Pregnancy-specific Beta 1-glycoprotein Research Articles

Human Chorionic Gonadotropin in Esophageal Carcinomas: An Immunohistochemical Study

We have examined immunohistochemically the presence of human chorionic gonadotrophin (hCG) in 29 esophageal carcinomas: 24 squamous cell carcinomas, 2 adenocarcinomas, 2 adenoid cystic carcinomas and 1 adenosquamous carcinoma. In hCG-positive tumors, the presence of human placental lactogen (hPL) and pregnancy-specific beta-1 glycoprotein (SP-1) was also assessed. HCG immunoreactive cells were found in S squamous cell carcinomas (21%) and in none of S non-squamous cell tumors. The hCG positive cells were found in the most infiltrating areas of the tumors where poorly differentiated and pleomorphic cells predominated. The positive tumors were 4 poorly differentiated (31%) and one moderately differentiated carcinoma (12%). Four out of 10 cases (40%) with lymph node metastases had hCG in the primary tumor, whereas only one out of 11 cases (9%) without metastases was hCG positive. HPL and SP-1 were found in two cases. These placental proteins were detected in similar areas than hCG but the number of hPL and SP-1 immunoreactive cells was lower than hCG positive cells. SP-1 was also seen in areas of squamous cell differentiation negative for hCG. None of these two cases showed trophoblastic differentiation.

Expression of the Pregnancy-Specific β1-Glycoprotein Gene in Cultured Human Trophoblasts

Human pregnancy-specific beta 1-glycoprotein (PS beta G), the major placental glycoprotein, shares strong sequence similarity with carcinoembryonic antigen and is a member of the immunoglobulin superfamily. To understand the role of PS beta G in placental ontogeny during pregnancy, we examined its synthesis and regulation in primary cultures of trophoblast cells. Freshly plated (12 h) cytotrophoblasts expressed little PS beta G transcripts; however, within 24 h of culture, PS beta G mRNAs became detectable. PS beta G synthesis and mRNA expression increased with time in culture, and maximal synthesis was achieved at 4 days, indicating that primary trophoblasts continue differentiating in vitro. Molecular cloning revealed that PS beta G is an extremely polymorphic protein. Most PS beta G cDNAs identified to date, including the three cDNAs (PSG16, PSG93, and PSG95) isolated in this laboratory, share strong sequence similarity in the 5' (designated PSG-5') and coding regions, but differ in sequences at the 3' region. The PSG-5', PSG93-specific, PSG16/PSG93-3', and PSG95-3' probes, which identify the majority of PS beta G mRNAs, hybridized with three mRNAs of 2.3, 2.2, and 1.7 kilobases in primary trophoblasts and human term placental tissue. Ribonuclease protection analysis demonstrated that primary trophoblasts expressed most of the placental PS beta G transcripts. However, culturing in vitro altered PS beta G gene expression, and the level of PS beta G transcripts containing a PSG95-3' sequence was preferentially increased in primary trophoblasts. Moreover, primary trophoblasts synthesized a 64K PS beta G polypeptide in variable amounts and three PS beta Gs of 72K, 62K, and 54K in roughly equal amounts, whereas purified human term placental PS beta G consists of a major polypeptide of 72K and two minor ones of 64K and 54K. PS beta G gene expression in primary trophoblasts was slightly reduced by 8-bromo-cAMP, but was markedly inhibited by sodium butyrate.

Second trimester maternal serum pregnancy specific beta‐1 glycoprotein (SP‐1) levels in normal and down syndrome pregnancies

