In search of specific cytotoxic T lymphocytes infiltrating or accompanying human ovarian carcinoma

Summary Lymphocytes infiltrating human ovarian carcinoma obtained directly from the tumour mass (tumour-infiltrating lymphocytes, TIL) or from the carcinomatous ascites (tumour-associated lymphocytes, TAL) were expanded in vitro in long-term cultures with interleukin-2 and tested for their specific cytolytic activity. Killing of the autologous tumour was detected only in a proportion of the patients, less frequently in TIL compared to TAL. In fact two out of ten TIL and four out of nine TAL cultures tested showed significant levels of lysis against the autologous tumour. This cytotoxic activity was not restricted to the autologous tumour, as other tumour cell lines, including non-ovarian ones, were lysed as well. The cultures that were not cytotoxic against the autologous tumour were in most cases able to lyse other tumour cell lines of ovarian or other histology. Cloning of TIL from one patient was performed: of 22 clones tested, 4 displayed higher cytotoxicity against the autologous tumour compared to the uncloned population and 3 out of these 4 did not kill an irrelevant carcinoma cell line. In order to stimulate the expansion of putative specific effectors we performed mixed lymphocyte/tumour cultures (MLTC) with autologous or allogeneic tumour cells. No stimulation of cytotoxicity against the autologous tumour was detected after MLTC in nine different TAL populations, using autologous or allogeneic tumours as stimulators. On the contrary, peripheral blood lymphocytes from two patients after MLTC with the autologous tumour showed increased killing of the autologous and decreased killing of an allogeneic target. In conclusion TIL and TAL from ovarian carcinoma expanded in vitro with interleukin-2 usually have non-MHC-restricted cytotoxicity and variable degrees of reactivity against the autologous tumour. A preferential killing for the autologous tumour was not observed even after MLTC. These results do not exclude the existence of tumour-specific cytotoxic T lymphocytes in ovarian carcinoma; nevertheless they suggest that putative specific effectors have very low frequency and that culture techniques for expanding their growth more selectively are still to be optimized.     

Functional aspects of the postovulatory follicle in the ovary of the African catfish,Clarias gariepinus,after induced ovulation

Summary In captive African catfish, Clarias gariepinus, ovulation was induced with human chorionic gonadotropin (HCG) 4 I.U./g body weight to study the function of postovulatory follicles (POFs). Ultrastructural and enzyme-histochemical data indicate that, apart from special theca cells, the granulosa of relative young POFs (i.e., from 16 h and 28 h after HCG-injection) is capable of producing steroids. Possible functions of the synthesized steroids are discussed. Histological comparison of POFs from stripped and from unstripped fish, as well as histochemical investigation of the contents of ovulated ova and granulosa of POFs at 48 h after HCG-injection, showed that the latter structure is involved in phagocytosis of the disintegrating ovulated eggs. The polysaccharide-lipid-protein material, initially taken up by heterophagolysosomes of the granulosa cells, subsequently undergoes fatty degeneration. The granulosa cells of the POFs showed strong acid phosphatase activity and abundant granular endoplasmic reticulum from 16 h after HCG-injection onward; heterophagolysosomes appeared at 32 h. These results indicate that after ovulation the phagocytotic function of the granulosa develops progressively. Autophagolysosomes, responsible for the final disintegration of POFs, become increasingly evident in the granulosa cells with increasing time after ovulation.     

Maturation of pig and rat oocytes transplanted into surrogate pig follicles in vitro

Pig oocytes obtained from slaughterhouse material and rat oocytes obtained from PMSG-treated immature females were incubated as isolated oocytes or injected into explanted pig follicles (5–8 mm). Free oocytes of both species, with or without their cumulus investment or gonadotropins during culture, matured at high rates after 30 hr or 9–10 hr of culture, respectively. Gonadotropic stimulation was necessary for maturation of both the native and injected cumulus-intact pig oocytes in follicle culture. Cumulus-free pig oocytes injected into follicle failed to mature in response to gonadotropic stimulation, suggesting an inability to perceive or respond to stimulation. Injected rat oocytes, however, matured irrespective of cumulus investment or gonadotropic stimulation. Their maturation was delayed and reduced at 9 hr. These results in the rat suggest that the pig follicular environment is incapable of regulating rat oocyte maturation but rather presents a permissive or supportive environment for their maturation. The explanted surrogate follicles from the pig or other species may provide a useful model for the study of oocyte-follicle interactions in oocyte maturation within or between species.     

Seasonal changes in ovarian state in a eusocial halictine bee,Lasioglossum duplex,based on stages of the oldest oocytes in each ovariole (Hymenoptera: Halictidae)

Summary The changes in ovarian activity in the life cycle of the eusocial halictine beeLasioglossum (Evylaeus) duplex (Dalla Torre) was studied in both queens and workers by examining the stages of the terminal follicle in each of the six ovarioles. By this method, ovarian activities of both queens and workers were more quantitatively determined than by observation of gross ovarian features as usually conducted. Queen ovaries clearly exhibited two active periods, corresponding to the spring solitary phase and the summer eusocial phase, with distinctly greater activity in the latter. In ovaries of overwintering queens oosorption of young follicle was observed. Worker ovaries were found more active in orphan than in queenright colonies. The order of ovarian activity obtained from pooled data, summer queens>spring queens>orphan workers>queenright workers, was also recognized by comparison of individual females. Bionomics of the eusocial halictine beeLasioglossum duplex IX. Contribution No. 3107 from the Inst. Low Temp. Sci.     

