Humangenetik 5, 294--304 (1968)

Genetic Studies on the Deficiency offi2-Glycoprotein I of Human Serum*

] -~ARTW I G C L E V E St"

Institut fiir Humangenetik (Direktor : Prof. Dr. G. G. WE~DT) der Universitgt Marburg/Lahn • dJ

Received January 29, 1968

Summary. fi2-glycoprotein I concentrations have been determined in human sera by the radial immunodiffusion technique. In 260 healthy individuals a bimodal distribution was observed. In 94% of sera concentrations ranged from 16 to 30 mg/100 ml with a mean value of 21.3 -E 3.6 mg/100 ml. In 6% of sera lower concentrations were found: ~ = 10.0 ± 1.3 mg per 100 ml. The results of a study of 38 families with 79 children suggest that fl2-glycoprotein I concentrations are controlled by a pair of autosomal co-dominant alleles called Bg N and Bg D. Individuals homozygous for the common allele Bg N have levels between 16 and 30 mg/100 ml, heterozygous individuals have lower concentrations of 6 to 14 mg/100 ml. Individuals homo- zygous for the rare allele Bg D have deficiency of fl2-glycoprotein I. The results in one family, however, did not conform to this simple genetic hypothesis. The concentration of fi2-glyco- protein I in serum is also influenced to some extent by non-genetic factors such as sex, age, and various pathologic conditions. The biological function of this glycoprotein is still unknown.

In the preceding repor t by HAUPT et al. (1968) a family wi th an apparen t ly

inher i ted defect of the/~2-glycoprotein I in two of its members is described. This

observat ion in i t i a ted the present study, fl~-glycoprotein I concentra t ions were

de te rmined in scra of hea l thy individuals , in sera of pa t ien ts wi th var ious patho-

logical conditions, and in sera of a family ma te r i a l by the radial immunodif fus ion

t echn ique of M A r t i n i et al. (1963). Fu r the r in format ion on the inher i tance of this

serum prote in deficiency has the reby been obtained. The s tudy provided also

informat ion on the influence of non-genet ic factors upon the serum concent ra t ion

of this protein.

Methods

Quantitative determinations of fl2-glycoprotein I were carried out with the radial immuno- diffusion technique of ~ANeI~I et al. (1963). The details of the procedure have been described previously (CLEvE, 1966). In this study prefabricated agar plates of the Behring-Werke Marburg/Lahn have been used (Partigen®-plates, Op. Nr. 80 and Op. Nr. 102). Standard curves were obtained with stabilized human sera (Op. Nr. 166F and 166L) which had a fl2-glyco- protein I concentration of 14 mg/100 ml. The concentration of fle-glycoprotein I in these standard sera was assayed by the Behring-Werke laboratories. Undiluted serum and 1:2, 1:3 and 1:4 dilutions were used for the standard curves. The r2-values of the precipitation halos were plotted against the concentrations of fl2-glycoprotein I. The assays were carried out with 1 : 3 dilutions of the sera to be tested. Sera with apparently very low concentrations

* Supported by a grant from the Deutsche Forschungsgemeinschaft, Bad Godesberg, and by U.S. PI-IS Grant AM 11796.

** Present address : Cornell University Medical College, Department of Medicine, Division of Human Genetics, New York, 10021, N.Y.

Genetic Studies on the Deficiency of/~2-Glycoprotein I of Human Serum 295

were also examined undiluted. The diameter of the precipitation halos were measured as described (CLErk, 1966). The r2-values were calculated and the fl2-glycoprotein I concen- trations were read from the standard curve.

Material

Blood samples were drawn by puncture of the cubital vein without addition of anti- coagulants. Sera were separated and stored in the deep-freeze at --20°C. The samples had been repeatedly thawed and frozen for other purposes prior to this investigation.

