Peripheral Blood Lymphocyte Count Research Articles

Mesenchymal Stem Cells Yield Transient Improvements in Motor Function in an Infant Rhesus Macaque with Severe Early-Onset Krabbe Disease.

Krabbe disease, or globoid cell leukodystrophy, is a rare disorder caused by deficient galactosylceramidase activity and loss of myelin‐forming oligodendrocytes, resulting in progressive demyelination and severely impaired motor function. Disease symptoms in humans appear within 3–6 months of age (early infantile) and manifest as marked irritability, spasticity, and seizures. The disease is often fatal by the second year of life, with few effective treatment options. Herein we evaluated the therapeutic potential of mesenchymal stem cells (MSCs) administered intracranially to a 1‐month‐old rhesus macaque diagnosed with severe early‐onset Krabbe disease that displayed neurologic and behavioral symptoms similar to those of human patients. The infant was subjected to physical and neurological behavior examinations and nerve conduction velocity tests to assess efficacy, and outcomes were compared with age‐matched normal infants and Krabbe‐affected rhesus monkeys with late‐onset disease. Changes in major blood lymphocyte populations were also monitored to assess host immune cell responses. MSC administration resulted in transient improvements in coordination, ambulation, cognition, and large motor skills, which correlated with increased peripheral nerve conduction velocities and decreased latencies. Improvements also corresponded to transient increases in peripheral blood lymphocyte counts, but secondary challenge failed to elicit allo‐antibody production. Nevertheless, white cell and neutrophil counts showed dramatic increases, and CD20+ B cell counts underwent a precipitous decline at late stages of disease progression. Correlative data linking MSC administration to transient improvements in motor function suggest that MSCs should be evaluated further as an experimental therapy for rare neurodegenerative diseases. Stem Cells Translational Medicine 2017;6:99–109

Change in peripheral blood lymphocyte count in dogs following adoptive immunotherapy using lymphokine-activated T killer cells combined with palliative tumor resection

We evaluated changes in peripheral blood lymphocyte (PBL) count in dogs following adoptive immunotherapy using lymphokine-activated T killer cells (T-LAK) in combination with surgery. Fifteen tumor-bearing dogs treated with T-LAK therapy combined with palliative resection of tumors were enrolled in the present study. T-LAK were generated from autologous peripheral blood mononuclear cells (PBMC) by culture with recombinant human interleukin −2 (rhIL-2) and solid phase anti-canine cluster of differentiation (CD)3 antibody. T-LAK were administrated intravenously at 2–4-week intervals. After the first administration of T-LAK, counts of PBL and T lymphocyte subsets (CD3+, CD4+ and CD8+ cells) increased and the CD4/CD8 ratio decreased, with significant increases in CD8+ cells (P<0.05). In 8 tumor-bearing dogs that were administered sequential T-LAK, available data on changes in PBL and T lymphocyte phenotypes until the fifth administration were also analyzed. In tumor-bearing dogs administered 5 rounds of T-LAK, CD8+ cell counts were maintained high until the fifth administration of T-LAK. Moreover, the CD4/CD8 ratio remained low until the fifth administration of T-LAK. These results indicate that T-LAK therapy combined with surgery may increase peripheral blood T lymphocytes, particularly CD8+ cells, in tumor-bearing dogs.

Sex-based differences in lymphocyte proliferation in the spleen after vanadium inhalation

Vanadium (V) is a transition metal often adhered to particulate matter and released into the atmosphere as vanadium pentoxide (V2O5) by the burning of fossil fuels. This air pollutant causes adverse effects in the immune system. Lymphocytosis and splenomegaly have been reported with increased white pulp in mice after V inhalation. The effect of V on the immune system as related to sex has been poorly investigated. This study sought to determine if V inhalation (a) produced lymphoproliferation that could explain the changes previously observed in the spleen and in peripheral blood lymphocyte counts and (b) whether any observed effects differed due to gender. Immunohistochemical analyses of Ki-67, a specific proliferation marker, was made in the spleens of CD-1 male and female mice exposed for 1 h, twice a week, over a 12-week period to V2O5 (at 1.4 mg V2O5/m3) by whole-body inhalation; similar analyses were performed on spleens of control mice exposed to vehicle (filtered air). The results showed that in male mice there was a significant increase in percentage of Ki-67 immunopositive lymphocytes starting from the second week and until the end of the exposure. The Ki-67 signal was cytoplasmic and nuclear in the exposed males, while in controls the signal was only nuclear. In female mice, V inhalation singificantly increased the percentage of proliferating lymphocytes only after 1 week of exposure. Ki-67 signal was observed only in the nucleus of lymphocytes from the control and exposed females. The results here help to explain the splenomegaly and lymphocytosis observed previously in male mice and support the lymphoproliferative effect induced by V. Lastly, the finding that there was a sex difference in the effect of vanadium on lymphocyte proliferation suggests a role for sex hormones in potential protection against V immunotoxicity; however, further studies are needed to support this hypothesis.

