In the last years, thanks to the improvement in the prognosis of cancer patients, a growing attention has been given to the fertility issues. International guidelines on fertility preservation in cancer patients recommend that physicians discuss, as early as possible, with all patients of reproductive age their risk of infertility from the disease and/or treatment and their interest in having children after cancer, and help with informed fertility preservation decisions. As recommended by the American Society of Clinical Oncology and the European Society for Medical Oncology, sperm cryopreservation and embryo/oocyte cryopreservation are standard strategies for fertility preservations in male and female patients, respectively; other strategies (e.g. pharmacological protection of the gonads and gonadal tissue cryopreservation) are considered experimental techniques. However, since then, new data have become available, and several issues in this field are still controversial and should be addressed by both patients and their treating physicians.In April 2015, physicians with expertise in the field of fertility preservation in cancer patients from several European countries were invited in Genova (Italy) to participate in a workshop on the topic of “cancer and fertility preservation”. A total of ten controversial issues were discussed at the conference. Experts were asked to present an up-to-date review of the literature published on these topics and the presentation of own unpublished data was encouraged. On the basis of the data presented, as well as the expertise of the invited speakers, a total of ten recommendations were discussed and prepared with the aim to help physicians in counseling their young patients interested in fertility preservation.Although there is a great interest in this field, due to the lack of large prospective cohort studies and randomized trials on these topics, the level of evidence is not higher than 3 for most of the recommendations highlighting the need of further research efforts in many areas of this field. The participation to the ongoing registries and prospective studies is crucial to acquire more robust information in order to provide evidence-based recommendations.

While during the first trimester of pregnancy natural killer (NK) cells represent the most abundant lymphocyte population in the decidua, their actual function at this site is still debated. In this study we analyzed NK cells isolated from decidual tissue for their surface phenotype and functional capability. We show that decidual NK (dNK) cells express normal surface levels of certain activating receptors, including NKp46, NKG2D, and 2B4, as well as of killer cell immunoglobulinlike receptors (KIRs) and CD94/NKG2A inhibitory receptor. In addition, they are characterized by high levels of cytoplasmic granules despite their CD56 bright CD16 ؊ surface phenotype. Moreover, we provide evidence that in dNK cells, activating NK receptors display normal triggering capability whereas 2B4 functions as an inhibitory receptor. Thus, cross-linking of 2B4 resulted in inhibition of both cytolytic activity and interferon-␥ (IFN-␥) production. Clonal analysis revealed that, in the majority of dNK cell clones, the 2B4 inhibitory function is related to the defi- IntroductionNatural killer (NK) cells represent a lymphocyte population capable of recognizing and lysing not only tumor cells but also virus-infected cells in the absence of previous sensitization. 1,2 NK-cell function is regulated by a balance between activating and inhibitory signals. 3 Among peripheral-blood NK (pNK) cells, 2 subsets can be identified on the basis of CD56 surface expression: those with low/intermediate levels of CD56 (CD56 dim ) and those with high levels of CD56 (CD56 bright ). 4 CD56 dim NK cells represent the majority (Ͼ 90%) of pNK cells, contain abundant cytoplasmic granules, are highly cytotoxic, and mediate antibody-dependent cell cytotoxicity (ADCC) against IgG-coated target cells through the engagement of CD16. 5 The phenotypically and functionally distinct CD56 bright subset, representing less than 10% of pNK cells, does not express CD16 molecules or expresses low-density CD16 molecules, does not contain cytoplasmic granules, and is poorly cytolytic but has greater cytokine-production capacity. 4 In addition, the 2 pNK-cell subsets differ in their expression of killer-cell immunoglobulin-like receptors (KIRs) and CD94/NKG2A receptor. Thus, CD56 bright pNK cells have low to absent expression of KIRs, whereas they express high levels of CD94/NKG2A inhibitory receptor. In contrast, most CD56 dim pNK cells are KIR positive and express low levels of CD94/NKG2A. 4 During the first trimester of pregnancy, NK cells constitute 50% to 90% of the lymphocytes present in the decidua, a tissue in which B and T lymphocytes are infrequent. It is currently believed that in the early stages of gestation, decidual NK (dNK) cells may play an important role in the control of trophoblastic growth, differentiation, and invasion. Decidual NK cells are phenotypically similar to the CD56 bright pNK cell subset. However, transcriptional geneexpression profiling has shown that dNK cells express both perforin and granzymes at a similar or even higher level than CD56 di...