The following specimen were included: 1. a) 100 sara from apparently healthy students, the blood was drawn in the morning

while students were fasting. b) 100 sara from apparently healthy members of the Bundes-Grenzschutz, this sample

will be referred to as guardsmen. c) 60 sera from blood donors. 2. Sera from 38 families with a total of 79 children. This material was provided by Doz. Dr. T. BE~R~D, Medizinische Poliklinik, University

of Marburg an der Lahn. The sera were collected during her survey of a rural village population on the prevalence of rheumatoid arthritis (BEm~END, 1966). The village is located near Mar- burg. Most of the persons included in this material were apparently healthy at the time the blood was obtained.

3. 38 cordsera from newborns. 4. 37 sera from pregnant women. 5. 198 sara from patients with various pathologic conditions.

Results and Discussion

Reproducibility o/Determinations

I n 20 de te rmina t ions of the s t andard h u m a n serum a mean value of 2 = id .8mg

per 100 ml and a s t andard devia t ion of s = ~ 0.877 mg/100 ml was found. The im- munodiffusion technique thus appears to provide re la t ively accurate results. The cons tancy of serum concentra t ions of fl2-glyeoprotein I in an ind iv idua l over a longer period of t ime was examined in 6 persons. F rom these 6 persons, blood samples were t aken twice with a 9 m o n t h interval . Table l i l lustrates t ha t over a period of 9 months , the serum concen-

t ra t ions of this protein remained con- s tant . Inf luence of storage was examined in 6 sera (Table 2). No signifieant ehanges were observed, when the results obta ined with fresh sera were compared with the values found after storage of sera in the deep-freezer at - - 2 0 ° C for 2 days to 4 weeks. Fur thermore , in a sample of sara

Table 1. Serum concentrations o/ fl2-glyco- protein I in 6 individuals

Concentration Concentration at 1. examination 9 months later mg/100 ml rag/100 ml

1. 19.2 22.2 2. 26.6 26.6 3. 12.6 10.2 4. 23.4 26.6 5. 20.4 20.4 6. 22.2 20.4

from 60 hea l thy blood donors similiar concentra t ions were found as in two other samples from s tudents and guardsmen, respectively (Table 4). The sara of blood donors had been stored in the deep-freezer for 2 years, the sera from guardsmen for 15 months , and the sera from s tudents for 9 months . Therefore, storage of sera frozen at - - 2 0 ° C apparen t ly does no t influence the concent ra t ion of fl2-glyeo- protein I. Also, the influence of enzymat ic cleavage of sialic acid by neuraminidase

296 H. CL~VE:

on the immunodif fus ion assay of th is p ro te in has been examined . F o r these experi- men t s a purif ied neuramin idase p r e p a r a t i o n f rom Vibrio cholerae (Behr ing-Werke) was employed . I n Table 3 is shown t h a t the diffusion ra te of fl~-glycoprotein I

Table 2. Serum concentrations ol fi~-glyeo- Table 3. Serum concentrations o/ fl2-glyco- protein I alter storage protein I belore and alter treatment o I sam-

ples with neuraminidase Fresh Serum fro- Storage at serum zen, kept - - 20 ° C for Concentrations Concentrations mg/100 m] at - -20°0 4 weeks, before treatment after incubation

for 2 days, frozen and mg/100 ml with neuramini- and thawed thawed two dase rag/100 ml times rag/100 ml

mg/100 ml 1. 23.4 26.6

1. 22.2 23.4 23.4 2. 19.2 20.4 2. 26.6 26.6 26.6 3. 26.5 23.4 3. 10.2 11.3 11.3 4. 22.2 20.4 4. 26.6 29.7 28.2 5. 24.9 26.6 5. 20.4 22.2 19.2 6. 20.4 22.2 23.4

is no t influenced to a no t icab le degree b y the sp l i t t ing off of sialic acid. No signifi- can t changes of the values were observed af te r t r e a t m e n t wi th the enzyme prepa- ra t ion .

