Alan E. Beer

Reproductive outcome in women with recurrent spontaneous abortions of alloimmune and autoimmune causes: Preconception versus postconception treatment

Objectives: The null hypothesis is that treatment of women with recurrent spontaneous abortions with anticoagulation and immunosuppression will not increase the reproductive outcome if it is started preconceptionally. Study design: Ninety-four women with recurrent spontaneous abortion with autoimmune abnormalities comprised the study group. Group I began autoimmune therapy 48 hours after ovulation: heparin 5000 U twice daily, aspirin 80 mg daily, and prednisone 5 mg twice daily, with an increase to 10 mg twice daily when pregnant. Group II started the same medication after a positive pregnancy test. Group III received no medication. Controls were 19 women with no autoimmune abnormalities. The frequency of reproductive outcome was subject to multiple comparison by the Duncan test. Results: The percentages of live-born children in groups I, II, and III were 74%, 44%, and 11%, respectively. Conclusions: Preconception diagnostic work-up and treatment of autoimmune abnormalities in women with histories of recurrent spontaneous abortion is advocated.

The alloantibody response of pregnant women and its suppression by soluble HLA antigens and anti-idiotypic antibodies

The aim of this study was to investigate the time course of maternal allosensitization to fetal HLA antigens during normal human pregnancy and to explore mechanisms of suppression of anti-HLA alloantibodies. We found that the mother produces antibodies against some but not all of the mismatched HLA antigens of the fetus as early as the 8th week of pregnancy. These antibodies (Ab1), however, are often complexed with soluble HLA alloantigens and become detectable when immune complexes are dissociated. Soluble HLA antigens of fetal origin are present in the maternal circulation throughout the entire pregnancy beginning at 8 weeks. In some women the production of anti-anti-HLA antibodies (Ab2) became evident as early as the first trimester, while in others Ab2 was documented during the second or third trimester. Analysis of antibody specificity showed that some healthy primipara develop antibodies reactive with self HLA antigens. Although the allo- and autoantibody responses appear to be modulated by soluble HLA antigens, cyclic variations in the level of alloantibodies, as well as the mothers selective response to some, but not all, paternal HLA antigens, are best explained by the development of anti-idiotypic antibodies.

Down-regulation of maternal antiphospholipid antibodies during early pregnancy and pregnancy outcome

OBJECTIVE We investigated the hypothesis that maternal autoimmune responses to phospholipid antigens measured before and during pregnancy are not related to successful pregnancy outcome. STUDY DESIGN One hundred twenty-three women with recurrent spontaneous abortions were serially tested for antiphospholipid antibodies during their pregnancies. RESULTS In 72 women with recurrent spontaneous abortions and without antiphospholipid antibodies before the pregnancy, the incidence of antiphospholipid antibody production at the time of pregnancy termination was significantly higher in those who miscarried the index pregnancy than those who were delivered of a live-born infant. In 51 antiphospholipid antibody-positive women with recurrent spontaneous abortions there were dramatic increases in titers of anticardiolipin antibody and antiphosphatidylserine antibody in those who miscarried the index pregnancy (p < 0.005). In women who were delivered of a live-born infant, the titers remained stable or decreased during pregnancy. CONCLUSIONS Down-regulation of antiphospholipid antibody production during early pregnancy is associated with favorable pregnancy outcome.

Antibodies to phospholipids and nuclear antigens in non-pregnant women with unexplained spontaneous recurrent abortions

