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PubMed1977

Discrimination of B, T and null lymphocytes by esterase cytochemistry.

A technique has been devised which optimally demonstrates non-specific esterase activity in human blood lymphocytes. The reaction is carried out on smears fixed with formalin vapour, using alpha-naphthol butyrate as substrate at pH 8 and at a low concentration for a short incubation period. The pattern of esterase activity revealed by this method provides a discriminating marker for mature T-lymphocytes, which show dense, localised, dot-like positivity, and a probable marker for 'null' cells in the form of scattered granular positivity. B cells appear to be negative. Monocytes show a clearly different pattern of granular positivity.

Acid Phosphatase↗
PubMed1980

Human peripheral null lymphocytes. II. Producers of type-1 interferon upon stimulation with tumor cells, Herpes simplex virus and Corynebacterium parvum.

Human blood lymphocytes, exposed for 6 to 24 h in vitro to tumor cells (K 562, IGR3, L1210), Herpes simplex virus type 1 (HSV) or Corynebacterium parvum (CP), produced high levels of anti-viral activity which was identified as type-1 interferon (IF). In mixed lymphocyte tumor cell cultures (MLTC), the generated type-1 IF was definitely shown to originate from the lymphocytes and not from the tumor cells. Supplementation of leukocyte cultures with 10% fetal calf serum instead 10% human AB serum had little influence on tumor cell-induced IF production, but strongly reduced CP-induced IF production. Lymphocyte fractionation procedures involving iron/plastic treatment, nylon wool columns, Ig-anti-Ig columns and rosette (E, EA) separation led to the identification of null cells as highly efficient producers of type-1 IF. T cells obtained by different ways (E-rosette sedimentation, passage through 1 nylon and 2 Ig-anti-Ig columns, or thoracic duct lymphocytes) were poor IF producers in response to tumor cells, HSV and CP, but secreted anti-viral activity when stimulated with phytohemagglutinin. In MLTC, the level of generated type-1 IF roughly stimulated with phytohemagglutinin. In MLTC, the level of generated type-1 IF roughly paralleled nautral killer (NK) cell activity. Evidence is presented that type-1 IF can be produced by an Fc receptor-negative null cell subset, whereas NK activity requires Fc receptor-positive cells. It is suggested that production of type-1 IF represents one of the earliest functions in the differentiation process of mononuclear phagocytes and is likely to develop before the appearance of Fc receptors, diffuse esterase staining and latex phagocytosis.

Adult↗
PubMed1977

Post-transplant monitoring of renal allograft recipients for T, B and null lymphocyte subpopulations.

Time-course studies in ten allograft recipients showed a significant reduction in the percentage of T cells in the immediate post-transplantation period. This was accompanied by an increase in the percentage of both B and 'null' cells. T, B and 'null' cell percentages reverted to normal by the 6th month after transplantation at a time when patients had achieved stable renal function and were receiving only maintenance immunosuppressive therapy. Similar trends were observed with regard to absolute number of T, B and null cells. Both pre- and post-transplant plasma samples failed to inhibit T-rosette formation. Transient B rosette inhibitory activity was, however, observed in the plasma samples of two patients but did not appear to correlate with the mixed leucocyte culture blocking factor activity (MLC-BFA) of these plasma samples. In the group of patients studied there was no apparent association between percent or absoulte T, B and 'null' cell determinations and the onset of rejection crises.

Adult↗
PubMed1980

Mitogen stimulation during rosette formation: a test to measure numbers and function of T, B, 'D' and 'null' lymphocytes in man.

A technically simple procedure is described for simultaneous determination of numbers and functions of human blood lymphocyte subpopulations. The method is based on stimulation with mitogens (PHA, Con A and PWM) with simultaneous rosetting to detect E receptors and C3 receptors. Con A and PWM activate the affinity of C3 receptors in normal lymphocytes increasing the number of cells rosetting with zymosan-complement particles, and also the number of cells giving rosettes with both zymosan-complement particles and sheep red blood cells (B cells and 'D' cells respectively). PHA activates E receptors, increasing the number of T cells detected. The lymphocytes of 33 patients with allergic diseases of the skin and respiratory tract showed a decrease in C3 rosettes during stimulation with Con A and PWM, which suggests a defect of helper and suppressor functions of lymphocytes in these patients.