Concentration o] fl~-Glycoprotein I in Healthy Individuals

The concen t ra t ion of fl~-glycoprotein I was de t e rmine d in 260 sera f rom ap- p a r e n t l y h e a l t h y persons. The d i s t r ibu t ion o£ the values is shown in Fig. 1. Two modes can clear ly be d is t inguished. I n the m a j o r i t y of sera the concen t ra t ion falls

n

50'

40'

3C

2C

1C

o lO

m

20 -trk,

30 mg°/o

Fig. 1. Distribution of fi2-glycoprotein I concentrations in sera of 260 healthy individuals

wi th in the range of 16 to 30 rag/100 ml , t he mean value was found to be 2 ~ 21.3mg per 100 ml, the s t a n d a r d dev ia t ion was s = ± 3.6 mg/100 ml (Table 4). The distr i- bu t ion in th is mode is s imil iar to a no rma l d i s t r i bu t ion curve, there is, however, a s l ight excess of h igher values. A second mode is cons t i t u t ed b y the values found in a p p r o x i m a t e l y 6 % of the sera, the mean value of th is mode is 2 = 10.0 mg per 100 ml, the s t a n d a r d dev ia t ion is s ~ -L 1.3 mg/100 ml (Table 4). I n th is sample the two modes are c lear ly s epa ra t ed wi thou t over lap (Fig. 1). Subdivis ion of the

Genetic Studies on the Deficiency of fl2-Giycoprotein I of Human Serum 297

t o t a l sample according to the or iginal sources of the sera shows s imil iar d i s t r ibu t ions of f l2-glycoprotein I concen t ra t ions in s tuden ts , guardsmen , and b lood donors. I n pa r t i cu la r , the f requency of persons wi th low se rum concent ra t ions of fl2-glyco- p ro te in I is s imi l iar in the th ree samples (Table 4).

Table 4. Concentration o/fi2-glyeoprotein I in sera o[ healthy individuals

Sample m . v . :t: s n n 1 ~Tb.V. -~ 8 112

t o t a l in rag/100 ml in mg/100 ml

Students 100 94 19.9 2.8 6 9.6 1.0 Guardsmen 100 95 21.6 3.3 5 9.2 1.0 Blood donors 60 55 23.2 4.3 5 11.3 0.9

Total 260 244 21.3 3.6 16 10.0 1.3 m.v. = mean value; 4-s ~ standard deviation.

Fami ly Studies

38 famil ies wi th a t o t a l of 79 chi ldren were examined , the resul ts are summar i zed in Table 5. Three classes were d i s t ingu ished according to the concen t ra t ion of f l2-glycoprotein I : INormal concen t ra t ion was defined as the range f rom 17 to 35 rag/100 ml. 64 pa ren t s a n d 71 chi ldren be longed to th is class N. Also 4 pa ren t s were la te r inc luded in th is group, one p a r e n t h a d on r e p e a t e d de t e rmina t ions a concen t ra t ion of 16.5 mg/100 ml, th ree pa r en t s h a d higher values of 36.6 to Table 5. f l2 -g lycopro te in I-co~centrations in 40.2 rag/100 ml. As i n t e r m e d i a t e concen- 38/amilies with 79 children

t r a t i o n the range f rom 6 to 14 rag/100 ml Parents n Children Total was defined. This class I inc luded 8 pa- N I D rents and 6 children. 2 chi ldren were ob- served wi th v e r y low concent ra t ions , t h e y N × N 30 64 0 0 64 were classified as deficient for fi2-glyco- N × I 8 7 6 2 15

p ro t e in I (D). I n the immunodi f fus ion 38 71 6 2 79

p la tes a fa in t p rec ip i t a t e was seen in the N = normal concentration sera f rom these 2 persons which indi- I = intermediate concentration cared the presence of a ve ry smal l a m o u n t D = fl2-glycoprotein I-deficiency

