Absolute Counts Of Lymphocyte Subsets Research Articles

Two nomograms constructed for predicting the efficacy and prognosis of advanced non‑small cell lung cancer patients treated with anti‑PD‑1 inhibitors based on the absolute counts of lymphocyte subsets

BackgroundPatients treated with immune checkpoint inhibitors (ICIs) are at risk of considerable adverse events, and the ongoing struggle is to accurately identify the subset of patients who will benefit. Lymphocyte subsets play a pivotal role in the antitumor response, this study attempted to combine the absolute counts of lymphocyte subsets (ACLS) with the clinicopathological parameters to construct nomograms to accurately predict the prognosis of advanced non-small cell lung cancer (aNSCLC) patients treated with anti-PD-1 inhibitors.MethodsThis retrospective study included a training cohort (n = 200) and validation cohort (n = 100) with aNSCLC patients treated with anti-PD-1 inhibitors. Logistic and Cox regression were conducted to identify factors associated with efficacy and progression-free survival (PFS) respectively. Nomograms were built based on independent influencing factors, and assessed by the concordance index (C-index), calibration curve and receiver operating characteristic (ROC) curve.ResultIn training cohort, lower baseline absolute counts of CD3+ (P < 0.001) and CD4+ (P < 0.001) were associated with for poorer efficacy. Hepatic metastases (P = 0.019) and lower baseline absolute counts of CD3+ (P < 0.001), CD4+ (P < 0.001), CD8+ (P < 0.001), and B cells (P = 0.042) were associated with shorter PFS. Two nomograms to predict efficacy at 6-week after treatment and PFS at 4-, 8- and 12-months were constructed, and validated in validation cohort. The area under the ROC curve (AUC-ROC) of nomogram to predict response was 0.908 in training cohort and 0.984 in validation cohort. The C-index of nomogram to predict PFS was 0.825 in training cohort and 0.832 in validation cohort. AUC-ROC illustrated the nomograms had excellent discriminative ability. Calibration curves showed a superior consistence between the nomogram predicted probability and actual observation.ConclusionWe constructed two nomogram based on ACLS to help clinicians screen of patients with possible benefit and make individualized treatment decisions by accurately predicting efficacy and PFS for advanced NSCLC patient treated with anti-PD-1 inhibitors.

The changes and its significance of peripheral blood NK cells in patients with tuberculous meningitis.

Tuberculous meningitis (TBM) is the most severe form of tuberculosis (TB). The purpose of this study was to explore the relationship between the number of natural killer (NK) cells and adaptive immune status, and disease severity in TBM patients. We conducted a retrospective study on 244 TB patients and 146 healthy control subjects in the 8th Medical Center of the PLA General Hospital from March 2018 and August 2023. The absolute count of NK cells in the peripheral blood of TBM patients was significantly lower than that in normal controls (NC), latent tuberculosis infection (LTBI), and non-severe TB (NSTB) patients (p < 0.05). The proportion of TBM patients (48.7%) with a lower absolute count of NK cells than the normal reference value was significantly higher than that in NC (5.2%) and LTBI groups (4.0%) (p < 0.05), and slightly higher than that in NSTB group (36.0%) (p > 0.05). The absolute counts of lymphocyte subsets in TBM combined with other active TB group, etiology (+) group, IGRA (-) group, and antibody (+) group were lower than that in simple TBM group, etiology (-) group, IGRA (+) group, and antibody (-) group, respectively. The CD3+ T, NK, and B cells in BMRC-stage III TBM patients were significantly lower than those in stage I and stage II patients (p < 0.05). The counts of CD3+ T, CD4+ T, and B cells in the etiology (+) group were significantly lower than those in the etiology (-) group (p < 0.05). The absolute counts of lymphocyte subsets in the peripheral blood of TBM patients were significantly decreased, especially in NK cells. The reduction of these immune cells was closely related to the disease severity and had a certain correlation with cellular and humoral immune responses. This study helps to better understand the immune mechanism of TBM and provides reliable indicators for evaluating the immune status of TBM patients in clinical practice.

Dynamic surveillance of lymphocyte subsets in patients with non-small cell lung cancer during chemotherapy or combination immunotherapy for early prediction of efficacy.

Non-small cell lung cancer (NSCLC) remains the leading cause of cancer-related deaths worldwide. Lymphocytes are the primary executors of the immune system and play essential roles in tumorigenesis and development. We investigated the dynamic changes in peripheral blood lymphocyte subsets to predict the efficacy of chemotherapy or combination immunotherapy in NSCLC. This retrospective study collected data from 81 patients with NSCLC who received treatments at the First Affiliated Hospital of Zhengzhou University from May 2021 to May 2023. Patients were divided into response and non-response groups, chemotherapy and combination immunotherapy groups, and first-line and multiline groups. We analyzed the absolute counts of each lymphocyte subset in the peripheral blood at baseline and after each treatment cycle. Within-group and between-group differences were analyzed using paired Wilcoxon signed-rank and Mann-Whitney U tests, respectively. The ability of lymphocyte subsets to predict treatment efficacy was analyzed using receiver operating characteristic curve and logistic regression. The absolute counts of lymphocyte subsets in the response group significantly increased after the first cycle of chemotherapy or combination immunotherapy, whereas those in the non-response group showed persistent decreases. Ratios of lymphocyte subsets after the first treatment cycle to those at baseline were able to predict treatment efficacy early. Combination immunotherapy could increase lymphocyte counts compared to chemotherapy alone. In addition, patients with NSCLC receiving chemotherapy or combination immunotherapy for the first time mainly presented with elevated lymphocyte levels, whereas multiline patients showed continuous reductions. Dynamic surveillance of lymphocyte subsets could reflect a more actual immune status and predict efficacy early. Combination immunotherapy protected lymphocyte levels from rapid decrease and patients undergoing multiline treatments were more prone to lymphopenia than those receiving first-line treatment. This study provides a reference for the early prediction of the efficacy of clinical tumor treatment for timely combination of immunotherapy or the improvement of immune status.

