Experimental anemias in the rat; II. The effect of various sulfonamides in producing a pteroylglutamic acid deficiency and the pteroylglutamic acid activity of test substances

1. Different sulfonamides were tested to ascertain their effect in producing thecharacteristic symptoms of acute PGA deficiency in rats fed on synthetic diets.Sulfasuxidine (1 per cent) and the less soluble phthalylsulfathiazole (1 per cent)were equally effective. Sulfathiazole in 1 per cent concentration produced a hemolytic anemia not reversible by PGA or whole liver powder. In a 0.5 per cent concentration it was also effective, but in view of its toxicity, the less soluble sulfonamides were to be preferred. A mixture of 0.5 per cent sulfathiazole and 0.5 percent sulfadiazine was extremely toxic and produced a hemolytic anemia. Sulfaguanidine was toxic at 1 per cent concentration.

2. When intermittent small doses of PGA were given to PGA-deficient rats toprolong their life from 45 up to 155 days, 1 per cent sulfasuxidine or phthalylsulfathiazole, or 0.5 per cent sulfathiazole were equally efficient in producing regularlya macrocytic normochromic anemia.

3. The response of PGA-deficient rats to single doses of PGA has been studiedand an assay procedure has been suggested which uses the weight increase andduration of cure as the measure of the response. The W.B.C. and reticulocyte response can also be used as a qualitative indication of PGA activity.

4. Of the substances tested by this procedure, vitamin B₁₂, purified perniciousanemia preparations, ascorbic acid and xanthopterin showed no PGA activity. Acommercial yeast preparation and Teropterin were found to possess biologic activity comparable with that found by other workers in assays on chicks.

Note: ACKNOWLEDGMENTSWe wish to thank Dr. E. Lester-Smith, of the Glaxo Laboratories, for the generous supply of vitaminB₁₂, Dr. T. H. Jukes, of the Lederle Laboratories, for the gift of PGA and Teropterin, and Dr. W. Jacobson for the samples of purified P. A. liver preparations and xanthopterin.

     

Hemolytic disease of the newborn due to anti-A1

1. The authors report clinical, hematological and serological data in a case ofA₁ iso-sensitization of pregnancy.

2. The mother’s serum displayed immune characters specific to A₁ cells immediately after delivery. On the 24th day post partum the specificity extendedto A₂ cells.

3. The disease exhibited by the infant was very mildly hemolytic. It wasmarked by a deep jaundice, repeated alimentary vomiting and a progressive stateof drowsiness. There was no anemia. The direct anti-globulin test was negative.

4. It is shown that the mildness of the hemolytic process in cases of placentaltransfer of immune anti-A or anti-B into the incompatible A or B fetus is probablydependent upon a peculiar "resistance" of the fetal cells. This may be demonstrated in vivo and in vitro. In the present case replacement transfusion withA₁ adult blood resulted in its in vivo sensitization, detectable by the antiglobulintest and eventually leading to hemolytic anemia.

Submitted on September 28, 1953 Accepted on August 15, 1954     

日本血吸虫可溶性虫卵抗原经两种免疫途径获得的鸡卵黄抗体IgY动态观察

目的对比两种免疫途径产生抗日本血吸虫可溶性虫卵抗原(SEA)的特异性IgY抗体水平动态变化。方法7只新西兰兔感染日本血吸虫尾蚴(1500/只)42d后解剖收集虫卵,制备SEA。将SEA分别经皮下和静脉注射免疫健康海蓝蛋鸡(3只/组),首次和第2次免疫间隔2周,之后每次间隔4周,共免疫5次,50μg/(只·次)。取两组免疫前,以及首次免疫后2～18周生产的鸡蛋,用水稀释法粗提IgY,ELISA检测每2周IgY的动态变化。IgY纯化试剂盒(EGGstract IgY Purifiction System)纯化静脉组首次免疫后8～18周的IgY,紫外吸收法检测抗体浓度,琼脂糖双扩散法和ELISA检测IgY峰值水平的抗体效价,十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹(Western blotting)分析比较免疫前后抗体特异性。结果静脉组和皮下组分别在首次免疫后8和12周IgY抗体水平达高峰,A492值分别为1.28和0.78,静脉组IgY水平至18周仍维持较高水平,皮下组抗体水平达到峰值后逐渐下降。纯化后IgY浓度约为6.5～9.0mg/ml,琼脂糖双扩散法和ELISA测得静脉注射组峰值水平抗体效价分别为1∶16和1∶51200。经SDS-PAGE和Western blotting分析,纯化后的IgY在相对分子质量(Mr)25000和68000处各有一条清晰条带,且免疫后IgY可与SEA发生特异性反应。结论静脉注射法比皮下注射法能获得更高水平的鸡抗日本血吸虫SEA特异性IgY抗体,且纯化后的IgY抗体具有较好的特异性。     

