Tumor-Infiltrating Lymphocyte Therapy or Ipilimumab in Advanced Melanoma.

BACKGROUND: Immune checkpoint inhibitors and targeted therapies have dramatically improved outcomes in patients with advanced melanoma, but approximately half these patients will not have a durable benefit. Phase 1-2 trials of adoptive cell therapy with tumor-infiltrating lymphocytes (TILs) have shown promising responses, but data from phase 3 trials are lacking to determine the role of TILs in treating advanced melanoma. METHODS: In this phase 3, multicenter, open-label trial, we randomly assigned patients with unresectable stage IIIC or IV melanoma in a 1:1 ratio to receive TIL or anti-cytotoxic T-lymphocyte antigen 4 therapy (ipilimumab at 3 mg per kilogram of body weight). Infusion of at least 5×109 TILs was preceded by nonmyeloablative, lymphodepleting chemotherapy (cyclophosphamide plus fludarabine) and followed by high-dose interleukin-2. The primary end point was progression-free survival. RESULTS: A total of 168 patients (86% with disease refractory to anti-programmed death 1 treatment) were assigned to receive TILs (84 patients) or ipilimumab (84 patients). In the intention-to-treat population, median progression-free survival was 7.2 months (95% confidence interval [CI], 4.2 to 13.1) in the TIL group and 3.1 months (95% CI, 3.0 to 4.3) in the ipilimumab group (hazard ratio for progression or death, 0.50; 95% CI, 0.35 to 0.72; P<0.001); 49% (95% CI, 38 to 60) and 21% (95% CI, 13 to 32) of the patients, respectively, had an objective response. Median overall survival was 25.8 months (95% CI, 18.2 to not reached) in the TIL group and 18.9 months (95% CI, 13.8 to 32.6) in the ipilimumab group. Treatment-related adverse events of grade 3 or higher occurred in all patients who received TILs and in 57% of those who received ipilimumab; in the TIL group, these events were mainly chemotherapy-related myelosuppression. CONCLUSIONS: In patients with advanced melanoma, progression-free survival was significantly longer among those who received TIL therapy than among those who received ipilimumab. (Funded by the Dutch Cancer Society and others; ClinicalTrials.gov number, NCT02278887.).

Circulating biomarkers, echocardiographic parameters, and incident subclinical atrial fibrillation in patients with diabetes and hypertension.

BACKGROUND: Long-term rhythm monitoring (LTRM) can detect undiagnosed atrial fibrillation (AF) in patients at high risk of AF and stroke. Biomarkers and echocardiographic parameters could, however, help identify patients benefitting most from LTRM. The aim of this study was to investigate, whether circulating biomarkers of cardiac and vascular function (brain natriuretic peptide (BNP), cardiac troponin I (cTnI), copeptin, and mid-regional proadrenomedullin (MR-proADM)) and echocardiographic parameters were associated with incident subclinical AF (SCAF) in a population at high risk of stroke in the presence of AF. For this purpose, we investigated individuals ≥65 years of age with hypertension and diabetes mellitus, but no history or symptoms of AF or other cardiovascular disease (CVD). METHODS: We included 82 consecutive patients (median age 71.3 years (IQR 67.4-75.1)). All patients received an insertable cardiac monitor (ICM) and were followed for a median of 588 days (IQR 453-712). On the day of ICM implantation, a comprehensive echocardiogram and blood samples were obtained. RESULTS: During a median follow-up of 588 days (IQR: 453-712 days), incident SCAF occurred in 17 patients (20.7%) with a median time to first-detected episode of 91 days (IQR 41-251 days). MR-proADM (median 0.87 nmol/L (IQR 0.76-1.02) vs 0.78 nmol/L (IQR 0.68-0.98)) and copeptin (median 13 pmol/L (IQR 9-17) vs 8 pmol/L (IQR 4-18)) levels were insignificantly higher in patients with incident SCAF. BNP and cTnI concentrations and echocardiographic parameters were similar in the two groups. CONCLUSIONS: MR-proADM, BNP, cTnI, copeptin, and several echocardiographic parameters were not associated with incident SCAF in this cohort of patients with hypertension and diabetes, but without any underlying CVD.