Different Sources of Hematopoietic Stem Cell Transplantation in Patients with Thalassemia Major: Relationship between Immune Reconstitution, Graft-Versus-Host Disease, and Viral Infections

Objective: To compare the relationship among immune reconstitution, graft-versus-host disease (GVHD), and viral infections in three kinds of hematopoietic stem cell transplantation(HSCT) using different host sources for thalassemia major(TM) patients: matched unrelated donor (MUD) HSCT, haploidentical (Haplo) HSCT with post-transplant cyclophosphamide (PTCy)-based prophylactic therapy, and TCRαβ-T cell-depleted HSCT (TDH). Methods: A total of 191 TM patients in our center between October 2018 and June 2022 were included in this study, comprising three transplant groups: MUD group (n=56), PTCy group (n=45), and TDH group (n=90). Four-color flow cytometry was used to detect the proportions and absolute counts of lymphocyte subsets at the 12th month after transplantation for the three different donor types. The immune reconstitution was analyzed and compared with the occurrence of GVHD and virus infections. Results: 1. Immune Reconstitution: (1) The ratio of helper cells (CD3 +CD4 +)/cytotoxic T cells (CD3 +CD8 +) at the 12 months after transplantation was compared among the three groups. There were statistically significant differences between the TDH and PTCy groups, as well as between the TDH and MUD groups(P=022 and 0.011, respectively). No statistical difference was found between the PTCy and MUD groups (P=0.932). The CD4 +/CD8 + ratio in the TDH group recovered the fastest and had already returned to the normal reference range (0.7-2.7). (2) The comparison of B cells (CD3 -CD19 +) at the 12 months after transplantation showed statistically significant differences between the TDH and PTCy groups, as well as between the MUD and PTCy groups (P=0.049 and 0.034, respectively). No statistical difference was found between the TDH and MUD groups (P=0.447). The recovery of B cells was faster in the TDH and MUD groups, while it was slowest in the PTCy group. (3) The comparison of natural killer cells (NK cells) (CD3 -CD56 +) at the 12 months after transplantation showed statistically significant differences between the TDH and PTCy groups, as well as between the TDH and MUD groups (P=0.048 and 0.000, respectively). No statistical difference was found between the PTCy and MUD groups (P=0.366). The TDH group exhibited the fastest recovery of NK cells. 2. Chronic GVHD (cGVHD): (1) 7/90 cases (7.78%) of cGVHD appeared in the TDH group, 19/45 cases (42.22%) in the PTCy group, and 2/56 cases (3.57%) n the MUD group. (2) No statistical difference showed in the incidence of cGVHD between the TDH and MUD groups (P=0.500). There was a statistical difference in the incidence of cGVHD between the TDH and PTCy groups (P=0.000), as well as between the MUD and PTCy groups (P=0.000). 3. Viral Infection: (1) 22/90 cases (24.44%) of viral infection occurred in the TDH group, 7/45 cases (15.56%) in the PTCy group, and 6/56 cases (10.71%) in the MUD group. (2) Among the 90 cases of TDH group, the CD4/CD8 ratio at the 6 months after transplantation was 0.41±0.24 in the 22 cases with viral infection and 0.69±0.40 in the 68 cases without that. There was a statistical difference in the mean CD4/CD8 ratio between the two divisions in the TDH group (P=0.013). Conclusion:The TDH group exhibited faster immune reconstitution compared to the other two groups, especially in terms of CD4/CD8 ratio, B cells, and NK cells. This may be related to two special factors for patients underwent TDH in our center: a rather high average infusion of CD34 cells (30×10 6/kg) , and the preservation of γδ cells, B cells, and NK cells with the removing of TCRαβ-T cells in the TDH treatment protocol. The TDH and MUD groups showed a lower incidence of cGVHD, while the PTCy group had a higher incidence. The occurrence of viral infection in the TDH group was 24.44%. Also, the lower a CD4/CD8 ratio at the 6 months after transplantation, the higher the incidence of viral infection. Table 1. The Absolute Counts of Lymphocyte Subsets at the 12th Month after Transplantation

Pre-Graft Immune Status Predicts Survivals and Relapse in Adults Receiving Matched Peripheral Blood Stem Cell Allotransplants for Myeloid Malignancies

