Production and characterization of monoclonal antibodies to guinea pig lymphoid differentiation antigens

We have prepared 2 mouse monoclonal antibodies which react with differentiation antigens on guinea pig lymphoid cells. Monoclone 5AB2 recognizes an antigen expressed on both T and B lymphocytes and absent on macrophages. It has proven useful in the preparation of populations of antigen presenting cells which are free of T and B lymphocytes. The second monoclonal, 8BE6, is specific for peripheral T cells and 10% of thymocytes. It reacts with a 68,000 dalton molecule which is also expressed on the guinea pig B cell leukemia, EN-L2C. 8BE6 has proven to be lytic for peripheral T cells in the presence of rabbit complement and has been used to deplete T cells from heterogenous cell populations.     

Interaction of immunoglobulin with actin

Actin can form specific, direct associations with immunoglobulin resulting in soluble complexes or cross-linked matrices. This interaction can be detected by four in vitro assays using purified components: (1) actin enhances the cytophilic activity of guinea pig IgG2; (2) in solutions of low ionic strength, actin and IgG2 co-precipitate: (3) soluble complexes exist in 0.1 M KCl as revealed by the displacement of actin from its expected sedimentation pattern in a gradient of sucrose when in the presence of IgG 1, IgG2, or IgM; (4) immunoglobulin (IgG1, IgG2, BGG)‡: increases the viscosity of F-actin solutions, presumably by crosslinking F-actin filaments. These data suggest that direct interaction of a cytoskeletal protein with a cell surface receptor is possible.     

Assessment of immunosuppression by serum inhibition of alloreaction and measurement of cyclosporin A (CyA) serum levels in kidney graft recipients under CyA

The immunosuppressive effect of kidney graft recipient sera was studied on T-lymphocyte alloreactive line (4H) proliferation and compared to native cyclosporin A (CyA) and CyA metabolite concentrations determined by radioimmunoassay (RIA) using specific or nonspecific monoclonal antibodies. Three clinical groups were studied: (1) patients experiencing acute renal rejection episodes (CyA-R), (2) patients experiencing CyA-dependent nephrotoxicity episodes (CyA-TOX) and (3) patients in a clinically steady state (CyA-ST), according to their therapeutic regimen i.e., monotherapy (CyA alone) or polytherapy (CyA associated with prednisolone and/or azathioprine). Regardless of the clinical state, sera of patients in polytherapy displayed more inhibitory activity than those of monotherapy patients (24% and 40% inhibition of 4H proliferation, respectively, at sera dilution of 1:2), something which was no doubt due to the inhibitory activity of prednisolone on T-lymphocyte growth. In the two therapeutic regimens, CyA-ST patient sera exhibited the lowest inhibitory activity on the 4H line (45% and 65% inhibition of 4H proliferation in mono-and polytherapy, respectively, at sera dilution of 1:2). Sera from CyA-TOX patients were highly inhibitory (74% and 86% inhibition of 4H proliferation in mono-and polytherapy, respectively, at sera dilution of 1:2), in agreement with RIA assays showing increased native circulating CyA and CyA metabolites and daily CyA intake in this group as compared to CyA-St. Surprisingly, CyA-R patient sera were no less inhibitory than those of CyA-ST patients on 4H-line, antigen-induced proliferation. This clinical group did not differ from others for CyA intake or level of circulating immunosuppressive molecules, suggesting that rejection could be associated with a state of interindividual variation in sensitivity to CyA. In addition, a polytherapeutic regimen seemed to modify CyA bioavailability in CyA-ST group patients, with a decreased CyA metabolite level as compared to their monotherapy counterparts (native CyA plus metabolite/native CyA ratio being 2.73 and 3.73, respectively). In contrast, in the CyA-R patient group, polytherapy appeared to be associated with an increase in CyA metabolite circulating levels (ratio 4.79). In view of the low inhibitory activity of CyA metabolites, this profile might lead to rejection.     

