The effect of lavender and bergamot oil applied via inhalation on the anxiety level and sleep quality of surgical intensive care unit patients.

OBJECTIVE: Due to numerous stressors in intensive care, common psychosocial problems arise in patients. Among these, decreased anxiety and sleep quality are observed. This study aims to determine the effect of lavender and bergamot oil applied by inhalation on anxiety and sleep quality in surgical intensive care unit patients. METHODS: Fifty-four patients hospitalized in the intensive care unit of a hospital in Istanbul, Turkey, were included in this study. They were randomly divided into three groups (Lavender, Bergamot, Control). Intervention groups were exposed to 3 drops of lavender oil or bergamot oil on pillows for 20 min, which were then placed 10 cm away from the patient's head. This intervention was applied for two nights. The patients' sleep quality and anxiety level were evaluated using the "State-Trait Anxiety Inventory (STAI)" and "Richard-Campbell Sleep Scale (RCSS)." Data were analyzed using Chi-square, independent t-test, One-way ANOVA tests. RESULTS: According to our results, anxiety scores significantly decreased in the lavender and bergamot groups. Sleep quality scores significantly increased (p = <0.001). The control group showed lower sleep quality scores. CONCLUSION: Lavender and bergamot oil inhalation appeared effective in reducing anxiety and improving sleep quality in surgical intensive care unit patients.

EBV and 1q Gains Affect Gene and miRNA Expression in Burkitt Lymphoma.

Abnormalities of the long arm of chromosome 1 (1q) represent the most frequent secondary chromosomal aberrations in Burkitt lymphoma (BL) and are observed almost exclusively in EBV-negative BL cell lines (BL-CLs). To verify chromosomal abnormalities, we cytogenetically investigated EBV-negative BL patient material, and to elucidate the 1q gain impact on gene expression, we performed qPCR with six 1q-resident genes and analyzed miRNA expression in BL-CLs. We observed 1q aberrations in the form of duplications, inverted duplications, isodicentric chromosome idic(1)(q10), and the accumulation of 1q12 breakpoints, and we assigned 1q21.2-q32 as a commonly gained region in EBV-negative BL patients. We detected MCL1, ARNT, MLLT11, PDBXIP1, and FCRL5, and 64 miRNAs, showing EBV- and 1q-gain-dependent dysregulation in BL-CLs. We observed MCL1, MLLT11, PDBXIP1, and 1q-resident miRNAs, hsa-miR-9, hsa-miR-9*, hsa-miR-92b, hsa-miR-181a, and hsa-miR-181b, showing copy-number-dependent upregulation in BL-CLs with 1q gains. MLLT11, hsa-miR-181a, hsa-miR-181b, and hsa-miR-183 showed exclusive 1q-gains-dependent and FCRL5, hsa-miR-21, hsa-miR-155, hsa-miR-155*, hsa-miR-221, and hsa-miR-222 showed exclusive EBV-dependent upregulation. We confirmed previous data, e.g., regarding the EBV dependence of hsa-miR-17-92 cluster members, and obtained detailed information considering 1q gains in EBV-negative and EBV-positive BL-CLs. Altogether, our data provide evidence for a non-random involvement of 1q gains in BL and contribute to enlightening and understanding the EBV-negative and EBV-positive BL pathogenesis.

Molecular monitoring of T-cell kinetics and migration in severe neurotoxicity after real-world CD19-specific chimeric antigen receptor T cell therapy.