Background The impact on outcomes of pre-graft immune status has not been explored so far in recipients of allogeneic stem cell transplant (allo-SCT). Patients and Methods This was a retrospective and monocentric study including all consecutive adults who underwent a peripheral blood stem cell (PB) allo-SCT for a hematological malignancy between May 2017 and December 2021 in our Hematology Department. Immune status was routinely evaluated before transplant in each patient. Absolute lymphocyte (ALC) and monocyte counts were obtained from complete blood counts, while absolute counts of lymphocyte subsets (CD3+, CD4+ and CD8+ T cells, B and NK cells) were assessed by multiparameter flow cytometry, and gamma globulin levels by standard serum electrophoresis. We evaluated the impact of these parameters, considered as continuous variables, on overall survival (OS), progression-free survival (PFS), graft-versus-host disease (GVHD)-free, progression-free survival (GRFS), non-relapse mortality (NRM), and relapse incidence (RI), according to the type of the disease (myeloid vs. lymphoid) and the type of the donor (matched vs. haploidentical). Results This study included 264 patients with a median age of 59 years old (range 20-73, Table). A majority had a myeloid malignancy (75%, n=196) and received a graft from a matched donor (64%, n=170, sibling n=46; matched unrelated n=124) and a reduced-intensity conditioning regimen (80%, n= 211). Almost all (96%) patients with a matched donor received anti-thymoglobulin as GVHD prophylaxis while it was the case for 79% of those receiving a graft from a haploidentical donor. With a median follow-up of 32 months estimated by reversed Kaplan Meier analysis, 3-year (y) OS, PFS, and GRFS were respectively 53%, 46% and 31%, while 3-y NRM and RI were 28% and 25%, for the whole cohort. Pre-graft immune status was evaluated at a median of 20 days before transplant, showing significant higher median T and B cell counts and gamma globulin level in patients with myeloid malignancy, reflecting probably the less immunosuppressive intensity of chemotherapies received before transplant ( Table). Considering myeloid malignancies, OS and NRM were not impacted by the pre-graft immune status. In multivariate analysis, adjusting for disease type, status, donor type (sibling versus [vs] unrelated), and conditioning (RIC vs MAC vs sequential), a higher ALC was associated with significant lower PFS (hazard ratio [HR] 1.39, 95%CI: 1.11-1.74, p=0.004) and GRFS (HR: 1.30, 1.07-1.59, p=0.009), and a significant higher risk of relapse (HR 1.85, 1.32-2.59, p=&amp;lt;0.001), but this was true only in case of a matched transplant. Interestingly, T-cell subsets were associated with relapse (CD3: HR 1.94, p&amp;lt;0.001; CD4: HR 2.34, p=0.004; CD8: HR 2.34, p&amp;lt;0.001), but not B-cells (HR 2.23, p=0.56), gamma-globulin levels (HR 0.98, p=0.68), nor NK-cells (HR 2.72, p=0.17). T cell subsets, B cells, NK cells, gamma-globulin and monocytes were not associated with PFS and GRFS. Also, to highlight the adverse effect of an elevated ALC, and using a ROC curve, a threshold of 1.23 Giga/L was retained as the best cut-off to predict relapse. This was then applied to create a visual comparison of PFS, GRFS and RI ( Figure), revealing an early increased RI in patients with ALC &amp;gt; 1.23 Giga/L (3-month relapse rate: 13% vs 2.3%, p=0.03). Finally, the pre-graft immune status did not impact the outcomes of patients receiving a PB haplotransplant for a myeloid malignancy. This was true also for those who received a PB matched or again a haploidentical transplant for a lymphoid disease. Conclusions This retrospective analysis suggests that higher ALC at transplant predicts lower PFS and GRFS due to higher RI in adults receiving a PB matched transplant for a myeloid malignancy. Patients with a myeloid malignancy had also a better immune status at transplant which may reflect less immunosuppressive treatments received before transplant. Paradoxically, this better immune status may interfere with allogeneic T lymphocytes by reducing the graft-versus-leukemia effect. Thus, increasing immunosuppression (via the conditioning or drugs used for GVHD prophylaxis) may improve outcome in some specific sub-groups of patients with higher autologous T lymphocytes at transplant.

The immunological response of Egyptians to coronavirus disease-19 infection: a cohort study of lymphocyte populations and peripheral blood counts

Background The worldwide pandemic of COVID-19 infection that started in 2019 still lays its shadows over all populations of the world. COVID-19 infection presented with a spectrum of symptoms that varied from wave to wave, and also led to a wide number of long-term sequelae. Many immune system cells and cytokines were implicated in COVID-19 pathophysiology. Thus, many immuno-modulator and immuno-suppressive drugs were used in the management of severe cases. Lymphocytes are the key players of immune system, the change in their count and different subsets is expected to vary with COVID-19 infection. Objective The current study aimed to evaluate the role of peripheral blood lymphocyte subsets in predicting the outcome of COVID-19 patients and to investigate their correlation with different clinical and laboratory variables. Materials and methods The study included 64 patients hospitalized with Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). They were stratified according to in-hospital mortality into survivors and nonsurvivors. Demographic, clinical and laboratory data were collected. Flowcytometric evaluation of lymphocyte subsets was done on admission. Results and conclusion Nonsurvivors showed lower relative lymphocyte count, higher absolute neutrophil count, and higher neutrophil to lymphocyte ratio (NLR) compared with survivors (P = 0.034, 0.006, 0.011; respectively). NLR at a cut off 15.3 had a sensitivity of 70.59% and specificity of 61.29% for predicting mortality in COVID-19 patients. The relative and absolute counts of lymphocyte subsets did not show a statistically significant difference between the two groups. Platelet count showed statistically significant positive correlation with absolute counts of total T lymphocytes, T helper, T cytotoxic, and B lymphocytes. The platelet to lymphocyte ratio (PLR), NLR and D-dimer results were negatively correlated with the total T lymphocytes, T helper, T cytotoxic, naïve T cytotoxic and B lymphocyte absolute counts. The NLR, absolute neutrophil count and platelet count may serve as adjuvant predictors of survival in COVID-19 disease. Although lymphocyte subsets did not differ statistically across survival groups, their correlation with other possible prognostic markers may justify further investigation on their role in COVID-19 pathophysiology.

Long-term analysis of cellular immunity in patients with RRMM treated with CAR-T cell therapy.

Chimeric antigen receptor T (CAR-T) cell therapy exhibits remarkable efficacy against refractory or relapsed multiple myeloma (RRMM); however, the immune deficiency following CAR-Ts infusion has not been well studied. In this study, 126 patients who achieved remission post-CAR-Ts infusion were evaluated for cellular immunity. Following lymphodepletion (LD) chemotherapy, the absolute lymphocyte count (ALC) and absolute counts of lymphocyte subsets were significantly lower than baseline at D0. Grade ≥ 3 lymphopenia occurred in 99% of patients within the first 30days, with most being resolved by 180days. The median CD4+ T-cell count was consistently below baseline and the lower limit of normal (LLN) levels at follow-up. Conversely, the median CD8+ T-cell count returned to the baseline and LLN levels by D30. The median B-cell count remained lower than baseline level at D60 and returned to baseline and LLN levels at D180. In the first 30days, 27 (21.4%) patients had 29 infections, with the majority being mild to moderate in severity (21/29; 72.4%). After day 30, 44 (34.9%) patients had 56 infections, including 20 severe infections. One patient died from bacteremia at 3.8months post-CAR-Ts infusion. In conclusion, most patients with RRMM experienced cellular immune deficiency caused by LD chemotherapy and CAR-Ts infusion. The ALC and most lymphocyte subsets gradually recovered after day 30 of CAR-Ts infusion, except for CD4+ T cells. Some patients experience prolonged CD4+ T-cell immunosuppression without severe infection.

