Lymphocyte Subpopulations In Bone Marrow Research Articles

Breast cancer molecular subtypes and bone marrow immune system in patient’s prognosis

Backgrоund. Breast cancer molecular subtypes serve as a basis for combined treatment of the disease. Some of them (e. g. triple negative subtype) are prognostically unfavorable. Recently, we’ve demonstrated role of bone marrow B-cells in breast cancer prognosis. Correlation between these parameters (molecular subtypes and bone marrow lymphocyte subpopulations) have not yet studied.Aim. To investigate prognostic role of breast cancer molecular subtypes and to see the relationships between these subtypes and subpopulational composition of bone marrow lymphocytes.Materials and methods. Detailed study of bone marrow has been performed in all 107 patients treated in breast cancer department of N.N. Blokhin National Medical Research Center of Oncology in 2013–2016 years. Distribution of patients according to molecular subtypes were as follows: Luminal A – 36 (33.6 %) pts, Luminal B HER2-negative – 38 (35.5 %), Luminal B HER2-positive – 15 (14 %), subtype with Erb-B2 hyper-expression – 5 (4.4 %), triple negative – 13 (12.1 %). Comparison of molecular subtypes with immunological parameters was done for the following bone marrow B-lymphocyte subpopulations: CD19+, CD19+CD10+, CD19+CD38+, CD19+CD5+. The following survival data were analyzed: overall survival, relapse-free survival, progression-free survival, metastasis-free survival.Results. Breast cancer molecular subtypes were not related to overall survival of patients but influenced relapse-free survival, progression-free survival and metastasis-free survival. There were no correlation between molecular subtypes and bone marrow B-lymphocyte subpopulations CD19, CD19+CD10+, CD19+CD38+. B1-lymphocytes were related to molecular subtypes, the levels of B1 cells were significantly higher in Erb-B2 subtype compared to triple-negative subtype (p = 0.039).Conclusion. Breast cancer molecular subtypes are different in prognosis – relapse-free survival, progression-free survival and metastasis-free survival. The most prognostically unfavorable is triple-negative subtype. There are some relationships of bone marrow subpopulations and molecular subtypes of breast cancer: levels of B1-cells are much higher in Erb-B2 subtype compared to triple negative subtype (p = 0.039).

Low Circulating B Lymphocytes in Newly Diagnosed Follicular and Diffuse Large B Cell Lymphoma: Differences Compared to Normal Subjects and Possible Prognostic Role

