A southern analysis of Rh blood group genes: association between restriction fragment length polymorphism patterns and Rh serotypes

Polymorphisms within the Rh blood group system have been defined by serologic agglutination methods, but have not yet been defined at the DNA level. Two closely related genes associated with the Rh D antigen and with the Rh C/c and E/e antigens have been cloned. We used a Southern analysis incorporating probes to the 5' and 3' regions of the Rh C, E gene and D gene to identify polymorphisms associated with Rh C/c and E/e antigens, respectively. The D gene dosage could be determined by comparing the relative intensities of the D bands with bands from the 5' and 3' region of the Rh C, E gene. The concordance between restriction fragment length polymorphism (RFLP) patterns and serologic phenotypes for 102 randomly selected blood donors was 100% for C, e, and D, 94.8% for c, and 94.3% for E. The data are consistent with the sequences encoding the C/c epitopes residing on the 5' side of those for the E/e epitopes. All samples discordant for the 3' probe and E had the cE (r") serotype. These data show that the gene coding for the cE serotype is different in Rh-positive and -negative individuals. The study demonstrates that Rh DNA typing, including D gene dosage measurements and Rh gene haplotyping, may supplement traditional serotyping methods in transfusion medicine.     

Hematopoiesis in vitro coexists with natural killer lymphocytes

The role of natural killer (NK) lymphocytes in the regulation of human hematopoiesis is controversial. NK-mediated inhibition of colony formation of hematopoietic progenitor cells has been irregularly reported for various cell lineages. In an effort to clarify such disparate findings, we studied the interaction of clearly defined NK and partially purified progenitor cell populations. Cell sorter purified CD16 positive blood NK cells and enriched autologous marrow progenitors were co-incubated at various lymphocyte to marrow cell ratios and then cultured in methylcellulose. There was no inhibition of myeloid, erythroid, or mixed colony formation. Similarly, activation of CD16 positive lymphocytes by interleukin-2 (IL-2) before co-incubation and co-culture did not result in inhibition of colony formation. Furthermore, in a newly designed assay system, we demonstrated that NK cells, which did not modulate colony-formation, remained capable of recognizing and killing rare K562 target cells seeded within the marrow cell population. Our results indicate that unstimulated and IL-2 activated isolated blood NK cells coexist with functioning autologous marrow progenitors in vitro.     

Interleukin-1 induces interleukin-8 secretion from endothelial cells by a juxtacrine mechanism

Inflammation is characterized by migration of neutrophils through the endothelium, and the chemokine interleukin-8 (IL-8) appears to be involved. We asked whether adherence of cells bearing a membrane-form of interleukin 1 (IL-1) induces IL-8 secretion from human umbilical vein endothelial cells (HUVEC) and fibroblasts. Human peripheral blood mononuclear cells (PBMC) were stimulated with endotoxin for 12 hours and then fixed for 4 hours with paraformaldehyde. When these cells were added to HUVEC or fibroblasts, IL-8 production was induced. This stimulation by fixed PBMC was attributed to IL-1, because pretreatment of HUVEC or fibroblasts with IL-1 receptor antagonist (IL-1Ra) reduced the induction by 95% and 80%, respectively, P < .005. Using anti-IL-1 alpha monoclonal antibodies, reduction was complete, whereas anti-IL-1 beta had no effect. IL-1 alpha was shown on the surface of monocytes by fluorescence-activated cell sorter (FACS) analysis. Blockade of IL-1 receptors on PBMC did not affect the activity of membrane-associated IL- 1 alpha, indicating that IL-1 is not anchored to the membrane through its receptors. However, PBMC treated with D-mannose before fixation resulted in a loss of activity; this loss of activity was associated with release of IL-1 alpha, not IL-1 beta, into the supernatant. Thus, anchoring of IL-1 alpha to the membrane may be via a lectin or mannose receptor-like interaction. Blockade of membrane IL-1 alpha required a 30-fold and fivefold excess of IL-1Ra compared with the amount required to block soluble IL-1 beta and IL-1 alpha, respectively. We conclude that the fixed PBMC IL-8 inducing activity is almost entirely caused by IL-1, that this represents IL-1 alpha bound to a surface lectin or mannose receptor on the monocyte, and that it functions in inflammation via juxtacrine interactions.     