Analysis of the Prognostic Effect of Peripheral Blood Lymphocyte and Monocyte Count after Autologous Hematopoietic Stem Cell Transplant in Refractory/Relapsed Hodgkin's Lymphoma in a Pediatric, Adolescent, and Young Adult Population

Analysis of the Prognostic Effect of Peripheral Blood Lymphocyte and Monocyte Count after Autologous Hematopoietic Stem Cell Transplant in Refractory/Relapsed Hodgkin's Lymphoma in a Pediatric, Adolescent, and Young Adult Population

Abstract P2-02-04: Distinct clinical and biological values of subpopulations of circulating tumor cells (CTCs) in primary breast cancer

Abstract Background: CTCs represent a heterogeneous population of cells with different phenotypes and biological values. Epithelial to mesenchymal transition (EMT) gives rise to cells with stem cell-like properties with increased resistance to chemotherapy that may be under detected by currently approved assays. The aim of this study was to characterize CTCs based on the expression of epithelial and mesenchymal markers in primary breast cancer (BC) and to correlate them with patients'/tumor characteristics. Methods: This prospective translational study included 422 patients with primary BC enrolled from March 2012 to February 2015. Blood for CTC detection was drawn before surgery (422 patients), before 1st cycle (95 patients) and before 2nd cycle (53 patients) of adjuvant therapy. Isolated peripheral blood mononuclear cells (PBMC) were depleted of cells of hematopoietic origin (CD45+) using RossetteSep kit (StemCell Technologies) negative selection with anti-CD45 antibody. RNA extracted from CD45-depleted (CD45) PBMC was interrogated for expression of EMT-inducing transcription factors (TWIST1, SNAIL1, SLUG, ZEB1) and epithelial (CK19) gene transcripts by quantitative reverse transcription-PCR. Expressions of gene transcripts in CD45- PBMC from patients were compared to those of CD45- PBMC of 60 healthy donors. Results: Totally, CTCs were detected in 116/422 (27.5%) patients before surgery, in 21/95 (22.1%) patients after surgery and before 1st cycle and in 19/53 (35.8%) of patients before 2nd cycle of adjuvant therapy. Before surgery, CTCs exhibited only epithelial markers in 38 (9.0%) patients, only EMT markers in 68 (16.1%) of patients, while in 10 (2.4%) patients CTCs with both epithelial and EMT markers were detected. Epithelial CTCs were more often detected before surgery compared to after surgery (11.4% vs. 2.1%; p = 0.003), while mesenchymal CTCs were more often detected after the 1st cycle of chemotherapy as opposed to detection before surgery (30.2% vs. 18.2%; p = 0.05). Patients with N2-3 disease had more often detectable CTCs compared to patients with N0-1 disease (41.4% vs. 24.9%, p = 0.01) and this was mainly driven by mesenchymal CTCs (31.0% for N2-3 vs. 16.0% for N0-1; p = 0.007). Similarly, patients that lacked p53 expression (wild type TP53) in primary tumor had more often CTCs with EMT phenotype opposite to patients with p53 expression (p = 0.02). Presence of epithelial CTCs was significantly associated with lower absolute lymphocyte (p = 0.02) and neutrophil (p = 0.02) counts in peripheral blood. Conclusions: Our results support the concept of CTCs phenotypic heterogeneity in breast cancer patients. These results support the role of EMT in cancer pathogenesis and suggest that CTCs with EMT phenotype are involved in tumor dissemination while their increase after chemotherapy might be a mechanism of treatment resistance. Moreover, these data suggest inverse relationship between immune cells and epithelial CTCs which stress the role of immune cells in tumor dissemination. Citation Format: Mego M, Jurisova S, Karaba M, Minarik G, Benca J, Sedlackova T, Manasova D, Malejcikova M, Sieberova G, Macuch J, Gronesova P, Sufliarsky J, Pindak D, Cristofanilli M, Reuben JN, Mardiak J. Distinct clinical and biological values of subpopulations of circulating tumor cells (CTCs) in primary breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-02-04.

Immune-Enhancing Effects of Taishan Pinus massoniana Pollen Polysaccharides on DNA Vaccine Expressing Bordetella avium ompA.

Bordetella avium is the causative agent of bordetellosis, which remains to be the cause of severe losses in the turkey industry. Given the lack of vaccines that can provide good protection, developing a novel vaccine against B. avium infection is crucial. In this study, we constructed a eukaryotic expression plasmid, which expressed the outer membrane protein A (ompA) of B. avium, to prepare a B. avium recombinant ompA-DNA vaccine. Three concentrations (low, middle, and high) of Taishan Pinus massoniana pollen polysaccharides (TPPPS), a known immunomodulator, were used as adjuvants, and their immune conditioning effects on the developed DNA vaccine were examined. The pure ompA-DNA vaccine, Freund’s incomplete adjuvant ompA-DNA vaccine, and the empty plasmid served as the controls. The chickens in each group were separately inoculated with these vaccines three times at 1, 7, and 14 days old. Dynamic changes in antibody production, cytokine secretion, and lymphocyte count were then determined from 7 to 49 days after the first inoculation. Protective rates of the vaccines were also determined after the third inoculation. Results showed that the pure DNA vaccine obviously induced the production of antibodies, the secretion of cytokines, and the increase in CD4+ and CD8+ T lymphocyte counts in peripheral blood, as well as provided a protective rate of 50% to the B. avium-challenged chickens. The chickens inoculated with the TPPPS adjuvant ompA-DNA vaccine and Freund’s adjuvant ompA-DNA vaccine demonstrated higher levels of immune responses than those inoculated with pure ompA-DNA vaccine, whereas only the ompA-DNA vaccine with 200 mg/mL TPPPS completely protected the chickens against B. avium infection. These findings indicate that the B. avium ompA-DNA vaccine combined with TPPPS is a potentially effective B. avium vaccine.