Introduction: Reduction of the lymphocyte to monocyte ratio (LMR) has an unfavorable impact on overall survival in Diffuse Large B Cell Lymphoma (DLBCL), and on treatment response in Follicular Lymphoma (FL). Abnormal LMR seems primarily due to a reduction in absolute value of lymphocytes. Based on these observations, we have prospectively investigated peripheral blood (PB) and Bone Marrow (BM) lymphocyte subpopulations in a series of previously untreated DLBCL and FL. Aim of the study was: i. to assess whether LMR alteration is linked to a distinct lymphocyte subset; ii. to correlate PB with BM cell subpopulations; iii. to compared DLBCL and FL with regards to PB and BM lymphocyte subpopulations. Methods: Overall, PB cells from 58 DLBCL and 27 FL at diagnosis were investigated; BM cell populations were analyzed in 77 DLBCL and 54 FL. None of the studied patients had BM involvement nor circulating lymphoma cells detectable by cytometry; in addition, none of the patients had been previously treated with immunosuppressive drugs. PB from 40 healthy subjects and BM samples from 69 subjects without primary BM dysfunctions were used as control. Multicolor flow cytometry (MFC) was used to evaluate percentages and absolute values of the following cell subsets: CD3+ T cell, CD3-CD56+ NK cells, CD19+CD20+ B cells. Among B cells, t he analysis included transitional (CD38+high/CD24+high/IgD+/CD27-), naive (CD38+/CD24+/IgD+/CD27-), memory IgD+ (CD38-/+CD24+/IgD+/CD27+), and memory IgD- (CD38-/CD24+high/IgD-/CD27+) subsets; B cell precursors were investigated as CD19+CD20-CD10+ and mature B cells as CD19+CD20+CD10-. Results: A significant reduction in median absolute lymphocyte count was seen in PB from lymphoma patients (median: 1,526/µL) compared to controls (median: 2,195/ µL). Moreover, absolute lymphocytes were more markedly reduced in DLBCL (median: 1,386 /µL) compared to FL (median: 1883/µl) (p<0.001). This resulted in a significantly lower median PB-LMR in DLBCL (median LMR: 3) compared to FL patients (median LMR: 5) (p=0.001). Among PB cell subpopulations, a marked reduction in circulating B cells was observed in lymphoma patients (median 96/µL) compared to controls (median 224/µL). Again, loss of circulating B-cells was more markedly pronounced in DLBCL (median: 81.5 cell/µL) compared to FL (median: 150 cell/µL) (p=0.013). Differences in circulating B-cells among DLBCL, FL and controls are shown in Figure 1A. At BM analysis, a reduction of B cell precursor (CD19+/CD20-/CD10+) could be documented only in DLBCL. A reduction in circulating T-lymphocytes in lymphoma patients compared to controls was detected as well, although at a lower extent compared to B-cells. No significant abnormalities in the amount of circulating NK-cells was seen in both DLBCL and FL patients. When these data were matched with treatment outcome, the reduction of circulating B cells proved to be associated with an unfavorable outcome in younger patients with DLBCL. Indeed, among DLBCL patients aged < 60 yrs, a poorer progression free survival was seen in those having less than 96 B-cells/µL compared to those with PB B-cells equal or above 96 B-cells/µl (Figure 1B). Conclusions: The study indicates that circulating B lymphocytes are significantly reduced in both DLBCL and FL compared to age-matched healthy controls; however, their loss is much more pronounced in DLBCL compared to FL. Indeed, abnormalities in B-cell progenitors are present in BM from DLCBL but not from FL. In addition, B-cell reduction may have an unfavorable impact in the long-term outcome of DLBCL, at least in younger patients. Lastly, both B and T-cell subsets are decreased in DLBCL and FL whereas circulating NK subset does not seem to be affected. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Correlation of Quantitative Changes in Bone Marrow Lymphocyte Subpopulations with Some Rhabdomyosarcoma Prognosis Factors in Children

Rationale: The known prognosis factors for rhabdomyosarcoma (RMS) in children do not always explain the unsatisfactory outcome of treatment. Changes in the subpopulation composition of Bone Marrow (BM) effector cells during the development of RMS may indicate new directions for the search for prognostic factors and points for the impact of targeted therapy. Purpose: To identify correlations between quantitative changes in the levels of subpopulations of T, B and NK-lymphocytes of BM and known risk factors for RMS in children. Objects: The study included 31 patients. The main group included 16 patients with RMS, average age—6.8 ± 1.0 years, while children 1 - 10 years old—13 (81.3%), over 10 years old—3 (18.8%) people, girls and boys were 8 people each. The embryonic variant of RMS was established in 10 (62.5%) cases, the alveolar variant—in 4 (25%) cases, in two patients (12.5%), the histological variant was not established. In 12 (75%) patients, an unfavorable localization of the RMS (parameningeal, extremities, prostate, bladder) was revealed, in 4 patients (25%), the localization of the tumor was regarded as favorable. Patients with T2b—13 (81.2%) and T2a—2 (12.5%) stages prevailed. Regional and distant metastases were detected in 10 (52.6%) patients. The comparison group included 15 children in whom the presence of malignant neoplasia was excluded, the average age was 8.4 ± 1.5 years, 11 boys (73.7%) and 4 girls (26.3%). Methods: All patients underwent morphological (myelogram counting) and immunological (quantitative analysis of lymphocytic subpopulations) bone marrow studies. Immunophenotyping in all patients was carried out by direct immunofluorescence using a triple fluorescent label. Results: Significant differences in the levels of subpopulations of BM T-lymphocytes were found when comparing the values of the main group, distributed by localization and histological variant, with the data obtained in the control group of patients. For example, the percentage of CD3+ T cells with the co-stimulatory molecule CD28+ was significantly higher in patients with parameningeal RMS (p = 0.010). Conclusion: Each clinical group of patients has its own individual immunological characteristics. The results obtained by us can be considered indicative and regarded as starting points for further study of the peculiarities of the subpopulation composition of BM in patients with RMS.

