The role of neutrophil membrane glycoprotein GP-150 in neutrophil adherence to endothelium in vitro

We have previously described two patients with a congenital defect in neutrophil function characterized by an inability to form pus. The patients' neutrophils lack a membrane glycoprotein of mol wt 150,000 daltons (GP-150) on analysis by SDS-PAGE. This glycoprotein is part of a membrane antigen complex recognized by the murine monoclonal antibody (MoAb) 60.3. Addition of MoAb 60.3 to normal neutrophils produces defects in chemotaxis and phagocytosis in vitro similar to those observed in the patients. Since neutrophil adherence to vascular endothelium is prerequisite to neutrophil emigration in vivo, we examined the interaction of the patients' neutrophils and normal neutrophils treated with MoAb 60.3 with cultured endothelium. Adherence was determined as the percentage of 51Cr-labeled purified peripheral blood neutrophils which remained adherent to plastic wells or endothelial monolayers after a 45-minute incubation at 37 degrees C. The percentage of neutrophils from patient 1 remaining adherent to uncoated, fibronectin-coated, or laminin-coated plastic was similar to that observed in normal neutrophils (55% to 84% adherence with normal neutrophils v 73% to 78% adherence with the patient's neutrophils and 63% to 82% adherence with MoAb 60.3-treated normal neutrophils). The adherence of the neutrophils from patient 1 and MoAb 60.3-treated normal neutrophils to human or bovine endothelium in serum-free medium was also not significantly different from that observed in normal neutrophils (less than 10% adherence with normal, MoAb 60.3-treated, and patient neutrophils). In medium containing 10% autologous or heterologous human plasma, however, the adherence of neutrophils from patient 1 or MoAb 60.3-treated normal neutrophils to endothelial monolayers was significantly reduced (35% +/- 7% of normal neutrophils in seven experiments). Although phorbol myristate acetate (PMA) (10 ng/mL) and calcium ionophore A23187 (10(-5) mol/L) markedly increased the adherence of normal neutrophils to endothelial monolayers in serum- free medium (40% to 85% adherence), neither agent increased the adherence of the neutrophils from patient 1 or normal neutrophils treated with MoAb 60.3 (less than 5% adherence). The adherence of PMA- activated neutrophils from patient 2 to endothelial monolayers was also markedly decreased when compared with that of normal neutrophils. Postsecretory cell-free supernatants from PMA-activated normal neutrophils failed to augment adherence of neutrophils from patient 1 (less than 5% adherence).(ABSTRACT TRUNCATED AT 400 WORDS)     

Occupational exposure to human immunodeficiency virus and hepatitis B virus: a comparative analysis of risk.

OBJECTIVE: To estimate the occupational risk from infection with the human immunodeficiency virus (HIV) in terms of loss of (quality-adjusted) life expectancy, and to compare that risk to those posed by other hazards faced by health care workers. DESIGN: Decision-analytic model. RESULTS: For a 30-year-old female health care worker (unvaccinated for hepatitis B virus [HBV]), the loss of life expectancy from a needlestick from a symptomatic HIV-positive (HIV+) patient is 39 days (range, 17 to 93 days), as compared with a loss of 17 days from a needlestick from a patient who is hepatitis-B-surface-antigen-positive (HBsAg+), and 38 days from a needlestick from a patient who is hepatitis-B-e-antigen-positive (HBeAg+). When morbidity is included in the analysis of risk (through calculation of the quality-adjusted loss of life expectancy), the risk from both HBV and HIV increases. The quality-adjusted loss of life expectancy due to a needlestick exposure from a symptomatic HIV+ patient is 45 days (range, 20 to 108 days), as compared with a quality-adjusted loss of life expectancy of 48 days from a needlestick from an HBsAg+ patient, and 109 days from a needlestick from a patient who is known to be HBeAg+. By comparison, a cross-country automobile trip is associated with a loss of life expectancy of approximately 1 day. The 45- to 50-day loss of quality-adjusted life expectancy from percutaneous exposures to HIV and HBV is approximately the same magnitude as the gain in life expectancy from 10 years of annual screening for breast cancer with mammography and physical examination. CONCLUSIONS: The risk associated with percutaneous exposures to symptomatic HIV+ patients is comparable to other risks that health care workers have faced knowingly and have accepted in the recent past. However, the loss of quality-adjusted life expectancy associated with a needlestick exposure is significant. Identification of cost-effective methods that increase the safety of medical personnel but also ensure full access to high-quality care for HIV+ patients should be a high priority.     