Dynamic changes in peripheral lymphocytes and antibody response following a third dose of SARS-CoV-2 mRNA-BNT162b2 vaccine in cancer patients

The aim of this study was to evaluate the association of circulating lymphocytes profiling with antibody response in cancer patients receiving the third dose of COVID-19 mRNA-BNT162b2 vaccine. Immunophenotyping of peripheral blood was used to determine absolute counts of lymphocyte subsets, alongside detection of IgG antibodies against receptor-binding-domain (RBD) of the SARS-CoV-2 Spike protein (S1) before booster dosing (timepoint-1) and four weeks afterward (timepoint-2). An IgG titer ≥ 50 AU/mL defined a positive seroconversion response. An IgG titer ≥ 4446 AU/mL was assumed as a correlate of 50% vaccine efficacy against symptomatic infections. A total of 258 patients on active treatment within the previous six months were enrolled between September 23 and October 7, 2021. The third dose resulted in an exponential increase in median anti-RBD-S1 IgG titer (P < 0.001), seroconversion rates (P < 0.001), and 50% vaccine efficacy rates (P < 0.001). According to ROC curve analysis, T helper and B cells were significantly associated with seroconversion responses at timepoint-1, whereas only B cells were relevant to 50% vaccine efficacy rates at timepoint-2. A positive linear correlation was shown between anti-RBD-S1 IgG titers and these lymphocyte subset counts. Multivariate analysis ruled out a potential role of T helper cells but confirmed a significant interaction between higher B cell levels and improved antibody response. These findings suggest that peripheral counts of B cells correlate with humoral response to the third dose of mRNA-BNT162b2 vaccine in actively treated cancer patients and could provide insights into a more comprehensive assessment of vaccination efficacy.

The relationship between absolute counts of lymphocyte subsets and clinical features in patients with pulmonary tuberculosis.

BackgroundThe aim of the present study is to investigate the clinical value and characteristics of peripheral blood lymphocyte subsets in patients with pulmonary tuberculosis (PTB) using flow cytometry.MethodsThe absolute counts of T, CD4+T, CD8+T, natural killer (NK), NKT and B lymphocytes in 217 cases of PTB were detected, and the variations in lymphocyte subset counts between different ages and genders and between aetiological detection results and chest radiography results were analysed.ResultsIn 75.3% of the patients with PTB, six subset counts were lower than the normal reference range, and 44% showed lower‐than‐normal CD4+T lymphocyte levels. The counts of T, CD4+T, CD8+T and B lymphocytes were significantly lower in patients aged >60 years, and the NKT cell counts were significantly lower in female patients than in male patients. Among the patients with positive aetiological results, 40.8% had reduced CD8+T counts; these were significantly lower than those in patients with negative aetiological results (P = 0.0295). The cell counts of T, CD4+T, CD8+T and B lymphocytes reduced as lesion lobe numbers increased. The counts of T, CD4+T and CD8+T lymphocytes were significantly higher in the group with lesions affecting one lobe than in the groups with two to three lobes or four to five lobes, and the counts of B lymphocytes were significantly higher in the group with one lobe and the group with two to three lobes than in the group with four to five lobes. The counts of CD4+T and CD8+T lymphocytes were highest in the no cavity group and showed a downward trend with the increase in cavities; the T lymphocyte count was significantly higher in the no cavity group than in the group with five or more cavities (P = 0.014), and the CD8+T lymphocyte count was significantly higher in the no cavity group than in the group with one to two cavities and the group with five or more cavities (P = 0.001 and 0.01, respectively).ConclusionsIn most patients with tuberculosis, immune function is impaired. The absolute counts of peripheral blood lymphocyte subsets are closely related to the aetiological results and lesion severity in patients with PTB; this could be used as evidence for immune intervention and monitoring curative effects.

The Predictive Value of Changes in the Absolute Counts of Peripheral Lymphocyte Subsets for Progression and Prognosis in Breast Cancer Patients.

Background As the number and proportion of lymphocyte subsets are an important indicator of the immune function, an in depth understanding of the immune function of patients with malignant tumor has important clinical values for the treatment, prognosis, and evaluation of the disease. This retrospective study was to evaluate the clinical value of the absolute counts of lymphocyte subsets as potential blood biomarkers for progression and prognosis in breast cancer patients. Methods A total of 237 BC patients and 55 age-matched female normal healthy donors were included in this study. Flow cytometry was used to determine the absolute counts and the percentages of CD3+, CD4+, CD8+, B, and NK cells. The receiver operating characteristic curve (ROC) was used to evaluate the accuracy of absolute count of lymphocyte subsets in the curative efficacy assessment. The clinicopathological parameters influencing the disease progression were determined by Cox proportional hazards regression. Progression-free survival (PFS) was estimated using the Kaplan–Meier method with the log-rank test. Results: Compared with the healthy donors, the absolute counts of lymphocyte subsets in patients decreased significantly. ROC analysis showed that the area under the curve of the CD4+ absolute count was 90% (95% confidence interval 0.859–0.940), and the sensitivity and specificity were 80.9% and 85.3%, respectively. The analysis of Cox regression showed that the cutoff value of the CD4+ absolute count ≥451 cells/μL might be a favorable prognostic factor. Multivariate analysis of prognostic factors of PFS showed that the CD4+ and CD8+ absolute count were independent factors for predicting PFS. Conclusions The remarkably impaired absolute counts of the CD3+, CD4+, CD8+, B, and NK cells in patients with breast cancer can be used as potential susceptible biomarkers to evaluate the patient's immune status. The higher level of CD4+ and CD8+ absolute counts probably contributed to the longer PFS and favorable outcome of BC patients.