Maternal serum pregnancy specific beta-1 glycoprotein (SP-1) levels in the second trimester may be predictive of Down syndrome (DS). An enzyme immunoassay was used to measure SP-1 sera from 46 DS pregnancies and 117 normal control women matched for maternal age, gestational age, and length of storage. In the normal control samples, there were slight correlations between the SP-1 concentration and maternal age. The maternal serum SP-1 levels increased with each week of gestation from 15 to 20 weeks. All but one of the DS sera had SP-1 levels greater than the normal median. Using a cutoff of 2.8 multiples of the median (MoM), 15.2% of the DS pregnancies were detected with a false-positive rate of 4.3%. A combinational logistic regression analysis of maternal age and pregnancy related serum proteins will detect additional DS pregnancies and decrease the false-positive rate. The combination of maternal age and SP-1 detected 33 (71.7%) of Down syndrome pregnancies. The addition of maternal serum alpha-fetoprotein (AFP) and human chorionic gonadotropin (hCG) levels allowed for the detection of 36 (78.3%) of the DS pregnancies with a decrease in the false-positive rate to 3.4%. The measurement of other serum constituents in conjunction with AFP appears to be a valuable addition to current screening programs, as this can increase the proportion of DS cases detected prenatally.

Linkage of two human pregnancy-specific beta 1-glycoprotein genes: one is associated with hydatidiform mole.

A genomic clone containing two linked human pregnancy-specific beta 1-glycoprotein (PS beta G) genes has been isolated and characterized. The two genes are arranged in the same 5'----3' orientation; the 3' region (including the A2 and B-C exons) of the upstream gene, PSGGA, is linked to the 5' region (including the 5'/L and L/N exons) of PSGGB, the downstream gene. Depending upon the domains compared, PSGGA and PSGGB share 92-98% nucleotide and 86-95% amino acid sequence identity with PSG93, the most abundant PS beta G transcript. The 3' exon (B-C) of PSGGA contains four alternative splice sites and three polyadenylylation sites, which account for the 3' heterogeneity previously reported in the PS beta G family. Each of the predicted PSGGA-encoded proteins would have a different carboxyl terminus. PSGGB corresponds to the previously identified cDNA PSG6, which encodes proteins containing a 34-amino acid leader peptide and a 108-amino acid N domain, which is one amino acid shorter than the majority of PS beta G N domains. Additionally, the PSGGB-encoded proteins contain the cell-surface recognition tripeptide Arg-Gly-Asp, shared by several previously reported PS beta Gs as deduced from cDNA sequences. Northern blot hybridization performed with a PSGGB-specific oligonucleotide probe to the N domain revealed that PSGGB or a PSGGB-like gene encodes a major 1.7-kilobase mRNA in hydatidiform mole tissues and a major 2.0-kilobase mRNA in term placenta tissues. Moreover, the PSGGB-specific probe hybridized most strongly with mRNA from molar trophoblastic tissue, suggesting that the PSGGB-like species may be the gene preferentially expressed in gestational trophoblastic disease. Additionally, the sequence of a 2315-base-pair PS beta G cDNA (PSG95) that contains an N-A1-A2-B2-C domain arrangement is reported. The coding region of PSG95 is identical to the previously reported cDNA clones PSG1d and FL-NCA, but PSG95 contains an additional 518 and 523 base pairs in the 3' end as compared with PSG1d and FL-NCA, respectively.

De NovoSynthesis and Secretion of Prolactin-Like Protein-B by Rat Placental Explants*

Mid- to late-gestation rat placenta expresses three PRL-related mRNAs, rat placental lactogen-II (rPL-II), rat PRL-like protein-A (rPLP-A), and rat PRL-like protein-B (rPLP-B). The protein product of rPL-II mRNA has been characterized, and the protein products of the rPLP-A mRNA were recently identified. The mol wt of a nonsecreted nonglycosylated rPLP-B protein would be 27,145 based on the mRNA sequence. The present study is the first to report the identification of the rPLP-B protein. Antiserum was generated against a chemically synthesized oligopeptide inferred from a specific region of the rPLP-B cDNA. Three or four distinct proteins synthesized and secreted by rat basal zone explants (day 15 gestation) showed cross-reactivity with the rPLP-B antiserum. The relative mol wt of these immunoreactive proteins is approximately 30,000, with a pI varying from 6.1-6.6. De novo synthesized rPLP-B proteins were not secreted by the explant tissue in the presence of tunicamycin, suggesting that the proteins are glycosylated. These data are consistent with the presence of one potential N-glycosylation site derived from the rPLP-B mRNA sequence. The rPLP-B antiserum showed no cross-reactivity with proteins identified using antisera against rPLP-A, rPL-II, or human pregnancy-specific beta 1-glycoprotein. Immunocytochemical studies were carried out using paraffin sections from placentas of day 14 and 17 pregnant rats which were treated with anti-rPLP-B, followed by avidin-biotin-peroxidase complex. These experiments show perinuclear staining, which was localized in basophilic cytotrophoblast cells, confirming previous in situ mRNA hybridization studies. Although no physiological role has been established for rPLP-B, the synthesis and secretion of this protein by cells in contact only with maternal circulation suggest a hormonal role.