CD19-specific chimeric antigen receptor (CD19-CAR) T-cell therapies mediate durable responses in late-stage B-cell malignancies, but can be complicated by a potentially severe immune effector cell-associated neurotoxicity syndrome (ICANS). Despite broad efforts, the precise mechanisms of ICANS are not entirely known, and resistance to current ICANSdirected therapies (especially corticosteroids) has been observed. Recent data suggest that inflammatory cytokines and/or targeting of cerebral CD19-expressing pericytes can disrupt the blood-brain barrier and facilitate influx of immune cells, including CAR T cells. However, specific tools for CD19-CAR T-cell analysis within often minute samples of cerebrospinal fluid (CSF) are not broadly available. Here, we applied our recently developed digital polymerase chain reaction assays to monitor CD19-CAR T-cell kinetics in CSF and blood in real-world patients with neurotoxicity. Consistently, we observed a CAR T-cell enrichment within CSF in ICANS patients with further progressive accumulation despite intense corticosteroid- containing immuno-chemotherapies in a subset of patients with prolonged and therapy-resistant grade 3-4 neurotoxicity. We used next-generation T-cell receptor-b sequencing to assess the repertoire of treatment-refractory cells. Longitudinal analysis revealed a profound skewing of the T-cell receptor repertoire, which at least partly reflected selective expansion of infused T-cell clones. Interestingly, a major fraction of eventually dominating hyperexpanded T-cell clones were of non-CAR T-cell derivation. These findings hint to a role of therapy-refractory T-cell clones in severe ICANS development and prompt future systematic research to determine if CAR T cells may serve as 'door openers' and to further characterize both CAR-positive and non-CAR T cells to interrogate the transcriptional signature of these possibly pathologic T cells.

Investigation of Preoperative Fasting Time and Preoperative and Postoperative Well-Being of Patients.

AIM: This study aimed to investigate the preoperative fasting time and preoperative and postoperative well-being of patients. METHOD: This is a descriptive study. In Istanbul, between January and June 2016, 130 patients from the Training and Research Hospital Urology Clinic were included in the study. In collecting the data of the research, the information form, which included the descriptive features and surgical intervention information of the patients, and together with the scale of "Quality of Recovery-40 Questions (QoR-40)" was used. Mean, Standard Deviation, Median, Frequency, Ratio, Minimum, Maximum, Students' t-Test, Mann Whitney U-test, Paired Samples T-Test, Pearson and Spearman's Correlation Analysis were used for data analysis. RESULTS: The mean age was 57.48 ± 11.12, 79.2% of the patients were male and 63.8% of them underwent robotic prostatectomy. It was observed that the mean of times for preoperative fasting, solid foods, liquid foods were 11.26 ± 2.17, 12.56 ± 2.47, 11.26 ± 2.17 hours. The percent score of patients who felt hungry, thirsty, and dryness of the mouth was 30.8, 48.5, and 40, respectively, and patients had complaints such as nausea, fatigue, and feeling anxious before surgery. CONCLUSION: It was determined that patients were still hungry after midnight until surgery and remained hungry for a long time. The preoperative and postoperative well-being was adversely affected; however, there was no statistically significant relationship between the preoperative fasting time and postoperative well-being of patients, duration of hospitalization (p > .05).

Sleep Quality and Factors Affecting Sleep in Individuals With an Intestinal Ostomy: A Descriptive Cross-Sectional Study.

BACKGROUND: Multiple factors affect the sleep quality of individuals with intestinal stomas. PURPOSE: This study sought to determine sleep quality and factors affecting sleep in individuals with intestinal ostomies. METHODS: A descriptive cross-sectional design was used. This study followed 68 individuals with intestinal stomas at the stoma therapy unit of a university hospital. A form was used to gather information about patient demographic and stoma-related data (age, sex, work status, stoma duration, cause and type of stoma, stoma care provider, sleep status during the day, daily coffee consumption, and stoma-related factors affecting sleep), and the Pittsburgh Sleep Quality Index was used to score patient sleep patterns. Descriptive statistics, t-test, chi-square test, Fisher's exact test, and logistic regression analysis were used for statistical analysis. RESULTS: The patients' mean age was 53.7 ± 13.8 years; 51.5% were male, and 66.2% were married. Of the 68 patients, 41.2% had a diagnosis of rectal cancer, and 55.9% had ileostomies. Mean stoma duration was 24.1 ± 5.8 months, and 57.4% of participants performed their own stoma care. On a scale of 0 to 21, the participants' mean sleep score was 9.08 ± 5.03, and 66.2% of patients were found to have poor sleep quality. High sleep quality was significantly positively associated with colostomy (odds ratio, 1.78; 95% confidence interval [CI], 1.18-2.69; P = .006) and self-performed stoma care (odds ratio, 1.54; 95% confidence interval, 1.03-2.30; P = .036). CONCLUSION: The results of the current study can provide reference data for future studies and highlight the importance of assessing sleep quality in persons with intestinal stomas.