Reference values for T and B lymphocyte subpopulations in Turkish children and adults

Background/aim Established reference values are critical for the interpretation of immunologic assessments. In particular, the proportion and absolute counts of T- and B- cell subpopulations are subject to change with age and ethnicity. We aimed to establish age-specific reference values for lymphocyte subsets using updated immunophenotyping panels.Materials and methods We studied a total of 297 healthy Turkish subjects aged 0 to 50 years, stratified into major age brackets in a cluster factor of 10 per age-group. The predetermined age intervals contained randomly allocated participants enrolled over a period of 6 months, who were homogenously distributed by sex. We analyzed a complete blood count test and simultaneously with detailed immunophenotyping enumerated the percent and absolute cell counts of lymphocyte subsets. Results The percentage and absolute counts of lymphocyte subsets show a marked surge across the age-span. T helper, T cytotoxic, and the natural killer cell numbers were increasing from birth until 6 months, followed by a gradual decrease thereafter. B cell numbers were rising until 2 years, followed by a gradual decrease for the upcoming years, accompanied by a steady expansion of unclass-switched- and class-switched- B cells.Conclusion We provide updated extensive reference intervals for lymphocyte subpopulations in Turkish people.

Abnormalities in subsets of B and T cells in Mexican patients with inborn errors of propionate metabolism: observations from a single-center case series.

Propionate inborn errors of metabolism (PIEM), including propionic (PA) and methylmalonic (MMA) acidemias, are inherited metabolic diseases characterized by toxic accumulation of propionic, 3-hydroxypropionic, methylcitric, and methylmalonic organic acids in biological fluids, causing recurrent acute metabolic acidosis events and encephalopathy, which can lead to fatal outcomes if managed inadequately. PIEM patients can develop hematological abnormalities and immunodeficiency, either as part of the initial clinical presentation or as chronic complications. The origin and characteristics of these abnormalities have been studied poorly. Thus, the aim of the present work was to evaluate and describe lymphoid, myeloid, and erythroid cell population profiles in a group of clinically stable PIEM patients. This was a retrospective study of 11 nonrelated Mexican PIEM patients. Clinical, biochemical, nutritional, hematological, and lymphocyte subsets were analyzed. Despite being considered clinically stable, 91% of patients had hematological or immunological abnormalities. The absolute lymphocyte subset counts were low in all patients but one, with CD4+ T-cell lymphopenia, being the most common one. Furthermore, of the 11 studied subjects, nine presented with a low CD4/CD8 ratio. Among the observed hematological alterations, bicytopenia was the most common (82%) one, followed by anemia (27%). Our results contribute to the landscape of immunological abnormalities observed previously in PIEM patients; these abnormalities can become a life-threatening chronic complications because of the increased risk of opportunistic diseases. These findings allow us to propose the inclusion of monitoring immune biomarkers, such as subsets of lymphocytes in the follow up of PIEM patients.

Absence of Influence of the Pre-Transplant Immune Status of Recipients on Survivals and Gvhd after Allogeneic Stem Cell Transplantation: A Retrospective Study of 195 Cases