Effects of sodium butyrate on the synthesis of human pregnancy-specific beta 1-glycoprotein.

Human pregnancy-specific beta 1-glycoprotein (PS beta G) is a polymorphic placental protein which shows strong sequence similarity with the oncofetal protein, carcinoembryonic antigen. To better understand the role of PS beta G in pregnancy, we examined its synthesis and regulation in placental fibroblasts, which had been shown to express the PS beta G gene. The major placental PS beta G is a 72-kDa glycoprotein, while the major fibroblast PS beta G is a 62-kDa species. Administration of sodium butyrate to these fibroblasts slightly stimulated the synthesis of the 62-kDa species but markedly increased the production of two additional PS beta Gs of 72 and 48 kDa. The similarity between the PS beta Gs synthesized by butyrate-treated fibroblasts and human placenta was confirmed by cell-free protein synthesis. Poly(A)+ RNA from butyrate-treated fibroblasts and placenta directed the synthesis of two polypeptides of 48 and 36 kDa, which form the polypeptide backbone of the 72- and 48-kDa glycoproteins. Moreover, the predicted molecular weights of PS beta Gs encoded by the two types of PS beta G cDNA clones were 48,000 and 36,000. Most PS beta G cDNAs identified to date, including the three cDNAs (PSG16, PSG93, and PSG95) isolated in this laboratory, share strong sequence similarity at the 5' region (designated PSG-5') but differ in sequences at their 3' regions. The PSG-5', PSG93-specific, PSG16/PSG93-3', and PSG95-3' probes, which identify the majority of PS beta G mRNAs, hybridized with three PS beta G mRNAs of 2.3, 2.2, and 1.7 kilobases from placental fibroblasts. Butyrate increased the steady-state levels of all three mRNAs. Ribonuclease protection analysis showed that butyrate increased the PS beta G mRNAs containing the PSG-5' or PSG93-specific sequence to approximately 20% of human placental levels. However, unlike human term placenta, which predominantly expressed PS beta G mRNAs with 3'-sequences similar to PSG16/PSG93, the butyrate-treated fibroblasts expressed roughly equal levels of PS beta G mRNAs with the PSG16/PSG93-3' and PSG95-3' ends. All PS beta G cDNAs identified encode proteins with distinct carboxyl termini, suggesting that the composition of the 72-kDa species in placenta and butyrate-treated fibroblasts is likely to be different. Placental fibroblasts provide a unique model for the study of the mechanisms responsible for the differential expression of the PS beta G gene.

First‐trimester maternal serum biochemical indicators in Down syndrome

A set of 21 early maternal serum samples (19 first-trimester and two at 14 weeks) from pregnancies resulting in a child with Down syndrome was matched for gestation and length of storage with 63 samples from unaffected pregnancies. The concentrations of alpha-fetoprotein (AFP), unconjugated oestriol (uE3), human chorionic gonadotrophin (hCG), pregnancy-specific beta 1-glycoprotein (SP1), and placental alkaline phosphatase (PALP) were measured. The ratios of the medians for Down syndrome pregnancies compared with the medians for controls were AFP 0.71, uE3 0.67, hCG 1.43, SP1 0.79, and PALP 0.92. Although the differences between the medians for affected and unaffected pregnancies were not significant, the trends for AFP, uE3, and hCG confirm earlier findings on first-trimester samples.

