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Immune abnormalities have been reported in recipients of cellular and plasma blood products. To document the effect of current transfusion practices, we performed ex vivo lymphocyte immunophenotypic studies on patients with thalassaemia major who had received multiple (leucocyte-depleted) transfusions and patients with haemophilia A and B who had received heat viral-inactivated factor concentrates. Patients with thalassaemia major showed a significant lymphocytosis, with mainly B-cell changes consistent with ongoing B-cell stimulation associated with chronic exposure to red cell antigens. Reduced T-cell IL-2Rα expression would be consistent with inhibition by desferrioxamine chelation therapy. In contrast, patients with haemophilia showed predominantly T-cell changes. Patients with haemophilia A showed significantly elevated activated CD8+ cytotoxic T lymphocytes whereas those with haemophilia B showed an increase in CD8+CD11adim and CD4+CD45RA+ suppressor T cells. Several of the immune abnormalities found may be due to the presence of cytokines not removed by leucocyte filtration or destroyed by factor concentrate production (e.g. TGF-β) causing a T-helper-2-like response. The extensive lymphocyte characterization in this study has not previously been performed and has enabled a closer examination of the functional lymphocyte immunophenotypes seen in patients treated according to current transfusion practices. [ABSTRACT FROM AUTHOR]

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Regional lymph node lymphocytes from five patients with primary lung cancer were analyzed for subset composition, and exposed in vitro to the polyclonal human B cell mitogen Staphylococcus aureus Cowan I (SACI) or the murine B cell mitogen lipopolysaccharide (LPS) and then fused with mouse myeloma cells for investigation at the clonal level of their antibody (Ab) production and its statistical relation to the original subset composition. No correlation was found between the proportion of CD19+, CD23+, or CD3+ cells in the lymphocyte sample prior to its exposure to either SACI or LPS, and the Ab production efficiency, defined as the ratio of the number of Ab producing wells to the total number of proliferating wells. For lymphocytes exposed to LPS, however, a strong correlation (r = 0.931, p = 0.02) was observed between the Ab production efficiency and the ratio of CD8+ to CD3+ cells (CD8/CD3) in the original sample at least within the ranges studied (CD8/CD3 = 0.216–0.288). For those exposed to SACI, no correlation was found between the Ab production efficiency and the CD8/CD3 ratio (r = 0.881, p = 0.12) or the proportion of CD8+ cells (r = 0.808, p = 0.19) in the original sample. These results suggest that the repertoire of B cells responsive to LPS is different at least in part from the repertoire responsive to SACI and that the ratio CD8/CD3 could serve as a practical predictor for Ab production by human lymphocytes stimulated with LPS. [ABSTRACT FROM AUTHOR]

The age-related changes in proportion of various subsets within lymphocytes were investigated in cord blood and peripheral blood from healthy children and adults. The percentages of T and B cells did not show age-related changes, whereas natural killer (NK) cells increased significantly with age. Within lymphocytes or the CD3+ T cell population the proportion of CD45RAbright+ lymphocytes decreased and that of CD45RO+ cells increased, while that of CD45RAdim+ cells showed no age-related change. Within lymphocytes, the percentage of CD45RAbright+ CD4+ cells decreased, together with a decline of that of CD4+ cells. The proportions of CD45RAbright+ CD8+ cells and S6F1bright+ CD8+ cells increased with age and the age-dependent increase of the proportion of CD8+ cells seems to be mainly attributable lo the increases in these subsets. The CD45RAdim+ CD4+ and CD45RAdim+ CD8+ cells co-expressing CD45RO at a low level nevertheless showed no age-related changes. In γδ T cells, both δTCSI+ and δTCSl- T cells increased with age, but the δTCSl- γδ T cells increased more than the δTCSl+ subset. Among lymphocytes, the percentages of CD20+, CD21+ and CD22+ cells remained similar, with no age-related changes, but the proportion of CD5+ cells within lymphocytes or B cells decreased. The proportions of CD16+ NK cells among lymphocytes increased with age, and this change was attributable to the increase of CD56+ cells. [ABSTRACT FROM AUTHOR]

