Lymphocytes In Peripheral Blood Smears Research Articles

Leukemic Involvement Is a Common Feature in Waldenström Macroglobulinemia at Diagnosis.

Waldenström Macroglobulinemia (WM) is a lymphoplasmacytic lymphoma with bone marrow (BM) involvement and IgM monoclonal gammopathy. To date, no studies have focused specifically on peripheral blood (PB) involvement. In this study, 100 patients diagnosed with WM according to the World Health Organization (WHO) criteria were included based on the demonstration of MYD88mut in BM and the availability of PB multiparametric flow cytometry (MFC) analysis. Leukemic involvement by MFC was detected in 50/100 patients. A low percentage of mature small lymphocytes in PB smears was observed in only 15 cases. MYD88mut by AS-qPCR was detected in PB in 65/100 cases. In cases with leukemic expression by MFC, MYD88mut was detected in all cases, and IGH was rearranged in 44/49 cases. In 21/50 patients without PB involvement by MFC, molecular data were consistent with circulating disease (MYD88mut by AS-qPCR 3/50, IGH rearranged 6/50, both 12/50). Therefore, PB involvement by standard techniques was detected in 71/100 patients. MYD88mut was detected in PB by dPCR in 9/29 triple negative cases. Overall, 80% of the patients presented PB involvement by any technique. Our findings support the role of PB MFC in the evaluation of patients with IgM monoclonal gammopathy and provide reliable information on correlation with molecular features. The development of a feasible MFC assay may stand as an objective tool in the classification of mature B cell neoplasms presenting with IgM monoclonal gammopathy.

Atypical Presentation of COVID-19 as Acute Leukemia: A Case Report

In the absence of respiratory system involvement, COVID-19 patients developing ARDS can clinically mimic other diseases including acute leukemia due to presence of atypical lymphocytes in peripheral blood smear and increased serum lactate dehydrogenase and serum uric acid levels. Herein, we report a case who was initially suspected to have acute leukemia based on his atypical symptoms without any respiratory system involvement and deranged laboratory parameters and finally, diagnosed with COVID-19. Our patient presented with fever, myalgia, gum bleed, and petechiae. On clinical and laboratory evaluation, he was suspected to have acute leukemia based on markedly deranged serum lactate dehydrogenase and serum uric acid and the presence of atypical cells in peripheral blood smear and bone marrow. On day 3 of hospitalization, he developed respiratory symptoms, breathing difficulty which progressed to ARDS, and subsequently, he succumbed to his illness. His real-time reverse transcriptase-polymerase chain reaction test for severe acute respiratory syndrome coronavirus-2 yielded positive results. Also, Flow cytometry and fluorescence in situ hybridization studies for leukemia workup did not show any abnormalities. Although we are reporting the findings of only a single case, we aim to enhance and contribute further to the understanding of this novel infection.

The frequency of occurrence of atypical lymphocytes in peripheral blood smears of natalizumab-treated patients with multiple sclerosis.

Aside from the extensive published data on immunophenotypic lymphocyte subsets in natalizumab-treated patients with multiple sclerosis (MS), an impact of natalizumab on lymphocyte morphology has rarely been studied. As patients treated with immunomodulating or immunosuppressive drugs are at risk for infectious disorders such as viral infections, knowledge of drug-derived changes in lymphocyte morphology may be beneficial in the diagnostic work-up in such clinical situations. This study aimed to determine the frequency of occurrence of atypical lymphocytes and defined subtypes of variant lymphocytes in natalizumab-treated patients with MS. We compared eight defined morphological lymphocyte subtypes in peripheral blood smears between 14 natalizumab-treated, 13 interferon-treated and 10 untreated subjects with relapse-remitting MS. Atypical lymphocytes were significantly enhanced in natalizumab-treated patients compared to the interferon and control group (P<.0001). Binucleated lymphocytes were restricted to the natalizumab group (P=.0058, P=.018), and plasmacytoid lymphocytes were more frequently found in the natalizumab group (P<.0001). Our data indicate that natalizumab enhances the fraction of atypical lymphocytes, and thereby especially the binucleated and plasmacytoid lymphocytes. Knowledge of these natalizumab-associated changes in lymphocyte morphology may be relevant in clinical routine, to avoid unnecessary diagnostic procedures or even a discontinuation of natalizumab treatment.