Asthma↗
PubMed1999

Quantification of B, T and null lymphocyte subpopulations in the blood and lymphoid organs of the pig.

Research on the pig's immune system is not only of general biological interest; the pig is also becoming more important as a large animal model in human biomedical research, e.g. as a donor for xeno-transplantation. With the increasing panel of monoclonal antibodies against porcine lymphocyte markers it is possible to gain more insight into the distribution and phenotype of lymphocyte subpopulations in the pig. In this study we investigated B cells (surface IgG: sIgG, sIgM and sIgA) and T cells (CD2, CD4, CD8, 8/1, MAC320) in the peripheral blood (pBL), thymus, spleen, tonsil, mesenteric and inguinal lymph nodes (mLN, iLN), jejunal and ileal Peyer's patches (jejPP, ilPP) in Göttingen minipigs. A flow cytometric technique was employed which enabled three color indirect immunofluorescence. B cell stained for surface IgG and surface IgA were found only in small percentages. Surface IgM positive cells were distributed at higher rates, with up to 24.9% in the iLN. Up to 64.2% of CD4+ and up to 73.1% of CD8+ cells were observed in the thymus. Most of the CD4+ cells were CD4/CD8 double positive cells. These cells were mostly triple positive in combination with CD2. A larger fraction of CD2- were CD8- which are taken to be NK cells. MAC320, a marker for a subtype of gamma/delta T cells, was predominantly found on cells in the pBL. The standardized flow cytometric technique produced comparable data on the distribution of major lymphocyte subpopulations in the blood and different lymphoid organs of the pig. The results provide a basis for future studies using the pig as animal model.

Animals↗
PubMed1980

Cytotoxic antibodies against surface immunoglobulin.

The reactivity of B lymphocytes, null lymphocytes, T lymphocytes, chronic lymphocytic leukemia (CLL) cells, B and T cultured cell lines, monocytes, and granulocytes was tested against a panel of antisera under cold conditions of 5 C incubation followed by 25 C incubation in complement. The degree of cold cytotoxic activity against these cell types was in the order given above, with the C cells being the most sensitive. Surface immunoglobulin (SIg) staining followed the same distribution sequence, with the B cells containing the most SIg. Human IgM inhibited cold cytotoxic antisera but human IgG or human serum albumin (HSA) did not. IgM did not inhibit HLA antisera. We conclude that at least some cold cytotoxic antibodies react against surface IgM.

Antilymphocyte Serum↗
PubMed1991

Analysis of workshop antibodies to null cells on a non-T/non-B lymphocyte population.

Non-T and non-B lymphocytes (null cells) were obtained by Sephadex G-10 depletion followed by treatment with mAbs to CD2 and MHC class II and complement. The enriched cells were principally CD5dim and contained greatly increased numbers of null lymphocytes. This methodology will be useful for null lymphocyte enrichment in examining cell surface molecules and functional attributes of null lymphocytes.

Animals↗
PubMed1983

[Role of T-, B-, null, suppressor lymphocytes and anti-brain antibodies in infectious-allergic disease of the nervous system].

Clinicoimmunologic studies involving 255 patients with infectious and allergic nervous disorders showed some alterations in their immune homeostasis in association with a certain imbalance in B-cell immunity. These changes were expressed in a decrease in the functional activity and absolute content of T-lymphocytes in the peripheral blood, in a steady increase in O-cells, and in insufficient lymphocyte T-suppressory control. The importance of elevated brain antibody levels for making the differential diagnosis of the autoallergic process in the nerve tissues is demonstrated.

Adult↗
PubMed1981

Ecto-adenosine triphosphatase deficiency in cultured human T and null leukemic lymphocytes. A biochemical basis for thymidine sensitivity.