of th is pro te in . The concen t ra t ion could no t be measu red wi th accuracy , i t was be low 1 rag/100 ml. Table 5 shows t h a t in the 30 famil ies in which bo th pa ren t s be longed to class N, also al l 64 chi ldren h a d no rma l concent ra t ions of f i2-glycoprotein I . I n 8 famil ies one paren t , e i ther the fa the r or the mother , h a d i n t e r m e d i a t e concent ra t ions . F r o m the i r 15 children, 7 h a d no rma l and 6 h a d i n t e rmed ia t e concent ra t ions , 2 chi ldren were deficient for f la-glycoprotein I . Pedigrees of these 8 famil ies are p re sen ted in Fig. 2. The d i s t r ibu t ion of fl2-glycoprotein I concent ra t ions in 7 of the 8 famil ies is in agree- m e n t wi th the a s sumpt ion of a MEI~DELIA~ mode of inher i tance . N o r m a l concen- t r a t ions would he reby cor respond to the p h e n o t y p e N of ind iv idua l s homozygous for an allele r egu la t ing the concen t ra t ion of f l2-glycoprotein I a t the level defined as t he no rma l range. I n t e r m e d i a t e levels would cor respond to the p h e n o t y p e I of ind iv idua l s he te rozygous for an allele, which in the homozygous t y p e would

21 Humangenetik, Bd. 5

298 H. CLnVE:

lead to a deficiency of this protein. The notations for the two alleles should be fin G I N and fl~ G I D. More simply, the abbreviations Bg N and Bg D may be employed, Bg thereby referring to Beta-2-glyeoprotein I. In this model the phenotypes N, I, and D correspond to the genotypes BgN/Bg N, Bg~C/Bg D, and BgD/Bg D, respectively. The results in 7 families of Fig. 2 and in the 30 families with normal concentrations support this interpretation. An autosomal co-dominant mode of inheritance of the two alleles may be assumed. Obviously, the results in family Nr. 16 do not conform

4. ~ 11.

19.2 19.2 12.6 11.3

28.2 ~3.8 0-t 0-1

17. ~ 27. ~ .

113 19,2 26.5 13.8

31. ~ 37.

26.6 12.6 23/-

Fig. 2. Segregation of fl~-glyeoprotein I deficiency in 8 families. Concentrations of/~2-glyeo- protein I are given in mg/lO0 ml

with this simple genetic hypothesis. In family Nr. 16 the father had normal and the mother had intermediate concentrations of fi~-glycoprotein I. The 2 children examined had both very low concentrations of this protein. The model of a two allelic system can only be applied to this family ff variation in the phenotypic expression in one of the genotypes is permitted. Either the 2 children deficient for fi~-glycoprotein I are to be classified as individuals heterozygous for the trait, or the father who has a normal level is a heterozygous carrier.

I t has to be pointed out, furthermore, tha t in this family material several individuals had relatively high concentrations of fl2-glycoprotein I. Also, in the 3 samples of normal, healthy individuals a slight excess of persons with high levels was noted. I t is possible, tha t these persons should not be classified as homozygous normals. Analysis of the family results does not exclude with certainty the ex- tension of the genetic model to a three allelic system with a third allele accounting for the high concentrations which were observed in these persons. However, no clear distinction of classes of persons with normal and with high levels can be

Genetic Studies on the Deficiency of fi2-Glycoprotein I of Human Serum 299

made in th is mate r ia l . This poss ib i l i ty , therefore , could no t be s u b m i t t e d to a cr i t ica l tes t . The resul t s of the q u a n t i t a t i v e de t e rmina t i ons of fl~-glycoprotein I in t he i nd iv idua l member s of the 38 famil ies wi th specif icat ion of age and sex are given in the a p p e n d i x to pe rmi t the inclusion of th is m a t e r i a l in the analys is of a larger s tudy .

Frequency o[ Ind iv idua l s wi th Intermediate Concentration

I n a sample of 260 h e a l t h y ind iv idua l s f rom Marburg (Germany) , 16 persons were found wi th i n t e rmed ia t e /~2-glyco- p ro te in I concen t ra t ions (Table 6). If , in accord wi th the genet ic hypothes i s , these ind iv idua l s would r ep resen t the he tero- zygotes wi th the geno type Bg~ /Bg D the ca lcu la ted gene frequencies would be 0.97 for Bg N and 0.03 for Bg D. The obse rved he te rozygo tes f requency was 6.15% or 1/16, the ca lcu la ted homozygo tes f ie- quency is 1/1056. The obse rved d i s t r ibu t ion of pheno types is in good ag reemen t wi th the d i s t r i bu t ion expec ted i f the genet ic berg equ i l ib r ium m a y be assumed.