Chronic Lymphocytic Leukemia with Bi-Nucleated Lymphocytes

Introduction: B-cell chronic lymphocytic leukemia is one of the common lymphoproliferative disorders in the adult patient population. It is very uncommon to find bi-nucleated lymphocytes as a morphological feature in this disorder. Our patient was diagnosed with CLL and was found to have bi-nucleated lymphocytes in the peripheral smear. The mechanism behind this type of morphological feature of lymphocytes is unknown in CLL, and whether it has prognostic value on disease outcome is undetermined.Case Description: 62 y/o man was referred to hematology oncology after diagnosis of small cell lymphocytic leukemia was made s/p a right inguinal lymph node biopsy. His CBC revealed a wbc count of 14,000, Rbc count of 4,360, Absolute lymphocyte count of 11,500 and Platelet count of 125,000. The patient did not have any B-symptoms. On physical exam, a pertinent finding was palpable right axillary adenopathy. The CT of abdomen /pelvic to evaluate these findings. This revealed extensive axillary, abdominal/pelvic lymphadenopathy, hepatosplenomegaly and cardio phrenic lymphadenopathy.The patient had a biopsy of the right inguinal lymph node as well as bone marrow biopsy. Biopsy results showed small lymphocytic cells, some of which show occasional large nucleoli were consistent with small lymphocytic lymphoma/chronic lymphocytic leukemia, and morphologic characteristics of the lymphocytes showed bi-nucleated lymphocytes in peripheral blood smear (figure A). Flow cytometric analysis confirmed a lymphocytic population with lambda light chain restriction, expressing CD5, CD19, CD20, and CD23 consistent with chronic lymphocytic leukemia/small lymphocytic lymphoma. Bone marrow biopsy showed a hypercellular marrow with 75 % cellularity mainly composed of mature lymphocytes with scattered macrophages and eosinophils, Flow cytometric analysis (Clarient FI11-041053) of the bone marrow is interpreted as chronic lymphocytic leukemia/small cell lymphoma with the abnormal B cells representing 56% of the viable white cells. FISH study showed deletion of the ATM gene (11q22-23), D13S319 (13q14) and TP53 (p53) were observed in 29%, 71% and 35.5% of the cells analyzed, respectively. A subset of cells with the 13q deletion (20.5% of the total cells) showed homozygous deletion of D13S319 (13q14). ATM deletion is associated with progressive disease and poor prognosis in cases of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL).He did not have any other previous history of malignancy or hematologic disorder.Discussion:B-cell chronic lymphocytic leukemia is one of the common lymphoproliferative disorders in the adult patient population. To make a diagnosis requires absolute lymphocyte count >4x10 9 and lymphoid cell morphology. In CLL, leukemic cells are small and mature appearing lymphocytes, which have regular nuclear and cytoplasmic outlines and scant weakly basophilic cytoplasm. Surface markers that define a CLL cell are proteins such as antibody light chains (kappa or lambda) and CD proteins (CD5, CD19, CD20, and CD23). In our patient absolute lymphocyte count was 11.5x109 and lymphocytic population showed surface marker lambda light chain and CD proteins CD5, CD19, CD20 and CD23 which was consistent with CLL/SLL on inguinal lymph node biopsy, but morphology of lymphocytes was small and mature bi-nucleated lymphocytes, which is very uncommon. Although bi-nucleated lymphocytes are described in a disorder "Polyclonal chronic B-cell lymphocytosis with bi-nucleated lymphocytes". Detection of an extra chromosome for the long arm of chromosome 3 +i(3)(q10) has been considered a specific marker of Polyclonal B-cell lymphocytosis with binucleated lymphocytes (PPBL),which was not present in our case. One case study by Amouroux et al, included four patients with B-cell CLL who were found to have bi-nucleated lymphocytes. Disease course was stable in one patient, one patient had an indolent course and only one required treatment due to rapid doubling time of lymphocytes. Our patient initiated chemotherapy with Rituxan and Fludara, as he had progressive disease with hepatosplenomegaly, lymph nodes and bone marrow involvement.Conclusion:Bi -nucleated lymphocytes in B-cell CLL are very rare. Explanations as to the etiology of this morphological feature in B-cell CLL is unknown. There is no sufficient evidence that bi-nucleated lymphocytes in CLL has any impact on disease progression. DisclosuresNo relevant conflicts of interest to declare.