Cultured leukemic T and null lymphocytes are highly sensitive to growth inhibition by thymidine, as well as the other deoxynucleosides, deoxyguanosine and deoxyadenosine. By contrast, Epstein-Barr virus-transformed B lymphocytes are relatively resistant to deoxynucleosides. Growth inhibition is associated with the development of high deoxyribotriphosphate pools after exposure to the respective deoxynucleotides. We show that malignant T and null lymphocytes are deficient in ecto-ATPase activity. We show this cell surface enzyme to be of broad specificity, capable of degrading both ribotriphosphates and deoxyribotriphosphates. High levels of this ecto-enzyme are found in deoxynucleoside-resistant, Epstein-Barr virus-transformed B lymphocytes. Ecto-ATPase deficiency may represent a mechanism for increased sensitivity to deoxynucleoside growth inhibition.

Adenosine Triphosphatases↗
PubMed1981

Studies of T, B, and "null" blood lymphocytes in normal persons of different age groups.

The numbers of T, B, and "null" blood lymphocytes were determined in 216 normal healthy persons, whose ages ranged from 20 to 94 years. No change was found in the numbers of T or B lymphocytes with increasing age of the subjects, whereas both the total lymphocytes and the number of "null" lymphocytes decreased just significantly with age (p less than 0.05). The response of blood lymphocytes to phytohaemagglutinin in vitro was measured in 25 subjects aged 20-30 years and compared with the response in 23 subjects aged over 60 years. There was no significant difference in the response of the two groups.

Adult↗
PubMed1988

Changes in subpopulations of lymphocytes in peripheral blood, and supramammary and prescapular lymph nodes of cows with mastitis and normal cows.

Direct immunofluorescence (IF) and indirect IF techniques were employed to analyze the distribution of B and T lymphocyte populations in peripheral blood, and in supramammary (draining), and prescapular (non-draining) lymph nodes of cows with mastitis and normal cows. In the peripheral blood there was a significant decrease in the percent and absolute number of B lymphocytes in mastitic cows (n = 29; 17.1 +/- 10.2%; 3.4 +/- 2.7 X 10(5) cells/ml) as compared to normal cows (n = 38; 25.2 +/- 7.8%; 9.3 +/- 5.4 X 10(5) cells/ml). The percent T lymphocyte count in mastitic cows (71.2 +/- 7.1%) was slightly increased over that of normals (65.8 +/- 7.2%), although the absolute number of T lymphocytes was decreased in mastitic cows (1.49 +/- 0.91 X 10(6) cells/ml vs. 2.47 +/- 1.28 X 10(6) cells/ml). In the prescapular lymph node the percent of B lymphocytes, but not T or "null lymphocytes", decreased significantly in mastitic cows as compared to that of normals. The decrease, i.e. 32%, paralleled the 32.1% decrease found in peripheral blood B lymphocytes. In contrast, in the supramammary lymph node of mastitic cows, the percent B lymphocytes increased over that of normals (35.1 +/- 2.0% vs. 20.4 +/- 9.4%), whereas the percent T lymphocytes decreased to 54.5 +/- 2.8% compared to 70.7 +/- 3.5% in normal cows. There was no significant change in percent "null lymphocytes". The weight of prescapular lymph nodes did not change in mastitic cows when compared to that of normals. As a result, the estimated number of B lymphocytes, but not of T and "null lymphocytes", decreased.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1981

Which Fc receptor-bearing cells are detected with the Ripley assay?

Mononuclear cells were isolated from peripheral blood and analysed with T and B markers (E rosettes and SIg) and on contaminating monocytes and PMN. The suspensions contained 63.3 +/- 4.8% T lymphocytes, 11.4 +/- 3.8% B lymphocytes, 9.0 +/- 5.5% null lymphocytes, 15.1 +/- 3.8% monocytes and 1.1 +/- 0.6% PMN. Of all cells, 27.6 +/- 12.1% formed EA rosettes with OR1R2 red cells coated with anti-CD Ripley. In preparations fixed after cytocentrifugation, the EA rosette-forming cells were studied with regard to esterase activity. The proportion of cells with detectable Fc receptors was further studied in purified T lymphocyte and in monocyte suspensions. Finally, EA rosettes were isolated by gradient centrifugation and the rosette forming cells studied with conventional T and B markers. All procedures gave corresponding results: on average 11-14% of the T lymphocytes and nearly 100% of the null cells formed EA rosettes, while only 2% of the B lymphocytes had detectable Fc receptors. Of the purified monocyte and PMN populations, on average 72 +/- 10.5 and 14.5 +/- 5.4%, respectively, formed EA rosettes. Thus, the Ripley assay, representing an important marker for null lymphocytes, cannot be regarded as specific for this population of white blood cells.