Granulocyte-macrophage colony-stimulating factor expression by human fibroblasts is both upregulated and subsequently downregulated by interleukin-1

Interleukin-1 (IL-1) treatment of human WI-38 lung fibroblasts results in granulocyte-macrophage colony-stimulating factor (GM-CSF) expression as well as a delayed increase in prostaglandin E2 (PGE2) production that closely correlates with the decline of GM-CSF mRNA levels. Pretreatment with PGE2 reduces the IL-1 induced GM-CSF mRNA and protein expression to 10% to 15% of control values at concentrations of PGE2 that are endogenously produced after IL-1 stimulation. Inhibition of PGE2 synthesis by indomethacin prolongs the IL-1 induced GM-CSF mRNA expression and increases the cumulative GM-CSF protein secretion. Exposure of WI-38 fibroblasts to PGE2 results in an increase in intracellular cyclic adenosine monophosphate (cAMP) levels. The inhibition of GM-CSF expression by PGE2 can be mimicked by stable cAMP analogs as well as cAMP elevating agents such as cholera toxin, forskolin, and isobutylmethylxanthine. Thus the inhibition exerted by PGE2 is mediated via cAMP. Taken together, these results suggest that IL-1 stimulation of human fibroblasts provides not only the upregulatory signal for GM-CSF expression but also a delayed and indirect downregulatory signal that serves to limit GM-CSF expression in the continued presence of IL-1.     

Bispecific-armed, interferon gamma-primed macrophage-mediated phagocytosis of malignant non-Hodgkin's lymphoma

To show that macrophages can be effectively targeted against malignant B cells, bispecific antibodies (BsAb) were constructed from two antibodies having specificity for the high-affinity Fc receptor for IgG (Fc gamma RI/CD64) and the B-cell differentiation antigens CD19 and CD37. Using a flow cytometry-based assay and confocal imaging, we show that these constructs mediated significant phagocytosis of B lymphocytes by macrophages that could be enhanced with interferon gamma (IFN gamma) and IFN gamma in combination with macrophage colony- stimulating factor. BsAb-dependent phagocytosis was triggered through Fc gamma RI and could be blocked only by using F(ab')2 fragments from the parent molecule or by cross-linking Fc gamma RI. BsAb-dependent phagocytosis was not blocked by antibodies to the other Fc receptors, Fc gamma RII and Fc gamma RIII. Because these antibody constructs bind to an epitope outside the Fc gamma RI ligand binding site, we show that autologous serum, polyclonal IgG, and monomeric IgG1 did not block BsAb- dependent phagocytosis, whereas autologous serum and the IgG fractions blocked parent molecule monoclonal antibody-dependent phagocytosis due to the avid binding of monomeric IgG to Fc gamma RI. Finally, BsAb- mediated phagocytosis was effective against the malignant B cells of patients with mantle cell lymphoma, prolymphocytic leukemia, and chronic lymphocytic leukemia. Based on these studies, we propose that BsAbs may provide an effective means of immunomodulation for patients with B-cell malignancies.     

Cutaneous lymphoblastic lymphoma with pre-B markers

Two children with cutaneous convoluted lymphoblastic lymphoma are reported. Malignant cells from both patients contained cytoplasmic Mu heavy chains characteristic of pre-B-cells and expressed CALLA and la antigens as well. Most cases of convoluted lymphoblastic lymphoma are T- cell-derived neoplasms. The non-T, non-B phenotype found in these two children demonstrates that histology does not necessarily predict immunophenotype. The association of the pre-B phenotype with cutaneous lymphoma has not been previously reported, but may represent a unique clinical-histopathologic-immunologic entity that occurs in young children.     

