Early ovarian pregnancy diagnosed by ultrasound and successfully treated with multidose methotrexate. A case report.

A case report of a primary interstitial ovarian pregnancy is presented. A 37-year-old married woman with two children after two Cesarean sections and a spontaneous abortion, with a contraceptive intrauterine device (IUD) inserted three years before, presented at five weeks plus five days amenorrhea with a positive pregnancy test and lower abdominal pain but with no vaginal bleeding. Her previous menstrual cycles had been regular. She was hemodynamically stable. On bimanual examination, the uterus was of normal size, and there was an approximate four-cm tender right adnexal mass. Serum beta-human chorionic gonadotropin (b-hCG) was confirmed positive. Ultrasound revealed a well-positioned IUD in the uterus and a right adnexal mass with normal vascular flow on Doppler, that contained a well-defined gestational sac, well-distinct from the quiescent hemorrhagic corpus luteum. There was no fetal node or cardiac activity or free fluid. The patient received four injections of methotrexate intramuscularly using the multidose regimen that involves the administration of methotrexate calculated according to body weight, alternated with 0.1 mg/kg of leucovorin calcium per os after 30 hours until the values of 3-hCG had decreased by 15%. The patient's post-treatment period was uneventful with a full restoration of ovarian morphology and the complete absorption of the gestational sac. This case is the first where diagnosis was made by endovaginal sonography and treatment was made by multidose methotrexate. Spiegelberg criteria for the diagnosis of ovarian pregnancy are obsolete; new ultrasound and laboratory criteria are needed for a diagnosis as early as possible without the need of surgery.

Cross-protection against lymphocytic choriomeningitis virus mediated by a CD4+ T-cell clone specific for an envelope glycoprotein epitope of Lassa virus.

Recombinant vaccinia virus expressing the Lassa virus (LV) envelope glycoprotein precursor, V-LSGPC, was used to study the basis of LV-induced cross-protective immunity against the closely related arenavirus lymphocytic choriomeningitis virus (LCMV). C3H/HeJ mice primed with V-LSGPC developed neither circulating antibodies nor CD8+ cytotoxic T cells specific for LCMV, yet they resisted a normally lethal LCMV challenge. Spleen cells from such mice gave a proliferative response to LCMV in vitro that was inhibitable by anti-CD4 antibody. Synthetic peptides corresponding to predicted T-cell sites common to the envelope glycoprotein precursor (GP-C) of LV and that of LCMV were used to map the specificity of the proliferative response to an epitope located between amino acids 403 and 417 of LV GP-C. Several CD4+ T-cell clones specific for the 403-417 peptide were isolated and found to produce gamma interferon in response to both the peptide and LCMV. One of these clones, C9, was selected for further study. C9 lysed I-AK-bearing target cells, and when adoptively transferred to C3H/HeJ mice, it was capable of mediating both a peptide-specific delayed hypersensitivity reaction and resistance to lethal LCMV challenge. These collective findings demonstrate, for the first time, that CD4+ T cells can play a major role in arenavirus-specific cross-protective immunity.

Differential expression of tumor target binding and cytolytic activities in bone marrow culture-derived macrophages.

The binding and killing of a number of murine (DBA/2J) leukemia cell lines by murine bone marrow-derived macrophages (M phi s) were studied under various conditions to provide clues to the nature and specificity of tumor recognition by M phi s. Under test was the hypothesis that tumor target binding by activated M phi s is effected by saturable, high-affinity, trypsin and low temperature-sensitive receptors, whereas target binding by normal M phi s is of low affinity, nonsaturable, insensitive to low temperature and trypsin, and is believed not to be mediated by conventional receptors. The results obtained showed that this scenario was realized only rarely and that each target cell line had its own distinctive binding characteristics. Thus target binding by both unactivated and activated M phi s can be sensitive or insensitive to trypsin and low temperature depending on the target in question. This finding suggests that the binding of different targets is mediated by distinct receptors or different combinations of receptors. Contrary to previous reports, the binding of all targets tested was found to decrease with M phi activation and induction of cytotoxicity, but whatever binding strength that remained was obviously adequate for the manifestation of cytotoxicity. Fractionation of M phi s according to size (and hence maturity) revealed that tumor binding capacity (per unit surface area) increased with M phi size. Although target binding decreased in all M phi fractions after activation, it still correlated with cytotoxic activity and cell size for a given activated M phi population. Thus target binding correlates well with antitumor potential in unactivated Møs and with cytotoxic activity in activated M phi s.

The interaction of activated macrophages with normal and neoplastic cells.

The interaction of various cell types with Salmonella enteritidis 11RX activated macrophages was examined. P815 tumour cells, Concanavalin A-activated blast cells and splenic lymphocytes were tested for their ability to bind to activated macrophages whilst the ability to inhibit macrophage-mediated lysis of 51Cr-labelled P815 tumour cells was tested with unlabelled tumour cells and blast cells. All three cell types were bound by activated macrophages but only P815 cells were lysed by the activated macrophages and only these cells could act as cold target inhibitors of cytolysis. This phenomenon could not be explained by the release of cytotoxic factors resulting from the interaction of blast cells with activated macrophages. The lack of cold target inhibition by blast cells may have been due to the detachment of initially adherent blast cells leaving the activated macrophages free to interact with tumour cells.

Characterization of the effector cells responsible for tumour resistance in Salmonella enteritidis 11RX-immunized mice.

In an attempt to characterize the effector cells responsible for tumour resistance in Salmonella enteritidis 11RX-immunized mice the anti-macrophage agent trypan blue was used in both in vivo and in vitro experiments. Resistance was measured in vivo by the clearance of 125I from the peritoneal cavity of mice injected intraperitoneally with 125I-5-iododeoxyuridine labelled Ehrlich Ascites Tumour (EAT) cells. The in vitro correlate was measured by lysis of 51Cr-labelled tumour cells by peritoneal cells (PC) from 11RX-immunized mice. Pre-treatment of resistant mice with trypan blue greatly reduced both 125I clearance and 51Cr release. The in vitro cytolytic activity was non-specific. Fractionation of cytotoxic PC on the basis of adherence to plastic or nylon wool and buoyant density, coupled with the use of appropriate cell targets, showed that the bulk of cytotoxic activity resided with macrophages, with some contribution from other cells such as natural killer cells. Killing of labelled tumour cells could be inhibited by competition with unlabelled cells or by separating the PC and tumour cells by a cell impermeable membrane. This showed that close association between the effector and target cells was necessary before killing could occur.