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Cellular composition including lymphocyte subsets in preterm and full term human colostrum and milk.

The cellular composition of colostrum (within 72 hours post partum) and mature milk samples (5th-7th day post partum) from 20 mothers delivering preterm babies and 20 mothers delivering full term babies was examined. Lymphocyte subsets including T cells, B cells, T4 and T8 cells were quantitated by indirect immunoperoxidase staining using specific monoclonal antibodies. The mean total cell count in preterm colostrum (9,338/mm3) was significantly higher than in full term colostrum (5,594/mm3). Similarly, counts for macrophages, neutrophils and lymphocytes were significantly higher in preterm colostrum and milk as compared to full term colostrum and milk. B and T lymphocytes including T4 and T8 cells were identified in both preterm and full term breast milk. The absolute count of T, B, T4 and T8 cells was significantly higher in preterm colostrum compared to term colostrum, though the relative percentage distribution of lymphocyte subsets showed no significant difference between the two groups. T and B cells constituted about 74% and 26% of total lymphocytes, respectively in preterm as well as full term colostrum and about 69% and 31% in preterm and full term milk. The mean T4/T8 ratio was higher in preterm colostrum than term colostrum, being 1.82 and 1.60 respectively. However, this difference was not significant statistically and did not change statistically in the milk sample.

B-Lymphocytes↗

Lymphoid organ development in rabbits: major lymphocyte subsets.

Although rabbits represent an important animal model, little is known about the lymphoid organ development in this species. In the present study, lymphocyte subsets in peripheral blood, spleen, mesenteric and popliteal lymph nodes in newborn and 2-, 4-, 6- and 8-week old and adult were characterized. Lymphocyte subsets were detected using flow cytometry and monoclonal antibodies against rabbit CD4, CD8, T-cell-specific antigen and cross-reactive antibody against B-cell antigen CD79alpha. In neonates, lower numbers of T cells were detected in both peripheral blood and spleen than in mesenteric lymph nodes. In comparison with other compartments, CD79alpha(+) cells prevailed in the spleen. Post-natal development was characterized by a decreased CD4(+)/CD8(+) ratio due to increasing frequency of CD8(+) lymphocytes in all organs but mesenteric lymph nodes, where it was due to decreased numbers of CD4(+) lymphocytes. Another significant feature was the increase of B cells in peripheral blood and mesenteric lymph nodes.

Animals↗

Lymphocyte subsets in bronchoalveolar lavage fluid and in circulating blood in epidermoid bronchogenic carcinoma.

Alveolar macrophages, lymphocyte and granulocyte percentages, together with OKT3+, OKT4+, OKT8+ lymphocyte subsets and OKT4+/OKT8+ ratio, were evaluated in bronchoalveolar lavage (BAL) fluid and in peripheral venous blood (PVB) of neoplastic and nonneoplastic subjects, in order to assess these aspects of immunity in neoplastic disease and to find out if the modifications in the bronchoalveolar environment are correlated to the ones in the circulation blood. BAL was performed in 30 patients undergoing fiberoptic bronchoscopy to ascertain the presence of lung cancer. Twelve of them had positive findings for epidermoid bronchogenic carcinoma, while in the remaining subjects the diagnosis was not confirmed. The 30 examined subjects were then grouped according to their smoking habit. In PVB, no significant difference was seen between neoplastic and nonneoplastic subjects, whereas in BAL the neoplastic patients showed a significant increase of lymphocytes OKT3+ and OKT8+. This tends to confirm that PVB is not a good indicator of organ immunity and may justify the reduced activity of alveolar macrophages in subjects affected by bronchogenic neoplasia. Between smokers and nonsmokers, lymphocyte subsets showed more significant differences than between neoplastic and nonneoplastic subjects (decrease of T4+ lymphocytes, increase of T8+ lymphocytes and, therefore, reduction of T4/T8 ratio); there were also scalar variations in the three groups (smokers with cancer, smokers without cancer and nonsmokers without cancer). Thus, the possible autonomous role of cigarette smoke and the presence of neoplasia in the immunity alterations of the alveolar environment with final joint effects were confirmed. These data may indicate a possible correlation between cigarette smoking, immunological alterations in BAL and the onset of bronchogenic carcinoma.