Sudden worsening of DRESS syndrome on tapering steroid dose with dramatic improvement on N-acetylcysteine and steroid dose escalation

Drug Reaction with Eosinophilia and Systemic Symptoms (DRESS) is a serious and life-threatening drug reaction characterized by fever, skin rash, peripheral eosinophilia, presence of atypical lymphocytes in peripheral blood smear, generalized lymphadenopathy and involvement of other body organs. The only agreed-upon therapy for DRESS in literature is withholding the culprit drug. There are no controlled trials on the use of steroids in this condition. Reports on the use of N-acetylcysteine in DRESS are very few. We report here a case of 36 year old female with severe DRESS that deteriorated rapidly upon tapering steroid therapy and dramatically improved on combination of pulse steroid therapy and N-acetylcysteine. This paper suggests that higher doses and gradual tapering of corticosteroid is a wise approach in treating patients with DRESS. It further emphasizes the possible beneficial role of N-acetylcysteine in this condition.

T-cell lymphoma presenting as drug rash with eosinophilia and systemic symptoms syndrome

Drug rash with eosinophilia and systemic symptoms (DRESS) syndrome is diagnosed by three criteria including cutaneous drug eruption, hematologic abnormalities, and systemic involvements. The hematologic abnormalities include presence of atypical lymphocytes or eosinophilia. The systemic involvements include lymphadenopathy, hepatitis, interstitial nephritis, interstitial pneumonia, or carditis. We experienced a 41-year-old female patient who presented DRESS syndrome at initial visit, but finally manifested with T-cell lymphoma. The patient complained of erythematous pruritic plaques with itching on her abdomen and thigh. There was no initiating factor and she was diagnosed with urticarial dermatitis. After treatment with antihistamine and systemic steroid, she recovered from skin lesion. However, 1 month later, she came to emergency department with aggravated skin lesion after taking nonsteroidal anti-inflammatory drug for 3 days. On admission, she showed a fever, skin rash, atypical lymphocytes in peripheral blood smear, and hepatitis. She was treated with systemic steroid under the impression of DRESS syndrome. Her symptoms began to improve, however, laboratory parameters were aggravated again. We performed bone-marrow biopsy because of her unusual progress. Finally she diagnosed with peripheral T-cell lymphoma and treated with allo-peripheral blood stem cell transplantation. In conclusion, we report a case of T-cell lymphoma which presented as DRESS syndrome. If patients with DRESS syndrome present lymphadenopathy and atypical lymphocytes, and do not respond to anti-inflammatory treatment, we should consider underlying lymphoproliferative disease. (Allergy Asthma Respir Dis 2013;1:280-283)

The Grade of Acute Thymus Involution in Neonates Correlates with the Duration of Acute Illness and with the Percentage of Lymphocytes in Peripheral Blood Smear

In various stressful conditions, the thymus is subjected to incidental involution, mostly due to the thymocytolytic effect of secreted glucocorticosteroids. The aim of this study was to examine acute thymic involution in sick neonates and to compare the morphological grade with some clinical and laboratorial parameters. The influence of the illness on thymus tissue was investigated in 100 neonates who were treated and died in a neonatology intensive care unit. The preterm infants (n = 73) were born before the 37th week of gestation. Analysis of 57 placentas showed inflammation in 32% and circulatory disturbances in 23% of the cases. The causes of death were confirmed by autopsy: 35 were preterm infants with respiratory distress syndrome without infection, 22 were malformed, and 10 had birth trauma or asphyxia. In contrast, 29 of the preterm infants had an infection, mostly pneumonia or sepsis, and 4 of the term infants had such infections. Acute thymus involution was histologically graded (0–4) according to the method of van Baarlen (see text). Resting state (grade 0) was found in 25 of 38 neonates who lived <12 h. In 13 of 38 neonates who lived <12 h, thymus involution suggested prenatal stress. The grade of thymus involution related to the duration of illness (p < 0.001). Placental inflammation was associated with features of thymus involution (p < 0.048). Infection as a cause of death was connected to advanced thymus involution (p < 0.001). In preterm newborns, infection was more often connected with acute thymus involution than was respiratory distress syndrome (p < 0.003). Among the parameters measured in all avaiable peripheral blood samples taken 24 h before death, only the lymphocyte count related to the grade of acute thymus involution (p < 0.05), with an increase in percentage of lymphocytes in peripheral blood smears from grade 0 to 2 and a decrease from grade 2 to 4. Although the white blood cell count is highly variable, a low percentage of lymphocytes might be a sign of advanced accidental thymus involution following acute stress.