Clinical characteristics of chronic lymphocytic leukemia occurring in chornobyl cleanup workers.

The recently demonstrated radiation-induction of chronic lymphocytic leukemia (CLL) raises the question as to whether the amount of radiation exposure influences any of the clinical characteristics of the disease. We evaluated the relationship between bone marrow radiation doses and clinical characteristics and survival of 79 CLL cases diagnosed during 1986-2006 in a cohort of 110 645 male workers who participated in the cleanup work of the Chornobyl nuclear accident in Ukraine in 1986. All diagnoses were confirmed by an independent International Hematology Panel. Patients were followed up to the date of death or end of follow-up on 31 October 2010. The median age at diagnosis was 57 years. Median bone marrow dose was 22.6 milligray (mGy) and was not associated with time between exposure and clinical diagnosis of CLL (latent period), age, peripheral blood lymphocyte count or clinical stage of disease in univariate and multivariate analyses. Latent period was significantly shorter among those older at first exposure, smokers and those with higher frequency of visits to the doctor prior to diagnosis. A significant increase in the risk of death with increasing radiation dose was observed (p = 0.03, hazard ratio = 2.38, 95% confidence interval: 1.11,5.08 comparing those with doses ≥22 mGy to doses <22 mGy). After adjustment for radiation dose, survival of CLL cases was significantly shorter among those with younger age at first exposure, higher peripheral blood lymphocyte count, more advanced clinical stage of disease and older age at diagnosis (all p < 0.05). This is the first study to examine association between bone marrow radiation doses from the Chornobyl accident and clinical manifestations of the CLL in Chornobyl cleanup workers. The current study provides new evidence on the association of radiation dose and younger age at first radiation exposure at Chornobyl with shorter survival after diagnosis. Future studies are necessary with more cases in order to improve the statistical power of these analyses and to determine their significance. Copyright © 2016 John Wiley & Sons, Ltd.

Assessment of bone marrow lymphocytic status during tyrosine kinase inhibitor therapy and its relation to therapy response in chronic myeloid leukaemia.

Tyrosine kinase inhibitors (TKIs) used in the treatment of chronic myeloid leukaemia have been reported to induce immunomodulatory effects. We aimed to assess peripheral blood (PB) and bone marrow (BM) lymphocyte status at the diagnosis and during different TKI therapies and correlate it with treatment responses. BM and PB samples were acquired from 105 first-line TKI-treated patients. Relative number of BM lymphocytes was evaluated from MGG-stained BM aspirates, and immunophenotypic analyses were performed with multicolour flow cytometry. Early 3-month expansion of BM lymphocytes was found during all different TKIs (imatinib n = 71, 20 %; dasatinib n = 25, 21 %; nilotinib n = 9, 22 %; healthy controls n = 14, 12 %, p < 0.0001). Increased PB lymphocyte count was only observed during dasatinib therapy. The BM lymphocyte expansion was associated with early molecular response; patients with 3-month BCR-ABL1 <10 % showed higher lymphocyte counts than patients with BCR-ABL1 >10 % (23 vs. 17 %, p < 0.05). Detailed phenotypic analysis showed that BM lymphocyte expansion consisted of various lymphocyte subclasses, but especially the proportion of CD19+ B cells and CD3negCD16/56+ NK cells increased from diagnostic values. During dasatinib treatment, the lymphocyte balance in both BM and PB was shifted more to cytotoxic direction (increased CD8+CD57+ and CD8+HLA-DR+ cells, and low T regulatory cells), whereas no major immunophenotypic differences were observed between imatinib and nilotinib patients. Early BM lymphocytosis occurs with all current first-line TKIs and is associated with better treatment responses. PB and BM immunoprofile during dasatinib treatment markedly differs from both imatinib- and nilotinib-treated patients.

Serum-soluble APRIL influences survival in patients with chronic lymphocytic leukemia

Background A proliferation-inducing ligand (APRIL) is a member of the tumor necrosis factor family. APRIL plays an important role in B-cell development; it activates chronic lymphocytic leukemia (CLL) cells by reacting with its receptors, enhancing immune recognition, proliferation, and survival of the leukemia cells. Aim The aim of this study was to determine the impact of serum-soluble APRIL levels detected at diagnosis in B-CLL patients, and their relationship with disease parameters and patient outcomes. Patients and methods Serum APRIL levels were estimated in 40 untreated B-CLL patients and 40 apparently healthy controls. Serum was withdrawn at diagnosis of untreated CLL disease and was tested by enzyme-linked immunosorbent assay. Results APRIL levels in peripheral blood samples were significantly higher in B-CLL patients than in normal participants. There were positive correlations between serum-soluble APRIL level, age, white blood cell count, peripheral blood lymphocyte count, CD38%, ZAP-70, and modified RAI staging. There were negative correlations between serum-soluble APRIL, hemoglobin level, and platelet count. During a 2-year follow-up, APRIL levels correlated with overall survival and disease-free survival. Conclusion APRIL is a powerful prognostic marker related to parameters of disease activity and staging and, more importantly, to overall survival and disease-free survival.