Detection of disseminated tumor cells and their relationship with a population of bone marrow lymphocytes in patients with non-small cell lung cancer

Introduction.Detection of disseminated tumor cells (DTC) in solid tumors is an important component of the assessment of disease prognosis. Bone marrow damage is common. There is evidence indicating an important role for bone marrow lymphocyte subpopulations in hematogenous metastasis.&#x0D; Aim.To evaluate the frequency of bone marrow damage in patients with non-small cell lung cancer (NSCLC) based on the detection of DTC by flow cytometry, as well as their effect on the population of bone marrow lymphocytes.&#x0D; Materials and methods.62 bone marrow samples of patients with a verified diagnosis of NSCLC: adenocarcinoma (33), squamous cell carcinoma (27), other types (2). Methods: morphological, multicolor flow cytometry. Studied DTC, lymphocyte populations CD3, CD4, CD8, CD19, CD20, CD16, CD27. Collection and analysis: FACS Canto II, USA, Kaluza Analysis v2.1.&#x0D; Results.In bone marrow, DTC (EPCAM+CD45-) were found in 43.5% of patients with NSCLC (1 cell per 10 million myelocariоcytes was taken as the threshold value). The presence of DTC did not correlate with the size of the tumor, the status of the lymph nodes, and the stage of the tumor process. DTC was more often observed in more differentiated tumors (p=0.023). A significant increase in the level of subpopulations of CD16+CD4-NK-cells (p=0.002), CD27+CD3+T-cells (p=0.015) with bone marrow damage was revealed.&#x0D; Conclusion.The possibility of detecting DTC in the bone marrow of patients with NSCLC was established, in 43.5% of patients with NSCLC in the bone marrow DTC was detected, and their presence was established even with a localized tumor process. More frequent bone marrow damage was observed with well-differentiated tumors. The relationship between DTC and bone marrow lymphocyte populations was revealed: subpopulations of CD16+CD4-, CD27+CD3+.

IMMUNOLOGICAL PARAMETERS OF BONE MARROW IN NON-SMALL CELL LUNG CANCER

Introduction Generation of most immunocompetent cells takes place in bone marrow Bone marrow. As well, bone marrow is a peripheral lymphoid organ where antitumor effector cells and memory cells are present. The aim of the work is to estimate peripheral lymphoid cell subpopulations in bone marrow of lung cancer patients. Materials and methods Study has been done in 68 pts with lung cancer: squamous cell cancer (n = 28), adenocarcinoma (n = 38), other forms (n = 2). In all cases standard diagnostic and staging procedures were performed, as well as morphological (myelogram) and immunological study of bone marrow lymphocyte subpopulations. Multicolor Flow cytomtry was used for study of bone marrow lymphocyte populations. We studied T-cells and its subpopulations, B-cells, NK-cells, perforin-positive cells, and CD27-positive cells. Results Squamous cell lung cancer was characterized by higher content of bone marrow mature T-cells (CD3), and CD8 lymphocytes. More typical for adenocarcinoma was mature B-cell reaction (CD20). Effector (perforin-positive) populations of lymphocytes also were related to histological type of cancer: for adenocarcinoma presence of CD4-positive cytotoxic lymphocytes and CD27-expression on effector cells. Perforin-containing lymphoid cells were in opposite correlation to erythrocaryocytes. Conclusion Subpopulational lymphocyte content of bone marrow is related to histological variant of cancer and erythropoiesis in lung cancer patients.

