The Rh Antigen D’ (Wiel) BRUCE CHOWN, M.D., MARION LEWIS, B.A., HIROKO KAITA. SYLVIA PHILIPPS, H.Sc.

From the Department of Pacdiatrirs, University of Mnnifoba nnd Rh Labornfory, ll’innipeg, Canada

Evidence is brought forward confirming that the antigen “Wid” (Transfusion 2: 150, 1962) is part ot the D segment of the Rh system; ia rare in Whites (none in 13,OOO) but not uncommon in Ncgroa (9 pOaitive in 253). Ita name is changed to Dw, Rh(23) or r h W O , the common gena designated cDWc or Rt, CDwc or Ry, and cDsFE or Ry, and a table of phenotypes and genotypes given. When Dw is not partnered with D it reacta as a Du; its specific nature is recognized by reaction with anti-Dw.

IN 1962 we reported a new antigen, Wiel, the facts then available indicating that it was probably:

1. in the Rh system: 2. a part of the D segment of that system; 3. rare in Whites but, based largely on

evidence obtained by Dr. Eloise Giblett, probably less rare in Negroes.

Further evidence, here to be presented, makes these probabilities certainties. Be- cause of this, and to put Wiel in its place in current nomenclatures, we have changed the name of the antibody to anti-D\v, anti- Rh(23) or anti-rhwo, and that of the anti- gen to DIV, of the antigenic determinant to Rh (23), of the factor to rhwo. To avoid confusion that arises from the use of more than one terminology in the same article we use the British nomenclature in its long and short forms throughout this paper with the following additions for D”+ genes: cDwe or Rr, CDwe or R Y and cDwE or R w o ; Rtno is used to distinguish CDwc from

a 1

Work aupported in part by the National Foun- dation and by N.I.H. Grant HD00527. Presented at the 16th Annual Meeting AABB. Detroit. Nov. 7, 1963.

Received for publication December 20, 1963: ac- ccnrc’rl lanuary 13, 1964.

C11’llc or Ry, and R.tflO to distinguish cDtuE from cDEw or Ry.

The Specificitj of Anti-@. The serum “Bill,” which is that of a R,r woman and contains anti-Dw, was absorbed on the cells

I

11

I

codrn H Due 1 0 0 D”- i Mt tamtad.

FIG. 1. Family 2 (top) is Family 2 of reference 1, now extended to include two earlier generations, one additional member in generation I11 (111-1) and a new baby in generation IV (IV-4) : Family 3 (bottom) is new.

It is to be noted that without anti-D” all blwda shown as R:r would be classified as R:r. Further, without this antibody it would be impossible to establish the shown genotypes in generations I and I1 of Family 2 and in generation I of Family 3 and hence the sources of R . and R:. In setting out the gcnotypa two assumptions have been made: (1) the usual one, that r represents genotype rr; (2) that since 1-2 of Family 2 and 1-2 of Family 3 both give normal reactions with anti-D they carry a D in addition to D”, and therefore carry, the one R,, the other R, .

1-1, 1-3 and 1-4 of Family 2 are written aa they are because it is known from the progeny that 1-1 carries R,, 1-3 carries R , and 1-4 R,. Although the resulting symbolization is contrary to genetic convention it is more informative than is the convention, and the purpose of a convention is to be informative.

All members of these pedigrees excepting 111-1 of Family 3 are V- ea-.

169

170 CHOWN, E T AL.

TAD1.F. 1. Renctions with Selected Saline-Active Anti-D Sera of Dw-Negative and Dm-Positive Bloods Heterozygous for D

Reaction Times in Minutes; Capillaty Technic ~ ~~

Sera Bloods Genotype .4nd Chi Pal Lei Fra Sho Faw

Dw-negatives 3-1-1 3-11-3

Dw-positives 1-111-4 2-IV-2 2-IV-4 3-11-2 3-111-2 3-111-6 3-111-8

All Rzr

4 4 4 3 5 2 3 9 7 9 4 7 4 6 4 4 4 4 6 6 4

The minus (-) sign indicates no reaction at 60 minutes. Dw + blood 2-IV-4 is from a three- month-old baby and 3-111-8 is cord blood: anti-D "And" always reacts more strongly with the blood of newborn and young infants than with that of adults. The sera are the same as those in Table 1 of reference 1.

TABLE 2. Reactions of Rh Antigens in Dw+ Bloods Hekrozygmcsfor the Giuen Antigen

A. Bloods of Proven Genotype

Reactions Antigen Genotype Normal Weak

C * 6 RIR: 6 0 D 7 Rzr 0 7 e * 2 RoR: 2 0 f 6 RIR:, 2 RaR: 8 0

B. Bloods of Given Phenotype, Assumed Genotype in Brackets

Reactions Antigen Phenotype Normal Weak .,

~~ ~

C 1 R;"r (R;Or) 1 0 D 4 RE (RE?), 1 R;"r (R7"r) 0 5

*Although the RIRZ bloods are heterozygous for C, and the RaRZ for E, C and E are not car- ried by the D"+ gene in them so that their normal mactions are immaterial to the argument.

TABLE 3. Frequency of Dw in 253 Negro Bloods of five D"+ people, three of phenotype R; and one each of R y r and R;o

229 9 220 (Harper), and eluates made. All bloods in D+ D- 1_4 0 24 the pedigrees were tested with at least one

253 of these eluates, while all five eluates were Phenotype of the 9 Dw+ tested against and reacted with ten Dw+

D' +

" . _ RE 5 R;OR 1 1 bloods, six of phenotype R; two of R y r , R;"r 2 R;O (Harper) 1 one R y R I and one Ry (Harper) ; none

THE Rh ANTIGEN Dw (WIEL) 171

Matings

Caucasian Negro Total

Dw+ x D1- 2' 5' 7

Type

6' 109

Total 105 1 1 116 - - Dw- x D"- 103 -

TABLE 4. LW Types in 116 Families

Children

Total D1- D"+ obs exp obs exP

23 12 1 I .5 1 1 11.5 350 350 350 n 0

373 -

reacted with any of a large panel of Dw- bloods.

