Prevalence of Non-Organ-Specific Autoantibodies in and Non-Pregnant Healthy Women


ANESTIS K. MAVRIDIS*, LIU XUE MING*, PETER HATZIPETROU**, GREGORY LENTZARIS**, NICOLAS G. PAPANIKOLAOU**, ATHANASIOS G. TZIOUFAS* and HARALAMPOS M. MOUTSOPOULOS* *Clinical Immunology Laboratory, Department of Internal Medicine, School of Medicine; **Department of Obstetrics and Gynecology, Hatzikosta General Hospital, Ioannina, Greece The

prevalence of autoantibodies during



studied. Sera from 568



pregnant and 365 non-pregnant) were tested for autoantibodies to double-stranded DNA, cardiolipin and extractable cellular antigens. Nineteen out of 203 pregnant women (9.4%) had one autoantibody against double-stranded DNA or cardiolipin, mainly of the IgM class. The non-pregnant control group had significantly higher incidence of autoantibodies P < 0.01). Sixty-six out of 568 women had had spontaneous abortions in (17.8%; X 2= 7.39, their past medical history, but there was no correlation between them and the presence of anticardiolipin antibodies. Two of the non-pregnant women had anti-Ro(SSA) antibodies. These findings suggest that (a) the prevalence of autoantibodies decreases during pregnancy; and (b) the presence of anticardiolipin antibodies in healthy pregnant women does not correlate with any pregnancy-related complications. However, our results indicated that autoimmunity is a rather common disorder in healthy women.

at least

Key Words: Autoantibodies

Pregnancy Autoimmunity


population and a non-pregnant control group. In addition, medical, family, gynecologic and reproductive nant

Autoimmune rheumatic diseases affect primarily women of childbearing age’ . Therefore, pregnancy in these patients can potentially be a clinical problem. In fact, previous studies have shown that spontaneous abortions and premature deliveries are common complications in pregnant women with systemic lupus erythematosus

histories were reviewed and correlated with the presence of these antibodies.

Materials and methods


Sera from 568 women who were followed in the Department of Obstetrics and Gynecology of the G. Hatzikosta General Hospital, Ioannina, Greece, were tested for the presence of aCL, double-stranded DNA (dsDNA) antibodies and autoantibodies to extractable cellular antigens. The study population consisted of 203 consecutive pregnant and 365 non-pregnant women visiting the out-patient gynecological clinic routinely (mean age 26.2 t 5.2 years and 27.6 =i= 6.5 years, respectively, with ages ranging from 20 to 40 years for both groups; Table I).

(SLE)2, 3. Medical complications related to pregnancy and the newborn have been attributed to certain autoantibodies, such as anticardiolipin (aCL) and anti-Ro(SSA) antibodies. The presence of aCL autoantibodies is associated with increased incidence of fetal loss in autoimmune patients’, while newborns of mothers with anti-Ro(SSA) antibodies can present with complete heart block and/or skin rashes5, 6. Previous studies have extensively examined the presence of these autoantibodies in pregnant women with autoimmune rheumatic diseases’. In contrast, few studies have been carried out on the prevalence of these particular autoantibodies in uncomplicated pregnancies of healthy women’. The aim of this study was to determine the frequency of non-organ-specific autoantibodies in a random preg-

Table I

A.G. Tzioufas, M.D., Department of Internal Medicine, School of Medicine, University of Ioannina, 451 10 Ioannina, Greece.


Biographical data of 568 pregnant and non-pregnant



NA: not


Histories of these women were carefully evaluated for significant medical illnesses (including systemic auto-


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Statistical analysis where indicated.

immune rheumatic diseases), gynecologic and reproductive disorders, family history and medications (such as oral contraceptives). In addition, past and current pregnancies were reviewed for the outcome (healthy newborns, abortions and stillbirths). All sera were kept at - 30 °C until tested. Autoantibodies to extractable cellular antigens Ro(SSA), La(SSB), Sm and U,NRNP were determined by counterimmunoelectrophoresis with the use of human spleen, calf thymus and rabbit thymus extracts8. Autoantibodies to CL and to dsDNA were detected by

performed by chi-square



Eighty-four out of 568 sera tested had at least one autoantibody (14.8%). Nineteen out of 203 pregnant women (9.4%) had anti-dsDNA antibodies and/or aCL (6/19 had both of these autoantibodies, 7/19 had only anti-dsDNA antibodies and 6/19 had aCL). The predominant isotype of the autoantibodies was the IgM class (Figure 1 a, b). Sixty-five out of 365 non-pregnant women in the control group (17.8%) presented also with serum autoantibodies. The prevalence of autoantibodies in nonpregnant women is statistically significantly higher compared with the corresponding prevalence of auto-

quantitative isotype-specific solid-phase enzyme-linked immunosorbent assays


(ELISA)9, 10.

antibodies in the pregnant women (X2 = 7.39, P < 0.01 ). Twenty-two out of 65 non-pregnant women had both anti-dsDNA antibodies and aCL, 14/65 had only anti-dsDNA antibodies and 21 /65 had aCL alone. Two women in the non-pregnancy group also had antibodies to ribonucleoprotein, Ro(SSA). None of the newborns of these two mothers presented with neonatal lupus. Sixty-six out of 568 women had a past history of spontaneous abortion, while one pregnant woman had abortion during this study. Interestingly, this woman presented with a four-fold increase in her level of aCL above the cut-off point. No correlation between the presence of aCL and past history of spontaneous abortions was noted (data not shown). Antibodies to extractable cellular antigens La(SSB), Sm and U,NRNP were not detected in any of the tested an


There was no correlation between the presence of aCL, anti-dsDNA and anti-Ro(SSA) antibodies and the gynecological reproductive past history of the women, past medical history, family history or any kind of treatment.



