, either by itself or combined with methotrexate. Cyclosporine was given either orally or intravenously at full doses from the day before transplantation until day 50; it was then tapered off and discontinued on day 180. Trough concentrations of serum cyclosporine were measured by radioimmunoassay. The relation between patients' characteristics and the risk of acute GVHD was analyzed with a relative-risk regression model. In 66 patients (37 percent), grades II to IV of acute GVHD developed 7 to 66 days (median, 13) after transplantation. The trough cyclosporine concentration for a given week was significantly associated with the risk that acute GVHD would develop during the following week. The relative risks were 0.7 (i.e., there was a 30 percent reduction in risk) for every increase of 100 ng per milliliter in cyclosporine concentration and 1.0, 0.60, and 0.20 for concentrations of less than 100, 100 to 199, and 200 or more ng per milliliter, respectively (P less than 0.01). A patient's age, prophylaxis regimen, and year of transplantation also influenced the risk of acute GVHD significantly. These data indicate that low cyclosporine concentrations can be a cause of treatment failure and that concentrations should be monitored in recipients of marrow transplants.     

Candida glabrata sepsis secondary to oral colonization in bone marrow transplantation.

Candida glabrata has emerged as a common cause of fungal sepsis in bone marrow transplant patients, particularly those receiving fluconazole prophylaxis. Colonization of the lower GI tract and indwelling catheters have been thought to be the primary sources of systemic infection with Candida. We report on a bone marrow transplant patient who developed Candida glabrata sepsis from pre-existing oral colonization.     

Foot-and-mouth disease virus-neutralizing antibodies induced in mice by anti-idiotypic antibodies.

A neutralizing monoclonal antibody (nmAb) to foot-and-mouth disease virus (FMDV) was used as antibody-1 (AB1) to induce anti-idiotypic antibodies (a-IdAb) in rabbits. The rabbit a-IdAb (AB2) were isolated on protein A-Sepharose, followed by cycles of separation on idiotype and isotype affinity columns. The specificity of the AB2 for the paratope of AB1 was determined by direct binding to AB1 in solid-phase radioimmunoassay (SP-RIA), and by competition RIA (C-RIA) with virus for binding to the AB1. The AB2, termed a-2PD11, was utilized to immunize six groups of female Swiss mice at weekly intervals with either one of three formulations, in doses of 50 micrograms or 5 micrograms, given in single subcutaneous (s.c.) spots. Anti-viral antibody (AB3) was first detected by RIA at the fifth week in the 50 micrograms/dose groups, and maximum levels were reached at the sixth week in the 50 and 5 micrograms/dose groups. The AB3 levels were at least three times higher for mice given 50 micrograms doses. In addition, the AB3 were also shown to neutralize FMDV infectivity in tissue culture and in a suckling mouse protection assay. Overall, mice exhibited variable responses to immunization with AB2. In a subsequent trial, mice received multispot s.c. and footpad injections of 50 micrograms of a-2PD11 coupled to keyhole limpet haemocyanin (KLH) on a weekly basis. In these mice, AB3 was detected earlier than in mice immunized with single s.c. injections. These results support the use of a-IdAb as potential surrogates of critical determinants for FMD vaccines.     

Graft-versus-host disease in allogeneic bone marrow transplantation: the role of monoclonal antibodies in prevention and treatment

Reconstitution of an individual's haemopoietic stem cells by bone marrow transplantation (BMT) is the recommended treatment for a number of haematological conditions, both malignant and non-malignant. Despite evolution in BMT technology over the past forty years, graft-versus-host disease (GvHD) remains a major, potentially lethal complication. GvHD normally affects the skin, liver and gastrointestinal tract, resulting in a high rate of morbidity. The standard prophylaxis for GvHD is a combination of methotrexate and cyclosporin A, but this is only partially effective. Acute GvHD is difficult to treat and many patients are resistant to steroid therapy. Alternative methods of prevention and treatment are now being sought, and include monoclonal antibodies (MAbs) which target T cells and cytokines. T-cell depletion of donor marrow using rat MAbs reduces the incidence of GvHD but can increase the chances of leukaemic relapse. Mouse MAbs also have been used but some produce severe side-effects. The most successful MAbs are those linked to toxins, and these immunotoxins (IT) have proved very effective in reversing steroid-resistant acute GvHD. MAb therapies are becoming increasingly important in the treatment and prevention of GvHD, and could replace steroids as the main treatment option in some situations. It is predicted that, ultimately, peripheral blood stem-cell transplantation will replace the use of BMT; however, this alternative stem-cell source will not remove the GvHD risks associated with allogeneic stem-cell transplantation. Therefore, reducing the risks will remain a major challenge in the successful allogeneic transplantation of haemopoeitic stem cells for the foreseeable future.     

Graft-versus-host disease in murine bone marrow transplantation. II. Modulation of acute and chronic GVHD in mice receiving bone marrow allografts pretreated with immunosuppressive factor derived from a human T cell line

BALB/c bone marrow treated with monoclonal anti-Thy 1.2 antibody and complement is unable to produce prolonged hemopoietic repopulation and survival when transplanted to lethally-irradiated allogeneic CBA recipient mice. Preincubation of the antibody treated bone marrow cells with an immunosuppressive factor (SAF) derived from a 6-thioguanine resistant cell line, itself derived from the human T cell line CEM, in contrast, allowed those bone marrow cells to produce a state of chimerism and long term survival. Parameters designed to gauge the degree of graft-versus-host reactivity (GVHR) in these animals suggested that acute GVHR was abolished with this procedure. Moreover, defining chronic GVHD as associated with abnormally high spontaneous proliferation of splenic cells, elevated anti-host mixed lymphocyte reactivity, or elevated serum immunoglobulin levels (in all cases when compared with the syngeneically repopulated BALB/c----BALB/c), our data suggest that preincubation with SAF modified chronic GVHD also.