Hemolytic reactions produced in dogs by transfusion of incompatible dog blood and plasma; serologic and hematologic aspects

1. Dogs injected intravenously with dog erythrocytes containing one or moreantigenic factors lacking in their own red cells developed iso-hemagglutinins andhemolysins exhibiting characteristics of immune antibodies.

2. Transfusions of incompatible whole dog blood and plasma were carried outunder controlled conditions. Pretransfusion observations were made and followedby closely spaced post-transfusion measurements of serologic and hematologicalterations.

3. The rate of destruction of incompatible donated corpuscles was determinedby tagging the cells with radioactive iron and also by employing the technique ofdifferential agglutination of erythrocytes. It was thereby shown that all of theincompatible donated cells disappeared from the recipient’s circulation withinthe first thirty to ninety minutes following transfusion. The probable mechanismsand relative importance of intra- and extravascular destruction of erythrocytes arebriefly discussed.

4. Destruction of recipient dogs’ corpuscles by donated immune plasma wasrelatively slow, and spherocytosis and increased osmotic fragility of the recipients’ cells were evident for periods as long as twenty days. These observationsare compared with those made in human beings after transfusions of plasma and ofblood from dangerous universal donors.

5. The titer of complement in the sera of recipient dogs was sharply reducedfor at least five hours after all transfusions of incompatible whole blood, but isoagglutinin titers were less regularly reduced after such transfusions.

6. Other notations of interest included estimates of the concentrations of serumbilirubin, sodium and potassium, determinations of clotting time, prothrombinconcentration, and observations on red cell morphology, intravascular erythrophagocytosis, and shifts in distribution of leukocytes and in the electrophoreticpatterns of plasma.

Note: ACKNOWLEDGMENTSIt is a pleasure to acknowledge the technical assistance of Mrs. Jane Peters, Miss Mary Jane Izzo andMiss Shirley Deshon.

     

Autoantibodies in Acquired Hemolytic Anemia With Special Reference to the LW System

Most autoantibodies in patients with warmantibody autoimmune hemolytic anemia(AIHA) have specificity within the Rh system. Using rare cells such as -D- andRh_null cells, Weiner and Vos (1963) described specificity against normal cells (nl),partially deleted cells (pdl), and deletedcells (dl). Recently, autoantibodies whichfailed to react with Rh_null cells that were ofanti-U specificity have been described. Itwas suggested that the "Rh related" autoantibodies that cannot be identified asspecific Rh antibodies may be anti-U. Inthe present study we examined eluatesfrom the red cells of eight patients withAIHA using a panel of extremely rare cellsand cross-absorption and elution techniques. We demonstrated autoantibodyspecificities not definable without the rarecells and, further, defined heterogeneity ofthe LW antigen. Autoantibodies with Uspecificity occurred in three eluates only.It was always present with an antibody ofanother specificity. Six of the eluates contained anti-LW, two anti-nl, five anti-pdl,three anti-dl, and one anti-e. Absorptionand elution studies using the rare Rh-positive LW-negative (Mrs. Bigelow)showed that anti-pdl may in fact representanti-LW + LW₁ and that Mrs. Bigelowmay represent a weak variant of LW. Injection of her red cells into guinea pigsproduced an anti-LW that reacted similarlyto the antibody produced by injecting Rh-positive LW-positive cells. An analogy tothe ABO is suggested that normal Rh-positive LW-positive cells represent LW₁,Rh-negative LW-positive cells representLW₂, Mrs. Bigelow represents LW₃ andRh_null cells represent the only true LW-negative (lw).