A case of OKT4 epitope deficiency referred for investigation with suspected immunodeficiency is described. Flow cytometry analysis of OKT4 epitope deficiency in a study group of healthy black children showed different manifestations of the lack of OKT4 epitope; a complete lack of OKT4+ CD4+ peripheral blood lymhocytes (PBL) with normal numbers of OKT4A+ and Leu-3a+ CD4+ PBL, decrease percentage OKT4+ CD4+ compared with OKT4A+ and Leu-3a+ CD4+ PBL, decrease fluorescent staining intensity with OKT4 and a biphasic OKT4 staining pattern associated with a reduced OKT4/Leu-3a ration. The percentage and fluorescent intensity of OKT4+ CD4+ PBL in the study group were significantly lower (P < 0.0001) than Leu-3a+ CD4+ and OKT4A+ CD4+ PBL. There is thus considerable risk of under-estimating the number of CD4+ cells in black South Africans if the OKT4 MoAb is used. [ABSTRACT FROM AUTHOR]

We examined sequential changes in the subsets of mononuclear cells infiltrating the pancreatic islets and splenic lymphocytes in pre-diabetic non-obese diabetic (NOD) mice, an animal model for type I diabetes. using immunofluorescent techniques. In the pancreas, a predominant infiltration by activated T lymphocytes, including helper inducer and cytotoxic suppressor T cells, was observed in the early stage of insulitis. Natural killer cells were also detected in the lesions, Immunoglobulin bearing cells tended to increase in number with the progression of insulitis. T lymphocytes were localized close to islet cells, while immunoglobulin bearing cells appeared adjacent to blood vessels and around T cell clusters. Immunoglobulin deposition or Ia expression on islet cells was not observed. The percentage of splenic T lymphocytes was markedly increased in the initial stage of insulitis as compared with control ICR mice and this elevated proportion of T cells continued throughout the observation period. As for splenic T cell subsets. cytotoxic suppressor T cells were increased in NOD mice. These results suggest that T lymphocytes play an important role in the initiation of insulitis long before the onset of overt diabetes. Moreover, NOD mice seem to have characteristic immunological features different from the RB rat or a reported case with human type 1 diabetes. [ABSTRACT FROM AUTHOR]

Marker expression was studied in rat lymphocyte populations isolated from parotid, submandibular and sublingual salivary glands. Comparative data were also obtained for lacrimal gland and spleen populations. Increased percentages of Thy-1+ cells were found in salivary gland populations when compared to spleen with the highest percentage noted for parotid gland. Thy-1 percentages in parotid gland were comparable to those obtained with lacrimal gland. Salivary gland sIg, CD5 and CD8 cell percentages were lower than those obtained for lacrimal gland and splenic populations. The percentages of CD4-bearing cells in submandibular and sublingual gland were lower than those found in parotid gland, lacrimal gland and spleen, and the CD4:CD8 ratios in parotid gland most closely approximated those in spleen. Increased percentages of Thy-1+ lymphocytes coexpressing sIg and CD5 were obtained for all salivary gland cell populations when compared to spleen; however, percentages of salivary gland cells bearing these 3 markers were lower than noted for lacrimal gland, which contained the highest percentage. With respect to adhesion molecules, lymph node homing receptor (LNHR) and Peyer's patch homing receptor (PPHR) bearing cells were found in all glandular populations with the percentages of LNHR+ exceeding PPHR+ lymphocytes. Homing receptor bearing populations were highest in spleen and were present in equal proportions. LFA-1 was expressed by all salivary gland cell populations in greater percentages than lacrimal gland, but lower than those in spleen. VLA-4 and CD44 expression was higher in parotid gland and spleen than in submandibular and lacrimal gland. These data show that the phenotypes of resident lymphocytes in salivary gland tissues differ from lacrimal gland and spleen, as well as each other, and indicate that Thy-1+ cells in glandular tissues bear both B and T cell markers. [ABSTRACT FROM AUTHOR]