Adult↗

Endometrial T-lymphocyte subset infiltration during the ovine estrous cycle and early pregnancy.

T-lymphocytes were quantitated within luminal, stromal and glandular areas of ovine endometrium. In experiment 1, ovariectomized (OVX), estrus (E) and day 13 (D13) ewes (six/group) received 500 micrograms of phytohemagglutinin (PHA) or vehicle in ligated right and left uterine horns, respectively. At 48 h, uteri were removed for the immunohistochemical evaluation of T-lymphocyte subsets. In experiment 2, T-lymphocytes were quantitated within non-pregnant and pregnant uterine horns on day 19. For experiment 1, mean numbers of T4 and T8 lymphocytes within luminal and stromal areas of PHA-treated horns were greatest (P less than 0.05) for D13 ewes and least (P less than 0.05) for E ewes. Numbers of T6 lymphocytes for these same areas were greatest (P less than 0.05) for PHA-treated horns of OVX ewes. Overall, the T4/T8 ratio (P less than 0.004) and mean number of T19 cells (P less than 0.009) were increased by PHA. Numbers of CD45R lymphocytes were not affected by PHA but were greater (P less than 0.05) in glandular and luminal than stromal areas. For experiment 2, mean numbers of endometrial T4, T6, T8 and T19 lymphocytes were similar (P greater than 0.05) between non-pregnant and pregnant horns; however, the number of CD45R lymphocytes was greater (P less than 0.05) in endometrial tissue of pregnant than non-pregnant horns. The data indicate that the in vivo response of specific ovine T-lymphocytes to PHA was generally dependent upon reproductive stage and the presence of conceptus tissue influenced the infiltration of CD45R lymphocytes.

Animals↗

Immunohistochemical study of T lymphocyte subsets on frozen substituted plastic embedding section of the bone marrow from patients with myelodysplastic syndrome and its clinical implication.

An immunohistochemical study of T lymphocyte subsets on frozen substituted plastic embedding bone marrow sections obtained from 10 patients with myelodysplastic syndrome (MDS) was presented. The results of qualitative and quantitative immunohistochemical analysis are as follows: (1) Labile antigens of T lymphocytes were well preserved, thus allowing analysis of distribution of T lymphocyte subsets in situ: (2) the average number of T3, T4 and T8 lymphocyte of the diffuse infiltrate was about 2%, 0.4%, 0.5%, respectively, of all nucleated cells in bone marrow, and T4/T8 of T cells were below 1.0 in patients with MDS; (3) there were cases of RAS showing T lymphocyte aggregation in bone marrow, but no patient exhibited progressive refractory anemia with excess of blasts (RAEB) and RAEB in transformation (RAEBT). These findings indicated that the immunological abnormalities are of importance in the evaluation of pathogenesis and prognosis of MDS.

Aged↗

Hierarchical and redundant lymphocyte subset control precludes cytomegalovirus replication during latent infection.

Reactivation from latent cytomegalovirus (CMV) infection is often associated with conditions of immunosuppression and can result in fatal disease. Whether the maintenance of systemic CMV latency is mainly governed by factors of the infected cell or by immune control functions is unknown. Likewise, the putative immune control mechanisms which could prevent the induction and spread of recurrent CMV infection are not clearly identified. We took advantage of latently infected B cell-deficient mice and a sensitive method for virus detection to study CMV reactivation after ablation of lymphocyte subsets. A crucial role of both T lymphocytes and natural killer (NK) cells was demonstrated. Within 5 d after depletion of lymphocytes, productive infection occurred in 50% of mice, and 14 d later 100% of mice exhibited recurrent infection. A hierarchy of immune control functions of CD8(+), NK, and CD4(+) cells was established. Reactivation was rare if only one of the lymphocyte subsets was depleted, but was evident after removal of a further subset, indicating a functional redundancy of control mechanisms. The salivary glands were identified as the site of most rapid virus shedding, followed by the detection of recurrent virus in the lungs, and eventually in the spleen. Our findings document a previously unknown propensity of latent CMV genomes to enter productive infection immediately and with a high frequency after immune cell depletion. The data indicate that only the sustained cellular immune control prevents CMV replication and restricts the viral genome to a systemic state of latency.