Development of a preliminary diagnostic measure for bovine leukosis in dairy cows using peripheral white blood cell and lymphocyte counts.

Analysis of the association between antibodies against bovine leukemia virus (BLV), BLV proviral load, and white blood cell (WBC) and lymphocyte counts was performed with 774 dairy cows. The average age, WBC counts and lymphoid cell counts tended to be higher in BLV antibody-positive cows than in antibody-negative cows. There was a similar trend in levels of proviral DNA. We analyzed age, WBC counts and lymphocyte counts by principal component analyses to create a distribution chart of the principle component scores. Using the chart, we categorized cows into four quadrants based on additional information, such as the presence of antibody and the levels of proviral DNA. Antibody-positive cows and cows with high BLV proviral load were found mostly in one quadrant of the chart, indicating that it is possible to predict the risk of infection without any knowledge on antibody status by using information, such as WBC counts as a biomarker. When only antibody-positive cows were included in the analysis, a characteristic distribution of different levels of proviral DNA was seen in the quadrants, suggesting that it is possible to estimate the extent of bovine leukosis infection by using this analysis. For this analysis and categorization of the cows into quadrants, we computed a mathematical formulation using discriminant analysis based on age and WBC and lymphocyte counts. This mathematical formulation for the hematological preliminary diagnosis of the disease is recommended as a screening tool to monitor bovine leukosis.

Influences of integrated nursing care on bad emotion and immune function of postoperative patients with esophageal cancer

Objective To explore the influences of integrated nursing care on bad emotion and immune function of postoperative patients with esophageal cancer. Methods A total of 102 postoperative patients selected from October 2012 to October 2013 were randomly divided into research group (52 cases, integrated nursing care) and control group(50 cases, routine nursing). Self rating anxiety scale (SAS) and self rating depression scale (SDS) were used to evaluate the anxiety and depression of patients before and after operations. We detected the contents of immune index such as immunoglobulin (IgA, IgG, IgM), peripheral blood lymphocyte count (LYM), cytokine and etc. before operation, the first and eighth days after operation. Results Patients in the research and the control group all had anxiety and depression symptoms before operation. After operation, the scores of SAS (34.6±6.7) and SDS (33.7±6.9) of patients in the research group were lower than that in the control group (P<0.05). Eighth days after operation, the contents of IgA and IgM rose again compared with that in the first postoperative day (t=5.174, 9.439; P<0.05). The contents of LYM and cytokine CD4/CD8 of patients in the research group were higher than that in the control group (P<0.05), and all of them were higher than that in the first postoperative day. Conclusions Integrated nursing care can relieve anxiety and depression of patients with esophageal cancer, enhance patients′ immune function, and benefit patients′ recovery. Key words: Esophageal neoplasms; Anxiety; Depression; Immune function; Integrated nursing care

Risk Factor Analysis for the Development of Large Granular Lymphocytosis after Allogeneic Hematopoietic Cell Transplantation: Randomized Stratification and Propensity Score Analysis