Non-Random Pattern of Integration for Epstein-Barr Virus with Preference for Gene-Poor Genomic Chromosomal Regions into the Genome of Burkitt Lymphoma Cell Lines.

BACKGROUND: Epstein-Barr virus (EBV) is an oncogenic virus found in about 95% of endemic Burkitt lymphoma (BL) cases. In latently infected cells, EBV DNA is mostly maintained in episomal form, but it can also be integrated into the host genome, or both forms can coexist in the infected cells. METHODS: In this study, we mapped the chromosomal integration sites of EBV (EBV-IS) into the genome of 21 EBV+ BL cell lines (BL-CL) using metaphase fluorescence in situ hybridization (FISH). The data were used to investigate the EBV-IS distribution pattern in BL-CL, its relation to the genome instability, and to assess its association to common fragile sites and episomes. RESULTS: We detected a total of 459 EBV-IS integrated into multiple genome localizations with a preference for gene-poor chromosomes. We did not observe any preferential affinity of EBV to integrate into common and rare fragile sites or enrichment of EBV-IS at the chromosomal breakpoints of the BL-CL analyzed here, as other DNA viruses do. CONCLUSIONS: We identified a non-random integration pattern into 13 cytobands, of which eight overlap with the EBV-IS in EBV-transformed lymphoblastoid cell lines and with a preference for gene- and CpGs-poor G-positive cytobands. Moreover, it has been demonstrated that the episomal form of EBV interacts in a non-random manner with gene-poor and AT-rich regions in EBV+ cell lines, which may explain the observed affinity for G-positive cytobands in the EBV integration process. Our results provide new insights into the patterns of EBV integration in BL-CL at the chromosomal level, revealing an unexpected connection between the episomal and integrated forms of EBV.

The Effect of Progressive Relaxation Exercise on Physiological Parameters, Pain and Anxiety Levels of Patients Undergoing Colorectal Cancer Surgery: A Randomized Controlled Study.

PURPOSE: This study aimed to determine the effect of progressive relaxation exercises on physiological parameters, pain, anxiety, and serum cortisol levels in patients undergoing colorectal cancer surgery. DESIGN: This study is a randomized controlled study and has been registered at the Clinical Trial Registry Center (ID: NCT04731428). METHODS: The study was conducted on 63 patients (experimental group = 31, control group = 32), who were scheduled for elective laparoscopic-colorectal surgery in the general surgery clinic of a university hospital in Istanbul between March 2018 and May 2019 and met the inclusion criteria. The patients in the experimental group were given progressive relaxation exercises for 15 minutes preoperatively and on postoperative days 1, 2, and 3 after breathing exercise training. The routine treatments and care process of the patients in the control group were maintained. The preoperative and postoperative pain and anxiety levels of both groups were evaluated. Blood pressure, heart rate and respiratory rate, oxygen saturation, and serum cortisol levels were measured at certain time intervals before and after the relaxation exercise. FINDINGS: The patients in the experimental group had lower postoperative pain and anxiety levels and a lower rate of using opioid analgesic on postoperative day 0 compared to the control group. Progressive relaxation exercise had no statistically significant effect on serum cortisol or physiological parameters (P > .05). CONCLUSIONS: Progressive relaxation exercise did not affect cortisol levels and vital signs but decreased pain and anxiety, and relatively increased tissue oxygenation, appearing an effective, safe, and practicable nursing intervention.

Local Intracerebral Immunomodulation Using Interleukin-Expressing Mesenchymal Stem Cells in Glioblastoma.