Table 6..Frequency o] intermediate concen- trations o/fl2-glycoprotein I in healthy persons

Sample n I

Students 100 6 Guardsmen 100 5 Blood donors 60 5

Total 260 16

n 37 I D

Obs. 260 244 16 0 Exp. 260 244.2 15.5 0.2 Gene frequencies : p = BgN = 0.9692

q = BgO = 0.0308

N = normal concentration I = intermediate concentration

D = fl~-glycoprotein I-deficiency

hypo thes i s is correct and H a r d y - W e i n -

Table 7. Concentrations o] fl2-glycoprotein I i~ sera o/males and ]emales

Sample n nl m.v. :J= s n~ m.v.

Family material, parents d" 38 32 28.1 4.3 6 11.3 38 36 26.9 5.2 2 13.2

Family material children d ~ 41 38 24.9 4.5 3 12.6 36 33 25.1 3.9 3 12.6

Students d t 58 54 20,5 3.1 4 9.3 0 42 40 19.2 2.2 2 10.2

m.v. = mean value in rag/100 ml j= s ~ standard deviation in mg/100 ml

Statistical analysis by t-test

Sample t 2 P

Family material, parents d ~ : ~ 1,0483 0.20--0.30 Family material, children ~ : ~ 0,1796 0.80--0.90 Students d ~ : 9 2,3225 0.02~0.025

Non-genet ic Factors which Inf luence the ~2-Glycoprotein I Concentration

Sex: Table 7 shows the ana lys i s of 3 samples according to sex. I n 2 samples s l igh t ly h igher values were observed in males t h a n in females. I n 1 sample th is difference was s t a t i s t i c a l l y s ignif icant a t the 5 % level. This sample was composed

21,

300 H. CL~W:

of s tuden t s , mos t of t h e m be tween 20 and 25 years of age. The 2 o ther samples showed no signif icant differences be tween males and females.

P r e g n a n c y : I n 37 p r e g n a n t women r e l a t i v e ly low concen t ra t ions of fi2-glyco- p ro t e in I were found. 2 women h a d i n t e r m e d i a t e levels of 7.8 a n d 9.0 rag/100 ml. I n 35 women concen t ra t ions r anged f rom 12.6 to 23.4 mg/100 ml. The mean va lue

Table 8. fi2-glycoprotein I concentrations in sera o] pregnant women

n rl I m.v. :J: s n 2 m.v.

in mg/100 ml in rag/100 ml

37 35 17.9 2.4 2 8.4

was 17.9 :L 2.4 mg/100 ml. Ana lys i s of 28 cases according to the m o n t h of preg- n a n c y d id no t r evea l charac te r i s t i c changes in the course of p regnancy , l~e la t ive ly low values were obse rved in mos t of the cases f rom the 5th to the 10th m o n t h of p regnancy .

A g e : Younger people appea r to have s l igh t ly lower fl~-glycoprotein I concen- t r a t i o n s t h a n older people (Table 9). A signif icant difference is found be tween the mean va lues of samples of younger b lood donors and of older b lood donors :

Table 9. Serum concentration o~ fl:-glycoprotein I in different age groups

Sample n n 1 m.v. ~ s n~ m.v. :J: s

l~ew borns 38 33 14.5 2.2 5 5.9 0.6 Blood donors, 20--30 years 30 27 22.0 4.0 3 10.9 - - Blood donors, 50--65 years 30 28 24.4 4.4 2 11.9 - - Family material, 10--19 years 23 22 25.3 4.5 1 13.8 - -

20--29 years 38 36 25.2 4.1 2 0--1.0 - - 30--39 years 14 11 23.4 3.9 3 12.6 - - 40--49 years 17 16 27.2 5.2 1 11.3 - - 50--59 years 35 31 27.9 4.7 4 12.3 - - 60~69 years 25 21 27.0 5.0 4 11.7 - -

m.v. ~ mean value in mg/lO0 ml; _u s ~ standard deviation in mg/lO0 ml.