Effects of Age, Level of Education and HIV Status on Cognitive Performance in West African Older Adults: The West Africa IeDEA Cohort Collaboration.

An in-depth understanding of the impact of aging, cognitive reserve, and HIV status on cognitive function is needed in older West African adults. Ninety-nine HIV-negative and 334 HIV-positive adults aged ≥ 50 years were enrolled in three clinics (Senegal and Côte d'Ivoire) participating in the IeDEA West Africa collaboration. All subjects underwent the Free and Cued Selective Reminding Test (FCSRT) and the Isaacs Set Test (IST). Age (both linear and quadratic), education level, and HIV status effects on Z-scores were assessed using multivariate linear regression models. Interactions between HIV status and age or educational level were tested. In the present cohort of older West African adults, the role of age and educational level on episodic memory and verbal fluency was observed without revealing an interaction between HIV status and age effect. As age had quadratic effects, older HIV-positive adults should not be considered as a unique group irrespective of their age. Low-educated HIV-positive patients had the lowest verbal fluency performance compared to others. Further studies are needed to duplicate these results. In clinical settings, screening and adapted programs focusing on improving cognition in those patients are needed.

Use of 'non-medical' ultrasound imaging before mid-pregnancy in Copenhagen.

We have investigated prospectively how many pregnant women purchase ultrasound imaging before week 20, why they purchase scans, and which professional skills and certifications women expect the person performing the scan to have. In addition, we wanted to investigate whether the women were aware of any professional authorization procedures. Women attending the second trimester malformation scan of the Danish Prenatal Screening Program (n = 645) filled in a questionnaire about their use of non-medical ultrasound scans. Of these women, 154 (24%) had bought ultrasound scans: 50% wanted to have the fetal health and development checked and 49% wanted to find out the gender of the fetus. In addition, 68% felt that they received an evaluation of the fetal health state and 58% believed that there was legislation demanding a professional authorization needed to perform ultrasound imaging. This study shows that there is a significant demand among pregnant women for commercial ultrasound imaging of their fetus for various reasons. Knowledge of certifications and requirements for legal authorization is, however, sparse.

[Evaluation of microalbuminuria and lipid profile among type 2 diabetics]. / Evaluation de la microalbuminurie et du profil lipidique chez les diabétiques de type 2.

OBJECTIVE: To establish the respective prevalence of microalbuminuria and dyslipidemia and to evaluate their association with diabetes type 2. ANALYZE DATA: Prospective study of 195 type 2 diabetic subjects (125 women and 70 men) from a hospital in the city of Dakar (Senegal) for a check-up of diabetes. Age and sex were determined ; fasting blood glucose, glycated hemoglobin, total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides and micro- albuminuria were measured. RESULTS: In this study, the mean age of patients was 57.9 ± 11.1 years. Age, glycated hemoglobin and microalbuminuria were significantly higher in women than in men (P < 0.01 ; P < 0.03 ; P < 0.01 respectively). The prevalence of microalbuminuria is 48.7% and that of dyslipidemia is 41.1%. Glycated hemoglobin is higher in subjects with microalbuminuria than in patients with normal microalbuminuria with a statistically significant difference (P < 0.001). There is a strong correlation (R = 0.82) between glycated hemoglobin and microalbuminuria, 1% increase in HbA1c corresponding approximately to an increase of 39.7 mg/I of microalbuminuria. CONCLUSION: Microalbuminuria and dyslipidemia are frequently found in type 2 diabetes, but the pathophysiological mechanisms of the association are not well known.

No Detectable Differences in microRNA Plasma Levels between Diabetic Hypertensive Patients with and without Incident Subclinical Atrial Fibrillation.