Characterization of cDNAs of the human pregnancy-specific .beta.1-glycoprotein family, a new subfamily of the immunoglobulin gene superfamily

Three highly homologous cDNAs encoding human pregnancy-specific beta 1-glycoprotein (SP1) were isolated from a human placental cDNA library. These cDNAs share greater than 90% nucleotide homology in their coding sequences, and greater than 79% of the encoded amino acids are homologous. Proteins encoded by these cDNAs are very similar to members of the carcinoembryonic antigen family and contain repeating domains, conserved disulfide bridges, and beta-sheet structure typical of the immunoglobulin gene superfamily. However, the high degree of sequence homology and relatively lesser degree of glycosylation among the SP1 proteins suggest that they exist as a unique family instead of being members of the CEA family. Both soluble and potentially membrane-bound forms of SP1 proteins were present in the placenta. Northern blot analysis using specific probes confirmed the expression of multiple mRNA species in human term placenta.

Expression of the pregnancy-specific beta 1-glycoprotein genes in human testis

Northern blot analysis with placental pregnancy-specific beta 1-glycoprotein (SP1) cDNA probe showed the presence of SP1 mRNAs in human testis. Presence of translational products of the mRNAs was demonstrated by Western blot analysis with anti-human SP1 antibodies albeit difference in mobilities between the testis and placental proteins was apparent. Screening of human testis cDNA library with placental SP1 probe yielded 4 groups of positive clones. Two groups were identical to human placental SP1 cDNAs previously reported. The other 2 groups consisted of cDNA of incompletely processed mRNAs. These 2 groups were present in high abundance. Sequence analysis suggested that the cDNAs were products of different genes.

Pregnancy-Specific βi-Glycoprotein Messenger Ribonucleic Acid and Immunoreactive Protein in the Rat Testis*

Human pregnancy-specific beta 1-glycoprotein (PS beta G) is synthesized in large quantities by syncytiotrophoblasts of the placenta. Recent studies using a partial cDNA of PS beta G isolated from human term placenta detected the presence of mRNA highly homologous to placental PS beta G in a number of extraplacental tissues, including human and rat testis. The present study determined in the rat that the amount of PS beta G mRNA based on percentage of total tissue RNA was greatest in the testis of mature rats, followed by senescent and prepubertal rats. In experiments using rats with testicular feminization syndrome (Tfm) which exhibit end-organ insensitivity to androgen stimulation, slot blot analysis of testicular RNA showed reduced levels of PS beta G mRNA in Tfm rats compared to normal rats. Northern blot analysis of rat testicular RNA probed with a human placental PS beta G cDNA demonstrated the presence of a single mRNA species of 1.65 kilobases. Subsequent studies investigated whether proteins immunologically similar to PS beta G were present in the testes from normal and Tfm rats. The rat testes were perfused with fixative, and sections from paraffin-embedded tissues were treated with rabbit anti-human PS beta G, followed by the avidin-biotin-peroxidase complex. Testes from the normal rat showed intense immunostaining in late spermatids (steps 16, 17, 18, and 19 of spermiogenesis), residual bodies, as well as cytoplasm of Leydig cells. Spermatozoa within the epididymis also demonstrated intense immunolabeling. Paraffin sections of the testes from the Tfm rat showed light diffuse cytoplasmic immunostaining in cells of the seminiferous tubules. Since spermiogenesis does not proceed normally, and no spermatids were seen, it was not possible to accurately stage the seminiferous tubules of the Tfm rat. The Leydig cells of the Tfm testes stained intensely, however, as was observed in the testes of the normal rat. These data suggest that the rat may provide an animal model for the investigation of the biological function and regulation of PS beta G in the testis.