In a collaborative study of 73 non-pregnant Kuwaiti women with unexplained spontaneous recurrent abortion (RSA), 30 control healthy non-pregnant multiparous Kuwaiti women and 20 North American women who received elective abortion(s), autoantibodies to 6 phospholipids and 9 nuclear antigens were measured. Women with recurrent spontaneous abortions demonstrated 3 times higher incidence of antibodies to phospholipids (30.1%) than controls (10% each) (P = 0.029). The incidence of both IgM and IgA class antiphospholipid antibodies were significantly higher than those of controls. The incidence of antibodies to cardiolipin in women with recurrent spontaneous abortions (12.3%) was significantly higher than those of controls (P = 0.035) and incidence of IgM but not IgG anticardiolipin antibody was significantly higher in women with RSAs than in controls (P = 0.053). The incidences of anti-polyinosinic acid (P = 0.035) and anti-histone 1 antibody (P = 0.052) were significantly higher in women with recurrent spontaneous abortions than controls. There was no significant difference in the incidence of autoantibodies between primary and secondary aborters. However, women with a history of second trimester abortions showed a higher incidence of antiphospholipid antibodies than women with first trimester abortions only. Recurrent spontaneous abortion is associated with autoantibodies to phospholipid epitopes including IgA antiphospholipid antibodies.

MHC class II compatibility in aborted fetuses and term infants of couples with recurrent spontaneous abortion

Maternal-fetal histocompatibility for alleles at HLA class II loci, HLA-DQA1 and HLA-DQB1, was examined in 40 abortuses and 31 liveborn children of 68 couples with a history of idiopathic recurrent spontaneous abortion (RSAB) who underwent leukocyte immunization prior to the index pregnancy. Significantly more couples with RSAB shared two HLA-DQA1 alleles as compared with fertile control couples (0.18 vs. 0.03, respectively; P = 0.031). There were no differences in HLA sharing between couples with RSAB who experienced a repeat abortion in the index pregnancy as compared with couples with RSAB who were delivered of a liveborn child. Non-significant deficits of abortuses who were compatible for alleles at the HLA-DQA1 (6 observed vs. 8.5 expected; P = 0.225) and the HLA-DQB1 (7 observed vs. 9.2 expected; P = 0.254) loci were observed. A significant deficit of HLA-DQA1 compatible liveborn children was observed (1 observed vs. 5.5 expected; P = 0.0069). The overall deficit of HLA-DQA1 compatible fetuses (7 observed vs. 14.0 expected; P = 0.0018) after approximately 8 weeks gestation suggests that HLA-DQA1 compatible fetuses may be aborted early in pregnancy, prior to the time when fetal tissue can be recovered for genetic studies.

Intravenous immunoglobulin infusion therapy in women with recurrent spontaneous abortions of immune etiologies

We have investigated clinical effectiveness of intravenous immunoglobulin G infusion (IVIg) on antiphospholipid antibody titers in five women with evidence of antiphospholipid antibody-associated recurrent spontaneous abortions and one with antinuclear antibody who became refractory to conventional autoimmune treatment during pregnancy and experienced pregnancy complications. Three women developed intrauterine growth retardation and three had complicated twin pregnancies with rising autoantibody titers. Antiphospholipid antibody and antinuclear antibody titers were tested pre and 2 weeks after each IVIg infusion. We report that: (i) IgG antiphospholipid antibody titers were significantly suppressed after each IVIg infusion (P < 0.05); (ii) IgM antiphospholipid antibody titers were also significantly suppressed after each IVIg infusion (P < 0.0001); (iii) decreased titers of autoantibodies paralleled increased levels of maternal IgG which lasted for at least 30 days; the autoantibodies showed a definite rise again prior to the next infusion; (iv) antinuclear antibody titers were effectively suppressed; and (v) rising autoantibody titers combined clinical manifestation of intrauterine growth retardation and women with complicated twin pregnancies. We conclude that IVIg infusion effectively suppresses IgM and IgG autoantibodies to phospholipids and antinuclear antibody in autoimmune women with a history of recurrent spontaneous abortions and refractory to conventional anticoagulation or immunosuppressive treatment.

The Biological Basis of Passage of Fetal Cellular Material into the Maternal Circulation

1. As a result of the apposition and subsequent union of the relatively fixed tissues of the host to those of the graft; 2. Through the escape of living cells, cell products, or cellular degradation products from the graft and their passage into the tissues, blood vessels, or lymphatic draining channels in the host. In this context, the contaminating donor leukocytes carried over in the vasculature and the tissue spaces of a graft may play a significant role; and finally 3. Passage of host immunocompetent cells through the vasculature of the graft and their return to the host via venous or lymphatic routes, providing an opportunity for the host cells to react with antigens and become peripherally sensitized.