Background: Long-term rhythm monitoring (LTRM) can detect undiagnosed atrial fibrillation (AF) in patients at risk of AF and stroke. Circulating microRNAs (miRNAs), which have been shown to play a role in atrial electrical and structural remodelling, could help to select patients who would benefit most from LTRM. The aim of this study was to investigate whether patients with diabetes mellitus (DM) and hypertension and screen-detected subclinical AF (SCAF) using an insertable cardiac monitor (ICM) have significantly different plasma baseline levels of five selected miRNAs playing a role in the modulation of atrial electrical and structural remodelling (miR-21-5p, miR-29b-3p, miR-150-5p, miR-328-3p, and miR-432-5p) compared to those without SCAF. Methods: This study was performed at the outpatient clinic of a secondary academic teaching hospital between December 2013 and November 2015. Eligible patients were ≥65 years of age with DM and hypertension but without known heart diseases. All patients received an ICM. On the day of ICM implantation, blood samples for the measurement of plasma levels of the five miRNAs were drawn. In this post hoc analysis, we investigated their expression by reverse transcription-quantitative polymerase chain reaction. MiRNA plasma levels in patients with and without newly detected SCAF were compared. Results: We included 82 consecutive patients (median age of 71.3 years (IQR 67.4-75.1)), who were followed for a median of 588 days (IQR: 453-712 days). Seventeen patients (20.7%) had ICM-detected SCAF. Plasma levels of miR-328-3p, miR-29b-3p, miR-21-5p, miR-432-5p, and miR-150-5p were slightly but not significantly different in patients with incident SCAF compared with patients without. Conclusions: In patients with hypertension and DM, newly detected SCAF was not significantly associated with changes in expression levels of miR-21-5p, miR-29b-3p, miR-150-5p, miR-328-3p, and miR-432-5p.

TIL Therapy: Facts and Hopes.

After a positive phase III trial, it is evident that treatment with tumor-infiltrating lymphocytes (TIL) is a safe, feasible, and effective treatment modality for patients with metastatic melanoma. Further, the treatment is safe and feasible in diverse solid tumors, regardless of the histologic type. Still, TIL treatment has not obtained the regulatory approvals to be implemented on a larger scale. Therefore, its availability is currently restricted to a few centers worldwide. In this review, we present the current knowledge of TIL therapy and discuss the practical, logistic, and economic challenges associated with implementing TIL therapy on a larger scale. Finally, we suggest strategies to facilitate the widespread implementation of TIL therapy and approaches to develop the next generation of TILs.

Cerebral alterations in West African HIV and non-HIV adults aged ≥50: An MRI study.

OBJECTIVES: To cross-sectionally describe brain alterations in PLHIV aged above 50 years old, receiving antiretroviral treatment (ART) and living in Senegal compared to HIV-negative subjects. METHODS: Twenty PLHIV and 26 HIV-negative subjects with comparable socio-demographic and clinical characteristics underwent an MRI exam (3D-T1 and FLAIR sequences). Global atrophy and White Matter Hyperintensities (WMH) were evaluated. After assessing the feasibility and acceptability of MRI scans in this population, we described atrophy and WHM prevalence and associated factors using logistic regressions. RESULTS: Overall, 43.5% of the study sample were aged ≥60 years, 58.7% were women, and 28.3% had hypertension. The overall prevalence of atrophy and WMH was 19.6% [95% CI: 8.1-31.1] and 30.4% [95% CI: 17.1-43.7]. HIV status had no significant effect on atrophy or WMH. Unemployment and hypertension were significantly associated with atrophy, whereas women were less likely to present atrophy. Aged ≥60 years was the only factor associated with WMH. CONCLUSIONS: A high prevalence of atrophy and WMH was observed in West African adults aged over 50 years without a clear HIV impact. As brain MRI studies are critical to better understand cognitive and emotional outcomes, we encourage those studies in older PLHIV in West Africa.

Detection of Subclinical Atrial Fibrillation in High-Risk Patients Using an Insertable Cardiac Monitor.