中脑导水管周围灰质内注射抗强啡肽血清对神经降压素镇痛作用的影响

目的：探讨大鼠中脑导水管周围灰质（PAG）内内源性强啡肽对神经降压素（NT）镇痛作用的影响。方法：以钾离子透入法引起大鼠甩尾反应的电流强度（mA)为痛行为反应的指标,观察向PAG内注入NT,抗神经降压素血清和抗强啡肽A1－13血清对动物痛阈的影响。结果：PAG内注入NT后,大鼠痛阈明显升高;注入抗神经降压素血清后,大鼠痛阈则明显降低,而注入抗强啡肽血清后,对NT的镇痛效应无显著影响,结论：PAG内NT在痛觉调制中发挥着重要的作用,且其作用不依赖于PAG内的内源性强啡肽。     

慢性特发性荨麻疹患者血清组胺释放活性的检测

目的 检测慢性特发性荨麻疹患者血中组胺释放活性。方法 自身血清皮肤试验和嗜好性粒细胞组胺释放试验。结果 自身血清皮肤试验和嗜碱性组胺释放试验的阳性率分别为３７．５％和４３．７５％。结论部分慢性特发性荨麻疹患者血中组胺释放活性增高,揭示该类病人具有特殊的肥大细胞和嗜碱性粒细胞激活机制,很可能与自身免疫有关。     

Treatment of Vitamin B12 Deficiency after Gastric Surgery for Severe Obesity

Background: Vitamin B₁₂ deficiency after gastric surgery for obesity is due to a failure of separation of vitamin B₁₂ from protein foodstuffs and to a failure of absorption of crystalline vitamin B₁₂ in the presence of intrinsic factor. The purpose of this study was to determine which of four oral doses of crystalline vitamin B₁₂ was most effective in treating vitamin B₁₂ deficiency in 102 patients. Methods and Results: At time of entry into the study, the patients had a serum vitamin B₁₂ < 100 pmol L ⁻¹, were 29.9 ± 21.7 months post-op, were 37 ± 8 years old and had a body mass index of 30 ± 6 kg m⁻². Eight (8%) had had a vertical banded gastroplasty and 94 (92%) a gastric bypass. For the first 3 months all patients received 350 μg per day of crystalline vitamin B₁₂ and all increased their serum vitamin B₁₂ levels to over 100 pmol L⁻¹. The patients were then assigned to receive for a further 3 month period one of four oral doses of crystalline vitamin B₁₂-100 μg, 250 μg, 350 μg and 600 μg. Serum vitamin B₁₂ levels were greater than 150 pmol L⁻¹ after 6 months in 83.3% of patients who received 100 μg; 92.3% of patients who received 250 μg; 94.7% after 350 μg and 95.2% after 600 μg (p%0.525). Conclusion: At least 350 μg per day is the appropriate oral dose of crystalline vitamin B₁₂ after gastric surgery for obesity to correct low serum vitamin B₁₂ levels in 95% of patients.     

Clonal growth of human acute leukemia cells in serum-free methylcellulose medium

Ｔｈｅｇｒｏｗｔｈｏｆｌｅｕｋｅｍｉａｉｎｓｅｍｉｓｏｌｉｄｃｕｌｔｕｒｅｉｎｖｉｔｒｏｗａｓｄｅｐｅｎｄｅｎｔｏｎｔｈｅｐｒｅｓ－ｅｎｃｅｏｆｅｘｏｇｅｎｏｕｓｓｅｒｕｍ．Ｓｅｒｕｍｉｓａｍｉｘ－ｔｕｒｅｃｏｎｔａｉｎｉｎｇｌａｒｇｅｎｕｍｂｅｒｏｆｕｎｉｄｅｎｔｉｆｉｅｄｃｈｅｍｉｃａｌｃｏｍｐｏｎｅｎｔｓａｎｄｕｎｉｄｅｎｔｉｆｉｅｄｂｉｏ－ｌｏｇｉｃａｌｌｙａｃｔｉｖｅｓｕｂｓｔａｎｃｅｓ［１］,ｗｈｉｃｈｐｒｏｖｉｄｅｔｈｅｎｕｔｒｉｅｎｔｓｎｅｃｅｓｓａｒｙｆｏｒｔｈｅｃｅｌｌｇｒｏ…     