The lymphocyte subsets in the peripheral blood and in liver biopsies from 4 patients with chronic hepatitis B obtained about 2-7 weeks before and after treatment with adenine arabinoside (Ara-A) were studied by a peroxidase-labeled antibody method using monoclonal antibodies against Leu-1, Leu-2a, Leu-3a, Leu-7 and Leu-10 antigens. In the peripheral blood, the percentage of Leu-2a + (cytotoxic/suppressor) cells was significantly reduced and the ratio of Leu-3a + (helper/inducer) to Leu-2a + cells was increased after the treatment with Ara-A. In the liver biopsies, the numbers of Leu-1 + (pan T) and Leu-2a + cells were significantly decreased after the treatment with Ara-A. As a result, the Leu-3a + /Leu-2a + ratio was significantly elevated in the liver after the therapy. The majority of lymphocytes distributed at sites of hepatocytic necrosis were positive for Leu-2a. The reduced numbers of Leu-1 + and Leu-2a + cells after the treatment were mainly due to the decrease of these cells infiltrating to the sites of hepatocytic necrosis. The numbers of other subsets (Leu-3a +, Leu-7 + and Leu-10 +) changed without any specific tendency both in the peripheral blood and in the liver biopsies after the treatment. With respect to viral replication, most of the patients showed a decrease of serum DNA polymerase activity or demonstrable intrahepatic HBsAg and HBcAg after the treatment. These data suggest that T cell-mediated cytotoxicity against HBV-infected hepatocytes is diminished after treatment with Ara-A. [ABSTRACT FROM AUTHOR]

To determine whether acute mental stress alters the composition of peripheral blood cells or components of the immune system, we determined blood cell counts and leukocyte differential counts, and examined lymphocyte subsets, before, during, and after 10 min. of mental arithmetic in 20 healthy female students. During mental stress the absolute number of leukocytes, lymphocytes, CD8+ cells, and CD16+ cells increased significantly, and the rate of CD8+ cell increase was higher than CD4+ cell increase. Therefore the CD4+/CD8+ ratio decreased significantly during mental stress. Erythrocyte counts, hemoglobin concentrations and packed cell volumes remained unchanged during and after mental stress. These results demonstrate that acute mental stress is associated mainly with rapid peripheral blood lymphocyte changes, including the release of CD8+ and CD 16+ cells into circulation. The absence of significant increases in erythrocyte counts, hemoglobin concentrations, and packed cell volumes during stress indicates that changes in lymphocyte composition cannot be explained by the hemoconcentration. Thus, acute mental stress has a detectable influence on peripheral blood cell counts and the immune system. [ABSTRACT FROM AUTHOR]

Lymphocyte subsets in the lymph nodes regional to papillary thyroid carcinoma were determined using flow cytometry to ascertain the differences in local immunological responses between elderly and young patients. Lymph nodes from age-matched patients with benign thyroid tumors were used as controls. No significant alterations in lymphocyte subsets were observed in the lymph nodes from the young patients regardless of whether metastasis was present, whereas those from the elderly patients showed significant decreases in pan T cell (CD2+, CD3+) and cytotoxic T cell (CD8+, CD8+CD11b−) populations, and a significant increase in B cells (CD19+) compared with those from both the young patients and the age-matched controls. These results indicate that local immunological alterations occur in elderly patients with papillary thyroid carcinoma, and we believe that immunological changes are one of the clinical characteristics of this tumor. [ABSTRACT FROM AUTHOR]

The effects of splenectomy on the development of cachexia, tumor growth and animal survival were studied in tumor-bearing CDF1 mice. Mice were inoculated with two subclones of colon 26 adenocarcinoma, clone 20 (with a potent capacity to induce cachexia) and clone 5 (without such activity), and underwent splenectomy before or after tumor inoculation. Splenectomy significantly prolonged the survival of mice bearing clone 20 when it was performed prior to tumor inoculation, although the progression of cachexia and tumor growth were not affected. The survival rate was higher in splenectomized than it was in nonsplenectomized mice 20-40 days after tumor inoculation. Such effects on survival were not observed, however, in mice splenectomized after inoculation with clone 20 or in mice that underwent splenectomy either before or after inoculation with clone 5. The decrease of peripheral blood lymphocyte count observed in mice bearing clone 20 was magnified when splenectomy was performed before tumor inoculation, but the serum levels of tumor necrosis factor and interleukin-6 were comparable. These results indicate that cancer death from cachexia is not directly attributable to enhanced catabolism. The mechanism by which splenectomy ameliorates the survival of cachectic mice remains to be studied, although several changes observed in the splenectomized mice after inoculation, including decreases in the peripheral blood L3T4 cells and Lyt-2 cells on the 9th day and 15th day respectively, and increase in the L3T4/Lyt-2 cell ratio on the 15th day suggest the involvement of the modified host's immune response. [ABSTRACT FROM AUTHOR]