Introduction: While many publications have studied immune reconstitution after allogeneic stem cell transplantation (allo-SCT), data exploring whether the pre-transplant immune status (IS) of the recipients has any impact after this procedure are still lacking. Material and Methods: Since May 2017, in our department, the IS of allo-SCT recipients has been systematically investigated at the time of pre-graft check-up. Data of this monocentric retrospective study on post-allo-SCT outcomes are reported here. Information was retrieved from complete blood counts (CBC) in terms of lymphocytes (normal range [NR] 1.5-4x109/L) and monocytes (NR: 0.15-0.9 x109/L). Classical multiparameter flow cytometry and a double platform were used to assess absolute counts of lymphocyte subsets, i.e. CD3+ (NR: 0.9-1.8 x109/L), CD4+ (NR: 0.5-1.2 x109/L), CD8+ (NR: 0.3-0.7 x109/L), CD19+ (NR: 0.1-0.4 x109/L) and CD56+ (NR: 0.1-0.4 x109/L). All patients also benefited from standard serum electrophoresis allowing to appreciate their global level of immunoglobulins (Ig) (NR: 8-13.6 g/L). The impact of pre-graft immune parameters was investigated for overall (OS) and disease-free (DFS) survivals as well as acute and chronic graft-versus-host disease (GVHD) incidence. Results: Between May 2017 and December 2019, 195 consecutive adults were checked for immune status before allo-SCT. They were 117 males and 78 females at a median age of 59 years-old (range: 23-71). The median follow-up for the whole cohort was 21.7 months (range: 5.77-37.5). The majority of patients had a myeloid disease (n=141) and received a reduced intensity (RIC) regimen (n=149, myeloablative [MAC] n=23; sequential n=23). The disease-risk index was low/intermediate and high/very-high for respectively 91 and 104 patients. GVHD prophylaxis included anti-thymocytes globulins (ATG) for 102, post-transplant cyclophosphamide (PTCy) for 54 and both ATG+PTCy for 39. Donors were siblings in 39 cases, matched-unrelated (MUD) in 86, haplo-identical in 66 and 9/10 mis-matched in 4. The IS was evaluated at a median of 20 days (range: 8-72) before allo-SCT. Except for monocytes (median: 0.48 x109/L, range: 0-26), all median immune cell populations were lower than the NR before allo-SCT: lymphocytes (0.84 x109/L, range: 0-8.4), CD3+ (0.7 x109/L, range: 0-3.5), CD4+ (0.37 x109/L, range: 0-2.37), CD8+ (0.25 x109/L, range: 0-2.56), CD19+ (0.005 x109/L, range: 0-0.56), CD56+ (0.097 x109/L, range: 0-0.77). Conversely, the Ig median level was in the NR (8.6 g/L, range: 1.4-28). The percentages of patients with immune cells and Ig above the lowest NR were as follows: lymphocytes: 14.8%, monocytes: 80%, CD3: 37.4%, CD4: 37.4%, CD8: 38.9%, CD19: 16.4%, CD56: 47.6%, Ig: 56.7%. Only 5 patients (2.5%) had a fully normal IS before transplant. Considering the whole cohort, 2-year OS and DFS were 55.6+3% and 50.4+3%, respectively, while incidences of day 100 grade 2-4 and 3-4 acute GVHD and 2-year moderate/severe chronic GVHD were 34.7%, 19.4% and 19.8%, respectively. The comparison of survivals between patients partitioned using either the median of the NR values for immune cells and Ig, showed no difference between subgroups. Similarly, IS had no impact on the incidence of acute nor chronic GVHD. Finally, no difference either was seen according to the conditioning regimen (RIC) or GVHD prophylaxis (ATG, PTCY or both). Conclusion: As expected, almost all pre-ASCT patients present with immune depression. However, pre-transplant IS does not appear to impact survival nor GVHD. This suggests that there is no need to decrease or replace immunosuppressive treatments commonly used in recipients before allo-SCT in the hope to provide better outcomes. Disclosures Touzeau: GlaxoSmithKline: Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Other: Travel, Accommodations, Expenses; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Amgen: Consultancy, Honoraria, Other: Travel, Accommodations, Expenses; Sanofi: Honoraria, Research Funding; Abbvie: Consultancy, Honoraria, Other: Travel, Accommodations, Expenses, Research Funding; Janssen: Consultancy, Honoraria, Other: Travel, Accommodations, Expenses. Le Gouill:Roche Genentech, Janssen-Cilag and Abbvie, Celgene, Jazz pharmaceutical, Gilead-kite, Loxo, Daiichi-Sankyo and Servier: Honoraria; Loxo Oncology at Lilly: Consultancy. Moreau:Abbvie: Consultancy, Honoraria; Novartis: Honoraria; Sanofi: Consultancy, Honoraria; Amgen: Consultancy, Honoraria; Janssen: Consultancy, Honoraria; Celgene/Bristol-Myers Squibb: Consultancy, Honoraria; Takeda: Honoraria.

Value of absolute counts of lymphocyte subsets in the early prediction of refractory Mycoplasma pneumoniae pneumonia in children

To study the value of absolute counts of lymphocyte subsets in the early prediction of refractory Mycoplasma pneumoniae pneumonia (RMPP) in children. A retrospective analysis was performed for the clinical data of 244 children with Mycoplasma pneumoniae pneumonia (MPP). Among these children, 166 had MPP, and 58 had RMPP. The two groups were compared in terms of clinical features and laboratory markers such as lymphocyte subsets, lactate dehydrogenase, C-reactive protein, procalcitonin and immunoglobulin E (IgE). The receiver operating characteristic (ROC) curve was used to evaluate the specific indices for predicting RMMP. There were significant differences between the two groups in the absolute counts of CD3+, CD4+, CD19+, and CD56+ lymphocytes and the serum levels of lactate dehydrogenase, C-reactive protein, and IgE (P<0.05). The ROC curve analysis showed that the absolute counts of CD3+, CD4+ and CD19+ lymphocytes had an area under the ROC curve (AUC) of 0.866, 0.900 and 0.842 respectively in the differential diagnosis of RMPP and MPP, with a sensitivity of 86%, 90% and 82% respectively and a specificity of 75%, 70% and 80% respectively. The absolute counts of CD3+, CD4+ and CD19+ lymphocytes can be used to predict RMPP in children.

Air pollution exposure and immunological and systemic inflammatory alterations among schoolchildren in China

Exposure to air pollution is associated with an increased risk of respiratory infection, to which children are more susceptible than adults. However, epidemiological evidence regarding the association of chronic exposure to air pollution with the immune and systemic inflammatory function of children is scarce, especially in the context of higher exposure levels. In this study, we included 163 chronically exposed schoolchildren from a polluted area and 110 schoolchildren from a control area in Licheng district, Jinan, China. Immune biomarkers, including the absolute counts of lymphocyte subsets and the levels of immunoglobulins G, A, and M, C3, and C4 were determined. To explore the related biological process of altered immune biomarkers, 2 systemic inflammatory biomarkers, including C-reactive protein and the neutrophil-to-lymphocyte ratio, were also determined. After adjusting for confounders, the decreased B lymphocyte count (p = 0.021) and C3 and C4 levels (both p < 0.001) and the increased monocyte count (p = 0.009) and CD8+ T lymphocyte proportion (p = 0.054) were associated with living in the polluted area. Significant differences in the C4 and C3 levels between the areas were only seen in male schoolchildren and in schoolchildren without passive smoking exposure (Pinteraction = 0.036 and 0.042, respectively). The alterations in immune biomarkers suggested that air pollution-induced immunotoxic effects and relevant adaptive responses were simultaneously present in schoolchildren exposed to a higher level of air pollution. Future studies investigating the temporal patterns of these biomarkers among children are warranted.