Evidence favoring lineage fidelity in acute nonlymphocytic leukemia: absence of immunoglobulin gene rearrangements in FAB types M4 and M5

The concept of lineage fidelity in acute leukemia has recently been challenged by the finding of rearrangements of the immunoglobulin heavy chain genes in a leukemic cell line and in a small number of sporadic cases of acute nonlymphocytic leukemia with a monocytic phenotype. We therefore screened leukemic blood or bone marrow samples of 33 adult patients with acute nonlymphocytic leukemia of FAB types M4 (23 patients) and M5 (10 patients); 28 were obtained at diagnosis and 5 at relapse. All cases were well characterized pathologically and histochemically. Cytogenetic analysis performed in each case demonstrated karyotypes that were representative of those generally seen in these types of leukemia, with a clonal abnormality present in all except 9 of 32 patients who were successfully studied. DNA prepared from each sample was digested with the restriction enzyme BamH1 and analyzed by Southern blot hybridization to probes for the JH region of the immunoglobulin heavy chain. All 33 cases had DNA retained in the germline configuration with no evidence of rearrangement. This finding supports the concept of lineage fidelity, and suggests that true interlineage infidelity, myeloid to lymphoid, is a rare occurrence in adult acute nonlymphocytic leukemia.     

The effects of three recombinant growth factors, IL-3, GM-CSF, and G- CSF, on the blast cells of acute myeloblastic leukemia maintained in short-term suspension culture

The blast stem cells of acute myeloblastic leukemia (AML) respond in cell culture to growth factors by both self-renewal and terminal divisions. Both of these functions have been shown to be stimulated by the recombinant growth factors granulocyte-macrophage colony- stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF). In this paper, recombinant gibbon interleukin-3 (IL-3), homologous to human IL-3, was tested on blast cells and compared with the effects of GM-CSF, G-CSF, and medium conditioned by the bladder cell line 5637 (5637-CM). We found that IL-3 was an effective stimulator of blast renewal and terminal divisions. However, great patient-to-patient variation was found. A graphic method of presenting complex comparisons between growth factors is also included.     

经心内膜右房线形消融治疗心房颤动的安全性评价

为探讨经心内膜右房线形消融治疗心房颤动（简称房颤）的安全性，１２只犬以乙酰胆碱静脉滴注和（或）电刺激建立房颤模型，观察射频导管消融前、后实验犬的病理生理变化。结果显示：①与消融前相比，消融后窦性心率（１５０．８２±３６．７１ｂｐｍｖｓ１６３．６７±３０．９９ｂｐｍ）、窦性Ｐ波时限（７３．６４±１６．８０ｍｓｖｓ６９．５８±１２．１４ｍｓ）、ＰＲ间期（１２０．７３±２６．２９ｍｓｖｓ１１４．０２±１９．２１ｍｓ）、校正窦房结恢复时间（７６．２５±１８．８７ｍｓｖｓ７２．５０±１１．９０ｍｓ）、右房压力（０．４９±０．０６ｋＰａｖｓ０．４６±０．０８ｋＰａ）以及血浆心钠素（０．４８±０．１１ｎｇ／ｍｌｖｓ０．５０±０．０７ｎｇ／ｍｌ）变化均无显著性差异（Ｐ均＞０．０５）。血清磷酸肌酸激酶于消融后即刻明显升高（５２５．９５±４２６．４９Ｕ／Ｌｖｓ１１５．２７±２８．７０Ｕ／Ｌ，Ｐ＜０．０１），但术后１４日与消融前相比已无显著性差异（１１４．０２±２３．３５Ｕ／Ｌｖｓ１１５．２７±２８．７０Ｕ／Ｌ，Ｐ＞０．０５）。②４只犬发生并发症，其中１只损伤窦房结，２只发现心脏巨大附壁血栓，另１只术后出现一过性房性早搏、短阵房?     

Quantitation of red cell-bound IgG by an enzyme-linked antiglobulin test in human immunodeficiency virus-infected persons

Anemia, thrombocytopenia, and neutropenia have been observed in patients with acquired immune deficiency syndrome (AIDS) and AIDS-related complex. To investigate whether red cells (RBCs) of patients with human immunodeficiency virus infection were coated with IgG and/or complement (C3), blood samples of 239 patients were tested. The prevalence of a positive direct antiglobulin test on RBCs was 16.7 percent. By use of an enzyme-linked antiglobulin test (ELAT) to measure more accurately the number of IgG molecules per RBC in a group of 67 patients, 30 of the 67 individuals were observed to have increased numbers (mean, 155) compared to normal controls and to patients with hypergammaglobulinemia due to multiple myeloma or chronic liver disease. Hemoglobin level was correlated with the number of IgG molecules per RBC (p = 0.008), but no correlation could be demonstrated between those numbers and serum immunoglobulin (p = 0.10) or circulating immune complexes (p = 0.38). Our results with ELAT suggest that some AIDS patients may have specific binding of IgG on the surface of their RBCs, rather than nonspecific uptake; further clinical correlations are necessary to confirm these findings.