Objectives: To explore the characteristics of immune function of healthy full-term infants at the age of 3 months, and to analyze the relationship of immune function with feeding pattern and sex., Methods: A total of 84 healthy full-term infants born in four hospitals in Beijing and Hohhot, China were prospectively recruited. Their feeding patterns remained unchanged within 4 months after birth. They were divided into a breast-feeding group and a milk powder feeding group according to their feeding patterns. At the age of 3 months after birth, peripheral venous blood samples of the two groups were collected to evaluate cellular immunity and humoral immunity and perform routine blood test. The laboratory indices were compared between infants with different feeding patterns and sexes., Results: Compared with the milk powder feeding group, the breast-feeding group had significantly lower proportion of T cell second signal receptor CD28, immunoglobulin M, and proportion and absolute count of neutrophils ( P <0.05) and significantly higher expression and proportion of HLA-DR, a surface activation marker of CD8 + T cells, and proportion of lymphocytes ( P <0.05). The male infants had a significantly lower white blood cell count and a significantly higher proportion of eosinophils compared with the female infants ( P <0.05)., Conclusions: Sex has no significant effect on the proportion of lymphocyte subsets in 3-month-old full-term infants, but feeding patterns are associated with the proportion of CD28 + T cells (lymphocyte functional subset) and HLA-DR + T cells (lymphocyte activation subset), suggesting that feeding patterns have a certain effect on the development of immune function in 3-month-old full-term infants.

Twenty-seven treatment-naïve patients with relapsing-remitting multiple sclerosis (MS) and 13 with neuromyelitis optica spectrum disorder (NMOSD) were enrolled during a time of acute flare-up. Common cerebrospinal fluid (CSF) features were increased CD29- and/or CD45RO-positive helper T cells capable of propagating inflammation in the central nervous system (CNS). B cell activation in the CSF was unique to MS, while an increase in CD4+CD192 (CCR2)+ cells in blood and breakdown of the blood-brain barrier (BBB) characterized NMOSD. Intravenous corticosteroid therapy suppressed neuroinflammation via modulation of cellular immunity in MS, as opposed to restoration of the BBB in NMOSD. [Display omitted] • B cell activation in cerebrospinal fluid (CSF) is unique to acute MS relapse. • Acute flare-up in NMOSD is characterized by breakdown of the blood-brain barrier. • Intravenous corticosteroids have effects on cellular immunity in the CSF in MS. • Intravenous corticosteroids improve NMOSD by restoring the blood-brain barrier. [ABSTRACT FROM AUTHOR]

Introduction  Acute myeloid leukemia (AML) is a heterogenous disorder consisting of clonal expansion of myeloblasts. Tumor immunity plays an important part in the pathobiology of AML. Understanding the components of tumor immunity is important for understanding tumor pathogenesis and the principles of immunotherapy. Methods  We studied 41 patients with AML, for total lymphocyte, CD4 positive helper T cells, CD8 positive cytotoxic T cells, and CD16/56 positive natural killer (NK) cells proportion. Quantification was done on bone marrow aspirate sample by flowcytometry. Whenever available, post induction bone marrow was also analyzed for the lymphocyte subset. Results  No significant difference was noted in the percentage of blasts among the three risk categories: favorable, intermediate, and adverse. However, there was significant difference in the total lymphocyte among the risk stratification groups, being highest in the favorable group and lowest in the adverse group. CD8 positive cytotoxic T cells were significantly less in Acute Promyelocytic Leukemia (APML) cases ( p  = 0.001). Total lymphocytes were, however, more numerous in APML ( p  = 0.005). NK cell proportion was not significantly different between APML and non-APML patients. On completion of induction chemotherapy, bone marrow samples for 12 patients could be processed for lymphocyte subset. On comparing the baseline against the post induction bone marrow, it was observed that there was significant increment in the proportion of CD4 positive T lymphocytes ( p  = 0.046). Conclusion  There is a difference in lymphocyte subset amongst patients with AML. Larger studies including functional aspects are needed to better define the role of lymphocytes in disease pathogenesis., Competing Interests: Conflict of Interest None declared., (The Indian Association of Laboratory Physicians. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial-License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).)