Biological variations of peripheral blood lymphocytesusing flow cytometric double-platform method

Objective To evaluate the biological variations of 8 lymphocyte subsets using flow cytometric double-platform method. Methods Twenty healthy adults were recruited from Peking Union Medical College Hospital in September 2013.At 8: 00 AM, 12: 00 PM, and 4: 00 PM on days 1, 3, and 5, venous blood was collected from the volunteers.The percent and absolute lymphocyte subset counts were measured using duel-platform method. The sample collection and handling techniques were standardized.Before each batch analysis, the instrument quality controls were performed using the same lots of reagents.The intra-individual coefficient of variation (CVI) and inter-individual coefficient of variation (CVG) were calculated by nested ANOVA with SPSS 13.0 software.The analytical coefficient of variation (CVA), index of individuality (II) and reference change value (RCV) were calculated by Excel2003.A mean pairwise comparison was determined by one-way ANOVA and variance analysis. The values between groups were analyzed by independent sample t test. Results For T cells (CD3+ ), helper T cells (CD3+ CD4+ CD8-), suppressor T cells (CD3+ CD4-CD8+ )and B cells (CD3-CD19+ ), the intra-individual coefficient of variation (CVI) and inter-individual coefficient of variation (CVG) were 0.03, 0.06, 0.05, 0.14 and 0.12, 0.16, 0.23, 0.31 respectively, which were all similar to those in previous studies.However, the II and RCV of the four lymphocyte subsets were very different from those in previous studies, which were 0.26, 0.40, 0.22, 0.44 and 8.77, 16.86, 14.93, 39.69, respectively.Moreover, variations in absolute count, CVI, CVG, and analysis coefficient of variation (CVA) of all 8 lymphocyte subsets were greater than those of relative count. Variations in the percent and absolute counts for the CD3+ CD4-CD8-, CD3+ CD4+ CD8+ , and CD3+ CD16+ CD56+ cell subsets were relatively high.The CVI, CVG and CVA for the cells of CD3+ CD4-CD8- were 0.12, 0.49 and 0.16.The CVI, CVG and CVA for the cells of CD3+ CD4+ CD8+ were 0.40, 0.93 and 0.55.The CVI, CVG and CVA for the cells of CD3+ CD16+ CD56+ were 0.28, 1.11 and 0.16. Conclusions Investigation on the CVI , CVG and CVA may allow us to obtain II and RCV, by which we can determine the utility of traditional population based reference ranges.Documentation of the RCV indices may be used as objective delta-check values in quality management and decide whether clinical significance existed in the continuously detected results.(Chin J Lab Med, 2016, 39: 350-355) Key words: Lymphocyte subsets; Flow cytometry; Immunophenotyping

Age- and sex-related reference intervals of lymphocyte subsets in healthy ethnic Han Chinese children.

Immunophenotyping of blood lymphocytes has become an important tool in the diagnosis of immunologic and hematologic disorders such as immunodeficiencies, lymphoproliferative and autoimmune diseases. Lymphocyte subsets include total T-cells (CD3(+)), TH (T helper, CD3(+) CD4(+)), TC (cytotoxic T cells, CD3(+) CD8(+)), B-cells (CD3(-) CD19(+)), and NK-cells (CD3(-) CD16(+) CD56(+)). Specific lymphocyte subset reference intervals should be locally established for meaningful comparison and to obtain an accurate interpretation of the results. Reference intervals of lymphocyte subsets for Chinese children are scarce. We performed dual-platform flow cytometry to determine the reference intervals of the percentages and absolute counts of lymphocyte subsets, including total T-cells, TH cells, TC cells, B-cells, and NK-cells in 1,027 ethnic Han children aged 4 months to 7 years in Henan, China. The children were divided into seven age groups. The percentages and absolute counts differed significantly with age, with the percentages of TH cells and B cells and the CD4/CD8 ratio peaking during the first year, while the percentages of total T cells, TC cells, and NK cells were obviously increased with age; girls showed a trend toward having a higher percentage of TH cells and a higher CD4/CD8 ratio than boys. The absolute counts of lymphocyte subsets peaked during first year and then decreased steadily with age. The reference intervals of lymphocyte subsets among children from China differed from the reported values in Hong Kong, the United States, Cameroon, and Italy. The differences observed could be due to genetic and environmental factors, coupled with the methodology used. The reference intervals of lymphocyte subsets could be used as initial national reference ranges in guidelines for children aged 4 months to 7 years.

Abstract 2536: Post-transplantation NK cell is a useful predictor of graft-versus-host-disease in allogeneic hematopoietic stem cell transplantation

Abstract Background: Reconstitution of the immune system following allogeneic hematopoietic stem cell transplantation (HSCT) is important for transplantation outcome. Quantitative measurement of the lymphocyte subset (LST) by flow cytometry has been used for analysis of the immune system. However, the association of each lymphocyte subset with transplantation has not been fully elucidated. Here, we investigated the relevance of each lymphocyte fraction as a predictor for HSCT outcome. Methods: A total of 70 patients who received allogeneic HSCT at national cancer center were included. The median age was 42 years (range 20-64) with 34 males (49%). The enrolled diseases included acute myeloid leukemia (n=33, 47%), acute lymphoblastic leukemia (n=13, 19%), myeloid dysplastic syndrome (n=9, 13%), non-hodgkin lymphoma (n=5, 7%), severe aplastic anemia (n=4, 6%), chronic myeloid leukemia (n=3, 4%), and multiple myeloma (n=3, 4%). LST measurements for CD3, CD4, CD8, CD19, and CD16/CD56-positive cells were performed at 7 days before and 30 days after HSCT. The absolute counts of lymphocyte subsets were evaluated for acute graft-versus-host disease (GVHD), infection (bacterial, viral, and fungal) and complications during 100 days post HSCT. GVHD was pathologically validated and determination of infection was based on microbiology results including culture, cytomegalovirus antigenemia and virus PCR. Moreover, the association between LST and treatment failure as relapse or death during any period was analyzed. Results: The number of each fraction of lymphocyte subsets at 7 days before and 30 days after HSCT were as following (mean ± SD/uL): CD3 (804.5 ± 564.6, 862.1 ±890.0, p =0.719), CD4 (339.74 ± 284.8, 223.5 ± 189.9, p =0.009, CD8 (430.6 ± 362.9, 576.9 ± 703.1, p = 0.169, CD19 (39.7 ± 72.3, 19.1 ± 22.9, p =0.035), NK (86.6 ± 86.8, 272.5 ± 192.2, p &amp;lt; 0.001). Each lymphocyte fraction does not show any association with complications and increased risk for infection. However, the decreased number of NK cells at 30 days after HSCT represented association with an increased risk of GVHD (n=21) (GVHD group vs. non-GVHD group: 206.4±195.2 vs. 310.1 ± 182.5, p = 0.033). When the NK cell level was divided as median value (219/µL), the group of lower level has correlation decreased overall survival (p = 0.018). Moreover, decreased number of NK cells at 90 days after HSCT was also associated with an increased risk of GVHD (145.3±107.9 vs. 370.2±265.8, p = 0.043). Conclusion: These results suggest that measuring the early recovery of NK cells at 30 days and analyzing the trend of LST can be a useful predictor of clinical outcome including GVHD in allogeneic HSCT. Citation Format: Seo Yeon Kim, Hyewon Lee, Hyoeun Shim, Hyeon-Seok Eom, Sun-Young Kong. Post-transplantation NK cell is a useful predictor of graft-versus-host-disease in allogeneic hematopoietic stem cell transplantation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2536. doi:10.1158/1538-7445.AM2014-2536