手术及经皮自体骨髓移植治疗骨延迟愈合和骨不连

观察手术及经皮自体骨髓移植治疗四肢骨折骨不连、骨延迟愈合的临床效果。选择1997-11/2004-02山东省聊城市第二人民医院应用经皮自体骨髓移植法治疗骨不连、骨延迟愈合的患者23例,患者均了解相关治疗目的、方法并同意治疗方案。①17例骨不连患者采用手术治疗。15例存在骨质缺损者在骨折端间隙及周围自体髂骨植骨10例,收集术中骨屑及骨髓原位植骨4例,异体植骨1例;2例单纯外固定架重新固定未植骨并行自体骨髓移植。术后2周抽取自体骨髓注入骨折部位,注射10～20mL/次,每2周1次,需注射2～4次。②6例骨延迟愈合患者仅行自体骨髓移植,未改变固定方式。术后定期随访,观察患者骨折愈合情况。骨折愈合标准:X射线片显示连续骨痂通过折线,骨折处无疼痛及压痛,能够全部负重。23例骨不连、骨延迟愈合患者全部进入结果分析,无脱落。23例患者术后平均随访12.5个月,其中随访7～9个月11例,10～12个月6例,13～15个月5例,16个月1例。23例患者均达到骨性愈合,愈合率100%,平均愈合时间为5.6个月。坚强固定加必要的植骨修复骨缺损是治疗骨折术后骨不连的基础和关键,自体骨髓具有成骨作用,可进一步促进骨折愈合。     

Approaches to preventing or reversing platelet alloimmunization using animal models

Animal transfusion models were established to assess treatment programs for preventing or reversing platelet alloimmunization. Five control baboons given weekly transfusions of radiolabeled platelets from a single unrelated donor became immunized after an average of 2.4 +/- 2.1 transfusions. Similarly, 18 of 21 (86%) dogs given up to eight platelet transfusions from a single unrelated donor became immunized after an average of 2.3 +/- 1.7 transfusions. In six of seven baboons, prednisone or antithymocyte globulin alone or in combination effectively delayed platelet alloimmunization. In contrast, only two of 12 (17%) dogs given prednisone or antithymocyte serum (ATS) resisted alloimmunization. Neither splenectomy nor cyclophosphamide prevented alloimmunization in the baboon. In addition, attempts to reduce the immunogenicity of transfused platelets by inactivating the contaminating leukocytes with gamma radiation or by giving leukocyte-poor platelets were of no benefit in dogs. Reversal of platelet alloimmunization was achieved in two of three dogs treated with ATS and procarbazine hydrochloride. However, neither splenectomy, cyclophosphamide, ATS plus prednisone, nor vincristine sulfate produced any improvement. These studies show that the highly immunogenic nature of platelet transfusions in animals makes feasible the study of the prevention and reversal of platelet alloimmunization.     

In vitro regulation of immunoglobulin synthesis after marrow transplantation. I. T-cell and B-cell deficiencies in patients with and without chronic graft-versus-host disease

Twenty-four patients with aplastic anemia or acute leukemia were treated by marrow grafts from HLA-identical donors after conditioning with high doses of cyclophosphamide and/or today body irradiation. They were studied between 4 and 63 mo (median 14.2) after transplantation. Seventeen patients had chronic graft-versus-host disease (C-GVHD) and 7 were healthy. They were studied for defects in their T- and B-cell function using and indirect hemolytic plaque assay for Ig production after 6 days of culture in the presence of pokeweek mitogen. T or B cells from the patients with or without C-GVHD were cocultured with T or B cells from their HLA-identical marrow donors or unrelated normal controls. Intrinsic B-cell defects, lack of helper T-cell activity, and suppressor T-cell activity were more frequently found in patients with C-GVHD than in healthy patients. Fifteen of the 17 patients with C-GVHD showed on or more defects in their T-and B-cell function compared to only 3 of the 7 patients without C-GVHD. None of the healthy controls, including the marrow donors, showed defects in their T- and B-cell functions. These in vitro findings may be helpful in assessing the process of immune reconstitution and the immunologic aberration found after human marrow transplantation.