Study of lymphocyte subpopulations in bone marrow in a model of protein–energy malnutrition

Objective Protein–energy malnutrition (PEM) is an important public health problem affecting millions of people worldwide. Hematopoietic tissue requires a high nutrient supply, and a reduction in leukocytes, especially lymphocytes, suggests that some nutritional deficiencies might be altering bone marrow function and decreasing its ability to produce lymphocytes. In this study, we evaluated the effect that PEM has on lymphocyte subtypes and the cell cycle of CD5 + cells. Methods Swiss mice were subjected to PEM using a low-protein diet containing 4% protein. When the experimental group had lost about 20% of their original body weight, we collected blood and bone marrow cells and evaluated the hemogram, the myelogram, bone marrow lymphoid markers using flow cytometry, and the cell cycle in CD5 + bone marrow. Results Malnourished animals presented anemia, reticulocytopenia, and leukopenia with lymphopenia. The bone marrow was hypocellular, and flow cytometric analyses of bone marrow cells showed cells that were CD45 + (91.2%), CD2 + (84.9%), CD5 + (37.3%), CD3 + (23.5%), CD19 + (43.3%), CD22 + (34.7%), CD19 +/CD2 + (51.2%), CD19 +/CD3 + (24.0%), CD19 +/CD5 + (13.2%), CD22 +/CD2 + (40.1%), CD22 +/CD3 + (30.3%), and CD22 +/CD5 + (1.1%) in malnourished animals and CD45 + (97.5%), CD2 + (42.9%), CD5 + (91.5%), CD3 + (92.0%), CD19 + (52.0%), CD22 + (75.6%), CD19 +/CD2 + (62.0%), CD19 +/CD3 + (55.4%), CD19 +/CD5 + (6.7%), CD22 +/CD2 + (70.3%), CD22 +/CD3 + (55.9%), and CD22 +/CD5 + (8.4%) in control animals. Malnourished animals also presented more CD5 + cells in the G0 phase of cell cycle development. Conclusion Malnourished animals presented bone marrow hypoplasia, maturation interruption, prominent lymphopenia with depletion in the lymphoid lineage, and changes in cellular development. We suggest that these changes are some of the primary causes of lymphopenia in cases of PEM and partly explain the increase in susceptibility to infections found in malnourished individuals.

Late-Onset Neutropenia in Rituximab-Treated Lymphoma Patients: Lymphocyte Subpopulation Imbalances, Bone Marrow Hematopoiesis and Immunohistology.

Late-onset neutropenia (LON) is a complication of Rituximab of yet unknown pathophysiology. We investigated potential underlying mechanisms in 12 patients with various non-Hodgkin lymphoma (NHL) subtypes who developed LON without identifiable causes at a median of 95 (range 67–420) days after completion of the intended treatment with Rituximab ± chemotherapy. The study also included two control groups:1.healthy donors (n=25) for comparative analysis of bone marrow (BM) functional parameters;2.NHL patients treated with similar Rituximab ± chemotherapy regimens without LON for comparative evaluation of peripheral blood and BM lymphocyte subpopulations (n=38) and BM pathology findings (n=27).Ten of 12 patients with LON and 33/38 NHL controls developed profound B-cell depletion. Inverted CD4/CD8 cell ratios were observed in 10/12 LON cases vs. 13/38 NHL controls (p<0.01). A rise in CD8 cell count >1.0x109/L was identified in 8/12 LON cases vs. 8/38 NHL controls (p<0.01). T-large granular lymphocyte proliferation (CD3+CD8+CD57+ cells >30%) was observed in 7/12 LON cases vs. 14/38 NHL controls (p=0.1885). BM biopsy samples from 10/12 LON cases were examined at onset of neutropenia. Mild to moderate lymphocytic infiltration with predominantly nodular and/or interstitial growth was observed in all cases. Lymphoid aggregates lacked CD20+ or CD79a+ B-cells and were composed entirely of CD3+CD45RO+CD43+ T-cells. In the control group, BM infiltration by T cells with similar features as above was observed in 18/27 cases. Seven of 10 LON patients showed moderate-to-significant hypoplasia of the granulocytic series; the remaining three exhibited hyperplasia. All ten LON patients had pronounced shift to the left, extending to maturation arrest in 6/10 cases. In contrast, 21/27 NHL control cases (78%) showed granulocytic hyperplasia; the remaining cases were either normal or showed granulocytic hypoplasia. As in the LON cases, the control NHL cases exhibited different degrees of shift to the left of the granulocytic series. Hyperplasia of the erythroid and megakaryocytic series with dyserythropoiesis and dysmegakaryopoeisis, respectively, was identified in all LON cases and most NHL controls. In all cases with repeat samples, MDS-like changes eventually resolved. Compared to healthy donors, LON patients displayed:1.low reserves of granulocytic progenitors associated with increased apoptosis and increased proportion of Fas-expressing cells within the CD34+/CD33+ cell compartment;2.Fas-Ligand and Interferon-γ mRNA expression within BM CD3+ cells;3.increased TNFα and IL-1β levels in long-term BM culture supernatants;4.accelerated apoptosis in all stages of BM granulocytic development.All LON patients were negative for granulocyte-reactive antibodies. We conclude that T-cell mediated autoimmune myelopathy/myelodysplasia associated with MDS-like changes of all myeloid series and suppression of predominantly granulocytic progenitor cell growth is critically implicated in the pathophysiology of Rituximab-related LON, at least in a subset of cases.