PURPOSE: Mesenchymal stem cells (MSCs) show an inherent brain tumor tropism that can be exploited for targeted delivery of therapeutic genes to invasive glioma. We assessed whether a motile MSC-based local immunomodulation is able to overcome the immunosuppressive glioblastoma microenvironment and to induce an antitumor immune response. EXPERIMENTAL DESIGN: We genetically modified MSCs to coexpress high levels of IL12 and IL7 (MSCIL7/12, Apceth-301). Therapeutic efficacy was assessed in two immunocompetent orthotopic C57BL/6 glioma models using GL261 and CT2A. Immunomodulatory effects were assessed by multicolor flow cytometry to profile immune activation and exhaustion of tumor-infiltrating immune cells. Diversity of the tumor-specific immune response as analyzed using T-cell receptor sequencing. RESULTS: Intratumoral administration of MSCIL7/12 induced significant tumor growth inhibition and remission of established intracranial tumors, as demonstrated by MR imaging. Notably, up to 50% of treated mice survived long-term. Rechallenging of survivors confirmed long-lasting tumor immunity. Local treatment with MSCIL7/12 was well tolerated and led to a significant inversion of the CD4+/CD8+ T-cell ratio with an intricate, predominantly CD8+ effector T-cell-mediated antitumor response. T-cell receptor sequencing demonstrated an increased diversity of TILs in MSCIL7/12-treated mice, indicating a broader tumor-specific immune response with subsequent oligoclonal specification during generation of long-term immunity. CONCLUSIONS: Local MSC-based immunomodulation is able to efficiently alter the immunosuppressive microenvironment in glioblastoma. The long-lasting therapeutic effects warrant a rapid clinical translation of this concept and have led to planning of a phase I/II study of apceth-301 in recurrent glioblastoma.

T cell receptor next-generation sequencing reveals cancer-associated repertoire metrics and reconstitution after chemotherapy in patients with hematological and solid tumors.

The dynamics of immunoaging and the onset of immunoparesis in healthy individuals and cancer patients has been controversially discussed. Moreover, the role of chemotherapy on T cell regeneration needs further elucidation in light of novel immunotherapies that have become standard of care for many elderly cancer patients. We used next-generation immunosequencing to study T cell receptor (TCR) repertoire metrics on 346 blood samples from healthy individuals and cancer patients producing a dataset with around 8.8 million TCR reads. This analysis showed that decline of T cell diversity and increase in T cell clonality is a continuous process beginning in healthy individuals over 40 years of age. Untreated patients with both hematological and solid tumors showed blood TCR repertoires with significantly lower diversity and higher clonality as compared to healthy individuals across all decades. Loss in T cell diversity was essentially driven by a loss in richness in aging healthy individuals, while in cancer patients a loss in repertoire evenness was an additional contributing factor. Interestingly, chemotherapy did not impair the regeneration of blood TCR repertoire diversity to pre-treatment age-specific levels. Surprisingly, even patients over the age of 70 years receiving highly T cell toxic therapies reestablished their pre-treatment T cell diversity suggesting rebound thymic activity rather than recovery of T cell counts by peripheral expansion only. Taken together, these data suggest that human TCR repertoire metrics gradually deteriorate in the aging individual, but age-specific TCR metrics are restored after T cell depleting therapy even in elderly cancer patients.

High-Throughput Immunogenetics Reveals a Lack of Physiological T Cell Clusters in Patients With Autoimmune Cytopenias.

