504

February Am. J, Obstet.

Correspondence

15, 1978 Gynecol.

Table I. Uptake of lz5 I -labeled hCG by human lymphocytes, fibroblasts, amniotic fluid cells, and rat testicular Leydig cells (number of cells used per experiment and mean values 2 standard errors)

a

0.021 25

is

31

33

35

q

39 40 4lrrc

weeks

gestation

Fig. 1. Logarithmic change in optical density of amniotic fluid at 450 ny~ prior to and following administration of hetamethasone to mother. (---) indicates time of betamethasone administration. to stimulate interest in revising the criteria to evaluate how much the fetus can be compromised by Rhhemolytic disease based on study of the amniotic fluid whenever corticosteroids are prescribed to the gravid patient. L. Pereira Leite Belmiro Patricia Services of Obstetrics Faculdade de Medicina do Port0 Portugal

Human lymphocytes, fibroblasts and amniotic fluld cells are not endowed with human chorlonlc gonadotropin receptors To the Editors: In contrast to purified human chorionic gonadotropin (hCG), crude hCG has been found to exert a modulating effect on the stimulation of cultured lymphocytes by mitogens. This effect was attributed to contamination of the hormone and not to the hCG itself. However, Beck and associates,’ using a phenol- and immunoglobulin-free hCG preparation, recently reported a modulating influence of the hormone on lymphocyte blastogenesis in culture. The authors speculated that the hCG effect on the lymphocytes is mediated by the activation of the adenyl cyclase-cyclic adenosine monophosphate system. The prerequisite for the activation of this system in a cell depends on the binding of the hormone to a specific receptor in the cell membrane. Thus, the question arises if lymphocytes are endowed with specific hCG receptors. Until the present time, hCG receptors have been demonstrated only in testicular and ovarian cells.2 Supported in part by the Deutsche Forschungsgemeinschaft (SFB 46). Reprint requests: Dr. Jan W. Siebers, Universitlts-Frauenklinik, D-7800 Freiburg, Hugstetterstrabe 55, FRG

Lymphocytes (6 x 106) Lymphocytes with PHA (2 x 106) Fibroblasts (2 x 106) Amniotic fluid cells (2 x 106) Leydig cells (2 x 106) Cultivated Leydig cells (2 x 106)

534 _’ 31

4952

471 t 25

435 7?r32

692 -c 36

691 + 40

832 2 20

796 i. 35

2.1962

13

155

644 2 65

2,214 + 132

1,005 ? 83

Human lymphocytes from adult male donors were isolated by Ficoll Isopaque gradient centrifugation and studied for hCG binding either immediately or after three days in culture with phytohemagglutinin* (PHA). PHA was reconstituted with 10 ml. of sterile distilled water and then used at a concentration of 10 ~1 per milliliter of culture. For comparison, single-cell suspensions of routinely cultured human fibroblasts and amniotic fluid cells as well as rat testicular Leydig cells were prepared. Leydig cells are endowed with hCG receptors.* The cells were obtained by rinsing the decapsulated adult testis in collagenase. They were used for the binding studies immediately after the preparation or after cultivation for three days in Eagle’s minimum essential medium supplemented with 10 per cent fetal calf serum.* For each experimental point, at least three preparations of 2 to 6 x lo6 cells were incubated in Tris-hydrochloric acid buffer (0.04 mole per liter, pH 7.4) containing magnesium sulfate (0.005 mole per liter), 0.1 per cent bovine serum albumin and 8 ng. of 1251-labeled hCG (biological activity 11.000 I.U. per milligram, specific radioactivity 30 to 50 &i per microgram, 1 ng. = 35,000 c.p.m.). The nonspecific binding was determined in the presence of a 1 ,OOO-fold excess of unlabeled hCG. The reaction was stopped by the addition of 1 ml. of ice-cold buffer and the incubates were washed three times. After the pellet was dissolved with Soluene-350,f the radioactivity was determined in a gamma counter.t As can be seen from Table I in contrast to Leydig cells, iz51-labeled hCG is not specifically bound by lymphocytes from the peripheral blood, cultured lymphocytes stimulated with PHA, fibroblasts, or amfiiotic fluid cells, clearly indicating that these cells are not en*Difco Labs., Detroit, Michigan. tPackard, Frankfurt, Germany.