Animals↗

Influence of bone marrow graft lymphocyte subsets on outcome after HLA-identical sibling transplants.

OBJECTIVE: The aim of this study was to analyze bone marrow lymphocyte subsets and CD34 cell dose and their influence on the outcomes of bone marrow transplantation. MATERIALS AND METHODS: Forty-eight patients (median age 30 years, range 5-54) receiving HLA-identical sibling bone marrow transplantation for hematologic malignancies were analyzed. RESULTS: Median number (range) of nucleated cells and CD34+ cells infused were 2.4 (0.4-6.0) x 10(8)/kg and 3.5 (0.5-13.0) x 10(6)/kg, respectively. Probability of neutrophil recovery was 97%. In a multivariate analysis, time to neutrophil recovery was shortened when a higher number of CD3/CD8 cells was infused (> or =1.0 x 10(7)/kg) (hazard ratio [HR] = 2.13, p = 0.018); when the patient was female or had negative cytomegalovirus serology (HR = 2.03, p = 0.03; HR = 0.41, p = 0.009; respectively). The incidence of grade II to IV acute graft-vs-host disease (GVHD) was 47%. Infusion of >1 x 10(7) CD4 infused/kg increased the risk of acute GVHD (HR = 2.86, p = 0.03). Nineteen of 40 patients at risk experienced chronic GVHD, the risk of which was increased by diagnosis of chronic leukemia (p = 0.03), <2.0 x 10(8) nucleated cells infused/kg (p = 0.05), and a low number of all lymphocyte subsets, except CD19. Estimated 3-year survival rate was 54%. Risk of death was increased in patients receiving <3.5 x 10(6)CD34 infused/kg (HR = 0.37, p = 0.02). Only six patients relapsed. CONCLUSIONS: A high cell dose of CD3/CD8 is associated with faster neutrophil recovery, whereas a high cell dose of CD4+ cells increases the incidence of acute GVHD. A high number of nucleated cells and CD34+ cells infused was associated with decreased risk of chronic GVHD and improved survival, respectively.

Acute Disease↗

Long-term effects of thyroid hormone on lymphocyte subsets in spleens and thymuses of mice.

We examined the effects of the long-term administration to mice of thyroid hormone or propylthiouracil (PTU) on lymphocyte subsets in spleens, and thymuses to clarify whether hyperthyroxinemia itself causes the changes in lymphocyte subsets, such as the marked increase in CD5+ B cells and decrease in natural killer (NK) cells, observed in hyperthyroid Graves' disease. Both the number and proportion of splenic NK (Thy-1+ asialo GM1+) cells were increased in hyperthyroxinemic mice treated with thyroxine (T4) for both short and long terms (8 and 32 weeks, respectively), those of splenic and thymic T (CD5+ sIgM-) cells and CD5-B cells were increased only in hypothyroxinemic mice treated with PTU for 32 weeks, compared with those in euthyroid mice. These data indicate that 1) long-term hyperthyroxinemia increases splenic and thymic T cells and splenic NK cells, but not CD5+ B cells, in mice, 2) long-term hypothyroxinemia induced by PTU treatment increases splenic B cells and CD5- B cells, and 3) hyperthyroxinemia itself does not cause the changes in CD5+ B cells and NK cells, which are observed in hyperthyroid Graves' disease, in mice.

Animals↗

Role of transfusion-mediated viral infections on the lymphocyte subset profile in multi-transfused children.