IntroductionLarge granular lymphocytes (LGL) are a morphologically recognizable subpopulation of lymphocytes comprising an immunophenotypically heterogeneous population of activated CD3+ T cells and CD3- natural killer (NK) cells that mediate non-MHC-restricted cytotoxicity. Increased number of circulating LGL can be found as a response to viral infections, autoimmune disease or malignant neoplasms, as a result of chronic antigenic stimulation. Of interest, LGL lymphocytosis has been reported to occur following hematopoietic cell transplantation (HCT), with a variable incidence of up to 20%. This population display improved transplant outcomes with a lower incidence of non-relapse mortality and relapse (Kim, BMT, 2013; Nann-Rütti, BBMT, 2012). The aim of the present study is to determine the risk factors associated with the development of LGL lymphocytosis after allogeneic HCT.MethodsA total of 826 patients who underwent an allogeneic HCT at Princess Margaret Cancer Centre, Toronto, Canada from 2000 to 2012 were retrospectively analyzed. LGL lymphocytosis was defined as the presence of at least two of the followings: 1) Sustained peripheral blood lymphocyte count ≥3.0 x 109/L observed in at least three consecutive determinations over a period of 2-3 months; 2) Predominance (≥30%) of LGL lymphocytes in the peripheral blood, as assessed by morphologic or immunophenotypic criteria; 3) T-cell receptor monoclonality assessed by PCR. The patient population was divided into discovery and replication sets using 2 different methods: stratified randomization and propensity score matching, using relevant baseline variables such as donor type, CMV serostatus, conditioning, T-cell depletion.ResultsNo significant imbalances were found between the discovery and replication sets in terms of relevant baseline characteristics and clinical outcomes, for both the randomly divided patients and the propensity score matched groups. The overall incidence of LGL lymphocytosis was 14.5% at 3 years. The incidence of LGL lymphocytosis was similar across all subgroups of patients, both for the randomly divided groups and the propensity score matching (P-value not significant).A multivariable analysis of the risk factors for the development of LGL lymphocytosis was performed, including the following variables: grades 3-4 acute graft-versus-host-disease (GVHD), chronic GVHD, CMV viremia, CMV serostatus of the recipient, donor type, transplant year and T-cell depletion (TCD) for GVHD prophylaxis.In the stratified randomization analysis, the following risk factors were identified: 1) Discovery set: chronic GVHD (Hazard Ratio: 8.3, 95% CI: 3.1-22.6, P<0.001), CMV viremia (HR: 2.7, 95% CI: 1.5-4.7, P<0.001) and use of an unrelated donor (HR: 2.1, 95% CI: 1.2-3.7, P =0.01). 2) Replication set: chronic GVHD (HR: 38.9, 95% CI: 5.3-284.9, P<0.001) and CMV viremia (HR 3.8, 95% CI: 2.2-6.6, P<0.001).For the propensity score matching analysis the risk factors for the development of LGL lymphocytosis were the following: 1) Discovery set: chronic GVHD (HR: 22.9, 95% CI: 3.2-169.3, P =0.002) and CMV viremia (HR: 2.2, 95% CI: 1.1-4.2, P =0.02). 2) Replication set: chronic GVHD (HR: 28.9, 95% CI: 3.9-212.5, P<0.001), CMV viremia (HR: 3.6, 95% CI: 1.9-6.7, P<0.001).ConclusionsChronic GVHD and CMV viremia are strongly associated with the development of LGL lymphocytosis following allogeneic HCT. This may reflect a chronic antigenic stimulation in the setting of GVHD and CMV infection, leading to the expansion of LGL. However, the underlying mechanisms of LGL activation and expansion in the allogeneic HCT setting still remain unclear. Thus, further investigations are needed to elucidate these mechanisms in particular in the setting of GVHD. DisclosuresLipton:Pfizer: Consultancy, Research Funding; Ariad: Consultancy, Research Funding; Teva: Consultancy, Research Funding; Bristol-Myers Squibb: Consultancy, Research Funding; Novartis Pharmaceuticals: Consultancy, Research Funding. Kim:Bristol-Myers Squibb: Consultancy, Research Funding; Novartis Pharmaceuticals: Consultancy, Research Funding.

Bone Marrow Lymphocytic Status during Tyrosine Kinase Inhibitor Therapy and Its Relation to Therapy Response in Chronic Phase Chronic Myeloid Leukemia Patients

IntroductionThe introduction of tyrosine kinase inhibitors (TKIs) has revolutionized the treatment of chronic myeloid leukemia (CML). In addition to blocking their main target kinase, the BCR-ABL oncoprotein, several studies have reported that TKIs could also have secondary effects on the immune system and lymphocyte behavior. The aim of this study was to assess the bone marrow (BM) lymphocyte status at diagnosis and during different first-line TKI therapies and correlate it with treatment responses.MethodsAltogether 105 first-line TKI treated patients were included in the study (imatinib n=71, dasatinib n=25 and nilotinib n=9) and samples from 14 healthy bone marrow donors served as controls. BM aspirate samples were taken from patients at the diagnosis and at 3, 6, 12 and 18 months after the TKI therapy start, and MGG-stained BM aspirate slides were examined for cellularity and individual cellular proportions. Treatment responses were evaluated with standard karyotyping and real-time quantitative PCR. Patients were divided in different groups according to ELN criteria based on their therapy response at 12 months. In addition, multi-color flow cytometry was done from both BM and peripheral blood (PB) samples using 5 different antibody panels including markers for T, B, NK and regulatory T cells.ResultsWe found an early (3 months) expansion of BM lymphocytes during all different TKI therapies (imatinib median lymphocyte count 20%; dasatinib 21%; nilotinib 22%; healthy controls 12%, p<0.0001). Increased PB lymphocyte count was only observed during dasatinib therapy, possibly due to mobilization phenomenon observed only in dasatinib, and not in imatinib or nilotinib treated patients. The BM lymphocytic expansion was associated with better early molecular response; patients with 3 months BCR-ABL<10% showed higher lymphocyte counts than patients with BCR-ABL>10% (median 23% and 17% respectively; p<0.05). Interestingly, patients with suboptimal (warning) response (no major molecular remission at 12 months) showed significantly higher lymphocyte counts at 3 months than patients with optimal response or failure (28%, 19.5%, and 18.5% respectively, p<0.0001). During the treatment at 12 and 18 months when patients were already in complete cytogenetic and molecular remission, BM lymphocytosis seemed to persist (median lymphocyte counts 19% and 18%), thus suggesting that the lymphocyte increase is TKI therapy induced and not only related to myeloablation and eradication of leukemia cells.Detailed phenotypic analysis showed that lymphocyte expansion consisted of various lymphocyte subclasses, but especially the proportion of CD19+ B cells and CD3negCD16/56+ NK cells increased from diagnostic values (3 months NK cells: 19.4% vs. 16.8 % at diagnosis, p<0.05; B cells 10.7% vs. 8.6%, p<0.05). Among different TKIs studied, the proportion of CD3+CD16/56+ NKT-like cells was higher in dasatinib treated patients, but no differences were observed in the amount of B, NK or T cells. However, within the T cell population dasatinib group had significantly more BM CD8+ T cells (48% and 51% at 3 and 6 months) when compared to imatinib (41% and 39%) and nilotinib treated patients (43% and 33%). The percentage of CD57+ expressing T cells was also clearly increased in dasatinib patients (40% vs. 21% and 25% in the imatinib and nilotinib groups, p=0.004) as well as the percentage of HLA-DR expressing cells (HLA-DR+ from CD4+ T cells 18% vs. 7% and 7% in imatinib and nilotinib groups, p<0.05 and from CD8+ T cells 41% vs. 22% vs. 15%, respectively, p<0.05). The proportion of regulatory T cells in the BM was significantly lower in dasatinib-treated patients (3.2%) than in the imatinib (5.7%, p<0.05) or nilotinib group (5%).ConclusionBM lymphocytosis occurs during all current first-line TKI treatments, and it is associated with early molecular response. On the contrary, PB lymphocytosis occurs only during dasatinib treatment, and the overall lymphocyte balance in dasatinib group is shifted more to cytotoxic direction (increased CD8+CD57+ and CD8+HLA-DR+ cells, and low T regulatory cells). In accordance with shared kinase inhibition spectrum, no marked changes in the immunoprofile are observed between imatinib- and nilotinib-treated patients. DisclosuresBjerrum:Bristoll Myers Squibb, Novartis and Pfizer: Other: educational activities. Porkka:Bristol-Myers Squibb: Honoraria; Celgene: Honoraria; Novartis: Honoraria; Pfizer: Honoraria. Hjorth-Hansen:Novartis: Honoraria; Ariad: Honoraria; Bristol-Myers Squibb: Research Funding; Pfizer: Honoraria, Research Funding. Mustjoki:Novartis: Honoraria, Research Funding; Academy of Finland: Research Funding; the Finnish Cancer Societies: Research Funding; Pfizer: Honoraria, Research Funding; Bristol-Myers Squibb: Honoraria, Research Funding; Sigrid Juselius Foundation: Research Funding; Finnish Cancer Institute: Research Funding; Signe and Ane Gyllenberg Foundation: Research Funding.