Volume Number

130 4

Correspondence

dowed with hCG receptors. Our results with lymphocytes explain the observation of other author? 4 that highly purified hCG does not impair lymphocyte stimulation. Furthermore, the modulating effect of phenoland immunoglobulin-free hCG on lymphocyte blastogenesis found by Beck and associates’ is not mediated by a specific receptor but might result from an unknown substance in the hCG preparation used or from a direct intracellular action of the hormone. It has been found that protein hormones such as parathyroid hormone, epidermal growth factor, prolactin, and insulin can enter target cells.” However, we have shown recently by cell fractionation of different tissues that hCG does not enter the cell but is bound exclusively to the membrane.* Jan W. Siebers Walther Vogel Wolfgang Engel

for

Institut Humungenetik University Gottingen Nikolausberger Weg 5 a D-3400 Gottingen, West Germany

of

*Unpublished

data.

REFERENCES

H., and Naot, Y.: AM. J. OBSTET. 1977. T., Mendelson, C., Ketelslegers, 2. Catt, K. J., Tsuruhara, J.-M., and Dufau, M. L.: Curr. Top. Mol. Endocrinol. 1: 1, 1974. 3. Caldwell, J. L., Stites, P., and Fudenberg, H. H.: J. Immunol. 115: 1249, 1975. 4. Pattillo, R. A., Shalaby, F. M. R., Hussa, R. O., Bahl, 0. P, and Mattingly, R. F.: Obstet. Gynecol. 47: 557, 1976. 5. Goldfine, I. D., Smith, G. J., Wong, K. Y., and Jones, A. L.: Proc. Natl. Acad. Sci. U. S. A. 74: 1368, 1977. 1. Beck, D., Ginsburg, GYNECOL.

129:

14,

Wlatation and cwettage for termination of seconbtrt~ pregnancy To the Editors:

It was with great interest that I read the article, “Dilatation and curettage for second-trimester abortions,” by Hodari and colleagues (AM. J. OBSTET. GYNECOL. 127: 850, 1977). However, I find some of the data slightly confusing. The authors stated that 99.2 per cent of the abortions were for gestations between menstrual weeks 15 and 18 of pregnancy. However, a quick glance at Table I reveals that 86.3 per cent of the abortions were done for menstrual weeks 13 and 14 of pregnancy and 97 per cent were done for menstrual weeks 13, 14, and 15 of pregnancy. It has been my experience, at several hospitals, that it is not unusual for trained gynecologists to perform second-trimester abortions up to 13, 14, or possibly even 15 weeks by dilatation and curettage; since 97 per cent of those done by Hodari and colleagues were within this gestational time period, I find this not to be very unusual.

505

In addition, it would be very helpful if the postabortion complications (Table III) could be listed not only by characteristic but also by gestational age in terms of menstrual weeks. Certainly, the total incidence of major complications numbering 35 is not very great, considering the total number of abortions done. However, if a great number of these occurred in those patients who were at 15 or more menstrual weeks of pregnancy, the incidence jumps to 10.2 per cent; if most of these occurred at 16 or more menstrual weeks, then the percentage is even higher (at the level of 45.5 per cent). The same holds true for the total incidence of 51 cases of morbidity where the changes in percentages would be 14.8 and 66.2 per cent, respectively. I would also be interested in knowing if any of these abortions were performed with the prior use of laminaria tents, which we have found to be very useful. Certainly, if the percentage of complications increases as cited above after the gestational age of the pregnancy is adjusted for, then there are an inordinate number of complications as compared to the figures given in Table V, which is a review of the literature. I would very much appreciate a reply and comments concerning the above, since it would be of great interest to know the specific results prior to promulgating the performance of second-trimester abortions by dilatation and curettage as a relatively safe procedure, safer than transabdominal amniocentesis with injection of hypertonic saline, prostaglandin, or another agent. Sylvain Fribowq. Department Obstettics and Gynecolq Southern California Permanente Medical Group 13652 Cantara Street Panorama City. California 91402

+?.I).

, F..4.C.O.C.

qf

Reply to Dr. Fribourg To the Editors:

There was a misprint in Table I under the heading entitled “Postoperative gestational evaluation” on page 851 of our article, “Dilatation and curettage for second-trimester abortions.” Originally the paper was written in the context of ovulatory weeks. When we revised it, we corrected for menstrual weeks. The correction for Table I was not incorporated in the published version of the paper. Also, as Dr. Fribourg requested, we have itemized the complications we observed in our study by gestational age in terms of menstrual weeks (Table I). The highest percentage of complications occurred at the sixteenth menstrual week of gestation, not at the more advanced gestational stages. We believe that the complications we saw were related to the skill of the physician who performed the procedure and the physical condition of’ the patient and were not directly associated with the stage of gesta-