RMI Nuclear Justice Documents

0 MARCH 25, 1961 ORIGINAL ARTICLES * Furthermore, when an Ag(a+) serum was submitted to starch-gel electrophoresis, the slow-«, fraction recovered . * by the technique of Gordon (1960) failed to show precipitat; ith pitation with th i-Acta- the anti-Ag(a-+) serum. The e furth further identification of the antigen present in the z-globulin is currently being undertaken. Characterisation of Antibody The patient’s serum was submitted to immunoelectrophoresis, and a serum giving a strong reaction in the Ouchterlony tests was placed in the side well. Within 18 hours a well-defined precipitation line appeared in the y-globulin region (fig. 4). This result, together with the shape of the precipitation curve in the Ouchterlonyplate, suggested that the antibody wasa relatively low-molecular- weight +-globulin. This interpretation was confirmed by an experiment in which the 7S- and 19S-+-globulin components in the patient’s serum were separated by density-gradient centrifugation. Precipitation lines in Ouchterlony plates were obtained with low-molecularweight fractions giving no reaction with immune-rabbit serum against 19S human y-globulin as described by. Franklin (1960b). It can, therefore, be concluded thar the antibody is a 7S-y-globulin. The serum of the patient’s daughter showed no reaction when tested against panel sera nor against her father’s serum. Stability of the Antigen The antigen is unaffected by heating at 57°C for 40 minutes, by hemolysis with its own or foreign human hemoglobin, or by storage at room temperature for up to 7 days. The antigen was not lost by dialysis or ultrafiltration. Sera of either type (reacting or non-reacting) TABLE I-—FREQUENCY OF POSITIVE REACTORS E Population 7 No. studied es o. ew Positing Parents of white U.S. families - U.S. Negroes. 1955, 1 was a reactor and the othernot; the reactions in sera withdrawn from the samesubjects in 1959 were unchanged. Variation in the position of the slow-«,-globulin after starchgel electrophoresis has been noted. By meansofvertical electrophoresis, using borate buffer at pH 8-6 (Smithies 1959), a slow-moving, a fast-moving, and a double band can be dis- tinguished. I: has been suggested that this variation may be due to storage (Smithies 1955); but this may not be the only explanation: Sera from different persons stored for the same length of time under identical conditions may show the variation, and not all stored sera show it (Blumberg 1961). In any event, this does not appearto affect the reaction with the patient’s serum. Persons of each of the three slow-a,-types were both positive and negative reactors. It appears, therefore, that the antigen is relatively stable and persistent in any one person. Distribution of the Antigen in Populations Because the supply of antibody serum is limited, extensive population surveys have not been possible. However, positive and negative sera were found in all * G12 719 F HF a the populations studied. On the basis of preliminary examinations, the frequency seems to vary in different populations. The frequencies of positive reactors in some C? mee ee et sistently gave the same reaction. Of 2 sera withdrawn in 36 31 Micronesians (Rongelap, Marshall Islands) - wari HI—-ORSERS 3205 10ih - 51 —————— earison of cb 50 (96%5 meespet of Agta - where there is ati =. e eee Tre More extensive stydi populations are shown in table 1. are planned when micromethods are developed. aie Inherttance of the Antigen a 126 sera from 29 American families (25 white,.4 Negro) were studied. The results are shown in table g. In addition, 38 sera from 8 Micronesian families wep tested. Since the frequency of positive reactors was in the Micronesians,all of these were Ag(a+-) x Agta+} matings, and nearly all the offspring were Ag(a+). Thee are also shown in table 11, but are not included in the. statistical evaluation, since they are not decisive in t the genetic hypothesis. In the families from the Uniteg States, among the offspring of positive X positive ang positive x negative matings, positive and negative og. spring were found, whereas among the offspring of Negative Xx negative matings only negative offspring appeared. This result suggests that the inheritance of the antigen follows mendelian segregation, with negative subjects homozygous for a recessive gene Ag, and positive subjects homozygous or heterozygous for theallelic gene Ag. It is suggested that the Ag4 gene controlsthe. TABLE II—-INHERITANCE OF THE ANTIGEN t Mating type No.of families stored for as long as 4 years at —20°C were consistently found to give the same reaction as fresh sera from the same subjects. The reaction in the fresh sera, however, was in some cases stronger and better defined than in the stored sera. Repeated freezing and thawing of the sera up to at least fifteem times did not affect the reaction, ‘Blood-samples were withdrawn twice weekly for 4 weeks from two normal volunteers whose sera reacted strongly with that of the patient. The reaction remained unchanged during this period. Sera withdrawn approximately twice yearly over a 2-year period from six subjects, two of whom reacted and four of whom did not, con- -- United States: Ag(at+)xAgiat) .. Agiat)xAgia—}.. Agla—)xAgla—).. Agta +) x Ag(at+) ae pees Ag(a—) 11 10 21 9 0 20 8 21 1 8 synthesis of the Ag(a+) antigen. + S jo lwo Offspring Ag(a+) , et te 5 13 Total.) ssa Mi “26. ee, cathe If a product of the . allelic gene is found, the gene could be termed Ag4 and . the antigen Ag(b +). ape The family data were analysed by the imethods sume marised by Smith (1956). In this, four types of compan-. son are made,iin each of which a correction is included for - family size. The comparisons of observed and expected — figures involve: SER. 1. The number of recessive Ag(—) children resulting fromAg(+) Ag(a— ) matings, given the number of families cam which there is at least one recessive child. 2. The numberofrecessives resulting from Ag(a+) x Agar) matings in which there is at least one recessive offspring, gives the total numberof families with at least one recessive offspring 3. In Ag(a-+-) x Agfa—) matings, the total numberof famibes with at least one recessive offspring, given the total number Ag(a+) x Ag(a—) matings and the gene frequencies 10 population from which the families were selected (q=frequest Ag= 0- 66). - Sah. : 4. In Ag(at+)x Ag(at+) matings, the total number oe families with at least one recessive offspring, given the number of Ag(a-+) x Ag(a*+) matings and the gene frequench The first two computations are independent of te gene frequencies in the population study. For the and fourth computations, however, an estimate is 0 of the gene frequency in the population from which families were drawn. We were unable to obtain a lant random sample from the population from which families were selected, and the gene frequency. was estimated from the frequency of the phenotypes in.oe a her of Agia ~ where there is ati ‘ parison of of Nerber of fam: Me3 . child, = aeney of Q- 6 eparben of ot he ther of fare child. - afeast of 06 mrarinanl of ot shee of rt \e a} matings ~arental mem ~7° were fou The results syole 1, whic \sa- ) maur -reliminaryfo -vpothesis. C ‘smilies, no & rilawing blo ABC, Rh, M°: surs to be re ‘he generic ho Independence 0. tariPPOTeD NS The reacti fractions Wer ~aproglobin, ceyregation 0% adependent . san x-globui focribed inh Aopartz (196: t Stockholm tmean Ge ty sectrophores: and negative types l-1, 2t the same wstem is in ‘“stum-protei: -stinct from Ticcipitate in «. 1900); th 2 serum and A factor is ‘“steactive p Tevion in j “inn 1959), The prec: » esence of a =9n5 so far w= the prec ‘tigen appe: a "morphic “tvealed, _ Besides be “Re finding ¢