Immunoturbidimetric determination of pregnancy-specific β1-glycoprotein (SP-1)

We describe a simple and rapid immunoturbidimetric end-point assay for measuring pregnancy-specific beta 1-glycoprotein (SP-1) in human serum with the Kone Progress clinical chemistry analyser, using a commercially available antiserum. The absorbance was measured at 340 nm 5 min after addition of antiserum. The assay range was 3-250 mg/l. Over this range the CV for between-day precision was below 6%. Bilirubin up to 160 mumol/l, haemoglobin up to 2 g/l and lipaemia up to 320 mg/l did not interfere. The reference interval in maternal serum at the 15th week of gestation, was 13 to 46 mg/l with a median of 25 mg/l (n = 78) and that of the 16th week was 6 to 60 mg/l with a median of 28 mg/l (n = 78). Comparison of results obtained by two commercially available radio-immunoassays and by the present method shows acceptable correlation. Rapid and reproducible, the immunoturbidimetric method is suitable for use in routine determination of SP-1.

Comparative study of placental protein 19, human chorionic gonadotrophin and pregnancy-specific ?1-glycoprotein as immunohistochemical markers for extravillous trophoblast in pregnancy and trophoblastic disease

PP19, a new placental tissue protein, has alpha 1-beta 1 electrophoretic mobility, a molecular weight of 36,500 and 3.9% carbohydrate. To study immunocytochemical PP19 localization in extravillous trophoblast, we obtained formalin-fixed specimens from extravillous tubal pregnancy at gestational weeks (GW) 7-9 (12 blocks); four early intrauterine pregnancies at GW 7-13 (12 blocks); four late pregnancies at GW 28-38 complicated with intramural uterine myoma, placenta increta and abruptio placenta (8 blocks); four invasive complete moles (9 blocks); and seven primary and metastatic gestational choriocarcinomas (12 blocks). Immunohistochemical staining was done for PP19, pregnancy-specific beta 1-glycoprotein (SP1) and human chorionic gonadotrophin (hCG) using the indirect-labeled antibody method [purified PP19 (Lot no. 225/242) and antibody against PP19 (Lot no. 632ZA) prepared by H. Bohn, antibodies against hCG (Behringwerke, Marburg, FRG) and SP1 (Dakopatts, Copenhagen, Denmark)]. In both early and late intrauterine pregnancies, the extravillous syncytiotrophoblastic cell (XST) showed positive staining for hCG and SP1 in the cytoplasm, as well as for PP19, which stained more intensively in the nucleus than in the cytoplasm. The three proteins were not seen in the evtravillous cytotrophoblastic cell (XCT) in the trophoblastic cell column and shell. The interstitial cytotrophoblast-like cell (ICT), which infiltrated into the decidua and myometrium, and their blood vessels, was immunoreactively positive for PP19 but negative for hCG and SP1 with the exception of SP1-positive ICT in the myometrium in late pregnancy. XST and ICT in the endosalpinx of tubal pregnancy stained for all three proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of Baboon Pregnancy-Specific α1-Glycoprotein1

Immunostaining of baboon placental tissues with anti-human pregnancy-specific beta 1-glycoprotein (SPI) antibodies demonstrated that an SP1-like molecule was present in the syncytiotrophoblasts. Staining was observed on the membrane and in the cytoplasm, but the nucleus was devoid of any staining. Western blot analysis further demonstrated the presence of five protein species in baboon placental extract, whereas four protein bands were detected in human placental extract. Culture medium of baboon placental villi also contained five SP1-like molecules with sizes slightly different from those present in the placental extract. Amniotic fluid and culture medium of decidua basalis and chorioamniotic tissue contained lesser quantities and fewer species of SP1-like molecules. However, an 87 kDa band was present in all samples examined. Northern blot analysis of baboon placenta with a human placental SP1 cDNA probe demonstrated the presence of a 1.65 Kb band, whereas two hybridizing bands (1.65 Kb and 2.25 Kb) were present in human placenta. Southern blot analysis of baboon genomic DNA further demonstrated the presence of multiple bands hybridizing with a human placental SP1 cDNA probe. These results showed the presence in baboons of multiple genes encoding mRNAs and proteins highly similar to human placental SP1.

Immunocytochemical staining patterns of placentas associated with hydrops fetalis.