OBJECTIVES: The study sought to determine the incidence of subclinical atrial fibrillation (AF) in high-risk patients and to compare the effect of continuous versus intermittent monitoring. BACKGROUND: AF often occurs in a subclinical form, which makes it difficult to detect. The authors do not know the incidence of subclinical AF among patients ≥65 years of age with hypertension and diabetes mellitus. This group of patients has increased risk of developing AF and in addition a high thromboembolic risk, if AF is present. METHODS: A total of 82 outpatients ≥65 years of age (median age 71.3 years [interquartile range [IQR]: 67.4 to 75.1 years]) with hypertension and diabetes mellitus, and no history of AF or any other cardiovascular disease, were consecutively included. All patients received an insertable cardiac monitor (ICM) and were followed for a median of 588 days (IQR: 453 to 712 days). We compared continuous monitoring with 72-h Holter monitoring 1 month after ICM insertion. The primary endpoint was AF ≥2 min for the ICM and AF ≥30 s for the Holter monitoring. RESULTS: During follow-up 17 (20.7%) patients were found to have subclinical AF detected by ICM with a median time to first detected episode of 91 days (IQR: 41 to 251 days) from inclusion. Only 2 (2.4%) patients also had AF episodes on the 72-h Holter monitoring. All detected episodes were completely asymptomatic. CONCLUSIONS: The incidence of subclinical AF in this group of patients was surprisingly high. Continuous monitoring with ICM detected significantly more AF episodes than 72-h Holter monitoring. (Detection of Subclinical Atrial Fibrillation in High Risk Patients Using Implantable Loop Recorder; NCT02041832).

ELISPOT assay for detection of peptide specific interferon-gamma secreting cells in rhesus macaques.

A reliable procedure to measure antigen specific T cell responses in rhesus macaques is required to determine the efficacy of vaccines and immunotherapies. The currently available T cell assays are poorly quantifiable or technically difficult to perform. Classical 51Cr-release cytotoxic T cell (CTL) assays are cumbersome and difficult to quantitate reproducibly. Detection of specific T-cell using MHC-peptide tetrameric complexes is highly sensitive, but requires knowledge of MHC type and prior identification of T cell epitopes. We therefore developed a rhesus interferon-gamma (IFN-gamma) ELISPOT assay capable of detecting IFN-gamma secretion in response to stimulation with pooled 20-mer peptides. Peripheral blood mononuclear cells (PBMCs) from rhesus monkeys immunized with a DNA vaccine and recombinant canary pox encoding the Plasmodium knowlesi circumsporozoite protein (PkCSP) were incubated with pools of peptides from PkCSP. Positive responses to peptide pools and individual peptides ranging from 100 to 450 spot forming cells (SFC)/10(6) PBMC were detected in four of four immunized monkeys and in zero of two control monkeys. In two monkeys studied in detail, the IFN-gamma response was focussed on a single 20-mer peptide, QGDGANAGQPQAQGDGANAG, and was dependent on CD4(+), but not CD8(+), T cells. Background responses in control monkeys and preimmunization PBMCs ranged from 10 to 50 SFC/10(6) PBMC. The average within assay and between assay coefficients of variation (CV) for this peptide ELISPOT were 21.9 and 24.7%, respectively. This peptide IFN-gamma assay will be a useful tool for evaluation of T cell responses in rhesus macaques.

In vitro expression and in vivo immunogenicity of Plasmodium falciparum pre-erythrocytic stage DNA vaccines.

DNA vaccine plasmids were constructed that encoded four pre-erythrocytic antigens from the human malaria parasite Plasmodium falciparum: circumsporozoite protein (PfCSP); sporozoite surface protein 2 (PfSSP2); carboxyl terminus of liver stage antigen 1 (PfLSA-1 c-term); and, exported protein 1 (PfExp-1). Antigen expression was evaluated in vitro by immunoblot analysis of tissue culture cells following transient transfection with each plasmid. Clearly detectable levels of expression depended upon, or were markedly enhanced by, fusion of the antigen encoding sequences in-frame with the initiation complex and peptide leader sequence of human tissue plasminogen activator protein. Mice injected with these plasmids produced antigen specific antibody and cytotoxic T lymphocyte responses. However, the magnitudes of the responses were not always predicted by the in vitro expression assay. The results of this study provided the basis for further testing of these plasmids in primates and the formulation of multi-component pre-erythrocytic DNA vaccines for efficacy testing in human volunteers.