Expression of S100 Protein in CD4-positive T-cell Lymphomas Is Often Associated With T-cell Prolymphocytic Leukemia.

S100 T-cell lymphomas are infrequent, and except 1 all have been CD4 negative. On the basis of an index case of CD4 S100 T-cell prolymphocytic leukemia (T-PLL), we studied S100 protein expression in 19 additional T-PLLs and 56 other T-cell lymphomas that are usually CD4, including 15 angioimmunoblastic T-cell lymphomas, 24 anaplastic large cell lymphomas (16 ALK and 8 ALK), 7 mycosis fungoides/Sézary syndrome, and 10 peripheral T-cell lymphoma, not otherwise specified (PTCL, NOS). Two additional S100 CD4 PTCL, NOS cases were also reviewed. Thirty percent (6/20) of T-PLLs were S100 compared with 0/56 other T-cell lymphomas with previously unstudied S100 reactivity (40 CD4, 2 CD8, 11 CD4/CD8, 3 unknown) (P=0.0007). There were no significant differences between the S100 and S100 T-PLLs with regard to the male:female ratio (2:1 vs. 1:1), age (71.6±7.7 vs. 65.4±9.3), peripheral blood lymphocyte count (67.2±116.6 vs. 101.1±159.7×10/L), or median survival (463 vs. 578 d, where known). The 2 S100 PTCL, NOS cases occurred in a 7-year-old boy and a 45-year-old woman. Both had involvement of the bone marrow and peripheral blood but were morphologically unlike T-PLL and lacked TCL1 gene rearrangement. These results demonstrate that S100 T-cell lymphomas include a subset that are CD4 and most often, but not exclusively, are T-PLL. Although having diagnostic implications, there were no documented clinical differences between the S100 and S100 T-PLLs.

Prognostic value of total lymphocyte count in cardiac surgery

The aim of this study retrospective cohort was to investigate the prognostic value of preoperative total lymphocyte count in peripheral blood as a predictor of postoperative complications and mortality in cardiac surgery. All adults undergoing cardiopulmonary bypass in 2009. The cohort size was 1 368 patients. Patient characteristics, hospital mortality, postoperative complications, ventilation period, intensive care unit and hospital stay were analysed. Preoperative total lymphocyte count &lt;1 611 cells/L was assotiated with significantly higher mortality by univariate (p&lt;0,0001) and multivariate (p&lt;0,022) analisys. Low preoperative total lymphocyte count was associated with more frequent inotropic support (p&lt;0,001); postoperative heart arrhythmia (p&lt;0,001); hemodialysis-dependent acute renal failure (p&lt;0,001); and a prolonged ventilation period (p=0,001), intensive care unit stay (p&lt;0,001), and hospital stay (p=0,007). Low preoperative total lymphocyte count in peripheral blood is a useful prognostic criterion for evaluation of a complicated postoperative period in cardiosurgical patients.

Thymocytes maintain immune activity through telomere elongation in rats under hypoxic conditions.

