Anne-Marie Rodriguez

A preoperative clinical prognostic model for non-metastatic renal cell carcinoma.

Authors from Naples, Paris and Rennes describe their efforts to develop a model for the preoperative prediction of outcome for non‐metastatic renal cancer. It is valuable to both urologist and patient to develop such a model, particularly so on preoperative criteria. The results of their study are interesting, leading to possibly helpful findings.

Placental Expression of HLA Class I Genes

This article presents an overview of the more recent data dealing with the constitutive, transcriptional, and translational expression of classical class la and nonclassical HLA‐E and ‐G class Ib products in the different trophoblast cell subpopulations that constitute the maternofetal interface during human pregnancy. Of particular interest is the expression of alternatively spliced HLA‐G transcriptional isoforms that may be translated in membrane‐bound or soluble protein products. Molecular regulatory mechanisms that may control the differential expression of class la and class lb molecules, according to the cell types, state of differentiation, and stages of gestation are also examined. They may operate at the levels of transcription, translation and/or transport of proteins to the cell surface. Functional significance of the absence of detectable cell surface expression of class Ia molecules in all trophoblast cell sub‐populations, and of the presence of membrane‐bound HLA‐G products in extravillous cytotrophoblast cells is finally questioned.

Safety of Intracavernous Bone Marrow-Mononuclear Cells for Postradical Prostatectomy Erectile Dysfunction: An Open Dose-Escalation Pilot Study

UNLABELLED Evidence from animal models replicating postradical prostatectomy erectile dysfunction (pRP-ED) suggests intracavernous injection of bone marrow-mononuclear cells (BM-MNCs) as a promising treatment approach for pRP-ED. We conducted a phase 1/2 pilot clinical trial of intracavernous autologous BM-MNC injection to treat pRP-ED (NCT01089387). Twelve patients with localized prostate cancer and vasculogenic pRP-ED refractory to maximal medical treatment were divided into four equal groups treated with escalating BM-MNC doses (2×10(7), 2×10(8), 1×10(9), 2×10(9)). Tolerance was the primary endpoint. Secondary endpoints were the effects on erectile function and penile vascularization at 6 mo, as assessed using the International Index of Erectile Function-15 and Erection Hardness Scale questionnaires, and color duplex Doppler ultrasound. We measured the peak systolic velocity in cavernous arteries and assessed endothelial function using the penile nitric oxide release test. No serious side effects occurred. At 6 mo versus baseline, significant improvements of intercourse satisfaction (6.8±3.6, 3.9±2.5, p=0.044) and erectile function (17.4±8.9, 7.3±4.5, p=0.006) domains of the International Index of Erectile Function-15 and Erection Hardness Scale (2.6±1.1, 1.3±0.8, p=0.008) were observed in the total population. Spontaneous erections showed significantly greater improvement with the higher doses. Clinical benefits were associated with improvement of peak systolic velocity and of % penile nitric oxide release test and sustained after 1 yr. Our results need to be confirmed by phase 2 clinical trials. PATIENT SUMMARY We report a phase 1/2 pilot clinical trial investigating cell therapy with injection of bone marrow mononucleated cells to treat postradical prostatectomy erectile dysfunction. No serious side effects occurred. Improvements of erectile function and penile vascularization were noted. Further studies are required to confirm these preliminary results.

ABSENCE OF IMPRINTING OF HLA CLASS IA GENES LEADS TO CO-EXPRESSION OF BIPARENTAL ALLELES ON TERM HUMAN TROPHOBLAST CELLS UPON IFN-GAMMA INDUCTION

Abstract Human trophoblast cells have developed various efficient regulatory mechanisms to prevent cell surface expression of the classical HLA-A, -B, and (but not always) -C class I molecules. This allows them to escape maternal alloimmune attack during pregnancy. However, recent results have demonstrated that such a lack of expression could be reversed in villous cytotrophoblast cells purified from term placenta by in vitro IFN-γ treatment. In this context, we investigated whether both maternal and paternal HLA class Ia antigens were co-dominantly expressed in such trophoblast cells. Using polymerase chain reaction sequence-specific primers for HLA-A and HLA-C alleles, we detected transcripts of both paternal and maternal origins, showing that these genes were not affected by genomic imprinting, at least in term placenta. After in vitro IFN-γ treatment, the polymorphic HLA-A and HLA-B antigens of both parental origins become detectable at the cell surface, as assessed by flow cytometry and/or complement-dependent microtoxicity test. Appearance of paternal antigens on trophoblast cells upon IFN-γ induction raises the question of the in vivo biological consequences of this phenomena, in term of materno-fetal tolerance and in particular of a potential allogeneic cytotoxic immune response.