This study examined the morphology and immunocytochemical staining patterns of distribution of beta-human chorionic gonadotropin (beta-HCG), human placental lactogen (HPL), placental alkaline phosphatase (PLAP), and pregnancy-specific beta-1 glycoprotein (SP1) in 13 third-trimester placentas associated with hydrops fetalis and six normal control placentas matched for gestational age (+/- 2 weeks). Seven placentas were hydropic (540-1,080 g) and demonstrated histologic immaturity with large edematous chorionic villi showing few blood vessels, most of which contained immature hematopoietic elements. These placentas showed consistently increased staining for beta-HCG and decreased staining for PLAP when compared with control placentas, a pattern reminiscent of less mature placentas. HPL and SP1 staining were similar to those of controls. Six placentas were either sclerotic (four) or histologically unremarkable (two), and these did not differ in their immunocytochemical staining properties from control placentas. We conclude that third-trimester hydropic placentas, in addition to showing histologic immaturity, exhibit an immunocytochemical staining pattern associated with first-trimester placentas.

A pregnancy-specific β1-glycoprotein, a CEA gene family member, expressed in a human promyelocytic leukemia cell line, HL-60: structures of protein, mRNA and gene

Both genomic and cDNA clones encoding a precursor for a pregnancy-specific beta 1-glycoprotein (PS beta G) belonging to the CEA family, expressed in a human promyelocytic leukemia cell line, HL-60, have been isolated and the entire primary structure of the precursor is deduced. The 335-AA precursor has a 34-AA signal peptide followed by domains of N, IIA, IIB and C, which are encoded by separate exons. The genomic sequence contains extra exons IA and IB between exons N and IIA. Apparently, exon IA is excluded from the mRNA by alternative splicing while IB is a pseudo-exon having a stop codon formed by a deletion of dinucleotide in the middle of the sequence. This provides another mechanism to render exon IB abortive and is different from that we reported for another PS beta G (Biochem. Biophys. Res. Comm. (1988) 156, 68-77).

Identification of proteins immunochemically related to human pregnancy-specific beta 1-glycoprotein in the rat placenta.

Human pregnancy-specific beta 1-glycoprotein (hPS beta G) consists of a set of glycoproteins present in placenta and maternal serum. This study characterized proteins in rat placenta that show immunological cross-reactivity with antisera to hPS beta G. Immunocytochemical studies using two independent preparations of anti-hPS beta G showed intense specific staining within basophilic cytotrophoblast cells of the basal zone of the gestation day 15 rat placenta. In contrast, basophilic cytotrophoblasts located in the labyrinth did not stain. Subsequent experiments used gel electrophoresis and immunoblot analysis to compare PS beta G in human placenta and serum with immunoreactive proteins in rat placenta and serum. A set of two or three proteins was detected in human villous tissue and pregnancy serum with apparent mol wt (Mr) ranging from 54,000-76,000. In contrast rat placenta showed a major immunoreactive protein with 120,000 Mr, while rat serum contained bands of 48,000 64,000 and 69,000 Mr. Explant cultures of rat basal zone tissue secreted two [35S]methionine-labeled proteins that were immunoreactive, a major 120,000 Mr species and a minor 76,000 Mr form, with pI values of 4.6-5.5; tunicamycin inhibited the secretion of both species. Thus, a 120,000 Mr glycoprotein appears to be the major tissue and secreted form of rat PS beta G analog in day 15 placenta. Finally, the cytochemical localization of PS beta G-like proteins in rat placenta showed a progressive gestational shift from giant trophoblast cells in the parietal yolk sac placenta on day 12 to the basal zone cytotrophoblast cells by day 15. Data indicate that the pregnant rat may provide an animal model for investigation of the biological function of PS beta G during late gestation.

Pregnancy-specific beta 1-glycoprotein: cDNA cloning, tissue expression, and species specificity of one member of the PS beta G family.

