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CHAPTER 80 MORPHOLOGY OF LYMPHOCYTES AND PLASMA CELLS

CHAPTER 80 MORPHOLOGY OF LYMPHOCYTES AND PLASMA CELLS
Williams Hematology

CHAPTER 80 MORPHOLOGY OF LYMPHOCYTES AND PLASMA CELLS

STEPHEN M. BAIRD

Definition and History
Microscopy and Histochemistry of Normal Blood Lymphocytes

Light Microscopy

Phase-Contrast Microscopy

Transmission Electron Microscopy and Cytochemistry

Scanning Electron Microscopy

Morphologic Changes Associated with Activation
Microscopy and Histochemistry of Plasma Cells

Morphologic Studies

Light Microscopy, Histochemistry, and Electron Microscopy
Antigens of Human Lymphocytes

B-Lymphocyte Antigens

T-Lymphocyte Antigens

Natural Killer Cells
Chapter References

Lymphocytes are a heterogeneous collection of cells that can be distinguished easily from other leukocytes by their characteristic morphology. However, this morphology is shared by all three major blood lymphocyte subsets, namely T cells, B cells, and natural killer (NK) cells. Although B cells can differentiate into plasma cells that have a distinctive morphology, most changes that occur in morphology during differentiation or activation are not unique to any one of the three major subgroups. Instead, other means are required to distinguish the major subsets and sub-subsets of lymphocytes. This has been achieved through the advent of monoclonal antibodies and the characterization of surface membrane antigens that are distinctive for each lymphocyte subset. This chapter describes the morphologic features that distinguish lymphocytes from other leukocytes and the membrane antigens that most commonly are used to distinguish the major lymphocyte subsets.

Acronyms and abbreviations that appear in this chapter include: ADCC, antibody-dependent cell-mediated cytotoxicity; CD, clusters of differentiation; LFA-3, lymphocyte-function–associated antigen-3; MHC, major histocompatibility complex; N-CAM, neural adhesion protein; NK, natural killer; PAS, periodic acid–Schiff; TCR, T-cell antigen receptor.