Summary:Knowledge of the impact of different conditioning regimens used in bone marrow transplantation on spermatogenesis is important in pre-BMT counselling for three reasons: (1) Most young patients who have not had children are concerned with their subsequent fertility; (2) For a number of diseases there are competing therapeutic options that may affect spermatogenesis more or less seriously; (3) Since spontaneous recovery of spermatogenesis is rare, it would be necessary to offer cryopreservation as soon as possible after diagnosis and prior to any treatment. This retrospective study evaluates 99 semen samples obtained in 64 patients who underwent BMT between 1982 and 1996. Recovery of spermatogenesis was observed in 90% of patients conditioned with cyclophosphamide (CY), in 50% of patients with CY plus busulphan (BU) or thiotepa and in 17% of patients with CY plus total body irradiation (TBI) or thoracoabdominal irradiation (TAI). Sperm quality following CY was within the normal range (WHO) in the majority of patients, whereas it was consistently severely impaired in patients who received irradiation or two alkylating agents. Following CY, spermatogenesis recovery was observed in 60% of patients tested 1 year post transplant and it was accomplished within the third year in 80% of cases. Keywords: bone marrow conditioning; spermatogenesis recovery; fertility counselling Due to improved survival following BMT for aplastic anemia or lymphohematological malignancies, there is a growing issue about counselling regarding subsequent fertility status. In addition, the introduction of new conditioning regimens could have a different impact on male reproduction. Although semen cryopreservation should always be suggested before starting any cytotoxic treatment, very often this procedure cannot be performed for various reasons. Firstly, when facing the diagnosis of cancer at a young age, patients and often their physicians, may not be prepared to plan for future fertility difficulties before starting therapy. Secondly, freezing may not readily be available in all oncological centers. Thirdly, in some patients poor sperm quality, due to the cancerous state itself, was previously considered to render semen unsuitable for freezing until the introduction of intracytoplasmic sperm injection (ICSI), a new technique which permits in vitro fertilization with few spermatozoa. The aim of the present study was to evaluate the degree, quality and kinetics of spermatogenesis recovery in 64 patients receiving various myeloablative regimens. Patients and methodsIn December 2000, the clinical charts of 64 males who underwent allogeneic BMT between 1982 and 1996, in the Department of Hematology, San Martino Hospital, Genoa, Italy who had had at least one semen analysis after treatment, were reviewed.Type of malignancy, previous chemotherapy treatment, conditioning regimen, age at BMT and previous reproductive history were obtained for each patient, as well as the fertility follow-up. Most semen analyses were performed in the Andrology ...

BackgroundThe issue of breaking bad news in assisted reproductive technology (ART) has been only partially explored by literature, and although some recommendations are available, specific guidelines are lacking. The present study aimed to explore the applicability of the oncologic SPIKES Protocol to the ART context.MethodsThirteen ART clinicians (7 gynecologists; 4 psychologists; 1 biologist; 1 obstetrician) completed the Critical Incidents Report (CIR) to describe the experience of delivering bad news in ART. The CIRs were first discussed with clinicians, then a focus group was created composed of 13 clinicians, one health communication expert and a patient to discuss the applicability of the six-step (SPIKES) Buckman Protocol to ART. The discussion was audiotaped, transcribed and analyzed with content analysis.ResultsThe SPIKES Protocol seems to fit ART consultations and participants found it practical and easy to understand. Some specificities were found for the ART context: the reiteration of bad news, the “patient” as a couple and the fact that ability to conceive is closely related to self-esteem, as well as to social and family identity. During the discussion of the SPIKES Protocol, participants highlighted the importance of: 1) providing a caring setting, by adding a reflection on the value of communication by phone; 2) exploring patients’ perceptions but also misinformation; 3) exploring patients’ desires and expectations, while balancing the need to be honest and clear; 4) applying Buckman’s suggestions for delivering information, and integrating clinical aspects with psychosocial ones; 5) managing and legitimizing patients’ emotions, in particular anger; 6) having a strategy for follow-up and supporting couples to make meaning of the ART experience.ConclusionThe proposal of a shared protocol for giving bad news in ART could be the starting point for training and experimental studies.

This study investigates the main functional features of subepithelial (SE) B cells and compares them with those of purified germinal center (GC) and follicular mantle (FM) B cells isolated from the same tonsils. Unlike FM B cells, SE B cells failed to produce polyspecific antibodies in vitro; unlike GC B cells, SE B cells expressed high levels of Bcl-2 and failed to undergo spontaneous apoptosis in vitro. The most striking function of SE B cells was their ability to produce IgM antibodies to T cell-independent type-2 (TI-2) (but not to TI-1) antigens (Ag). These antibodies could not be detected when both FM and GC B cells were stimulated with TI-2 Ag in vitro. Moreover, B cells isolated from peripheral blood were unable to mount a response to TI-2 Ag. The latter finding is consistent with the observation that B cells with the phenotypic features of SE B cells were virtually absent in the peripheral blood and emphasizes the notion that SE B cells belong to a subset of non-recirculating B cells. SE B cells were by far superior to FM B cells in mixed lymphocyte reaction (MLR) stimulation of allogeneic T cells in vitro, although they were not as efficient as dendritic cells (DC). In order to stimulate T cells efficiently, SE B cells had to be exposed to anti-mu antibody, a treatment which induced expression of activation markers such as CD80, CD86, CD69 and CD39, usually absent in resting SE B cells. CD80 and CD86 molecules expressed by SE B cells participated in the chain of events required to promote the proliferation of allogeneic T cells as demonstrated by inhibition tests with the appropriate mAb. The expression of CD80 and CD86 by anti-mu-treated SE B cells was not, however, the sole explanation for their good antigen presenting capacities since the exposure of FM B cells to anti-mu antibody also induced expression of these surface structures. Nevertheless, these cells failed to become good MLR stimulators. Collectively, the above data contribute further to the characterization of a distinct subset of tonsillar B cells which resemble, both phenotypically and functionally, the B cells of the splenic marginal zone.