Evidence for Du' Being in the Rh System and Part of the D Segment. The first evi- dence that Dw is in the Rh system comes from a study of its transmission in families. One large family (Family 1) and part of a second (Family 2) were reported in our first paper.1 In Figure 1 Family 2 has been extended and a third family (Family 3) added.

In Family 1 Dw was traced through three generations; in Family 2 it is now traced through four, and in Family 3 through three. Clearly Dw is a dominant character fully expressed in the heterozygote. In all three families it is transmitted by the gene Ro ( R ; ) , so placing it firmly in the R h system.

The first evidence suggesting that Dn7 is part of the D segment comes from the reactions of Ror bloods with anti-D sera. In all three families all Ror bloods that are Dw+ react weakly or not at all with se- lected anti-D sera and therefore may be classed as Rtr, whereas those that are Dr- react normally with these sera and are classed as R,r; the reactions with anti-D sera are set out in Table 1. In addition to the seven Dw+ bloods of known genotype in Table 5 other Dw+ bloods have been found that give similar reactions with these anti-D sera; four are of phenotype R; and are assumed to be Ry, the fifth is assumed to be Ryor (Mrs. J.N. of ref. 1 ) . DW prob- ably always manifests as a sort of D".

If Dw is a product of the gene RO in these families, i t could be a part of the c, of the

TABLE 5. Exampler oj the Reactiom, Phenotyficr and Genorypcs o j Some D" + Bloods

Reactions Phenotypes Genotypes

C c D D " E e f I cD"c .cdc

RE1 CDIe. Cde

R;"R' CD"e,cDe; CDc.cD"e; CD"e.cD"c CD"e.cdc; Cde.cD"c ! R;"R. R I R : R;"RE I R;% R'R: + + (+) + - + + R7"r

cD"E.cdE 1 R';"R" cD'E . cDt; cDE . cD"c; cD"E. c D"e cD"E. cdc; cdE , c D"c 1 R;"R, R,R: R;''R; I R;"r; R"R; - + (+) 4- + + + R;"r

D is bracketed because the reactions will vary with the phenotype. Blooda to the right of the line dividing the genotypes will probably all react i~ll DYs; all cD"e.cdc bloods encountered and probable CD"c.cdc have so reacted. How a homozygoua D" will react we do not know, nor how R' and R, will affect the expression of D or D" in a D"+ blood.

172 CHOWN. ET AL.

D, or of the e segment. The reactions of the antigens c, D, e and f in Dw+ bloods heterozygous for these antigens are sum- marized in section A of Table ‘2 and those of the above-mentioned five bloods in sec- tion B. It may be seen that only the D antigen gives reactions out of the usual.

That Dw can occur in the absence of c and of f is proved by the finding of one Dw+ blood of phenotype KyR1 The blood referred to above as R? (Harper) has been studied in the laboratories of Drs. Race and Sanger and Dr. Cahan and in our own. It gives a number of irregular reac- tions and is thought possibly to be R 2 F . Perhaps in it DW is a product of R ,.

From all the above facts and obsektions we conclude that the D, and the D alone, is anomalous in Dw+ bloods and that Dw is therefore a part of the D segment.

The Racial and Family Distribution of Dw. Although both the original proposita and Mrs. J.N., referred to above, were Whites we have not found another positive among 13,000 random bloods from Whites. In contrast nine of 253 Negro bloods sent us from Nova Scotia, Virginia and New York have been positive. The phenotypes of these nine are set out in Table 3. The inheritance of Dw in 116 families is set out in Table 4.

Anti-Dw. Dw in Negroes has roughly the same frequency as CW in Whites. If they are about equally antigenic, anti-DW should not be a very rare antibody in the blood of Rh-positive patients transfused with blood from Negroes, and should be a rare cause

of hemolytic disease of the newborn of Negro fathers: in the latter case the antigen might easily pass at first for a “private” or “family” antigen. At times anti-Dw will be associated with other Rh antibodies: the present serum was that of a woman of phenotype R,r and contained anti-C, anti- Ce, separable anti-Cw, and anti-V as well as anti-Dw. Anti-Dw in association with anti-D would not be recognizable. We are anxious to identify further examples of anti-Dw and would be glad to examine suspect sera.

Terminology. The superscript-w, whether it refer to Cw, DW or Ew, is perfectly clear in the CDE nomenclature but can be con- fusing for shorthand phenotype and geno- type descriptions. We have given our sug- gestions earlier in this paper for the commoner D w + genes in the CDE and short British symbols. Examples of the resulting phenotypes and genotypes are set out in Table 5.

Acknowledgments \Vc arc grateful to the families for their generous

cooperation, to Drs. Herbert Read, Bruce Morton and Amos Cahan and Mr. Bartholomew Picone for seiitling US tlic Negro bloods, and to Miss Catherine Anderson for the random blood of Whites.

Addendum A recent study of a Caucasian family in collabora-

tion with Dr. C. R. Macphenon, of Columbus, Ohio has confirmed the existence of Ilh gene complex CDwe (R1w0).

Reference 1. Chown, B., M. Lewis and H. Kaita: A new Rh

antigen and antibody. Transfusion 2: 150, 1963.