Submitted on January 19, 1973 Revised on March 5, 1973 Accepted on March 6, 1973     

Anti-A agglutinins in pooled plasma as a cause of hemolytic anemia

Five sets of observations are presented showing the development of hemolyticanemia in group A patients as the result of prolonged administration of relatively large amounts of pooled plasma. During the hemolytic episodes, spherocytosis, increase in osmotic fragility, rising bilirubin levels and sometimesclinical jaundice and splenomegaly were observed. Antibody attached to therecipients’ erythrocytes was demonstrated by means of the direct Coombs test,and in several instances free antibody of anti-A specificity was found in therecipient’s plasma during the hemolytic episode.

Random testing of commercial pooled plasma revealed a number of instancesof astonishingly high anti-A titers, indicating that neither dilution in the poolnor neutralization by natural A substance in the component A plasmas can berelied upon.

The pools implicated in the production of hemolytic reactions had anti-A₁titers of 1:64 in saline or higher, and with the usual technics showed no clear-cut"immune" characteristics except in one instance. The administration of substantial amounts of plasma pools of low titer (1:16) was not followed by adversereactions in several instances. On the other hand, the administration of originallyhigh titered plasmas, neutralized in vitro to a saline titer of 1:16 resulted inblood destruction, indicating that under conditions of prolonged massive administration, specific soluble substance, though effective in vitro, is inadequatein vivo. Whether the adverse effects of the moderately high and high titeredplasma pools were the result of purely quantitative factors connected with theintroduction of large amounts of natural anti-A agglutinins or whether specialimmune isoantibodies not detected by the methods used were responsible couldnot be determined by the data at hand.

Prolonged administration of untitered pooled plasma to recipients of bloodgroups A and probably B is potentially dangerous.

Submitted on August 3, 1955 Accepted on November 9, 1955     

Cell Antigens and Cell Specialization. III. On the H Antigen Receptors of Human Epidermal Cells

This article describes the application of the mixed agglutination method todemonstrate the H antigen receptors on human A₁, A₂, B and O epidermalcells, and also the presence of A and B antigen receptors in different spatiallocations than the H receptors in A and B epidermal cells.

Submitted on April 16, 1963 Accepted on June 18, 1963     

Effect of Anticoagulant and ABO Incompatibility on Recovery of Transfused Human Platelets

The effects of anticoagulant solutions on the recovery of transfused plateletswere studied. Citrate anticoagulants at pH 7.4 or pH 6.5 were found to beequally effective in preserving the viability of platelets when centrifugationof the cells was not required. When centrifugation is required, as in mostplatelet survival studies, citrate at pH 6.5 gives maximum recoveries. Ethylenediamine tetraacetate (EDTA) caused temporary sequestration of nearly alltransfused platelets and reduced maximum recoveries by about 50 per cent.

Platelet recovery was lowered by ABO-incompatibility between donor platelets and recipient serum, but survival time of remaining platelets was notaltered. Lowest platelet recoveries resulted when group A₁ or A₁ B plateletswere given to group O recipients with high isoagglutinin titers.

It is suggested that ABO-compatible platelets prepared in citrate should beused where possible in the treatment of thromboctyopenic disorders.

Submitted on January 5, 1965 Accepted on March 13, 1965     

A Quantitative Immunogenetic Study of Gene Suppression Involving A1 and H Antigens of the Erythrocyte without Affecting Secreted Blood Group Substances. The ABH Phenotypes Ahm and Ohm

The A^h_m phenotype is distinguished by the absence of H from the cells andanti-H from the serum, the presence of A_x antigen on the cells and A₁ and Hantigens in the secretions. Similarly, O^h_m has no cellular H or serum anti-Hand normal quantities of H substances are found in the saliva.

Both phenotypes appear to result from the action of a recessive gene, possibly sex-linked, suppressing H and A₁ on the cells but with no effect on theA_x antigen. A_x is considered to be largely independent of the biosyntheticpathway that proposes H substance as a precursor to A and B antigens.

Quantitative hemagglutination studies also disclosed evidence of othergenes modifying the quantitative expression of cellular A and H.