[Effect of gamma irradiation on the production and degradation system of the second hormonal messenger (author's transl)]. / Effets de l'irradiation gamma sur les systèmes de production et de dégradation du 2e messager hormonal

Gamma irradiation of rat liver plasma membranes leads to a decrease of the adenylcyclase activities stimulated by glucagon and fluoride. The observed inhibition is more important for the activity stimulated with glucagon. The 5'-nucleotisade activity is not changed by irradiation. When the phosphodiesterase system is submitted to gamma irradiation, the radiosensibility of enzymatic complex is more important.

[Variation of plasmatic adenosine 3'', 5-cyclic monophosphate in irradiated-enterectomised dogs (author's transl)]. / Variations du taux d'adénosine 3',5-monophosphate cyclique (ampc) chez des chienx irradiés entérectomisés

In the irradiated dog at different doses 250-400 R., a level plasma AMPc rise has been shown since the fourth hour with a maximum at 24 hours, returning to normal values about the thirtieth day. In irradiated at 250-300 R and secondarily enterectomised animals, nucleotide level rises 24 hours later, however this phenomenon is less intensive and later irradiated at 350 and 400 R.

Effects of butalamine on oxydative phosphorylations in the rat liver.

The effect of 3-phenyl-4-dibutylaminoethyl-5-inimino-1,2,4-oxadiazole (butalamine) was investigated in vivo and in vitro on oxydophosphorylation of mitochondria isolated from rat liver. In vitro with succinate and glutamate-malate as oxydable substrates, butalamine stimulates respiration at lower concentrations (100% at 3 X 10(-4) M for succinate and 40% at 4 X 10(-4) M for glutamate-malate) and inhibits it at higher ones (50% at 1 mM). Butalamine inhibits State 3 respiration and decreases ADP/O. Data on respiration blocked by rutamycine and on the effects of DNP, show that butalamine affects oxydophosphorylation in vitro, probably before DNP reactions are involved. In contrast to DNP, butalamine is an uncompetitive inhibitor when succinate is used as substrate in State 3. The potent lipophilic properties of butalamine should be responsible for these unspecific effects. In vivo, at a dosage of 50 mg/kg p.o. over 8 days, no alteration of State 3, State 4, ADP/O and respiratory control ratio (RC) was found.

Immunogenicity of the Plasmodium falciparum serine repeat antigen (p126) expressed by vaccinia virus.

cDNA encoding the serine repeat antigen (SERA) (also called p126) of Plasmodium falciparum has been isolated from the FCR3 strain and inserted into a recombinant vaccinia virus designated vP870. Expression analysis of vP870-infected Vero cells by immunoprecipitation has demonstrated several intracellular forms of SERA and a single secreted SERA peptide. Endoglycosidase digestion of these immunoprecipitated SERA peptides indicated that the intracellular SERA peptides contain simple, high-mannose N-linked oligosaccharides and that the secreted SERA peptide contains complex N-linked oligosaccharides. Pulse-chase experiments indicate that the multiple intracellular SERA peptides in infected Vero cells represent a trafficking pathway whereby the smallest SERA peptide is converted into larger peptides by co- and posttranslational modifications, including glycosylation, and eventually secreted from the cell with complex N-linked oligosaccharides. To study the immunogenicity of vaccinia virus-expressed SERA, rabbits were immunized with vP870 and their sera were analyzed for reactivity with authentic, parasite-derived SERA protein. The anti-vP870 rabbit sera reacted with P. falciparum-infected erythrocytes by immunofluorescence analysis, recognized authentic SERA from schizonts by both immunoprecipitation and Western blot (immunoblot) analyses, and recognized proteolytically processed fragments of SERA secreted into the culture medium by Western blot analysis. These results indicate that when expressed by vaccinia virus, SERA is glycosylated and secreted from infected cells and that in immunized rabbits, vaccinia virus-expressed SERA can stimulate a humoral immune response against SERA derived from blood-stage parasites.