A letter to the editor is presented in response to the article about lymphocyte subsets by R. Schmidmaier and colleagues in the June 24, 2011 issue.

Background Rituximab is an anti-CD20 chimeric monoclonal antibody that depletes pre-plasma B cells. It is used in paediatric rheumatology for conditions including juvenile systemic lupus erythematosus (jSLE), juvenile dermatomyositis (JDM), vasculitides and refractory juvenile idiopathic arthritis (JIA). We aim to describe the cohort of patients treated with rituximab in the Scottish Paediatric and Adolescent Rheumatology Network (SPARN) in a 5-year period, and assess the need for a network guideline. Methods A review of paper and electronic case records. Patients seen within the SPARN network who received Rituximab between 2014 and 2018 were included. Results Vasculitis Rituximab was given to 16 patients in 5 different SPARN centres between 2014 and 2018. Male: Female ratio was 1:4.3. Mean age at first dose of rituximab was 12.75 years. Mean disease duration pre rituximab was 1 year. 8 (50%) patients had jSLE, 7 multisystem and 1 isolated lupus nephritis. 3 had ANCA-positive vasculitis and 5 had other conditions. (1 had RF+ve JIA; 1 severe panuveitis; 1 had an unspecified autoinflammatory condition; 1 had COPA syndrome with severe interstitial lung disease and 1 had autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED). Documentation of pre-rituximab checks was poor. Baseline bloods were done in all 16 patients. 11 patients had lymphocyte subsets and 13 immunoglobulins documented. Disease-specific antibodies were measured in all patients in whom applicable (11). 11 had a chest x-ray prior to rituximab. In 9 cases a pre-rituximab pregnancy test was indicated: none were documented. In only 3 patients was a pre-rituximab history of recent infections documented. In 8 patients a discussion regarding treatment risks and benefits was documented. Post-infusion lymphocyte subset monitoring was variable but documented in 15. 1 patient was never B cell deplete. 9 never fully reconstituted. RTX dose information was available for 14. All received doses of 750mg/m2 with 6 having the dose capped at 1g. Dose frequency information was available for all. The majority (9) received 2 doses. Of the rest, 1 patient received a single dose. 3 received 3 doses, 2 had 4 and 1 had 5. All patients in whom information was available, received pre-medication with paracetamol, chlorphenamine and methylprednisolone (n = 14). 4 patients had side effects noted (rash/itch in 1, anaphylaxis in 1, facial flushing in 1 and post-infusion infection in 1). Clinical response was clearly documented in 8 patients, 7 of whom showed clinical improvement (2 had systemic jSLE, 2 had ANCA +ve vasculitis, 1 had isolated lupus nephritis, 1 had COPA syndrome and 1 had RF+ JIA). Conclusion Rituximab is used for a range of conditions and in a number of centres within SPARN. This review highlights variation in and poor documentation of pre-rituximab discussions and investigations, prescribing, monitoring and response. A SPARN rituximab protocol is being developed to address this. Conflicts of Interest The authors declare no conflicts of interest. [ABSTRACT FROM AUTHOR]