A total of 74 multitransfused (MT) children of beta thalassaemia major were analysed for prevalent viral markers transmitted through transfusion. A higher incidence of serological markers for Hepatitis B virus (HBV), cytomegalovirus (CMV) could be observed in the group of MT children compared to control group. There was a significant trend (chi 2 = 33.4; P < 0.001) in the increase in prevalence of viral markers along with the increase in the number of transfusions. MT children receiving more than 50 transfusions were found to have evidence for at least one or multiple viral infections transmitted through blood. Children receiving more than 50 transfusions were characterized by marked alteration of T3, T8, and B cells while T4/T8 ratio was found to be significantly decreased (P < 0.001) only in the group of children receiving more than 100 transfusions. Relative assessment of the alteration of lymphocyte subsets in various groups of viral infection showed that cases with CMV IgM to have more marked influence on the alteration of T8 cells, T4/T8 ratio, and B cells compared to other groups of viral infections. Reassessment of the lymphocyte subset profile in MT children in the light of CMV IgM positive cases revealed that in children receiving more than 50 transfusions significant alterations of lymphocyte subjects were influenced by the presence of CMV IgM positive cases in these groups. Our study points out that the correlation between the alteration of lymphocyte subset profile and number of transfusion in MT children need to be reassessed in the light of acute CMV infection in the form of CMV IgM.

Acquired Immunodeficiency Syndrome↗

Elevated levels of serum soluble E-selectin in patients with chronic hepatitis B: Correlation with T lymphocyte subsets, NK cells and liver inflammation.

BACKGROUND: It has been reported that serum soluble E-selectin (sE-selectin) may be increased in some inflammatory liver diseases. The purpose of our study was to investigate changes of sE-selectin, T lymphocyte subsets, natural killer (NK) cells, and HBV load in patients with chronic hepatitis B (CHB), analyze the relationship between them, and discuss their significances. METHODS: Fifty-four patients with chronic hepatitis B and fourteen controls were studied. Serum sE-selectin, T lymphocyte subsets, NK cells, and hepatitis B virus (HBV) load were detected by enzyme-linked immunosorbent assay (ELISA), flow cytometry (FCM), and fluorescence quantitative polymerase chain reaction (FQ-PCR), respectively. RESULTS: sE-selectin level in patients with CHB increased significantly than that in controls (p<0.01). The percentages of CD4 positive cells and NK cells decreased while the percentage of CD8 positive cells increased significantly in CHB patients than in controls (p<0.01, respectively). sE-selectin level significantly related to levels of T lymphocyte subsets, NK cells, serum alanine aminotransferase (sALT) and aspartate aminotransferase (sAST) (p<0.01, respectively), but had no relationship with HBV level. CONCLUSION: The sE-selectin levels in patients with chronic hepatitis B increase significantly and correlate to liver inflammation, suggesting that sE-selectin can be considered as an additional useful marker of CHB inflammatory activity, and E-selectin may play part of role in pathogenesis of chronic hepatitis B.

Journal Article↗

Hyperglobulinemia and lymphocyte subset changes in naturally infected, inapparent carriers of equine infectious anemia virus.

OBJECTIVE: To determine blood protein concentration, immunoglobulin concentration, and lymphocyte profiles in equine infectious anemia virus (EIAV) seropositive, naturally infected horses without clinical signs of disease. ANIMALS: 26 clinically normal seropositive horses, 6 febrile ponies with experimentally induced EIA, and 52 clinically normal seronegative horses and ponies. PROCEDURE: Serum and EDTA-anticoagulated blood were obtained from all horses and ponies, and total serum protein and albumin concentrations, immunoglobulin concentrations, and blood lymphocyte subset counts were determined. RESULTS: Compared with seronegative horses, EIAV seropositive inapparent carrier horses had no significant difference in serum reverse transcriptase activity, PCV, or platelet count. Inapparent carrier horses had increased plasma total solids and serum globulin concentrations and decreased serum albumin concentration and albumin-to-globulin ratio. Total serum immunoglobulin and serum IgM concentrations were increased. Inapparent carrier horses had significantly decreased percentages of CD5+ and CD4+ blood lymphocytes. CONCLUSIONS: Serum protein and lymphocyte subset changes in EIAV-infected inapparent carrier horses are consistent with immune activation or chronic inflammation, both of which may, in part, be the result of virus-induced polyclonal B-cell activation. CLINICAL RELEVANCE: EIAV seropositive horses have immune-related abnormalities consistent with ongoing viral activity regardless of the duration they have been infected, even when the usual signs of disease (anemia, fever, weight loss) are not apparent.