Autoimmune cytopenias (AIC) such as immune thrombocytopenia or autoimmune hemolytic anemia are claimed to be essentially driven by a dysregulated immune system. Using next-generation immunosequencing we profiled 59 T and B cell repertoires (TRB and IGH) of 25 newly diagnosed patients with primary or secondary (lymphoma-associated) AIC to test the hypothesis if these patients present a disease-specific immunological signature that could reveal pathophysiological clues and eventually be exploited as blood-based biomarker. Global TRB and IGH repertoire metrics as well as VJ gene usage distribution showed uniform characteristics for all lymphoma patients (high clonality and preferential usage of specific TRBV- and TRBJ genes), but no AIC-specific signature. Since T cell immune reactions toward antigens are unique and polyclonal, we clustered TCRß clones in-silico based on target recognition using the GLIPH (grouping of lymphocyte interactions by paratope hotspots) algorithm. This analysis revealed a considerable lack of physiological T cell clusters in patients with primary AIC. Interestingly, this signature did not discriminate between the different subentities of AIC and was also found in an independent cohort of 23 patients with active autoimmune hepatitis. Taken together, our data suggests that the identified T cell cluster signature could represent a blood biomarker of autoimmune conditions in general and should be functionally validated in future studies.

T-cell repertoire profiling by next-generation sequencing reveals tissue migration dynamics of TRBV13-family clonotypes in a common experimental autoimmune encephalomyelitis mouse model.

The experimental autoimmune encephalomyelitis (EAE) model is indispensable for autoimmunity research, but model-specific T cell dynamics are sparsely studied. We used next-generation immunosequencing across lymphoid organs, blood and spinal cord in response to immunization with myelin basic protein (MBP) to study T cell repertoires and migration patterns. Surprisingly, most spinal cord T cells were unique to the individual animal despite the existence of shared MBP-specific clones, suggesting a previously underestimated T cell diversity. Almost complete emigration of pathogenic clones from blood to spinal cord indicates that blood is not a suitable compartment to study EAE-mediating T cells.

Deep sequencing of bone marrow microenvironments of patients with del(5q) myelodysplastic syndrome reveals imprints of antigenic selection as well as generation of novel T-cell clusters as a response pattern to lenalidomide.

In myelodysplastic syndromes with a partial deletion of the long arm of chromosome 5, del(5q), lenalidomide is believed to reverse anergic T-cell immunity in the bone marrow resulting in suppression of the del(5q) clone. In this study we used next-generation sequencing of immunoglobulin heavy chain (IGH) and T-cell receptor beta (TRB) rearrangements in bone marrow-residing and peripheral blood-circulating lymphocytes of patients with del(5q) myelodysplastic syndromes to assess the immune architecture and track adaptive immune responses during treatment with lenalidomide. The baseline bone marrow B-cell space in patients was comparable to that of age-matched healthy controls in terms of gene usage and IGH clonality, but showed a higher percentage of hypermutated IGH sequences, indicating an expanded number of antigen-experienced B lineage cells. Bone marrow B lineage clonality decreased significantly and hypermutated IGH clones normalized upon lenalidomide treatment, well in line with the proliferative effect on healthy antigen-inexperienced B-cell precursors previously described for this drug. The T-cell space in bone marrow of patients with del(5q) myelodysplastic syndromes showed higher TRB clonality compared to that of healthy controls. Upon lenalidomide treatment, myelodysplastic syndrome-specific T-cell clusters with low to medium spontaneous generation probabilities emerged; these clusters were shared across patients, indicating a common antigen-driven T-cell response pattern. Hence, we observed B lineage diversification and generation of new, antigen-dependent T-cell clusters, compatible with a model of adaptive immunity induced against the del(5q) clone by lenalidomide. Overall, this supports the concept that lenalidomide not only alters the functional T-cell state, but also the composition of the T- and B-cell repertoires in del(5q) myelodysplastic syndromes.

Determining the incidence of postoperative delirium in elderly patients who undergo orthopaedic surgical interventions in Turkey.

OBJECTIVE: To determine the incidence of postoperative delirium in elderly patients having undergone orthopaedic surgical interventions. METHODS: The cross-sectional study was conducted at the traumatology clinic of GATA Haydarpasa Training and Research Hospital in Istanbul, Turkey, from April 2014 to April 2015 and comprised patients who underwent orthopaedic surgical interventions. The subjects included were aged >65 years, had no mental disorders, no acute cerebrovascular disease, no known history of delirium and/or dementia. Data was collected using a self-generated questionnaire, mini mental state examination and delirium rating scale. SPSS 18 was used for data analysis. RESULTS: Of the 60 participants, 39(65%) were female and 21(35%) were male. The overall mean age was 77.07±8.66 years. Besides, 22(36.7%) patients hadmoderate cognitive impairment preoperatively, and 51(85%) had no delirium postoperatively while 9(15%) had delirium. CONCLUSIONS: Degree of cognitive impairment,advanced age and type of surgery were determined to be risk factors for delirium.