The effect of pregnancy in the function of the immune system and the emergence of autoimmune phenomena remains controversial. Clinical studies have suggested that pregnancy influences the immune system&dquo;. The preponderance of autoimmune disorders in women and the exacerbation of certain autoimmune rheumatic diseases during pregnancy imply that sex hormones or other factors closely related with pregnancy have a direct effect on the immune system. Previous studies on the presence of autoantibodies in pregnant women with a well-documented autoimmune disease yielded conflicting results. In fact, autoantibody titers have been reported to increase’2, decrease&dquo; or remain unchanged&dquo;

Figure 1 (a) The incidence of anti-dsDNA antibodies (IgG and IgM classes) in pregnant and non-pregnant women. (b) The incidence of aCL (IgG and IgM classes) in both groups. In both isotype-specific assays (anti-dsDNA and aCL) the cut-off point (indicated by the dashed line) was calculated as the mean optical density (OD) plus three standard deviations of sera from normal healthy blood donors as previously described9, 10.


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pregnancy, depending on the autoimmune disorder studied. Therefore, conclusive generalizations on the effect of pregnancy on autoantibody production are difficult to draw. The purpose of this study was to compare the prevalence of autoantibodies in a healthy pregnant population to that of a control non-pregnant group and to determine whether pregnancy influences the autoantibody production. Our results showed that pregnant women have a significantly lower prevalence of autoantibodies compared to a healthy matched control population. These autoantibodies are mainly of the IgM class, found in low titers without any clinical importance. These are probably natural autoantibodies with low affinity to the antigen against which they are directed&dquo;. In a recent study 16 it was shown that 11.8% of healthy non-pregnant women had sera containing natural autoantibodies that did not necessarily indicate a pre-autoimmune disease state, as was demonstrated after a 5-year follow-up period. The decreased prevalence of the natural autoantibodies during a normal pregnancy can be explained by the following observations. Firstly, immunoglobulin levels are generally decreased in normal pregnancy and show a rebound increase postpartum&dquo;. Secondly, in a normal pregnancy there is an increase in maternal suppressor T cells with a concomitant decrease in helper T cells&dquo;. Thirdly, lymphocyte responses to antigens such as purified protein derivative (PPD) and polyclonal stimulators such as phytohemagglutinin (PHA) are depressed in normal pregnant women 19.20 due to lymphocyte-depressing factor, an alpha-2 macroglobulin, which increases in pregnancy2’ . Finally, an important element for consideration is the role of steroid hormones found in high levels in the sera of pregnant women. Cortisol is present in four-fold concentrations22 and, although pregnant women do not appear immunosuppressed by cortisol, this elevation may influence natural autoantibody production. Estrogens and progesterone are also found in high concentrations during pregnancy. Pharmacological doses of either suppress inflammation, delay allograft rejection and decrease in vitro blastogenic responses23. Therefore, they can also have a potential effect on normal autoantibody production. Anti-Ro(SSA) antibodies occur predominantly in patients with primary Sj6gren’s syndrome (SS) and SLE, particularly in those with seronegative lupus, subacute lupus and histological manifestations of SS24. This antibody has also been described in rheumatoid arthritis patients with sicca syndrome25. Antibodies to Ro(SSA) cellular antigen were detected in two non-pregnant women. Although no association between the presence of these antibodies and neonatal lupus was observed, the detection of anti-Ro(SSA) antibodies in sera of healthy women, by a specific method such as counterimmuno-


electrophoresis, implies that a considerable percentage (0.35%) of healthy women have at least laboratory manifestations of an autoimmune process. This finding may suggest




’hidden’ autoimmune dis-

order in these two ’healthy’ women. Our results are in contrast with previous reports that showed an increased prevalence of autoantibody detection in a ’normal’ healthy pregnant population26, 27. This discrepancy can be attributed to the lack of an adequate control group in these studies. Additional factors that may contribute to this difference are the small sample population tested and the type of the assays used’. In our study highly sensitive and specific ELISA methods and a widely approved counterimmunoelectrophoresis method were used to detect autoantibodies to CL, to dsDNA and to extractable cellular antigens in sera from

large sample population’-’O. In the present study a complete gynecological reproductive history of the women was evaluated compared with the presence of autoantibodies. a

and and Our results indicated that the incidence of spontaneous abortions or complicated pregnancies had no correlation with the presence of autoantibodies and did not differ from that reported previously in healthy Greek women3.


Dr Liu Xueming was on sabbatical leave from the Third Teaching Hospital, Normal Bethune University of Medical Sciences, Changchun Jilin, People’s Republic of China, supported by a fellowship from the Epirotic Society ’Friends for Education’. The authors wish to thank Mr G.E. Papanikolaou for

secretarial assistance.

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sopoulos mune


11. 12. 13.






(Received 1 October 1991) (Accepted 17 December 1991)


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Prevalence of non-organ-specific autoantibodies in pregnant and non-pregnant healthy women.

The prevalence of autoantibodies during pregnancy was studied. Sera from 568 women (203 pregnant and 365 non-pregnant) were tested for autoantibodies ...
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