Animals↗

Age-related reference intervals for lymphocyte subsets in whole blood of healthy individuals.

Enumeration of various lymphocyte subsets is used widely in the diagnosis and monitoring of various disease states. With the development of flow cytometric technology and whole blood analysis, methodologies have become more sensitive. It is therefore important to establish reference intervals in normal, healthy individuals using these techniques to give a better indication of the border between health and disease. Since some lymphocyte subpopulations are known to change with age, we have enumerated common subsets in healthy individuals from all decades of adult life, including nonagenarian subjects. We report reference intervals for these subsets in each age group, which will be of use in diagnosis and disease monitoring, particularly in elderly subjects, the most rapidly expanding group within the population today.

Adolescent↗

Restraint stress-induced changes in lymphocyte subsets and the expression of adhesion molecules.

The effect of restraint stress on the distribution of lymphocyte subsets were studied in young BALB/c male mice. Loss of whole body weight, a reduction in the weights of spleen and lymph nodes, and higher levels of serum corticosterone were evident after a single continuous restraint period of 16-18 h. Tissue sections of spleens from restrained animals revealed erythrocyte depletion in the contracted red pulp. Furthermore, there was a significantly higher proportion of CD4+, but not of CD8+, lymphocytes in the spleen. The proportion of the CD4+ subset was markedly diminished in peripheral blood, whereas no changes were detected in lymph nodes. Restraint resulted in enhanced allogeneic mixed lymphocyte reactivity and in altered expression of some CD4+, but not CD8+, splenocyte adhesion molecules (CD44, LFA-1 and VLA-4). Removal of circulating corticosteroids by surgical adrenalectomy abolished the restraint-induced changes in lymphocyte adhesion molecule expression. The findings suggest that the observed differences in lymphocytes subset distribution of lymphoid organs may be due to changes in the pattern of adhesion molecule expression.

Adrenalectomy↗

[Effect of Astragalus membranaceus on T-lymphocyte subsets in patients with viral myocarditis].

The efficacy of Astragalus membranaceus (AM) oral liquor combined with routine therapy and routine therapy alone on T-lymphocyte subsets of peripheral blood in viral myocarditis patients have been studied. The results showed that the T-lymphocyte subsets profile and OKT4/OKT8 ratio of peripheral blood were significantly lower in viral myocarditis patients than that in healthy control (P < 0.05, 0.01). Routine therapy combined with AM could significantly enhance OKT3, OKT4 and OKT4/OKT8 ratio in the above-mentioned patients (P < 0.05, 0.01). The possible pharmacodynamic mechanism of improved cell immunity in viral myocarditis patients by combining routine therapy with AM was discussed.

Adolescent↗

Alterations of lymphocyte subsets in children of diabetic mothers.