The main purpose of the present study was to investigate the change in thymocyte telomere length of rats exposed to different hypoxic conditions for different periods of time, as well as its effect on the immune system. A total of 110 male Wistar rats were randomly assigned to one of the three following groups: i) Sea level (SL) group, in which 10 rats were maintained at an altitude of 10 m; ii) moderate altitude (MA) group, in which 50 rats were maintained at an altitude of 2,260 m and then randomly sacrificed on days 1, 3, 7, 15 and 30 (n=10 each); and iii) simulated high altitude (SHA) group, in which 50 rats were maintained at a simulated altitude of 5,000 m, and then randomly sacrificed on days 1, 3, 7, 15 and 30 (n=10 each). The morphological changes of the thymus were observed, while the telomere length, the mRNA and protein expression levels of telomerase reverse transcriptase (TERT), and the peripheral blood lymphocyte count were measured. The results indicated that hypoxia induced morphological changes and apoptosis in thymocytes, as well as atrophy of the thymus tissue, and resulted in a significant increase in telomere length and TERT mRNA and protein expression levels. This effect appeared to be more pronounced in the SHA group compared with that in the MA group; however, no statistically significant changes were observed in the peripheral blood lymphocyte count. Based on these findings, the hypoxia-associated loss of thymic function appears to be only quantitative and not qualitative, and the thymus may be able to maintain its immune function even under hypoxic conditions.

Mortality prediction to hospitalized patients with influenza pneumonia: PO2/FiO2 combined lymphocyte count is the answer

IntroductionCommunity‐acquired pneumonia (CAP) severity scores perform well in predicting mortality of CAP patients, but their applicability in influenza pneumonia is powerless.ObjectivesThe aim of our research was to test the efficiency of PO2/FiO2 and CAP severity scores in predicting mortality and intensive care unit (ICU) admission with influenza pneumonia patients.MethodsWe reviewed all patients with positive influenza virus RNA detection in Beijing Chao‐Yang Hospital during the 2009–2014 influenza seasons. Outpatients, inpatients with no pneumonia and incomplete data were excluded. We used receiver operating characteristic curves (ROCs) to verify the accuracy of severity scores or indices as mortality predictors in the study patients.ResultsAmong 170 hospitalized patients with influenza pneumonia, 30 (17.6%) died. Among those who were classified as low‐risk (predicted mortality 0.1%–2.1%) by pneumonia severity index (PSI) or confusion, urea, respiratory rate, blood pressure, age ≥65 year (CURB‐65), the actual mortality ranged from 5.9 to 22.1%. Multivariate logistic regression indicated that hypoxia (PO2/FiO2 ≤ 250) and lymphopenia (peripheral blood lymphocyte count <0.8 × 109/L) were independent risk factors for mortality, with OR value of 22.483 (95% confidence interval 4.927–102.598) and 5.853 (95% confidence interval 1.887–18.152), respectively. PO2/FiO2 combined lymphocyte count performed well for mortality prediction with area under the curve (AUC) of 0.945, which was significantly better than current CAP severity scores of PSI, CURB‐65 and confusion, respiratory rate, blood pressure, age ≥65 years for mortality prediction (P < 0.001). The scores or indices for ICU admission prediction to hospitalized patients with influenza pneumonia confirmed a similar pattern and PO2/FiO2 combined lymphocyte count was also the best predictor for predicting ICU admission.ConclusionIn conclusion, we found that PO2/FiO2 combined lymphocyte count is simple and reliable predictor of hospitalized patients with influenza pneumonia in predicting mortality and ICU admission. When PO2/FiO2 ≤ 250 or peripheral blood lymphocyte count <0.8 × 109/L, the clinician should pay great attention to the possibility of severe influenza pneumonia.

Dynamic monitoring of the neutrophil/lymphocyte ratio could predict the prognosis of patients with bloodstream infection

