The differentiation and function of human T lymphocytes

Reinherz, Ellis L.; Schlossman, Stuart F.

doi:10.1016/0092-8674(80)90072-0

Cited by 1,417 publications

(394 citation statements)

References 28 publications

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“…The results were expressed as the percentage positive cells of the total number of cells counted. The following monoclonal antibodies were employed: OKT 3, OKT 4 and OKT 8, described to label all peripheral T lymphocytes, helper/inducer T cells and suppressor/cytotoxic T cells, respectively [17]. The OKT antibodies were obtained by fusion of non-secreting myeloma cells with spleen cells from mice immunized with human peripheral T cells or thymocytes [18].…”

Section: T Cell Preparation and Analysmentioning

confidence: 99%

T lymphocyte subsets in patients with newly diagnosed Type 1 (insulin-dependent) diabetes: A prospective study

Buschard¹,

Røpke

Madsbad³

et al. 1983

Diabetologia

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Summary. T lymphocyte subsets in peripheral blood from 11 newly diagnosed Type 1 (insulin-dependent) diabetic patients were studied prospectively at three time intervals: as soon as possible after diagnosis, 3 weeks and 5 months later. Lymphocytes were marked with monoclonal OKT antibodies and examined in a fluorescence-activated cell sorter. The percentage of T lymphocytes (OKT 3) did not change significantly at the three study times. The percentage of helper/inducer T cells (OKT 4) was high the first week after diagnosis, but decreased at the 5-month examination (p< 0.05). The percentage of suppressor/cytotoxic T cells (OKT 8) was low at diagnosis but increased at 3 weeks (p< 0.02) and 5 months (p< 0.01). The ratio OKT 4/OKT 8 lymphocytes was 2.28 at diagnosis, decreasing to 1.77 at 3 weeks and 1.87 at 5 months, compared with 1.46 for 16 age-matched control subjects. There was no significant change in the absolute number of lymphocytes. It is concluded that the distribution of T cell subsets was abnormal at the time of diagnosis, but changed towards normal within a few weeks, after which there was no significant change at 5 months. It is as yet unknown whether the high proportion of helper/inducer T cells and/or the low percentage of suppressor/cytotoxic T cells at diagnosis favour immune reactions involved in the pathogenesis of Type 1 diabetes.

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Section: T Cell Preparation and Analysmentioning

confidence: 99%

T lymphocyte subsets in patients with newly diagnosed Type 1 (insulin-dependent) diabetes: A prospective study

Buschard¹,

Røpke

Madsbad³

et al. 1983

Diabetologia

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“…The lymphocyte population has been shown to be heterogeneous with respect to cell surface antigens (21). The functional properties of cells correlate strongly with these lineage-specific differentiation antigens (14).…”

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confidence: 99%

Fixation and long‐term storage of human lymphocytes for surface marker analysis by flow cytometry

Lal¹,

Edison²,

Chused³

1988

Cytometry

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A method to preserve stained human lymphocytes for subsequent cell surface analysis by flow cytometry (FCM) is described. Cells stained with fluorescein isothiocyanate (FITC) and phycoerythrin (€'E)-conjugated monoclonal antibodies and then fixed in 1% paraformaldehyde, followed by extensive washing and resuspension in 1% BSA medium, could be stored at 4°C for at least 2 weeks prior to FCM analysis without significant alteration in the light scatter or fluorescence properties of the cells. Furthermore, the method was also suitable for analyzing lymphocytes that express T-cell activation markers in certain disease conditions. In addition, we have identified monoclonal antibody combinations that discriminate different lymphocyte subsets that are satisfactory for multiparameter analysis after 2 weeks of storage. This method should prove useful for enumerating lymphocyte subsets in field study sites remote from flow cytometry laboratories.Key terms: Paraformaldehyde, filariasis, systemic lupus erythematosusThe lymphocyte population has been shown to be heterogeneous with respect to cell surface antigens (21). The functional properties of cells correlate strongly with these lineage-specific differentiation antigens (14). The subpopulations of B cells (1) and T cells (10) can be quantitated by flow cytometry (FCM) based on the relative expression of two or more cell-surface markers (8), and this approach has been used to define abnormal lymphocyte subsets in a variety of clinical conditions (9, 11).For such clinical studies, fresh lymphocytes (7) have generally had t o be used, since conflicting results have sometimes been observed with cryopreserved cells (31, an observation suggesting that certain differentiation antigens may be cryolabile. Thus, in most studies, cells are stained by conventional immunofluorescence techniques and analyzed either fresh or shortly after paraformaldehyde fixation (13). For studies carried out at sites remote from a flow cytometer, this protocol, however, creates practical problems, since the effect of longterm fixation on FCM analyses has not been well defined.Therefore the purpose of this study was to develop the optimal method of preserving stained cells for as long a time as possible. The basic requirement of such a technique would be that it permit 2-3 weeks storage of the cells prior to analysis without significant alteration of the light scatter and fluorescence properties of the cells. In this report, we describe such a method and define monoclonal antibody reagents satisfactory for studies requiring prolonged storage of FCM samples prior to analysis. MATERIALS AND METHODS Study PopulationThirteen individuals were studied. Eight were healthy North Americans, two were normal volunteers from New Delhi, India, two were patients with systemic lupus erythematosus, and one was an Indian patient with tropical pulmonary eosinophilia.Preparation of Fixative A 1.0% (w/v) solution of paraformaldehyde (Eastman Kodak Co., Rochester, NY) was prepared by dissolving paraformaldehyde in phosphat...

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“…From the discovery of CD4 + T cells (T helper cells) in the early 1980s31, 32 with the advent of monoclonal antibody technology and the initial distinction between Th1 and Th2 from Tim Mosmann and Bob Coffman33, 34 the T helper subtypes organization has become far more complex. Starting from the two main Th1 and Th2 groups, five other new members have now joined the CD4 + T‐cell panorama, such as Th17,35, 36 the regulatory T cells,37 the follicular helper T cells38 and, most recently, the Th939 and Th2240 subsets.…”

Section: Heterogeneity In the Immune Systemmentioning

confidence: 99%

Single‐cell technologies to study the immune system

Proserpio

Mahata

2015

Immunology

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SummaryThe immune system is composed of a variety of cells that act in a coordinated fashion to protect the organism against a multitude of different pathogens. The great variability of existing pathogens corresponds to a similar high heterogeneity of the immune cells. The study of individual immune cells, the fundamental unit of immunity, has recently transformed from a qualitative microscopic imaging to a nearly complete quantitative transcriptomic analysis. This shift has been driven by the rapid development of multiple single‐cell technologies. These new advances are expected to boost the detection of less frequent cell types and transient or intermediate cell states. They will highlight the individuality of each single cell and greatly expand the resolution of current available classifications and differentiation trajectories. In this review we discuss the recent advancement and application of single‐cell technologies, their limitations and future applications to study the immune system.

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The differentiation and function of human T lymphocytes

Cited by 1,417 publications

References 28 publications

T lymphocyte subsets in patients with newly diagnosed Type 1 (insulin-dependent) diabetes: A prospective study

T lymphocyte subsets in patients with newly diagnosed Type 1 (insulin-dependent) diabetes: A prospective study

Fixation and long‐term storage of human lymphocytes for surface marker analysis by flow cytometry

Single‐cell technologies to study the immune system

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