t = 2.072, 2 P ~- 0 .025--0.05. Also, the analys is of the f ami ly ma te r i a l reveals s l igh t ly h igher levels in older people, t he difference is aga in s t a t i s t i ca l ly signif icant : F = 2.603, F5,1~ 5 = 2.29 for P ~ 0.05. R e l a t i v e l y low concent ra t ions of fl~-glyco- p ro te in I are found a t b i r th . I n 38 newborns , 5 h a d concen t ra t ions of 5.4 to 6.6 mg per 100 ml, t h e y cor responded p r o b a b l y to t he p h e n o t y p e I . I n the o ther 33 newborns a range f rom 10.2 to 19.2 mg/100 ml and a mean va lue of 14.5 -b 2.2 mg/100 ml was found. The difference to t he m e a n va lue of h e a l t h y adu l t s is s t a t i s t i ca l ly h igh ly significant .

D i e t : Since in a sample of 100 s tuden t s who h a d fas ted s l igh t ly lower concen- t r a t i ons were obse rved t h a n in two samples f rom non-fas t ing h e a l t h y individuals , t he influence of food i n t ake upon the se rum concen t ra t ion of th is p ro te in was examined (Table 10). I n 3 ind iv idua l s concent ra t ions were assayed in t he fas t ing

Genetic Studies on the Deficiency of fl2-Glycoprotein I of Human Serum 301

s ta te and 4, 8, and 12 hours af ter in take of a f a t t y meal. No significant changes

were observed, the concent ra t ion of /~2-glycoprotein I appears to be unaffected

by food in take.

Table 10. Concentrations o/fl2-glycoprotei~ I in sera o/three persons aJter /cod intake (in mg]lO0 m 0

l~ast After After After 4hours 8 hours 12 hours

I. 19.2 i9.2 19.2 19.2 2. 22.2 20.4 19.2 19.2 3. 22.2 26.6 24.9 26.6

Diseases: fi~-glycoprotein I concent ra t ions were de te rmined in sera of 198 pa-

t ients wi th var ious pathologic conditions, the resul ts are summar ized in Table 11.

25 pa t ien ts had chronic l iver diseases. I n mos t of these pa t i en t s pro te in syn-

thesis was severely impa i red as ev idenced by significant decreases of serum con-

centra t ions of cer ta in p lasma proteins which are synthes ized by the liver. These

Table 11. Serum concentrations o] fl2-glycoprotein I in various pathologic conditions

Diagnosis n n 1 range m.v. =~ s n 2 range m.v. -4- s

ia mg/100 ml in mg/100 ml

Liver cirrhosis with porto- caval anastomosis 5 0 - - - - - - 5 3.3--15.0 9.0 4.5

Liver cirrhosis 19 12 16.5--24.9 19.5 2.7 7 0.6--11.3 6.2 3.8

Liver metastasis 1 - - - - 1 - - 1 1 . 3 - -

Stomach cancer 32 26 15.0--,94.9 19.6 3.0 6 11.3--13.8 12.3 1.2

Lymphogranulomatosis 10 8 16.5--23.4 19.5 2.3 2 13.8 13.8 - -

Reticulo-sarcoma 2 2 16.5--24.9 20.7 - - - -

Multiple myeloma 6 4 17.7--23.4 30.6 - - 2 9.0---12.6 10.8 - -

Macroglobulinemia Waldenstr6m 2 2 16.5--26.6 21.6 - - - -

Chronic progressive 25 23 17.7--33.0 23.5 4.7 2 10.2--12.6 11.4 - - polyarthritis

Spondylitis ankylopoietica 3 3 19.2--24.9 22.5 - - - -

Erythema nodosum 1 1 - - 23.4 - - - -

Colitis ulcerosa 1 1 - - 28.2 - - - -

Diabetes melli~us, pat. with retinopathia 46 41 19.2--53.4 28.4 6.8 5 9.0---13.8 11.4 1.9

Diabetes mellitus, pat. with- out retinopathia 45 42 16.5--31.5 23.8 4.0 3 9.0--12.6 11.0 - -

changes have been repor ted earl ier and inc luded decrease of the concentra t ions

of acid a l -g lycopro te in and of the group-specific componen t (CLErk. and STgog-