Cell Separation↗
PubMed1978

Maturation of bone marrow lymphocytes. II. Development of Fc and complement receptors and surface immunoglobulin studied by rosetting and radioautography.

Radioautographic DNA labeling and rosetting techniques were combined to study the development of surface IgM, Fc, and complement receptors (FcR, CR) on small lymphocyte populations in mouse bone marrow. [3H]thymidine was either infused continuously to label newly formed cells for periods up to 4 days, or injected daily, 21--35 days before use, to label a sample of long-lived cells. Bone marrow cells were incubated with sensitized sheep erythrocytes to detect surface IgM, FcR, and CR, respectively, and examined radioautographically after cytocentrifugation. During [3H]thymidine infusion, marrow small lymphocytes lacking surface markers were the first to show [3H]thymidine labeling. Most of these cells became labeled by 4 days (IgM--ve, 89%; FcR--ve, 92%; Cr--ve, 88%). Labeling of small lymphocytes bearing surface IgM, FcR, and Cr began after an initial lag and increased to high values by 4 days (IgM + ve, 73%; FcR + ve, 82%; CR + ve, 83%). Labeled IgM + ve small lymphocytes formed progressively larger rosettes as cell age increased. Some proliferating large lymphoid cells formed rosettes for IgM, FcR, and CR. Labeled long-lived small lymphocytes expressed surface IgM, FcR, and CR, the incidence of each receptor being uniformly high (38--43%) and the rosettes tending to be larger than those formed by newly formed lymphocytes. In double-surface marker studies, FcR and CR rosettes were formed by some IgM--ve small lymphocytes as well as IgM + ve cells in the marrow. After transfusion of marrow cells from donor mice infused with [3H]thymidine for 24 h, many labeled newly formed lymphocytes homed into the splenic red pulp of unlabeled syngeneic recipients. Subsequently, these cells showed a rapid increase in the incidence of rosettes for surface IgM, FcR, and CR, together with a progressive enlargement of each type of rosette. Although all the labeled small lymphocytes recovered from the spleen developed both surface IgM and FcR by 3 days, only approximately one-half developed CR. The results demonstrate that most of the small lymphocytes bearing FcR, CR, and surface IgM in mouse bone marrow are newly formed indigenous cells. Each receptor becomes detectable by rosetting soon after the small lymphocytes are first produced. The newly formed, marrow-derived small lymphocytes are able to continue their development of surface IgM, FcR, and CR after migrating into the spleen, consistent with a maturation of primary B lymphocytes. In addition, the data indicate the genesis in mouse marrow of a non-B lineage of lymphocytes (notably, IgM--ve FcR + ve cells.). A minority of small lymphocytes bearing IgM, FcR, and CR in mouse marrow are long-lived cells, presumptive recirculating immigrants, differing in receptor status from the newly formed cells. The results are discussed with regard to the heterogeneity of marrow lymphocytes and proposed models of primary B lymphocyte and null lymphocyte production.

Animals↗
PubMed1982

Membrane marker characterization of the eosinophil colony-forming cell.

We investigated the membrane markers of the eosinophil colony-forming cell (EO-CFC). Eosinophil colonies were identified by their ability to show densely packed cell growth and by staining for cyanide-resistant peroxidase activity. Bone marrow cells were characterized prior to soft agar culture by one of two methods: (1) standard erythrocyte (E) and erythrocyte-antibody-complement (EAC) rosetting, or (2) by the human T- and B-cell surface markers, Leu 1 and HLA-DR, using the fluorescence-activated cell sorter. Eosinophil colonies developed mainly from the nonrosetting, null cell population. EO-CFC lacked the human T-cell marker Leu 1, but demonstrated the human B-cell marker, HLA-DR. EO-CFC were also present in peripheral blood and possessed similar properties to those in the bone marrow. The results indicate that EO-CFC are derived from the null lymphocyte cell population and possess HLA-DR markers.

Antibodies, Monoclonal↗