HLA-DQA1 and HLA-DQB1 haplotypes in aborted fetuses and couples with recurrent spontaneous abortion

HLA haplotypes may be associated with spontaneous abortion through a variety of mechanisms, including maternal hyporesponsiveness to fetal alloantigens, maternal autoimmunity, and HLA-linked t-locus homologues. HLA-DQA1 and HLA-DQB1 haplotypes were determined in 37 couples with a history of recurrent spontaneous abortion (RSAB), 40 of their abortuses, and 20 fertile control couples. The distribution of haplotype frequencies did not differ between control subjects and RSAB wives, RSAB husbands, or abortuses. The frequency of the HLA-DR5-linked haplotype, DQA1*501/DQB1*301, which was considered a marker for immune hyporesponsiveness, did not differ between RSAB wives and control subjects (P = 0.353). The frequency of the autoimmune-associated HLA-DR3-linked haplotype, DQA1*0501/DQB1*0201, did not differ significantly between RSAB wives and control subjects (P = 0.103). The frequency of the DQA1*0201/DQB1*0201 haplotype in RSAB husbands was greater than the 95th percentile confidence limit of the frequency of this haplotype in control subjects. Among seven RSAB husbands who were heterozygous for this haplotype and did not share a DQA1*0201 allele with his wife, the haplotype was transmitted to 6 of 7 abortuses (3.5 expected). Although the small size of this sample precludes drawing conclusions regarding HLA transmission biases in RSAB couples, these data have generated a specific hypothesis regarding the DQA1*0201/DQB1*0201 haplotype that can be investigated in future studies.

Characterization of antibodies induced by paternal lymphocyte immunization in couples with recurrent spontaneous abortion

This study was designed to identify and characterize the allo- and autoantibodies induced following successful paternal lymphocyte immunization to prevent recurrent spontaneous abortion. Firstly the titers of maternal anti-paternal antibodies in women with successful pregnancies as determined by the flow cytometry crossmatch (FCXM) were highly variable; however, in all cases, the initial pre-immunization titers were negative and the post-immunization titers were positive by the FCXM in successfully treated women. Secondly, the specificities of maternal alloantibodies to paternal HLA antigens (immunogen) were evaluated. No all predicted antibodies to mismatched paternal HLA antigens were found by microlymphocytotoxicity (MCX) assays and the specificities varied. Thirdly, antibodies in post- but not preimmunization sera reacted with two lymphoid cell lines, SupT1 and SB; in addition, the rise and fall of the titers of these sera with paternal cells seemed to be reflected with the cell lines by the FCXM. Fourthly, autoantibodies to activated lymphocytes were detected and seemed to correlate with successful immunization since women who had another abortion following immunotherapy lacked these autoantibodies. These findings suggest that the antibody response following successful immunotherapy is complex and needs to be studied further to understand the mechanism of this treatment.

New Horizons in the Evaluation and Treatment of Recurrent Pregnancy Loss

During the past 15 years we have evaluated couples with unexplained recurrent spontaneous abortion (RSA), couples with normal pregnancies, and women who electively terminate their pregnancies early in gestation and have found major differences in the alloimmune and autoimmune status in women with unexplained RSA (1–3). In studies of over 1500 couples in this latter category we have shown (i) a higher incidence of HLA-DR and HLA-DQ antigen sharing compared to fertile controls (4); (ii) a higher incidence of HLA-DR and -DQ homozygosity in the male partners (5); (iii) a markedly decreased level of maternal alloantibody to paternal T and B lymphocytes compared to fertile control couples (2, 6); (iv) a strikingly higher incidence of antiphospholipid antibodies to phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylglycerol (PG), and cardiolipin (CL) that increases in incidence and titer with each subsequent pregnancy loss in three distinct populations of women studied (United States, Kuwait, and Colombia) (3, 7); and (v) an 89% incidence of subsequent pregnancy loss following lymphocyte immune therapy in autoimmune women untreated for the autoimmune abnormalities (2).