Studies on Protein Fractions Isolated from Human Plasma

S ummary . Fractions of human plasma separated by precipitation with cold ether and ethanol were examined for a number of protease and esterase activities. Plasminogen, kallikrein, increased vascular permeability, thrombin and TAMe esterase were concentrated in fraction G 2 / i R, a lipo-protein fraction containing a mixture of alpha and beta globulins and a gamma-M globulin, which also contained activated coagulation contact factors. ATEe esterase was concentrated in G 2 / i R and fibrinogen. Kinin-ase and TAMe esterase were also found in G 2 / 2 a mixture of alpha, beta and gamma globulins.
Tests of fractions of G 2 / i R separated by chromatography confirmed the findings of Kagen et al , (1963) and showed that kallikrein was a gamma-M globulin. Vascular permeability activity, but not kininogenase activity was removed by high-speed centrifugation from this fraction.
Fractions G 2 / i R, G 2 / 2 and to a lesser extent G 3 contained a substance which induced kininogenase activation in intact plasma, but was not identified as PF, plasmin or activated contact factor and had no esterase activity.
Kininogenase activity was associated with contaminants of fibrinogen and gamma globulin concentrates and absent from albumin. In fractions prepared by large-scale methods, only those made from G 2 are likely to contain sufficient kininogenase to cause a clinical reaction. Kininogenase was not adsorbed by Al(OH)³ gel or C^a3(PO⁴)² and would not contaminate clotting factors prepared from G².     

Blood Group A Antigen in Human Erythrocytic Membranes and Membrane Fractions1

Abstract. Erythrocytic membranes from blood group A individuals were assayed for A antigen using a quantitative hemagglutination inhibition technique. The membranes were then extracted for lipid and glycoprotein. Although some A antigen was usually found in the glycoprotein fraction, most of the activity was in the lipid fraction. The sum of A antigen activity in the lipid, glycoprotein, and membrane residue fractions only occasionally was equal to the A activity in the erythrocytic ghosts. However, when certain lipid preparations with little or no A antigen (enhancement factors) were added to the glycolipid fractions, the amount of A antigen demonstrated was usually greatly increased. Under these conditions, the sum of the fractions often was much greater than the A antigen demonstrated in erythrocytic membranes. This suggests that the organization or arrangement of A antigenic determinants in the red cell membrane may not always permit a stoichiometric reaction with anti-A molecules.     

几种动物的抗—HBs应答观察

许多动物种属(包括一些家畜和家禽)中存在低滴度抗-HBs,推测其产生原因可能与HBAg有关,或由于动物暴露于与 HBsAg 有共同表位的环境抗原,诱导可引起交叉反应的抗体。本文用 HBsAg 阳性、HBeAg 阴性的 HBV 感染患者血清免疫豚鼠、鸡和鹅各 2只,均产生抗-HBs应答,其抗-HBs 活性可被纯化 HBsAg 中和,证明此抗-HBs 是特异性的。采用 SPRIA 检测,豚鼠抗-HBs 滴度高于 1:800,而鸡和鹅虽经多次加强免疫,滴度仅达 1:10～1:100,表明鸡、鹅对HBsAg 的反应性低,仅引起较弱的抗-HBs 应答,从而证实动物 HBsAg 暴露是产生低滴度抗-HBs的原因之一。另外,观察免疫后动物血清,除 HBsAg 阳性(RPHA≥1:128)外,未出现其它 HBV感染标志,无显著的生化和病理改变,肝脏 HBsAg 阴性,提示动物未经历 HBV 活性复制。     

Further Data on HBs Antigen Subtypes – Geographical Distribution

Abstract. 605 HB antigens from silent carriers of different geographic areas were subtyped. Guinea pigs and goats were immunized with selected HB antigens. Subtyping was performed by CIEP using monospecific or duospecific antisera, and by ID with unabsorbed antisera if the antigen was strong enough. Eight categories of HB antigens previously described were confirmed: a₁y(w), a₂Iy(w), a₂3y(w), a₃(w), ayr, a₂Id(w,) a₃d(w) and adr. The relations between the four categories of a subdeterminants and ‘w’ are analysed. All the antigens with ‘w’ determinant are either a₁ or a₂l or a₂3 or a₃. The ‘w’ determinant can be subdivided into the system of a subdeterminants which are mutually exclusive among themselves. r appears to be allelic with the group of the four a subdeterminants. It cannot yet be stated if r may be considered as a fifth subdeterminant of a or if it is independant of a subdeterminant. For the present time we use the following terminology a₁(w), a₂l(w₂), a₂3(w₃), a₃(w₄) and r. By combining a(w) subdeterminants, r, d and y determinants, it is possible to greatly increase our knowledge about the geographical distribution of HB antigen: ay subtype is predominant in Africa and in Mediterranean countries but three areas can be distinguished by means of a subdeterminants: a₃(w₄)y being mainly found in West and Central Africa (87%) and a₂1(w₂)y in Mediterranean countries (86%) except for Greece where a same ratio of a₂1(w₂)y and a₂3(w₃)y is encountered. This a₂3y subtype is the most frequent of ay subtypes in France and in Hungary. In these two countries, the subtypes ad(w) are predominant: 67% of a₂1(w₂)d and 1.3% of a₃(w₄)d in France. adr is widespread in Asia and especially in the Thai area (Thailand and Laos), but we have found a₁(w₁)y in Vietnam. Many examples are given which show that the subtype of HB antigen is related to the country of origin and does not seem to depend of the host. These findings illustrate the importance of HB antigen subtyping for epidemiology and for study of populations.     