Distinct lymphocyte subpopulations in bone marrow from rheumatoid arthritis and osteoarthritis patients: a role for IL-15?

Recent data indicate that bone marrow plays an important role not only as a primary lymphoid organ responsible for haemopoiesis, but also as a secondary lymphoid organ with capability of antigen presentation exceeding that of lymph nodes. Although in chronic inflammatory/immune disease, like rheumatoid arthritis (RA), bone marrow participates in the initiation and/or perpetuation of the disease, there is little information about the real number of lymphocyte subpopulations in bone marrow of these patients and how they can be modulated by T-cell growth factors. IL-15 acting through IL-15 receptors (including the high-affinity IL-15R alpha chain) is a key cytokine influencing the development of natural killer cells in bone marrow, and proliferation and maintenance of the memory T-cell pool. However, there is no information about the levels of IL-15 in bone marrow.

Immune dysfunction following infection with chicken anemia agent and infectious bursal disease virus. I. Kinetic alterations of avian lymphocyte subpopulations

The potential effect of chicken anemia agent (CAA) alone or in combination with infectious bursal disease virus (IBDV) on the immune system of young chickens was determined by measuring alterations in hematocrit values, lymphoid organ-to-body weight ratios and lymphoid cell concentrations at 4, 7, 10, 14, 17, 21, 28 and 42 days post-inoculation (PI). Lymphocyte subpopulations were identified and counted by flow cytometry using cell suspensions stained with monoclonal antibodies (Mabs) for panlymphocytes (K55), cytotoxic T-cells (CTLA3), T-helper cells (CT3), Ia-expressing cells (P2M11) and macrophages (P7). Chicken anemia agent induced a substantial but transient decrease in hematocrit value, thymus-to-body weight ratio and bursa-to-body weight ratio between 7 and 21 days PI corresponding to a generalized lymphocytopenia in the thymus, bursa and spleen. However, cytotoxic T-cell, T-helper cell and Ia-expressing cell concentrations increased in the bone marrow of birds inoculated with CAA alone or in combination with IBDV during the same time period. T-helper-to-cytotoxic T-cell ratios increased in the thymus and spleen during severe lymphocytopenia, indicating a selective decrease in cytotoxic T-cells. T-helper-to-cytotoxic T-cells ratios increased in the bone marrow, indicating a selective increase in T-helper cell concentrations. The increase in Ia-expressing cells in the bone marrow may be a reflection of increased number of activated T-cells which express Ia antigen. Infectious bursal disease virus alone induced a persistent depression of Ia-expressing cells in the bursa and the spleen and no measurable change in the bone marrow lymphocyte subpopulations. Chickens inoculated simultaneously with CAA and IBDV experienced clinical signs observed in chickens inoculated with each virus separately with a prolonged acute phase prior to recovery or mortality.