American journal of reproductive immunology : AJRI : official journal of the American Society for the Immunology of Reproduction and the International Coordination Committee for Immunology of Reproduction最新文献

To assess the presence of human leukocyte antigens (HLA) on first trimester human trophoblast cells, frozen sections of villous trophoblast and monolayer cultures of isolated cells from placental villi were prepared and exposed to a mouse monoclonal antibody directed against HLA-DR and then incubated with fluorescein-conjugated goat antimouse antibodies. Fluorescence microscopy demonstrated that HLA-DR antigens were present on only the small polygonal epithelioid cells of the monolayer culture. The crescentic staining pattern was consistent with widespread distribution of antigen on the cell membrane. There was no staining over giant multinucleated structures or on fibroblasts of such cell cultures. No HLA-DR was detected when this indirect immunofluorescent technique was applied to tissue sections of villous trophoblast. Existence of high concentrations of hCG in culture supernatants and coincident localization of both hCG and HLA-DR using antibodies conjugated with rhodamine or fluorescein on the polygonal epithelioid cells indicate the trophoblastic origin and expression of HLA-DR antigen under in vitro monolayer culture conditions.

We have successfully applied SDS (sodium dodecyl sulfate) gel/protein blot radioimmunobinding method to identify the molecular size of sperm antigens that elicit antisperm antibodies from patients with unexplained infertility. Following the transfer of renatured proteins from SDS gel of human sperm extract onto nitrocellulose strips, the radioimmunobinding was performed by incubating the strips with patients' sera at 1:100 dilution and then with I125-labeled goat antihuman immunoglobulin G (IgG) or protein A as detecting probes. Unique sperm antigens that reacted with some patients' sera were identified following the autoradiography of the incubated paper strips. Among the fifty-nine standard serum samples from the Reference Bank of the World Health Organization, about one-fourth of them were found to react predominantly with a sperm protein band having the reference value (Rf value) of 0.2 and the approximate molecular weight of 90,000 dalton. A similar analysis was also performed with serum samples from vasectomized patients. Some of them also revealed a specific binding with the sperm antigen(s) of similar molecular weight. The results of this analysis were also compared with those of conventional tests for sperm antibodies as well as those of microplate radioimmunoassays and enzyme-linked immunoassays. This study suggests that SDS gel/protein blot radioimmunobinding method can be a useful tool for the molecular identification of unique human sperm antigen(s) that elicit naturally occurring antisperm antibodies in patients with unexplained infertility.

Using monoclonal antibodies, indirect immunofluorescence, and flow cytometry, the proportions and absolute numbers of various lymphocyte subsets in peripheral blood have been measured in normal human pregnancy. Groups of ten women were studied at 12, 28, and 36 weeks of gestation and compared with 16 nonpregnant control women. The percentage of T cells (OKT3+) was constant throughout pregnancy, and this was confirmed in three women studied serially prior to and throughout early pregnancy. A slight fall in the proportion of helper cells (OKT4+) and rise in the proportion of suppressor cells (OKT8+) was observed at 12 and 28 weeks, but these changes, and the resulting fall in helper/suppressor ratio, were not statistically significant. Absolute lymphocyte counts determined by white cell count and differential were lower during pregnancy. The absolute numbers of T cells, helper cells, suppressor cells, and Ia-bearing cells (mainly B cells) were significantly lower at 36 weeks' gestation. T cells and helper cells were significantly reduced in absolute number at 12 weeks' gestation. There was no change in the ratio of T cells to B cells at any stage of gestation. The lack of any significant change in the balance between helper and suppressor cells in peripheral blood suggests that these cells are not important in the immune adaptation to pregnancy.

Leukocyte adherence inhibition (LAI) assay was used to evaluate cell-mediated immunity in patients with invasive squamous cell carcinoma of the cervix. The reactivity of the peripheral blood leukocytes of these patients was evaluated after incubation with pooled extracts of allogeneic squamous cell carcinoma of the cervix. One hundred sixty-seven sets of LAI assays were performed on 54 individuals, including 23 patients with Stage I squamous cell carcinoma of the cervix, 9 patients with other stages of this tumor, 9 patients with unrelated tumors and 13 normal healthy volunteers. A protein concentration of one milligram per milliliter in the tumor extract and 10% fetal bovine serum in the feeding media gave the best results. Eighty-seven percent (28/32) of patients with squamous cell carcinoma of the cervix showed marked specific reactivity. No difference was found in the LAI indices of different stages of the disease.

Autoimmune thrombocytopenic purpura (ATP) is an antibody-mediated autoimmune disease of platelets. Transplacental passage of the antibody during pregnancy can result in transient neonatal thrombocytopenia, but it is not known why some infants of antibody-positive, thrombocytopenic mothers are not affected. We have developed a competitive enzyme-linked immunosorbent assay (ELISA) to measure circulating antiplatelet antibody and have used this technique to investigate the influence of maternal titers on the occurrence of neonatal thrombocytopenia. The assay is sensitive over a range of 12.5 to 800 ng of immunoglobulin G (IgG) per microtiter well and closely correlates with the complement lysis inhibition assay (CLIA) for antiplatelet antibody (correlation coefficient = 0.726). In many instances, the level of circulating antiplatelet antibody in maternal and cord bloods reflected the degree of maternal and neonatal thrombocytopenia, but several important exceptions were observed. We suggest that levels of antiplatelet antibody in the maternal blood and cord blood are not always predictive of the degree of neonatal thrombocytopenia.