Submitted on April 29, 1964 Accepted on June 7, 1964     

Intracellular protein resembling Russell bodies in malignant lymphomas associated with acquired hemolytic anemia

Large amounts of intracellular periodic acid-Schiff positive protein, closelyresembling Russell bodies tinctorially and histochemically and, to varying degrees, morphologically, were found in the neoplastic cells of three patients withmalignant lymphoma associated with acquired hemolytic anemia. Direct Coombstests were positive in all three of these patients, and in two of them, immuneautoantibodies against red cells were observed in unusually high titers. One ofthe latter also had abnormally high serum globulin levels with cryoglobulinemia.

A similar intracellular protein was present in the neoplastic tissue of seven of acontrol group of 65 unselected cases of malignant lymphoma. The serum globulinwas elevated in four of the seven patients and one of these also had cryoglobulinemia. Definite evidence of hemolytic anemia was noted in one, and suggestive evidence in two of these four patients. A lymph node removed from afifth patient, on whom no serum protein determinations had been made, showedhemosiderosis and marked erythrophagocytosis. This patient had not receivedblood transfusions or other treatment before biopsy was performed.

On the basis of our observations, it is suggested (1) that under certain conditions the neoplastic cells of malignant lymphomas are capable of synthesizingabnormal proteins; (2) that the presence of Russell body-like intracellular PAS-positive material in the neoplastic cells may be the morphologic manifestation ofthis protein synthesis; (3) that these abnormal proteins may play an importantpart in the immunological mechanisms responsible for the development of acquired hemolytic anemia in association with malignant lymphomas.

Submitted on May 20, 1954 Accepted on June 22, 1954     

Starch block electrophoretic studies of human hemoglobin solutions. I. Technic and results in the normal adult

1. Erythrocyte hemolysates from hematologically normal, adult individuals were studied by means of starch block electrophoresis.

2. The electrophoretic patterns obtained indicated the presence, in addition to the main component of A hemoglobin A₁, of three other hemoglobinfractions designated as A₂, A₃ and A₄, A₂ and A₄ move slower, while A₃moves faster in alkaline buffer. These components were isolated by elutionfrom the starch after separation by electrophoresis.

3. The concentrations of the various fractions in the hemolysates wereestimated from a sample of normal individuals investigated.

4. In our experiments, electrophoretic patterns obtained on starch thathas been re-used as a supportive medium for electrophoresis were somewhatdifferent but comparable to those obtained on starch that was used for thefirst time as a supportive medium; these differences are discussed.

5. Some characterizations of the isolated components are made.

Submitted on September 7, 1957 Accepted on December 15, 1957     

Microscopic and histochemical studies on the Auer bodies in leukemic cells

(1) Seventeen cases of acute leukemia with Auer bodies have been reported.Studies were carried out on 7 cases of acute monocytic leukemia and 3 cases ofsubacute myelogenous leukemia.

(2) Histochemical studies showed the Auer bodies to be oxidase, peroxidase,and periodic acid-Schiff positive; sudanophilic, slightly metachromatic and to givepositive tests for acetal lipids and ribonucleic acid.

(3) The Auer bodies were negative for acid and alkaline phosphatase, lipase,glycogen, desoxyribonucleic acid and were non-birefringent.

(4) A change in the chemical nature of the Auer body from an acid conditionto a more neutral state was noted. This change corresponded with the changesof the normal cytoplasmic granulation of the myelocytes and monocytes duringmaturation.

(5) The effects of cellular movements, trauma, and temperature changes uponthe Auer bodies were studied.

(6) Several leukemic cells, containing Auer bodies, were studied during theprocess of mitosis.

(7) A theory as to the formation and disintegration of the Auer bodies hasbeen presented.

Note: ACKNOWLEDGMENTSThe author wishes to express his thanks to Dr. B. K. Wiseman, Dr. B. C. Houghton, Dr. R. A. Knouff,and Dr. E. R. Hayes for their help and suggestions, and to Dr. J. D. Thomas, Dr.J. F. Gamble, Dr. R. J.Rohn, Dr. H. Schiro, Mrs. Jo Myers, M.Sc., Miss Georgia Gwinner, M.T., Mrs. Susan Ragsdale, M.T.and Mrs. Jeanne Marie Willison, M.T., for their help in securing the experimental material.