To investigate the impact of diabetic mothers on the maturation of the immune system in their offspring, immunophenotypic markers of major lymphocyte subpopulations were evaluated by two-colour flow cytometric analysis in 160 healthy children of diabetic mothers (100 with insulin-dependent diabetes mellitus (IDDM): 48 with gestational diabetes), including 22 neonates, 45 infants aged 8-12 months, 46 children aged 1-2 years, 29 children aged 3-6 years and 18 children aged 7-17 years. Results were compared with 21 neonates of healthy mothers from our hospital and with 110 paediatric subjects of a reference population. In neonates of diabetic mothers, percentages of total lymphocytes (p = 0.044), T and B lymphocytes (p = 0.004, respectively) were significantly decreased compared to our neonates of healthy mothers. By subdividing the group of neonates in offspring of mothers with IDDM (n = 15) or gestational diabetes (n = 7), differences compared to normal neonates were mainly observed in neonates of mothers with IDDM (T lymphocytes: p = 0.006; B lymphocytes: p = 0.008). In cord blood, 45.5% of neonates had antibodies to islet cells, insulin or glutamic acid decarboxylase, most likely transmitted through the placenta of the diabetic mother. No association was found between alterations of lymphocyte subsets and antibody-positivity in cord blood, nor was there any correlation of lymphocyte counts and mean HbA1 during pregnancy, maternal age at delivery, diabetes duration, or neonatal birth weight, respectively. Comparisons among age groups from newborn infants through adolescents revealed higher percentages of total lymphocytes and lower percentages of activated T cells in children of diabetic mothers compared to children of the reference population between the age of 1 to 6 years (67-73% of the cases above and 62-77% below the interquartiles of the reference range, respectively). No significant differences in lymphocyte subpopulations between children of mothers with IDDM diabetes and gestational diabetes have been detected. In addition, there were no abnormalities of lymphocyte subsets in children who are at high risk for the development of IDDM. In summary, we suggest that the observed changes in children of diabetic mothers may reflect a cellular immune reaction to the particular maternal environment, characterized by both an abnormal metabolic state and persisting autoimmunity in the affected mother.

Adolescent↗

Age-related quantitative alterations in lymphocyte subsets and immunoglobulin isotypes in healthy horses.

OBJECTIVE: To characterize age-associated changes in lymphocyte population subsets and immunoglobulin isotypes. ANIMALS: 30 healthy young light-breed horses (5 to 12 years old) and 30 healthy aged light-breed horses (> 20 years old). PROCEDURE: Lymphocyte subset populations were identified, using monoclonal antibodies to cell surface markers CD5, CD4, CD8, and IgG. Subset populations were quantitated by use of flow cytometric analysis of antibody-stained cells. Serum immunoglobulin concentration was determined using single radial immunodiffusion. RESULTS: Absolute cell counts of total lymphocytes, T cells, CD4+ and CD8+ T cells, and B cells were decreased in aged horses, compared with young horses. There was a significant decrease in the percentage of CD8+ cells and an increase in the CD4+-to-CD8+ cell ratio in the aged population, compared with young horses. However, serum concentration of IgG, IgG(T), IgM, or IgA did not differ with age. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, total lymphocyte count and lymphocyte subset cell counts decrease with age. Age-matched control values are necessary for optimal evaluation of hematologic variables in aged horses. The decrease in lymphocyte subset cell counts in healthy aged horses mimics that seen in other species and may contribute to an age-associated decrease in immunocompetency.

Age Factors↗

Characterization of circulating T-lymphocyte subsets in alcoholic liver disease by monoclonal antibodies: correlation with the absolute lymphocyte count.

Peripheral blood T-lymphocytes from 27 patients with alcoholic liver disease were assessed on the basis of their ability to react with OKT monoclonal antibodies. A significant positive correlation was observed between the absolute numbers of T-lymphocytes and suppressor/cytotoxic T cells and the absolute lymphocyte count. These data suggest that quantitative changes in T-lymphocyte subsets in alcoholic liver disease do not necessarily reflect an imbalance in T immunoregulatory mechanisms.

Antibodies, Monoclonal↗

Characterization of peripheral blood T lymphocyte subsets in pulmonary sarcoidosis.

The peripheral blood of sarcoidosis patients has been shown to have abnormalities in the T Lymphocyte subsets. There is a consistent reduction in the helper/suppressor T Lymphocyte ratio caused by an increase in suppressor cell percentage as determined by monoclonal antibodies. These findings suggest that the peripheral blood T Lymphocyte subsets may be a useful adjunct for determining the underlying immunological cellular dysfunction in sarcoidosis patients.

Adult↗