Background IgG4-related disease (IgG4RD) is an immune-mediated fibroinflammatory condition characterized by the infiltration of IgG4-carrying plasma cells and storiform fibrosis in most of the tissues. The condition is reported to cause multisystem involvement, however salivary gland is the most commonly affected organ with IgG4-related sialadenitis. Raised IgG4 concentrations in the serum and prominent infiltration by plasmacytes expressing IgG4 in the lacrimal and salivary glands have been confirmed. Methods We conducted a retrospective case review of a 10 year-old girl who was diagnosed with IgG4-RD at Great Ormond Street Hospital for Children in 2017. Patient's clinical, laboratory and imaging data was extracted from our database and literature search was done to find out different manifestations of the IgG4-RD in Children. Results 10 year-old-girl with lacrimal and salivary gland swelling, sicca symptoms and fatigue. Ultrasound scan neck revealed multiple small lymph nodes and enlargement of both submandibular glands (Picture1) Salivary glands also appeared bulky and heterogenous with multiple small hypoechoic focci. Appearances likely to represent sialoadenitis and there was no evidence of malignancy or lymphoma. USS abdomen normal. Full blood count, routine biochemistry and urine microscopy normal. Autoantibodies came back negative (ANA: Negative, ANCA: Negative, Anti-Ro and Anti-La: Negative, RF:Negative, Thyroid autoantibodies:Negative) IgG level elevated (in repeated samples). IgA, IgM and IgE levels normal. IgG subgroups revealed significantly elevated IgG4 levels (21.49 Normal range: 0-1.1) IgG1, IgG2 and Ig G3 levels elevated as well. Lymphocyte subsets were normal. The biopsy of salivary gland: Chronic inflammation with IgG4 staining, suggestive of IgG4 related disorder. Lung function test showed decreased DLCO at 76 %, FEV1 at 82% and FVC at 89% suggestive of interstitial lung involvement. CT Thorax: Multiple abnormalities including moderate lymphadenopathy, renal parenchymal lesions and interstitial lung abnormality. MRI Abdomen: T2 hypointense renal foci, suggestive of infiltration as part of the IgG4 related disease. Diagnosed with IgG4 related disease. Treatment started with intravenous methylprednisolone followed by Anti-CD20 (Rituximab) therapy and a weaning plan for steroids. Mycophenolate mofetil commenced for the maintenance therapy. Patient has shown good response with clinical and lung function improvement. Conclusion IgG4-RD is a rare condition which can cause multisystem involvement with the infiltration of IgG4-bearing plasma cells in the tissues. We wanted to emphasize that this condition could also be seen in the paediatric population. We also wanted to highlight that after further investigations multisystem involvement can be discovered. Steroids are mainstay of the treatment with Anti-CD20 medication (Rituximab) and steroid sparing agents such as mycophenolate mofetil could be the choice for maintenance therapy. However, further studies need to be done in paediatric age group. Conflicts of Interest The authors declare no conflicts of interest. [ABSTRACT FROM AUTHOR]

Background: The prognostic role of peripheral lymphocyte subsets in early stage of acute pancreatitis (AP) is unknown., Methods: After enrollment, blood samples were collected in the first 24 hours of the onset of AP in 164 patients. The lymphocyte count and the percentage of CD3 + , CD3 + CD4 + , CD3 + CD8 + , CD3 - CD19 + and CD3 - CD (16 + 56) + cells were assessed., Results: Reduction of the lymphocyte count and the percentage of CD3 + CD8 + and CD3 - CD (16 + 56) + cells within 24 hours of the onset of AP as well as an increase in the percentage of CD3 + and CD3 + CD4 + cells were observed. The percentage of CD3 - CD (16 + 56) + cells had the highest accuracy in prediction of AP severity., Conclusions: Marked changes of the percentages of lymphocyte subset were found in peripheral blood in the early stage of AP. The percentage of CD3 - CD (16 + 56) + cells in peripheral blood could be a potent prognostic predictor., (Copyright © 2019. Published by Elsevier Inc.)

In this report we have analysed the peripheral blood lymphocyte of several patients with chronic hepatitis B virus infection with flow cytometry. Based on the presence and absence of the HBeAb, patients were divided into two groups. In both, all the patients were HBsAg positive with normal range of serum alanine aminotranferase (23.9 ± 17.8). We have found that the immunophenotypic profiles of patients were different from healthy donors with significant decrease in CD3+ T cells, specially CD8+ T cells and a siginificant increase in the CD19+ B cells. The differences were seen in other subset of T cells (CD4+) or NK cells (CD56+/CD16+ ) and HLA-DR markers were not significant. When the phenotypic profiles of both groups were compared with each other, such changes were more dominant in group II, with HBeAb positive than in group I, with HBeAb negative. Also, we have seen a correlation between the increase of CD19+ B cells and the decrease of CD3+ T cells. No such correlation was observed with other cells.