     

Experimental production of a nutritional macrocytic anemia in swine

1. A deficiency of pteroylglutamic acid has been produced in 32 swine fed apurified diet containing casein and supplemented with seven B vitamins, sulfasuxidine and a folic acid antagonist. The casein was fed at two levels, 10 and 26per cent. Two types of casein were used: a crude preparation possessing significant"extrinsic factor" activity and a purified casein with little activity.

2. The hematologic manifestations observed were (a) severe macrocytic anemia,(b) leukopenia, due to a proportionately greater reduction in polymorphonuclearthan in mononuclear cells, (3) slight thrombocytopenia, and (4) hyperplastic bonemarrow with an increase in immature nucleated red cells which resemble themegaloblasts seen in the bone marrow of patients with pernicious anemia.

3. The feeding of a 26 per cent rather than a 10 per cent crude casein diet did notprevent but did delay the onset of the blood changes. Anemia developed mostrapidly in the animals receiving 10 per cent purified casein.

4. The group receiving 26 per cent casein developed a greater degree of macrocytosis in the same period of time than did the group receiving 10 per cent casein.In all groups the degree of macrocytosis increased as the duration of the anemiaincreased.

5. The hematologic manifestations were not delayed nor was their developmentprevented by the intramuscular administration of 15 U.S.P. units of liver extractevery 15 days.

6. The blood and bone marrow returned rapidly to normal following the administration of pteroylglutamic acid, pteroyldiglutamic acid, pteroyltriglutamicand pteroylheptaglutamic acid. Thymine and xanthopterin had little or no activity. Tyrosine, adenine and uracil were inactive.

7. Purified liver extracts and crystalline vitamin B₁₂ were found to possess somehemopoietic activity in several animals but the activity was considerably less thanthat of the pteroylglutamic acid compounds.

8. The urinary excretion of "tyrosyl" (hydroxphenyl compounds) was notabnormal in the pteroylglutamic acid deficient pigs and was not altered by eitherpteroylglutamic acid or liver extract therapy.

9. The urinary excretion of allantoin and uric acid did not differ significantlyfrom the normal. Immediately following therapy with pteroylglutamic acid,however, in association with the reticulocytosis and lasting for the same period,there was a marked increase in the excretion of allantoin.

10. The results suggest that both pteroylglutamic acid and a factor in liverextract similar to or identical with vitamin B₁₂ are required for normal hemopoiesisin the pig.

Note: ACKNOWLEDGEMENTSThe crude methylfolic acid antagonist, xanthopterin, and the pteroylglutamic acid compounds, withthe exception of pteroylheptaglutamic acid, were kindly furnished by the Lederle Laboratories, PearlRiver, New York, through the courtesy of Dr. T. H. Jukes and Dr. S. M. Hardy.Sulfasuxidine was generously furnished by Sharp & Dohme, Inc., Philadelphia, Pa., through thecourtesy of Dr. W. A. Feirer.Pteroylheptaglutamic acid and Natola were supplied by Parke, Davis & Company, Detroit, Mich.,through the courtesy of Dr. A. E. Sharp and Dr. J. J. Pfiffner.Biotin was obtained from Hoffmann-LaRoche, Inc., Nutley, N. J., through the courtesy of Dr. E. L.Sevringhaus.The vitamins, with the exception of pteroylglutamic acid and biotin and including vitamin B₁₂ werekindly furnished by Merck and Company, Inc., Rahway, N. J., through the courtesy of Dr. A. Gibsonand the late Dr. D. F. Robertson.Experimental liver extracts (No. 1124, 1063, 1066 and 1067) were generously furnished by Armour andCompany, Chicago, Illinois through the courtesy of Dr. E. E. Hays.We are indebted to Mrs. Darlene Kehl, Mr. George Trappett, and Mr. Ocie Hadley for technicalassistance.

     

Fast Hemoglobin in Lead Poisoning

1. An electrophoretically fast hemoglobin was found in approximately 40per cent of preschool children with elevated blood lead levels.

2. Fast hemoglobin was found more often in lead-poisoned patients withhypochromic anemia than in patients with normochromic red cells.