To explore the value of dynamic monitoring of the neutrophils/lymphocyte ratio (NLR) in peripheral blood for the prognosis of patients with bloodstream infection (BSI). A retrospective study was conducted. 205 patients who were ≥18 years old, their length of hospital stay>24 hours, and they were treated in the China-Japanese Friendship Hospital from January 2013 to October 2014 were enrolled. According to the 28-day survival, the patients were divided into survival group (n=160) and death group (n=45). The white blood cell (WBC), neutrophils count (NEU), neutrophils ratio (Neut%), lymphocyte count (LYM), lymphocyte ratio (Lym%), and NLR in peripheral blood were recorded at 1, 3, 7 days after admission. Receiver-operating characteristic curve (ROC) was plotted for evaluating the value of these factors on the 28-day prognosis, and logistic regression analysis was used to evaluate the risk factors for predicting the outcome. (1) On the 1st day, WBC, NEU, Neut%, NLR, and procalcitonin (PCT) in the death group were significantly higher than those in the survival group [WBC (×10(9)/L): 15.28±8.23 vs. 11.58±6.55, NEU (×10(9)/L): 13.34±7.53 vs. 10.03±5.31, Neut%: 0.886±0.076 vs. 0.845±0.102, NLR: 21.20 ( 13.10, 28.80) vs. 12.08 (6.81, 20.47), PCT (μg/L): 3.13 (0.85, 10.12) vs. 1.34 (0.36, 5.81), P<0.05 or P<0.01], while hemoglobin (Hb), platelet count (PLT), albumin (ALB) content were significantly lower than those of the survival group [Hb (g/L): 86.09±19.83 vs. 107.89±22.82, PLT (×10(9)/L): 157.51±117.81 vs. 195.44±97.28, ALB (g/L): 24.11±6.94 vs. 31.99±6.89, P<0.05 or P<0.01]. On the 3rd day and 7th day, WBC, NEU and NLR in the death group were significantly higher than those of the survival group [WBC (×10(9)/L): 16.61±10.25 vs. 8.91±4.93, 16.05±9.46 vs. 8.79±4.45; NEU (×10(9)/L): 14.15±9.98 vs. 6.97±4.64, 14.36±9.03 vs. 6.59±4.07; NLR: 24.13 (8.49, 38.26) vs. 5.52 (3.58, 8.87), 17.74 (10.74, 32.85) vs. 4.35 (2.78, 7.27), all P<0.01 ], and the LYM and Lym% were significantly lower than those in the survival group [LYM (×10(9)/L): 0.61 (0.38, 1.04) vs. 1.05 (0.78, 1.43), 0.69 (0.35, 0.92) vs. 1.37 (0.93, 1.76); Lym%: 0.039 (0.024, 0.101) vs. 0.135 (0.094, 0.186), 0.056 (0.033, 0.082) vs. 0.170 (0.108, 0.237), all P<0.01]. (2) It was shown by ROC curve that the maximum area under the ROC curve (AUC) of WBC, NEU, Neut%, LYM, Lym%, and NLR about prognosis of BSI were observed on 7 days (0.777, 0.819, 0.905, 0.755, 0.880, 0.887). Based on Neut%>0.855 on the 7th day as a predictor of cut-off value of death in 28 days, the sensitivity was 78.8%, specificity 89.1%, respectively. When Lym%<0.088 on the 7th day as a predictor of cut-off value of death on 28 days, the sensitivity was 89.5%, and specificity was 83.9%. When NLR>10.34 on the 7th day as a predictor of cut-off value of death in 28 days, the sensitivity was 81.8%, and specificity was 91.0%. (3) Survival analysis showed that the 28-day survival rate in the patients with 7-day NLR<10.34 was significantly higher than that in those with 7-day NLR>10.34 (95.0% vs. 34.1%, χ2=82.650, P=0.000). (4) It was shown by multi-factor logistic regression analysis that the levels of 1-day Hb and 7-day NLR were the independent prognostic predictors of 28-day mortality [Hb: odds ratio (OR)=0.946, 95% confidence interval (95%CI)=0.913-0.981, P=0.003; 7-day NLR: OR=34.941, 95%CI=8.728-139.884, P=0.000]. The trend of changes in NEU, LYM and NLR as shown by repeated routine blood examinations contributes to prediction of the outcome of patients with BSI. The levels of 1-day Hb and 7-day NLR are the independent prognostic predictors for 28-day mortality.

THE NUTRITIONAL INDEX ‘CONUT’ IS USEFUL FOR PREDICTING PROGNOSIS OF ACUTE DECOMPENSATED HEART FAILURE

Controlling Nutritional Status (CONUT), an easy-to-use nutrition assessment system which scores laboratory data including serum albumin, total cholesterol level and peripheral blood lymphocyte count, has been considered to be efficient for early detection of hospital undernutrition. Prior studies

Ovarian hormone level alterations during rat post-reproductive life-span influence CD8 + T-cell homeostasis.

The study examined the putative role of ovarian hormones in shaping of rat peripheral T-cell compartment during post-reproductive period. In 20-month-old rats ovariectomized (Ox) at the very end of reproductive period, thymic output, cellularity and composition of major TCRαβ + peripheral blood lymphocyte and splenocyte subsets were analyzed. Ovariectomy led to the enlargement of CD8 + peripheral blood lymphocyte and splenocyte subpopulations. This reflected: (i) a more efficient thymic generation of CD8 + cells as indicated by increased number of CD4+CD8 + double positive and the most mature CD4-CD8+TCRαβ(high) thymocytes and CD8 + recent thymic emigrants (RTEs) in peripheral blood, but not in the spleen of Ox rats, and (ii) the expansion of CD8 + memory/activated peripheral blood lymphocytes and splenocytes. The latter was consistent with a greater frequency of proliferating cells among freshly isolated memory/activated CD8 + peripheral blood lymphocytes and splenocytes and increased proliferative response of CD8 + splenocytes to stimulation with plate-bound anti-CD3 antibody. The former could be related to the rise in splenic IL-7 and IL-15 mRNA expression. Although ovariectomy affected the overall number of CD4 + T cells in none of the examined compartments, it increased CD4+FoxP3 + peripheral blood lymphocyte and splenocyte counts by enhancing their generation in periphery. Collectively, the results suggest that ovariectomy-induced long-lasting disturbances in ovarian hormone levels (mirrored in diminished progesterone serum level in 20-month-old rats) affects both thymic CD8 + cell generation and peripheral homeostasis and leads to the expansion of CD4+FoxP3 + cells in the periphery, thereby enhancing autoreactive cell control on account of immune system efficacy to combat infections and tumors.