Effect of interferon γ and tumour necrosis factor α on sensitivity to cisplatin in ovarian carcinoma cell lines

This study aimed to investigate whether the biological response modifiers (BRM) interferon (IFN) and tumour necrosis factor (TNF) could enhance the cytotoxic action of cisplatin on ovarian tumour cells in vitro. The sensitivity of four cell lines (OAW42, GG, JAM and PE01) to drugs and drug combinations was tested by a radiolabelled-thymidine incorporation assay. Cell lines demonstrated a range of sensitivity to cisplatin and the innate cytotoxic effect of each of the BRM. When IFN was used in combination with cisplatin, a significant enhancement of cisplatin toxicity occurred in three of four cell lines. TNF demonstrated such an effect in two cell lines but diminished the toxicity of cisplatin in one cell line. A purely additive effect of the agents may explain the enhanced toxicity of cisplatin in some of these cases. However, in one cell line at least (PEO1), both TNF and IFN demonstrated a clearly synergistic effect with cisplatin. These BRM used in conjunction with cisplatin may provide better antitumour regimen than cisplatin alone in some patients with ovarian cancer, but the response is likely to be heterogeneous between patients.     

Circ_0021573 acts as a competing endogenous RNA to promote the malignant phenotypes of human ovarian cancer cells

Circular RNAs (circRNAs) have been reported to be implicated in the tumorigenesis and progression of ovarian cancer. Here, the study was designed to explore the activity of human circ_0021573 in ovarian cancer pathogenesis and its regulation through the competing endogenous RNA (ceRNA) crosstalk. Circ_0021573, microRNA (miR)? 936, and cullin 4B (CUL4B) were quantified by qRT-PCR and western blot. Cell proliferation ability was detected by XTT, 5-Ethynyl-2′-Deoxyuridine (EdU), and colony formation assays. Cell apoptosis, migration, and invasion were assessed by flow cytometry, wound-healing, and transwell assays, respectively. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were used to evaluate the direct relationship between miR-936 and circ_0021573 or CUL4B 3′UTR. Xenograft studies were applied to assess the role of circ_0021573 in tumor growth. Our data showed that circ_0021573 expression is enhanced in human ovarian cancer. Inhibition of circ_0021573 impedes cell proliferation, migration, and invasion and promotes apoptosis in vitro, as well as diminishes tumor growth in vivo. Mechanistically, circ_0021573 contains a miR-936 binding site, and miR-936 is a relevant mediator of circ_0021573 regulation. MiR-936 direct targets and inhibits CUL4B. MiR-936-mediated suppression of CUL4B hinders cell proliferation, migration, and invasion and accelerates apoptosis in vitro.. These data suggested that circ_0021573 might promote the malignant phenotypes of ovarian cancer cells by functioning as a ceRNA for miR-936 to induce CUL4B, which provided a promising target for the prevention and inhibition of ovarian cancer.     

Ultrastructure of the theca interna of ovarian follicles in sheep

Summary The theca interna of non-atretic ovarian follicles from 2.0 mm in diameter up to the stage shortly following ovulation was studied by light and electron microscopy.In follicles <3.0mm in diameter, the theca interna consisted of about 8–12 layers of flattened cells, together with many capillaries and small bundles of collagen. Two main forms of cellular differentiation were seen. These were towards either fibroblast-like cells or presumed steroidogenic cells whose cytoplasm contained large amounts of predominantly smooth tubular endoplasmic reticulum, to which some ribosomes were attached. The majority of cells were of relatively undifferentiated or intermediate structure.In larger follicles up to the early stages of oestrus the theca interna cells became larger and less flattened, and cells rich in tubular endoplasmic reticulum became proportionately more numerous. By 18 h after the onset of oestrus the theca interna was oedematous, and many cells possessed pseudopodia. Many cells also contained numerous lipid droplets, but there were no signs of thecal cell degeneration or death. Shortly after ovulation the basal lamina of the membrana granulosa was incomplete, and it became more difficult to distinguish between theca and granulosa layers. Structural heterogeneity, with two major cell types and cells of intermediate structure, was present at all stages.It was concluded that: (1) the theca interna of 2.0–2.9 mm follicles contained many cells whose structure was compatible with a steroidogenic capacity; (2) changes in the differentiated thecal cells up to the early stages of oestrus were quantitative rather than qualitative, and suggestive of an increased steroidogenic capacity; (3) the accumulation of lipid in many cells of the theca interna by 18 h after the onset of oestrus probably reflected a reduction in steroidogenic activity; and (4) there was no evidence of any structural specialization to facilitate the transport of steroids from the theca interna to the membrana granulosa.     

Ultrastructural comparison of smooth muscle from ovarian follicle and oviduct of the golden hamster

Summary Ultrastructural characteristics of smooth muscle taken from ovarian follicles and oviducts of hamsters are compared. Differences between the two muscle types are more quantitative than qualitative, thus confirming that follicular muscle is a true smooth muscle with no unique characteristics. While both muscle types contain 50–80 Å filaments, -glycogen deposits, and organelles characteristically found in smooth muscle, the oviductal cells have substantially more sacs, tubular structures, sarcoplasmic reticulum, and mitochondria. Another difference concerns the cellular junctions; the oviductal cells exhibit nexuses, whereas the follicular cells show desmosomelike junctions. Based on ultrastructural differences, follicular smooth muscle seems to be a relatively toneless muscle suited for short, infrequent contractions, whereas oviductal smooth muscle is probably involved in more active tonic contractions.Supported by an Institutional Research Grant from Texas Women's University, by NIH Grant HD 12988, and by the Department of Anatomy at Wright State University