3. Fast hemoglobin differed from hemoglobins produced in vitro by incubation with chromate or oxidized glutathione. It had electrophoretic propertiessimilar to that found in a few patients receiving tolbutamide.

4. Fast hemoglobin could not be differentiated from normal hemoglobin A₃by any technic utilized.

5. Both lead and A₃ hemoglobins were heterogeneous molecular species.

6. The mechanisms leading to the production of hemoglobin A₃ and leadhemoglobin remain unknown.

Submitted on October 7, 1965 Accepted on December 10, 1965     

A and B and A1Leb Substances in Glycosphingolipid Fractions of Human Serum1

Abstract. A and B and A₁Le^b substances were adsorbed onto red cells exposed to glycosphingolipid fractions prepared from the serum of group A and B and A₁, Le(a-b+) donors. Group O cells exposed to fractions prepared from the serum of group A or B donors were agglutinated by an IgM cross-reacting antibody present in some group O sera. Cells exposed to fractions from A₁, Le(a-b+) serum were agglutinated by anti-A₁Le^b. The amount of A substance in the fractions was related to the A subtype (A₁ or A₂) and to the Lewis and secretor phenotype of the donor. The uptake of blood-group substances from the lipid fractions was inhibited by the addition of whole serum to the fractions.     

The Lutheran blood groups: a second example of anti-Lu b and three further examples of anti-Lu a

1. An example of the blood group antibody, anti-Lu^b, was found in a patientwho had a mild hemolytic transfusion reaction. It was shown to possess thecharacteristics of an immune antibody and to be able to distinguish between asingle dose and a double dose of the Lu^b gene.

2. Three new examples of the antibody, anti-Lu^a, are presented. All of themwere found in normal blood donors and have properties which indicate that theyare naturally occurring antibodies.

Dr. R. R. Race and Dr. R. Sanger confirmed the presence of anti-Lu^b in Mrs. S.’sserum, and studied other members of her family and the three anti-Lu^a sera. We aregrateful to them for many favors and their kind encouragement.

We are obligated to Miss Marie Cutbush for making available the Lu^aLu^a cells fromMrs. R. and her sister, and for a supply of anti-Lu^b serum.

Thanks are due to Dr. A. E. Mourant who furnished our original supply of anti-Lu^aserum and to Dr. Philip Levine for the anti-Tj^a and anti-Vel sera.

We are indebted to Dr. J. M. Fine of Milwaukee for permission to study Mrs. S. andto the patient and her family for their cooperation.

The sera from 18,613 blood donors were studied by Betty McCarthy, Rosemary Polka,Pearl Lemke, Agnes Molnar, Jeannette Flagstadt and Betty Hutter.

Submitted on March 20, 1957 Accepted on July 1, 1957     

Changes in the Group A Antigen in a Case of Acute Myeloblastic Leukemia

1. The authors describe a case of acute leukemia in a man of 36 years. During the course of his illness, a change in the agglutinability of the red cellswas observed. The cells, originally A₁, changed continuously so that in somesamples it was possible to detect as many as three populations of red cells,namely, A₁, weak A, and so-called non-A. In the course of the illness, fluctuations in the numbers of these cells in the whole population could be observed.

2. The clinical course and the results of detailed isoserologic examinationsare presented.

3. Possible causes of these changes are discussed: (a) Somatic mutation;(b) chromosome abnormalities; (c) metabolic changes of the hemopoietictissue.