Discovery of a Lectin with Affinity for the HBs Antigen

An extract of marguerite leaves was found to contain an agglutinin which has affinity for HBs (adr type) antigen-sensitized sheep erythrocytes and for pure HBs antigen. This agglutinin activity was inhibited by the plasma of hepatitis patients who were known to have circulating HBs antigen. The agglutinin activity was also inhibited by 0.03M N-acetylneuraminic acid.     

弓形虫抗原组分提取及免疫原性观察

以不同剂量的弓形虫粗制与纯化抗原经皮下注入小白鼠，３５天后检查血清，在高剂量粗制抗原组显示有免疫学改变。以０．２ｍｌ弓形虫感染小鼠后新鲜腹水感染试验鼠，亦显示试组具保护力，可延长小鼠的存活时间。但总的看来，保护力不强，尤其是低剂量粗制抗原及纯化抗原均无保护力。     

HBs Antigen Subtypes among Acute Hepatitis Patients in Japan: Evidence of Imported Hepatitis

HBs antigen subtypes were determined among 137 acute hepatitis patients in Japan. The distribution among the 99 male patients was 40 adr, 37 adw, one ayw, and 21 undetermined while that among the 38 female patients was 19 adr, 11 adw, and eight undetermined. The distribution among the male patients was different from that among HBs antigen carriers in Japan, suggesting that they were exposed not only to domestic HBs antigen carriers, but also to other sources. Thirty-one (29 males and two females) of the 137 patients (22.6%) were found to have been abroad at some time during the incubation period. the distribution among them was seven adr, 21 adw, and three undetermined. the majority bad the same subtypes that are prevalent in the countries they visited, supporting the hypothesis that most of them bad contracted the disease overseas. The remaining 106 patients bad a distribution of the subtypes similar to that of domestic HBs antigen carriers. "Imported hepatitis" would be an important cause of acute type B hepatitis in Japan.     

Recovery of Australia Antigen from Human Plasma Products Separated by a Modified Cohn Fractionation

Abstract. One liter of strongly Australia antigen positive plasma was subjected to a modified Cohn fractionation and samples from the various steps of the process were assayed quantitatively for Australia antigen. More than 85% of the original quantity of Australia antigen was recovered from fraction IV. Intermediate amounts, about 2%, were found in fraction III and in the washing fluids after the fibrinogen wash. The final fibrinogen product had no detectable Australia antigen. About 1 % of the antigen was recovered from the albumin. γ-Globulin before treatment with DEAE-Sephadex had a low content, about 0.1% of the original amount of antigen, whereas DEAE-Sephadex treated material (final gammaglobulin) was free of demonstrable Au-antigen.
The findings are discussed in relation to clinical reports on hepatitis risks connected with the various products.     

A Tumor-Associated Antigen in Human Nephroblastomas

Antisera prepared against extracts of human nephroblastomas (Wilms' tumors) allowed the characterization in these extracts, and in a few extracts of tumors of other organs, of a tumor-associated antigen. This antigen is different from all previously described antigens, and was named W antigen (W for Wilms). It does not seem to be a carcinoembryonic antigen, and hence it could be of viral origin.     

Separation of Hepatitis-Associated Antigen (HAA) from Human Plasma for the Production of Rabbit Anti-HAA

Abstract. A method is described for obtaining hepatitis-associated antigen (HAA) from the acid citrate dextrose plasma of routine blood donations found to be HAA-positive. The separation consisted of the following steps: ammonium sulphate precipitation, absorption on and elution from DEAE-Sephadex, and gel filtration on a Sepharose-6B column. HAA was demonstrated by cross-over immuno-electrophoresis and lines of identity shown by the Ouchterlony double diffusion technique. Freedom from contaminating protein was shown by immuno-electrophoresis. This material, when injected into rabbits, produced an anti-HAA reagent suitable for use cross-over immuno-electrophoresis without absorption.     

I-Active Antigen of Human Erythrocyte Membrane

Abstract. An I-active substance was isolated from the aqueous layer obtained after extraction of human erythrocyte membranes with n -butanol. This substance was found to be identical with the antigens with A, B, and H blood group specificity which we had isolated previously. Thus, all A, B, H and I blood group activity in aqueous extracts of erythrocyte membranes is dependent upon the same kind of antigenic material which may carry different blood group specificities. The material is sialic acid free and comprises about 90% carbohydrate.     

Delta infection in chronic HBs Ag carriers in Tunisia: high prevalence in chronic asymptomatic HBs Ag carriers and in HBs Ag positive cirrhosis

The presence of hepatitis B virus DNA, delta antigen and anti-delta antibodies was examined in 159 Tunisian chronic HBs Ag carriers: 45 were asymptomatic and 114 suffered from cirrhosis. Serum hepatitis B virus DNA was detected in two (4.5%) asymptomatic HBs Ag carriers and in 11 (10%) HBs Ag positive cirrhosis patients. The prevalence of HDV infection determined by the presence of anti-delta was relatively high in asymptomatic HBs Ag carriers (33%) and in HBs Ag positive cirrhosis patients (21%). Active ongoing HDV infection, detected by serum HD Ag and anti-delta IgM, was shown in five patients with cirrhosis and active hepatitis B virus replication. We conclude that hepatitis delta virus may be endemic in Tunisia and does not always inhibit hepatitis B virus replication.     

Primary Amyloidosis with Increased Plasma Carcinoembryonic Antigen Concentration

ABSTRACT A patient with suspected malignant disease had increased concentration of plasma carcinoembryonic antigen (CEA). Amyloidosis was demonstrated at autopsy. The amyloid fibril composition was characterized by immunohistochemical and immunochemical techniques and proved to be of the lambda light chain (AL) type. CEA was demonstrated in the liver parenchyma by using antihuman CEA antiserum. Increased plasma CEA concentration in a patient with primary amyloidosis has, to our knowledge, not been reported before.