Intra-day and inter-day biological variations of peripheral blood lymphocytes

BackgroundThe proportion and absolute numbers of lymphocyte subsets are important indices that reflect the immunological status of the body. Few studies on biological variations of absolute lymphocyte subset counts and no study in Asian population are currently available. Furthermore, there have been few reports on the biological variation of these indices in the short term. MethodsAt 8:00AM, 12:00PM, and 4:00PM on days 1, 3, and 5, venous blood was collected from 20 healthy volunteers. All participants maintained their normal lifestyles. The percent and absolute lymphocyte subset counts were measured using single -platform method. ResultsIntraday and interday biological variations for the absolute and percent counts of lymphocyte subsets were relatively constant. The intra-individual coefficient of variation (CVI) and inter-individual coefficient of variation (CVG) were similar to those in previous studies. Biological variations in the percent and absolute counts for the CD3+CD4−CD8−, CD3+CD4+CD8+, and CD3+CD16+CD56+ subsets were relatively high. ConclusionsThese observations are clinically valuable. Investigation on the CVI and CVG may allow us to determine the utility of traditional population based reference ranges. Documentation of the reference change values may be used as objective delta-check values in quality management and decide whether the change that occur in an individual's serial results before the change is significant. The present study also enriched the database regarding the biological variations of lymphocyte subsets in Asian population.

Patterns of Immune Reconstitution in Patients with Acute Promyelocytic Leukemia Treated with Single Agent Arsenic Trioxide and Its Impact On Time to Molecular Remission

AbstractAbstract ▪3552▪This icon denotes a clinically relevant abstractThere is a substantial data to suggest that immune response to acute promyelocytic leukemia (APL) contributes significantly towards achieving durable remission. The impact of arsenic trioxide (ATO) on immune reconstitution, when used to treat APL, has not been studied. Single agent ATO is able to induce complete molecular remission in practically all patients who complete an induction and consolidation course. However, while ATO is well tolerated approximately 20% of newly diagnosed cases will relapse with such a regimen. We undertook a prospective study to evaluate the pattern of immune reconstitution in patients with newly diagnosed APL treated at our center with a single agent ATO regimen.Peripheral blood samples were obtained from patients before treatment, on day 15 after starting ATO, post induction, pre consolidation, maintenance cycle 2 (6 months from diagnosis) and maintenance cycle 6 (10 months from diagnosis) for flow cytometry analysis. Briefly cells were labeled using a panel of monoclonal antibodies to CD3, CD4, CD8, CD56, CD16, CD19, CD45RO, CD45RA directly conjugated with FITC, PE, PerCP or APC followed by red cell lysis and cells were then washed and analyzed using FACS Calibur. Cells were gated using forward versus side scatter dot-plots. Lymphocyte subset percentages were calculated from the lymphocyte gate and absolute lymphocyte subset counts were calculated for the analysis.Analysis of lymphocyte subset reconstitution patterns have shown that all the subsets were below the normal range at diagnosis. Following treatment there was differential pattern of immune reconstitution in different lymphocyte subsets. The earliest recovery to normal range was seen in the CD8 subset (Figure 1) while the mean CD4 subset reached the normal range only at 3 months from diagnosis. A normal CD4 to CD8 ratio had not been reached even at the last follow up in this study (10 months from diagnosis). There was a significant delay in the immune reconstitution of NK cells (CD56+CD3-) and naïve T cells (CD4+CD45RA+). The pattern of immune reconstitution of some of the lymphocyte subsets that were evaluated is summarized in Figure 1.The number of relapses in this cohort was too few to make any association between relapse and the pattern of immune reconstitution. However, we had previously reported that RT-PCR positivity at the end of induction was a significant independent risk factor for subsequent relapses with this regimen. Among the 29 cases available for evaluation at the time of completion of induction and documentation of hematological remission 14 (48.2%) were RT-PCR negative. At this time point there was a significantly higher number of NK-T cells (CD56+CD3+) in those that were RT-PCR negative versus those that were positive (6.1±4.5 × 107/Lt Vs. 2.9±4.1 × 107/Lt; P=0.040). There was also a trend to lower CD3 and a lower CD56br/CD16dim subset in the cases that were RT-PCR positive at the end of induction.This study demonstrates that there is significant heterogeneity in the pattern of immune reconstitution in different lymphocyte subsets post treatment of APL with ATO. Modulation of immune recovery and response could potentially improve leukemia clearance and maintenance of remission. [Display omitted] Disclosures:No relevant conflicts of interest to declare.