Precautions Taken by Nurses about the Prevention of Hospital-Acquired Infections in Intensive Care Units.

OBJECTIVE: To determine the precautions that nurses take for avoiding hospital-acquired infections in intensive care units of a State University Medical Faculty Hospital in Istanbul. METHODS: The research data were collected by a questionnaire developed by the authors. The study was conducted in intensive care units of a medical faculty hospital of a state university in Istanbul province. 85 nurses working in different various intensive care units and providing informed consent participated in the study. RESULTS: Intravenous catheterization, urinary catheterization, ventilator-associated infections and surgical site infections were assessed. The questionnaire was scored by applying a conversion of 100 to the total scores obtained, with the highest score being 100 and lowest score being 0. The percentage of nurses that practised all of the approaches about preventing hospital-acquired infections was estimated to be 8.2% for catheter-related bloodstream infections, 67.1% for surgical site infections, 72.9% for catheter-associated urinary tract infections, 27.1% for ventilator-associated infections, 29.4% for isolation preventions and 62.5% for attempts related to sterilization/disinfection of the medical devices. CONCLUSION: It was seen that nurses use most of the effective measures in order to prevent hospital-acquired infections. The guidelines generated for intensive care units should be updated according to international standards as needed. These guidelines should be used effectively; the differences between intensive care units should be resolved and all nurses should be trained at certain intervals.

Dynamic changes of the normal B lymphocyte repertoire in CLL in response to ibrutinib or FCR chemo-immunotherapy.

Using next-generation immunoglobulin (IGH) sequencing and flow cytometry, we characterized the composition, diversity and dynamics of non-malignant B cells in patients undergoing treatment with the Bruton tyrosine kinase (BTK) inhibitor ibrutinib or chemo-immunotherapy with fludarabine, cyclophosphamide, and rituximab (FCR). During ibrutinib therapy, non-malignant B cell numbers declined, but patients maintained stable IGH diversity and constant fractions of IGH-mutated B cells. This indicates partial preservation of antigen-experienced B cells during ibrutinib therapy, but impaired replenishment of the normal B cell pool with naïve B cells. In contrast, after FCR we noted a recovery of normal B cells with a marked predominance of B cells with unmutated IGH. This pattern is compatible with a deletion of pre-existing antigen-experienced B cells followed by repertoire renewal with antigen-naïve B cells. These opposite patterns in B cell dynamics may result in different responses towards neoantigens versus recall antigens, which need to be further defined.

Immunophenotyping of Newly Diagnosed and Recurrent Glioblastoma Defines Distinct Immune Exhaustion Profiles in Peripheral and Tumor-infiltrating Lymphocytes.