Pregnancy-specific beta 1-glycoprotein (PS beta G) is a heterogeneous product of the human syncytiotrophoblast, closely related to the CEA-NCA multigene family. In the present study, immunoscreening was carried out with anti-PS beta G antibodies to isolate cDNA sequences from a placental lambda gt11 expression library. One 1847-bp cDNA clone comprising the major portion of the coding sequence of a putative 48-kd peptide was sequenced and characterized. Hybridization of human genomic DNA to the PS beta G sequence revealed a complex pattern of restriction fragments, a finding well in agreement with the assumption that there are several independent PS beta G genes. A variable PstI band was found in human DNA. Transfer blot analysis of human placental RNA identified transcript of 2.2kb and 1.7kb that appear transiently with increasing levels of expression during gestation. No hybridization of PS beta G cDNA to human RNA from liver, kidney, heart, thyroid, and ovary was observed. In analyses of placental RNA from mouse, goat, sheep, and cow, no corresponding transcripts could be detected, and DNA hybridization under low-stringency hybridization conditions resulted in very faint cross-reacting bands, presumably indicating sequences that were scarcely related. However, PS beta G-specific DNA sequencies with similar restriction patterns were found in primates. These results are compatible with the assumption of late evolutionary development of certain PS beta G sequences.

Molecular cloning and expression of cDNA for a carcinoembryonic antigen-related fetal liver glycoprotein.

Carcinoembryonic antigen (CEA) is considered to be an embryonic antigen that is reexpressed in carcinomas. However, at the molecular level little is known about fetal forms of CEA. We have studied fetal liver, which was originally considered to contain CEA. A first-trimester cDNA library from fetal liver was screened with CEA-specific probes, and a dominant cDNA clone was identified and sequenced. This 1.7-kilobase cDNA codes for a complete protein of 426 amino acids, of which 34 constitute a leader peptide. Structurally, it can be divided into four immunoglobulin-like domains homologous to CEA (N-A1-A2-B2) and a hydrophobic tail (12 residues). The A and B domains each contain two cysteines; the N domain has none. The protein has seven potential sites for asparagine-linked glycosylation. It is a form of pregnancy-specific beta 1-glycoprotein (PS beta G) but differs from other PS beta G species at the C terminus. The N and A1 domains show 45% and 51% amino acid sequence identity with the corresponding domains of the three CEA family members whose sequences have been determined. Expression studies showed that the cDNA codes for a 72-kDa glycoprotein that reacts immunologically with antisera to CEA, biliary glycoprotein I, and PS beta G. The 72-kDa glycoprotein was released from the transfected cells. At least six mRNA species were identified in human tissues by using this cDNA as a probe. Genomic DNA analysis with an N-domain-specific probe indicated that the number of genes is relatively small.

Detection of Immunoregulatory Factors in Retroplacental Serum in Human Pregnancy

Retroplacental serum (RPS) obtained from pregnant women at term deliveries was studied for regulatory effects on T-lymphocyte proliferation and for pregnancy-associated substances and compared with peripheral serum (PS) of the same donor. Proliferative response to phytohemagglutinin and alloantigens in RPS was lower than that in PS. RPS contained higher levels of human placental lactogen, progesterone, estradiol, and prostaglandin E2 than of PS. However, there were no differences in concentrations of pregnancy-associated alpha 2-glycoprotein, pregnancy-specific beta 1-glycoprotein, prostaglandin F2 alpha, alpha-fetoprotein, human chorionic gonadotropin, cortisol, carcinoembryonic antigen, and immunoglobulins between RPS and PS. The amounts of human placental lactogen, progesterone, or prostaglandin E2 seen in RPS did not inhibit T-cell proliferation. Mixtures of various doses of these three substances were still not inhibitory. Thus, the suppressive activity of RPS could not be explained by these pregnancy-associated substances, but a possible involvement of unknown immunoregulatory factors at fetomaternal interface might be suggested.

Regional localization of the gene encoding pregnancy specific beta-1-glycoprotein 1 (PSBG1) to human chromosome 19q13.1.

Pregnancy specific beta-1-glycoprotein (PSBG), a major product of the human placenta, is encoded by multiple genes. Southern blot hybridization of human x rodent somatic cell hybrid DNAs with a cDNA specific for one member of the PSBG gene family allowed us to map this gene to human chromosome 19. Further analysis using hybrids with subchromosomal segments of 19q revealed that the gene maps to the distal segment of region 19q13.1.