Differences Between Human Sperm and Somatic Cell DNA in CpG Methylation Within the HLA Class I Chromosomal Region

PROBLEM: We investigated the possible negative regulatory mechanisms that repress classical human leukocyte antigen (HLA) class I gene expression in human spermatozoa and searched for novel testis‐specific coding sequences that might be present in the MHC class I chromosomal region.

Intracavernous Injections of Bone Marrow Mononucleated Cells for Postradical Prostatectomy Erectile Dysfunction: Final Results of the INSTIN Clinical Trial

We recently reported stage I of a phase 1/2 clinical trial of cell therapy to treat postradical prostatectomy erectile dysfunction (INSTIN, INtra-cavernous STem-cell INjection clinical trial, NCT01089387). In this first stage, four doses of intracavernous autologous bone marrow mononuclear cells (BM-MNCs) were tested in 12 patients. Here, we report the results of stage II, in which six additional patients received the optimal dose identified in stage I (109 BM-MNCs), and the long-term results in the 12 patients included in stage I. The objectives were to assess the safety and efficacy of this new treatment. In stage II, no patients had side effects, and the erectile function improvements were similar to those seen in stage I: after 6 months, significant improvements versus baseline were noted in International Index of Erectile Function-15 intercourse satisfaction (7.8±3.1 vs 2.2±3.4, p=0.033) and erectile function (18±8.3 vs 3.7±4.1, p=0.035) domains. In stage I patients, after a mean follow-up of 62.1±11.7 mo, there were no prostate cancer recurrences, and erectile function scores were somewhat lower compared with the 1-yr time point. These findings suggest that intracavernous BM-MNC injections are safe and improve erectile function. The decline in erectile function over time suggests a need for assessing repeated injections. PATIENT SUMMARY We report a phase 1/2 pilot clinical trial of cell therapy consisting in intracavernous injection of bone marrow mononuclear cells to treat postradical prostatectomy erectile dysfunction. Erectile function was improved after 6 mo in the patients given 1×109 cells. No serious side effects (life threatening or requiring hospitalisation) occurred after a mean follow-up of 62.1 mo in the first 12 patients.

Fine regulation of HLA class Ia gene expression in term human villous trophoblast cells

Summary Human trophoblast cells have developed various regulatory mechanisms to prevent cell surface expression of the classical HLA-A,-B, and (but not always)-C class I molecules, allowing them to escape maternal alloimmune attack during pregnancy. However, we have demonstrated that such a lack of expression could be reversed in villous trophoblast cells purified from term placenta after in vitro IFN-γ treatment and/or after human cytomegalovirus infection. In this context, we investigated whether both maternal and paternal HLA class Ia antigens were co-dominantly expressed in such trophoblast cells. Using PCR-sequence-specific primers for HLA-A and HLA-C alleles, we detected transcripts from both paternally and maternally inherited alleles, showing that these genes were not affected by genomic imprinting. Following in vitro IFN-γ treatment, the polymorphic HLA-A and HLA-B antigens of both parental origins become detectable at the cell surface, as assessed by flow cytometry and/or complement-dependent microcytotoxicity test. These results demonstrate that genomic imprinting of the classical Major Histocompatibility Complex class I genes is a non-conserved process between species, as some of these genes have been found to be paternally expressed in the rat placenta. Appearance of paternal antigens on human trophoblast upon IFN-γ induction raises the question of the in vivo biological consequences in terms of materno-fetal tolerance and, in particular, of potential allogenic cytotoxic immune responses and anti-viral immune reactions.

Molecular Regulatory Mechanisms That Repress Classical HLA Class I Gene Expression in Human Placenta

The human major histocompatibility complex (MHC) class I genomic region, located on the short arm of chromosome 6 (6p21.3 position), is composed of about 2000 kb of DNA. Besides pseudogenes and recently evidenced non-HLA class I genes, such as P5, R1, B30 (1), OTF3 (2), and S (3), this region contains both classical and nonclassical HLA class I genes (4). Polymorphic classical HLA-A, -B, -C class I genes are constitutively expressed (or inducible) on most somatic tissues, although at different levels, and play a critical role in cancer and viral immunity. Expression of these genes is developmentally regulated:They are not expressed on the cell surface of either male or female germinal cells (5, 6) or trophoblast cells (7). Current evidence favors repression of their transcription in all subpopulations of trophoblast cells in situ:Neither cytotrophoblast (from both term and first-trimester placentas) nor syncytiotrophoblast expresses these molecules (8). This suggests that such a repression plays a role in the maintenance of pregnancy. In contrast, the nonclassical HLA-E, -F, -G genes have a more restricted somatic tissue distribution, exhibit a low polymorphism, and are not yet associated with a known immunological or other biological function (7). Although nothing is known yet about expression of these genes in germinal cells, it is now clear that cytotrophoblast expresses HLA-G (8), and HLA-E and -F transcripts have been reported in the human placenta (9).