Purpose: Immunotherapeutic treatment strategies for glioblastoma (GBM) are under investigation in clinical trials. However, our understanding of the immune phenotype of GBM-infiltrating T cells (tumor-infiltrating lymphocytes; TILs) and changes during disease progression is limited. Deeper insight is urgently needed to therapeutically overcome tumor-induced immune exhaustion.Experimental Design: We used flow cytometry and cytokine assays to profile TILs and peripheral blood lymphocytes (PBLs) from patients with GBM, comparing newly diagnosed or recurrent GBM to long-term survivors (LTS) and healthy donors. TCR sequencing was performed on paired samples of newly diagnosed and recurrent GBM.Results: We identified a clear immune signature of exhaustion and clonal restriction in the TILs of patients with GBM. Exhaustion of CD8+ TILs was defined by an increased prevalence of PD-1+, CD39+, Tim-3+, CD45RO+, HLA-DR+ marker expression, and exhibition of an effector-/transitional memory differentiation phenotype, whereas KLRG1 and CD57 were underrepresented. Immune signatures were similar in primary and recurrent tumors; however, restricted TCR repertoire clonality and a more activated memory phenotype were observed in TILs from recurrent tumors. Moreover, a reduced cytokine response to PHA stimulation in the blood compartment indicates a dysfunctional peripheral T-cell response in patients with GBM. LTS displayed a distinct profile, with abundant naïve and less exhausted CD8+ T cells.Conclusions: TILs and PBLs exhibit contrasting immune profiles, with a distinct exhaustion signature present in TILs. While the exhaustion profiles of primary and recurrent GBM are comparable, TCR sequencing demonstrated a contracted repertoire in recurrent GBM, concomitant with an increased frequency of activated memory T cells in recurrent tumors. Clin Cancer Res; 24(17); 4187-200. ©2018 AACRSee related commentary by Jackson and Lim, p. 4059.

Monitoring multiple myeloma by next-generation sequencing of V(D)J rearrangements from circulating myeloma cells and cell-free myeloma DNA.

Recent studies suggest that circulating tumor cells and cell-free DNA may represent powerful non-invasive tools for monitoring disease in patients with solid and hematologic malignancies. Here, we conducted a pilot study in 27 myeloma patients to explore the clonotypic V(D)J rearrangement for monitoring circulating myeloma cells and cell-free myeloma DNA. Next-generation sequencing was used to define the myeloma V(D)J rearrangement and for subsequent peripheral blood tracking after treatment initiation. Positivity for circulating myeloma cells/cell-free myeloma was associated with conventional remission status (P<0.001) and 91% of non-responders/progressors versus 41% of responders had evidence of persistent circulating myeloma cells/cell-free myeloma DNA (P<0.001). About half of the partial responders showed complete clearance of circulating myeloma cells/cell-free myeloma DNA despite persistent M-protein, suggesting that these markers are less inert than the M-protein, rely more on cell turnover and, therefore, decline more rapidly after initiation of effective treatment. Positivity for circulating myeloma cells and for cell-free myeloma DNA were associated with each other (P=0.042), but discordant in 30% of cases. This indicates that cell-free myeloma DNA may not be generated entirely by circulating myeloma cells and may reflect overall tumor burden. Prospective studies need to define the predictive potential of high-sensitivity determination of circulating myeloma cells and DNA in the monitoring of multiple myeloma.

T-cell diversification reflects antigen selection in the blood of patients on immune checkpoint inhibition and may be exploited as liquid biopsy biomarker.

Cancer immunotherapy with antibodies targeting immune checkpoints, such as programmed cell death protein 1 (PD-1), shows encouraging results, but reliable biomarkers predicting response to this costly and potentially toxic treatment approach are still lacking. To explore an immune signature predictive for response, we performed liquid biopsy immunoprofiling in 18 cancer patients undergoing PD-1 inhibition before and shortly after initiation of treatment by multicolor flow cytometry and next-generation T- and B-cell immunosequencing (TCRß/IGH). Findings were correlated with clinical outcomes. We found almost complete saturation of surface PD-1 on all T-cell subsets after the first dose of the antibody. Both T- and B-cell compartments quantitatively expanded during treatment. These expansions were mainly driven by an increase in the activated T-cell compartments, as well as of naïve B- and plasma cells. Deep immunosequencing revealed a clear diversification pattern of the clonal T-cell space indicative of antigenic selection in 47% of patients, while the remaining patients showed stable repertoires. 43% of the patients with a diversification pattern showed disease control in response to the PD-1 inhibitor. No disease stabilizations were observed without clonal T-cell space diversification. Our data show for the first time a clear impact of PD-1 targeting not only on circulating T-cells, but also on B-lineage cells, shedding light on the complexity of the anti-tumor immune response. Liquid biopsy T-cell next-generation immunosequencing should be prospectively evaluated as part of a composite response prediction biomarker panel in the context of clinical studies.