聚丙烯酰胺凝胶电泳分析615纯系小鼠血清蛋白

目的：根据各种蛋白质在一定的泳动条件下,都有一定的相对迁移率不同的特点,分析６１５ 纯系小鼠血清中的蛋白质在聚丙烯酰胺凝胶上的分布,进行了定位分析。方法：采用高ｐＨ 不连续性聚丙烯酰胺凝胶电泳及蛋白质特殊染色法。结果：６１５ 鼠血清蛋白质在聚丙烯酰胺凝胶上可分离出１３ ～１５ 条区带,脂蛋白３ 条、白蛋白２ 条、糖蛋白５ 条、铜蓝蛋白１ 条、血红蛋白１ 条、结合珠蛋白３ 条。根据每种蛋白质的Ｒｍ 值定位顺序从阳极到阴极依次为１ ．糖蛋白、２．脂蛋白、３ ．白蛋白、４．脂蛋白、５ ．糖蛋白、６ ．巨球蛋白（ 根据其它资料） 、７ ．铜蓝蛋白、８ ．结合珠蛋白、９ ．Ｈｂ、１０ ．运铁蛋白（ 根据其它资料）、１１ ．脂蛋白、１２ ～１４ ．均为糖蛋白。光密度扫描６１５ 鼠血清蛋白质相对面积比分别为白蛋白为４８－４、α１ 球蛋白为６－９、α２ 球蛋白２３－８ 、β球蛋白为５－０ 、γ球蛋白为１３－８ 。结论：６１５ 鼠血清蛋白质的定位分析对于６１５ 鼠可移植性淋巴细胞型白血病鼠的蛋白质研究具有一定的实际意义,利用Ｒｍ 值来作蛋白质定位分析简单易行,关键在于掌握实验条件的恒定。     

Extracellular fluid and its proteins: dehydration, shock, and recovery

This review highlights characteristics of extracellular fluid (ECF) that are often overlooked. ECF has, in addition to plasma and interstitial fluid (ISF) surrounding cells, a third large compartment, the ISF of skin and connective tissue. This acts as a reservoir that gives up ECF to plasma volume (PV) in order to sustain circulation in the event of either shock or dehydration. While Starling forces drive filtration, ECF is returned to PV more by lymph and less by Starling forces than previously appreciated. Lymph return to PV is dependent on physical activity and muscle contraction to overcome gravity. Regional change in metabolic rate alters the need for oxygen and nutrients that is met by a regional increase in capillary blood flow. Blood flow is controlled by vasoactive compounds released in response to a drop in PO₂; these relax capillary smooth muscle to increase blood flow and delivery of oxygen and nutrients. Plasma proteins, including albumin, are filtered into the interstitium through larger pores than those filtering ECF. The rate of protein filtration is set by size and charge of these larger endothelial pores and by size and charge of proteins. Charge of these pores, hence albumin permeability, is regulated by many of the same vasoactive compounds that control capillary flow. As a consequence, in response to gravitational stress and other forms of shock that reduce effective circulation, albumin as well as ECF is rapidly shifted from plasma and sequestered in ISF. When this has occurred, as in burn shock, restoration is better effected by generous expansion of ECF with Ringer’s solution alone, rather than with Ringer’s solution supplemented with human serum albumin or other colloid. Restoring both PV and ISF volume restores lymph circulation and returns sequestered albumin to PV. Received: 12 November 1998 / Revised: 30 March 1999 / Accepted: 2 April 1999