Platelet Satellitism as Presenting Finding in Mantle Cell Lymphoma

Platelet satellitism surrounding polymorphonuclear neutrophils has been observed almost exclusively in EDTA-treated blood at room temperature. The mechanism underlying this phenomenon is not understood fully. We report a case of platelet rosetting around atypical lymphocytes in peripheral blood smears made from EDTA-treated and untreated blood. Flow cytometry of the peripheral blood sample and immunohistochemical stains of the subsequent bone marrow biopsy specimen revealed a monoclonal B-cell population positive for CD5, CD20, and cyclin D1 and negative for CD3 and CD23; cytogenetic findings revealed a complex karyotype that included t(11;14). These findings were consistent with mantle cell lymphoma. To our knowledge, the finding of platelet satellitism involving mantle cell lymphoma cells in peripheral blood has not been reported previously.

Image processing in pathology: XI. Light microscopic discrimination of human peripheral blood lymphocyte subpopulations

Non-labelled lymphocytes in peripheral blood smears and in smears of purified lymphocyte suspensions were differentiated in T- and non-T-cells (controlled by E-rosetting assay and surface immunoglobulin detection) by automatic image processing at the light microscopic level. By various classification methods it was found that these lymphocyte subpopulations differ in several visual and subvisual morphologic features. The morphologic differences between subpopulations were superimposed by individual deviations as well as by influences from the preparation. Single features were strongly affected by exogenic factors. Most of the different features are comparable with fine structural differences, as previously reported.

The Use of Salmonella Schottmulleri for Mapping and Separation of Human Lymphocyte Subpopulations

Human lymphocyte subpopulations (B cells, B 1, B 2, T 1, T 2, T 3, and T 4 cells; our denomination) have been previously identified and isolated by bacterial adherence and functional differences between them have been demonstrated. Here we examined the binding properties of Salmonella schottmulleri to human lymphocytes in peripheral blood smears and found that it binds to more lymphocyte subpopulations, namely B, T 1, T 2 and T 3 cells, than any bacteria previously tested. Thus, using only four bacteria: Salmonella schottmulleri, Brucella melitensis, Arizona hinshawii and Bacillus globigii we identified in blood smears B cells, two B and four T cell subpopulations. When we used gelatin-coupled monolayers of Sal. schottmulleri to isolate lymphocyte subpopulations, we showed that the nonadherent (T 4) cells could be efficiently separated from the adherent cells. Furthermore, we tested the isolated subpopulations for natural killing (NK) activity and for antibody-dependent cell-mediated cytotoxicity (ADCC). Using both NK and ADCC assays, we observed a significantly higher cytotoxic activity in the nonadherent cell population than in the unseparated or adherent cell populations. Also the nonadherent cells contained most of the lymphocytes that have receptors for the Fc portion of IgG and those cells described as large granular lymphocytes. We concluded that Sal. schottmulleri is a valuable new reagent for the identification and separation of human lymphocyte subpopulations.

'Salla Disease'

Severe mental retardation, coarse facial features, clumsiness, and speech failure were common findings in three brothers and one female third-cousin of a family from northern Finland. All the patients had vacuolated lymphocytes in peripheral blood smears, and electron microscopy of fresh skin biopsy specimens showed abundant cytoplasmic inclusions in various types of cells of the skin. Eight lysosomal hydrolases were assayed in peripheral blood lymphocytes and cultured skin fibroblasts, but no enzyme deficiency was detected. Urinary excretion of mucopolysaccharides, amino acids, glycoasparagines, and oligosaccharides was normal. Clinical findings, course of the disease, and the presence of cytoplasmic inclusions, indicating lysosomal storage phenomenon, suggest that the patients suffer from a genetic lysosomal storage disorder not described earlier. The eponym "Salla disease" was introduced, referring to the geographically restricted area where the family resides.