A MAGE-A1 peptide presented to cytolytic T lymphocytes by both HLA-B35 and HLA-A1 molecules.

Antigens encoded by melanoma antigen (MAGE) genes are of particular interest for cancer immunotherapy because of their strict tumoral specificity and because they are shared by many tumors. Antigenic peptide EADPTGHSY encoded by MAGE-A1 and known to be presented by HLA-A1 is currently being used in therapeutic vaccination trials. We report here that a cytotoxic T-lymphocyte (CTL) clone, which is restricted by HLA-B35, recognizes the same peptide and, importantly, lyses HLA-B35 tumor cells expressing MAGE-A1. This peptide can be presented to CTL by both HLA-B*3501 and HLA-B*3503 molecules, which are expressed by approximately 19% of Caucasians. These results infer that the current clinical use of peptide EADPTGHSY can now be extended to HLA-B35 patients.

Induction of murine cytotoxic T lymphocytes against Plasmodium falciparum sporozoite surface protein 2.

Sporozoite surface protein 2 has been identified as a target of malaria vaccines designed to produce protective CD8+ cytotoxic T lymphocytes (CTL) because mice immunized with mastocytoma cells expressing a fragment of Plasmodium yoelii sporozoite surface protein 2 (PySSP2) are protected against malaria by an immune response that requires CD8+ CTL. To define CTL epitopes in the Plasmodium falciparum sporozoite surface protein 2 (PfSSP2), spleen cells (SC) from mice immunized with irradiated sporozoites (irr spz) were stimulated with synthetic peptides, and these effectors were tested for cytolytic activity against peptide-pulsed, major histocompatibility complex (MHC)-matched targets. Two peptides containing CTL epitopes, A6 (Pf SSP2 3D7 214-233) and BH1 (Pf SSP2 3D7 3-11) were identified in bulk cultures of SC from immune C57BL/6 mice, and by production of CTL lines. Immunization with recombinant vaccinia expressing the full length PfSSP2 induced antigen specific, MHC-restricted, CD8+ T cell-dependent cytolytic activity against these two peptides. Finally, CTL were induced by immunization with a bacteria-derived recombinant fragment of PfSSP2 (rPfSSP2) mixed with a liposomal formulation containing a cationic lipid (Lipofectin Reagent, LPF). Induced CTL lysed target cells pulsed with peptide A6 or with LPF/rPfSSP2, but not targets pulsed with only rPfSSP2. These studies demonstrate that CTL specific to PfSSP2 are present in C57BL/6 mice and that immunization with purified rPfSSP2 delivered with LPF induces a cytotoxic T cell response.

Genetic polymorphisms of Q region genes from wild-derived mice: implications for Q region evolution.

Both serological and DNA sequence analyses were performed to determine the extent of genetic polymorphism in Q region genes. A panel of Qa-2-specific monoclonal antibodies (mAbs) was tested on 35 wild-derived and inbred mouse strains. Members of this reagent panel recognize multiple and distinct epitopes on the Qa-2-bearing molecule(s). Although quantitative variations in Qa-2 levels were observed, no structural polymorphisms were detected. All strains were either entirely positive or entirely negative with the complete set of reagents. Moreover, cell surface Qa-2 expression was not significantly affected by differences in age or sex of the mouse or cell cycle status. To confirm this apparent lack of genetic polymorphism, the polymerase chain reaction (PCR) technique was used to amplify a portion of the 3' end of the Q region genes, Q4 to Q9, from several independent wild-derived strains of mice. Sequence analysis of the amplified material revealed very little evidence of nucleotide divergence. All strains tested had a Q even DNA sequence identical to that of Q6/Q8 in the B10 strain. Likewise, all tested strains had a Q odd DNA sequence identical to Q7/Q9 in the B10 strain. Two strains showed additional Q even sequences, while all strains tested possessed additional Q odd sequences. The observed lack of polymorphism suggests that the Q genes have evolved in a different manner from H-2K and H-2D. Moreover, duplications of these genes appear to have arisen prior to nucleotide sequence divergence.