MrYEg, 1967). 13 pa t i en t s showed decreased levels of fi~-glycoprotein I, the values

ranged f rom 0.6 to 15.0 rag/100 ml. I n the remain ing 12 pa t ien ts a range f rom

16.5 to 24.9 mg/100 ml was observed. I n par t icular , all the pa t i en t s wi th severe

l iver cirrhosis had a low fi2-glycoprotein I concentra t ion. I n pa t ien ts wi th chronic

302 H. CL~vE:

liver diseases, therefore, classification of individuals with genetically determined fl~-glycoprotein I deficiency is not possible. These findings indicate, furthermore, that the liver is probably the site of synthesis of this glycoprotein.

32 patients with stomach cancer were examined. 6 patients had intermediate concentrations, the frequency of 6/32 appears to be relatively high. Furthermore, there was no clear distinction between normal and intermediate levels in this group. I t is possible, therefore, tha t stomach cancer causes in some patients a decrease of the serum concentration of this protein. In a ]imited number of patients with lymphogranulomatosis, reticulosarcoma, multiple myeloma, and macroglobulinemia WaldenstrSm no significant changes were observed.

25 patients with active rheumatoid arthritis had values corresponding to the normal range. I t is noteworthy, tha t this glyeoprotein does not participate in the increase of several of the known glycoproteins of human serum which occurs during inflammatory reactions or in neoplastic diseases.

91 sera of patients with diabetes mellitus were studied. The distribution of fl~-glycoprotein I concentrations in 45 diabetics without retinopathia corresponded to the distribution in the normal control. In 46 diabetics with retinopathia, how- ever, a significant increase was observed. The mean value in this sample was 28.4 ± 6.8 mg/100 ml. Some patients in this group had concentrations as high as the two fold of the normal mean concentration.

The data summarized in Table 11 provide no information on the possible biological function of this glycoprotein.

Acknowledgements. The author would like to thank Drs. H. G. SCWCTICK, K. STSRIKO and Mr. H. HAurT from the Behring-Werke Marburg an der Lahn for their kind cooperation. Thanks are due to I)oz. Dr. T. BE]tl~END, Medizinische Poliklinik, Universit~it Marburg an der Lalm, for providing the family material. The expert technical assistance of Mrs. G. MSLLE~- SCOFFER is gratefully acknowledged.

R e l e r e n e e s

BEn~END, T. : Epidemiologische Untersuehungen in einer l~ndlichen BevSlkerung und in Sippen yon Probanden mit chronischer Polyarthritis. Habilitationssehrift, Marburg, 1966.

CLEVE, H. : Quantitative immunologische Bestimmung yon saurem al-Glykoprotein und ax- Antitrypsin; Serumkonzentrationen bei gesunden Blutspendern. Klin. Wschr. 44, 1256-- 1260 (1966).

- - u. G. STROm~I~Y]~R: Quantitative Variationen yon Serumglykoproteinen bei pathologischen Prozessen; Bestimmung yon saurem al-Glykoprotein, Ge und a~-Makroglobulin mit der radialen Immunodiftusion. Klin. Wschr. 45, 1051--1054 (1967).

ttAU]~T, H., H. G. ScnwieK u. K. STORIXO: t~ber einen erblichen ~-Glykoprotein-I-Mangel. Humangenetik (ira Druck).

MANCl~I, G., J.-P. VA]mMA~L A. O. CXRBONA~A, and g. F. HER~rANS : A single radial diffusion method for the immunological quantitation of proteins. (H. PE]~T]~RS, Ed.) Prot. Biol. Fluids, Proe. XIth Coll. Bruges, 1963, p. 370--373. Amsterdam: Elsevier 1964.