Submitted on October 25, 1960 Accepted on January 17, 1962     

华支睾吸虫ATP合酶b亚基模拟亚细胞定位与细胞周期的关系

目的了解华支睾吸虫ATP合酶b亚基(CsATP-synt_B)核定位序列(NLS)介导该蛋白进入细胞核与细胞周期的关系,及该蛋白对自身和宿主同源基因表达的影响。方法将已构建的CsATP-synt_B与绿色荧光蛋白(GFP)融合表达质粒(pEGFP-N1-CsATP-synt_B)转染Hela细胞,转染细胞经同步化处理后,在激光共聚焦显微镜下观察在不同细胞周期相,细胞不同部位的荧光分布。流式细胞技术检测该融合蛋白表达量的变化,并通过半定量RT-PCR分析不同细胞周期相CsATP-synt_B和Hela细胞同源基因的表达。结果转染细胞经同步化处理后,流式细胞检测结果显示,空载体pEGFP-N1在G0/G1期Hela细胞中的表达量明显下降,而pEGFP-N1-CsATP-synt_B在该时期表达量呈上升趋势;在其他细胞周期时相,两者的荧光表达量的变化趋势相似。显微镜观察发现,在G0/G1期、S期和G2/M期CsATP-synt_B集中在线粒体表达,G1/S期细胞核内见绿色荧光,且多数细胞的绿色荧光集中见于核仁。在不同的细胞周期,半定量RT-PCR分析结果显示,该重组蛋白进入细胞核后,目的基因CsATP-synt_B和人源性ATP-synt_B(Ho-moATP-synt_B)mRNA的表达均呈上升趋势,CsATP-synt_B上升幅度较大,但两者的表达变化趋势一致。结论 CsATP-synt_B在G1/S期进入细胞核,受细胞能量需求和细胞周期的调控。     

A study of sickling of young erythrocytes in sickle cell anemia

1. Data have been presented to show that most reticulocytes from patients withthe sickle cell trait or sickle cell anemia sickle as readily as do more mature redblood cells. The most immature reticulocytes and normoblasts tend to sickle moreslowly.

2. Orthochromatic normoblasts were the only type of normoblasts whichsickled; the basophilic and polychromatophilic types could not be sickled.

3. It is suggested that the sickle cell forms seen in ordinary stained smears represent old cells which have lost their "elasticity" while stagnating in the sickleshape, and are unable to revert to a biconcave disk. This would explain the factthat these forms are so rarely found to be reticulated when stained with brilliantcresyl blue.

Note: ACKNOWLEDGMENTI wish to express my gratitude to Dr. William B. Castle, Dr. William Dock (Department of Medicine)for many stimulating suggestions, to Dr. John M. Pearce (Department of Pathology) for assistance inpreparing this report and to Mrs. Muriel MacDowell (Department of Pathology) for the photomicrography.

     

Specific and cross-reacting antibodies in ABO heterospecific twin pregnancy

A "study in depth" is reported concerning the case of hemolytic disease of agroup B infant born to a group O mother, whose group A twin was apparentlyunaffected. It was shown that the hemolytic disease of the affected infant wasdue to a specific anti-B antibody. The study included parallel examinations ofthe antibodies in the sera of the three individuals.

The specificity of the B-anti-B reaction was demonstrated in vitro and in vivo.The powerful anti-B antibody of the mother had no effect on the group A twin,in whose serum anti-B was present in large amounts.

In vitro, studied by the usual technics of cross absorption the maternaland the fetal anti-A and anti-B serum antibodies behaved as if strictly specific.By applying successive absorption, elution and neutralization techniques, however, it was possible to demonstrate additional cross-reacting antibodies in thematernal serum which could be separated from one another and from the specificanti-A and anti-B antibodies. From the erythrocytes of the normal group Atwin such cross-reacting antibody could be eluted.

The cross-reacting anti-B antibody, isolated in pure form in eluates, could beshown to be loosely attached to group A erythrocytes without producing visibleagglutination reactions while after elution from A cells it did visibly agglutinategroup B cells. It could be eluted from A cells, absorbed by fresh A cells and reeluted while retaining its anti-B effect. It was neutralized by group B saliva only.A separate anti-A antibody with similar properties was eluted from B cells andspecifically neutralized by group A saliva.

A partial affinity of these antibodies for heterologous erythrocytes but not forspecific soluble substances was thus demonstrated. These findings support neitherthe linkage hypothesis of cross reactions between anti-A and anti-B nor theC-anti-C hypothesis of hemolytic disease. They are in keeping with the view thatgroup O sera contain variable complexes of anti-A and anti-B antibodies, composed of multiple fractions with different partial specificities. It is suggested thatthe occurrence or non-occurrence of cross-reacting antibodies found in sera ofgroup O mothers whose infants develop hemolytic disease is best explained onthis basis. It is further stressed that the demonstration of an antibody in motheror child in ABO hemolytic disease does not necessarily indicate its pathologicsignificance.

Submitted on January 11, 1957 Accepted on April 19, 1957