Genetic Studies on the Deficiency of fl2-Glycoprotein I of Human Serum 303

Appendix fl2-glyeoprotei~ I concentrations in sera o/the members o/38 ]amilies with 79 childreu

Number Re]ation- Age in Concentration Number Relation- Age in Concentration of family ship years in mg/100 m] of family ship years in rag/100 ml

1 father 42 29.7 11 father 83 11.3 mother 42 28.2 mother 78 26.6 son 18 29.7 son 53 12.6 son 17 26.6 daughter 5i 11.3

2 father 56 33.0 12 father 58 26.6 mother 51 19.2 mother 60 20.4 daughter 25 22.2 son 32 20.4 daughter 21 22.2 daughter 31 26.6 daughter 15 28.2 son 29 20.4

son 25 20.4 3 father 52 26.6 13 father 50 11.3

mother 46 24.9 mother 49 28.2 daughter 17 33.0 son 20 28.2 daughter 15 23.4 daughter 14 13.8

4 father 48 11.3 14 father 65 24.9 mother 46 36.6 mother 65 26.6 son 20 19.2 daughter 35 22.2 son 18 19.2 daughter 32 20.4

5 father 61 24.9 15 father 62 31.5 mother 61 34.8 mother 55 23.4 daughter 32 31.5 son 28 23.4

daughter 24 24.9 6 father 60 31.5

mother 55 34.8 16 father 59 26.6 mother 55 13.8 son 29 22.2

daughter 26 26.6 son 26 0--1 daughter 22 33.0 daughter 22 0--1

son 18 19.2 17 father 60 29.7 daughter 17 22.7 mother 65 12.6 daughter 15 26.6 son 32 11.3

son 30 19.2 7 father 56 26.6

18 father 57 28.2 mother 51 22.2

mother 54 29.7 son 22 20.4 son 20 28.2 daughter 28 28.2 son 17 24.9 daughter 25 29.7

19 father 55 28.2 8 father 48 29.7 mother 48 28.2

mother 47 20.4 daughter 19 20.4 son 21 29.7 son 16 23.4 daughter 18 28.2

son 15 28.2

9 father 55 26.6 20 father 56 31.5 mother 54 28.2 mother 42 22.2 son 24 26.6 daughter 22 23.4 son 17 19.2 son 17 34.8

10 father 61 26.6 21 father 55 28.2 mother 51 23.4 mother 54 28.2 son 29 31.5 son 27 26.6 son 27 31.5 daughter 24 28.2 son 22 29.7 son 17 26.6

304 H. CLEVE: Genetic Studies on the Deficiency of fl~-Glyeoprotein I of H u m a n Serum

Appendix (Continuation)

Number Relation- Age in Concentrat ion Number Relation- Age in Concentrat ion of family ship years in rag/100 ml of family ship years in rag/100 ml

22 father 50 22.2 30 father 49 37.5 mother 48 20.4 mother 41 29.7 daughter 17 19.2 son 15 28.2

23 father 61 19.2 31 father 66 10.2 mother 56 26.6 mother 67 26.6 daughter 38 23.4 son 38 26.6

daughter 20 26.6 32 father 66 34.8 mother 63 28.2 24 father 66 26.6

mother 65 26.6 son 28 19.2

son 42 29.7 33 father 65 20.4 mother 62 26.6

25 father 62 34.8 daughter 33 20.4 mother 57 31.5 daughter 31 20.4 daughter 27 29.7

34 father 59 24.9 26 fa ther 51 34.8 mother 60 26.6

mother 41 23.4 daughter 27 22.2 son 14 23.4

35 father 53 23.4 27 father 69 12.6 mother 54 40.2

mother 73 28.2 daughter 25 22.2 daughter 41 26.6 daughter 17 23.4

son 36 13.8 36 father 58 29.7

28 father 57 26.6 mother 57 26.6 mother 57 34.8 daughter 24 20.4

son 29 22.2 37 father 61 11.3 son 28 26.6 mother 54 28.2 son 23 20.4 daughter 30 12.6

29 father 52 23.4 daughter 28 23.4

mother 49 20.4 38 father 66 29.7 son 24 20.4 mother 65 16.5 daughter 20 23.4 son 39 26.6 son 16 28.2 son 28 33.0

Dr. HARTWIG CLEVE Cornell University Medical College Depar tment of Medicine 525 E 68th Street New York, N.Y. 10021 (U.S.A.)