DEFINITION AND HISTORY
Lymphocytes and plasma cells first were described morphologically in 1774 and 1875, respectively.1,2 Investigations of these cells from then until the 1960s primarily were to further define their morphology. Subsequently, lymphocytes were found to make immunoglobulins and to be necessary for cell-mediated immunity.3,4,5,6 and 7 With the advent of monoclonal antibodies and flow cytometry, the refinement of in vitro functional assays, and the application of molecular techniques, there have been major advances in the understanding of lymphocytes. Membrane antigens have been identified that assist in the designation of lymphocyte subsets and function. Three major blood lymphocyte subsets have been identified: T lymphocytes, B lymphocytes, and NK cells. Further, there are small numbers of circulating hematopoietic stem cells that resemble lymphocytes and that are capable of differentiating into any one of the various lymphocyte subsets.
MICROSCOPY AND HISTOCHEMISTRY OF NORMAL BLOOD LYMPHOCYTES
LIGHT MICROSCOPY
Classic studies of blood and tissues have demonstrated populations of spherical and/or ovoid cells that are from 6 to 15 µm in diameter when flattened on glass slides.4 Some of these studies described small lymphocytes, which are 6 to 9 µm in diameter, and large lymphocytes, which have a diameter of 9 to 15 µm. There are increased numbers of circulating large lymphocytes in patients with acute viral illnesses and in certain genetic immunologic deficiencies, particularly the Wiskott-Aldrich syndrome. Normal adults have a mean absolute number of circulating small lymphocytes of 2.5 × 109/liter (range of 1.5 to 4.0), or 35 percent of the total leukocytes (with a range of 20 to 50 percent).
The typical small lymphocyte as observed with Romanovsky polychromatic stains (e.g., Giemsa or Wright) has an ovoid or kidney-shaped nucleus that stains purple, has densely packed nuclear chromatin, and occupies about 90 percent of the cell area (see Plate XX-4). There is a small rim of cytoplasm that stains light blue. Although nucleoli rarely are observed in Giemsa-stained films, they can be demonstrated with methyl green-pyronine stains. Cytoplasmic basophilia is related to RNA content. The cytoplasm of some lymphocytes, particularly large lymphocytes, contains a number of coarse pink granules, usually 5 to 15 per cell, and occasional clear vacuoles. Cytoplasmic glycogen is detected with periodic acid–Schiff (PAS) and methenamine-silver techniques. A number of enzymes, including phosphorylase, acid hydrolases, nucleases, and mitochondrial enzymes, are in the lymphocyte cytoplasm.8 Peroxidase reactions are negative in lymphocytes.9
In a normal adult about 3 percent of blood lymphocytes are large granular lymphocytes10 (see Plate XX-3). These cells are a mixed population consisting of NK cells and some of the CD8 subset of mature T cells. The majority of mature T lymphocytes, however, show a localized “dot” staining pattern for acid phosphatase, acid and neutral nonspecific esterases, b-glucuronidase, and N-acetyl-b-glucosaminidase.11,12 and 13 B lymphocytes either lack esterase and acid phosphatase or show scattered granular staining.
Enzymes in the purine salvage pathways are expressed differently in lymphocyte subsets. The enzyme 5′-nucleotidase is detectable on plasma membranes of both B and T cells. In contrast, more adenosine deaminase and purine nucleoside phosphorylase are present in the cytoplasm of T cells than in the cytoplasm of B cells.14,15 Terminal deoxynucleotidyl transferase is present in cortical thymocytes, undifferentiated stem cells, and the malignant cells of acute lymphoid leukemias.16,17
PHASE-CONTRAST MICROSCOPY
Active movement of lymphocytes is studied by phase-contrast, or interference-contrast, microscopy. Lymphocytes move slowly with a “hand mirror” appearance. Cytoplasmic spreading does not occur. However, during cell movement a thickening occurs in the cytoplasmic rim (the Hof region), a region that houses most of the cell’s organelles, including the Golgi. Lymphocytes from patients with chronic lymphocyte leukemia have decreased movement.18
TRANSMISSION ELECTRON MICROSCOPY AND CYTOCHEMISTRY
As visualized by transmission electron microscopy,19,20,21 and 22 the circulating lymphocyte measures about 5 µm in diameter. The nucleus has an abundance of electron-dense, condensed heterochromatin, a feature characteristic of nonproliferating cells. The nucleoli are round in section, about 1.0 to 1.5 µm in diameter, and composed of three distinct and concentrically arranged structural units: the central region or agranular zone; the middle, fibrillar region; and the granular zone, which contains intranucleolar chromatin. The lymphocyte’s nuclear membrane contains nuclear pores and a perinuclear space.
The cytoplasmic organelles of the lymphocytes are characteristic of eukaryotic cells.19,20,21 and 22 Some organelles, like the Golgi zone, are poorly developed. The cytoplasm contains free ribosomes, occasional ribosome clusters, and strands of rough-surfaced endoplasmic reticulum (Fig. 80-1). Centrioles, mitochondria, microtubules (diameter of approximately 0.25 µm), and microfilaments (diameter of about 0.07 µm) are present in the cytoplasm adjacent to the cell membrane. The cytoplasm also contains lysosomes, which are about 0.4 µm in diameter, are electron-opaque, and contain classic lysosomal enzymes (e.g., acid phosphatase, b-glucuronidase, and acid ribonuclease).8 The lymphocyte plasma membrane stains with colloidal iron, a marker for membrane sialic acid. Lymphocyte cell membranes and cell coat glycoproteins are shown with other electron-dense markers including phosphotungstic acid, lanthanum colloid, and ruthenium red.

FIGURE 80-1 (A) Electron micrograph of normal human blood lymphocytes. Organelles are labeled in (B). (×12,000) (b) Diagrammatic representation of normal blood lymphocyte.

SCANNING ELECTRON MICROSCOPY
Scanning electron microscopy provides three-dimensional information.23 However, the resolution achieved with scanning electron microscopy, about 0.1 µm, is considerably less than that possible with transmission electron microscopy, generally 0.002 to 0.0039 µm. Normal blood lymphocytes, washed and collected onto silver membranes and fixed in glutaraldehyde, have a spherical topography with varying numbers of stubby or fingerlike microvilli (Fig. 80-2).24,25 In contrast, monocytes are much larger, have few microvilli, and display ruffled membranes and ridgelike profiles. T lymphocytes have smaller numbers of microvilli than B lymphocytes.24,25 However, the surface morphology of B lymphocytes is heterogeneous. Many B cells have moderate to markedly villous surfaces, but about 10 to 20 percent of B cells are smooth with few microvilli and thus are indistinguishable from most T lymphocytes.26 Furthermore, human blood lymphocytes fixed in suspension appear uniformly covered with short microvilli, and no differences between T and B cells are demonstrable.

FIGURE 80-2 Scanning electron micrograph of normal blood lymphocytes separated by the Ficoll-Hypaque method. Cells show varying numbers of microvilli. (×5000) (Fig. 80-3, Fig. 80-4, and Fig. 80-5 provided by Dr. Aaron Polliack of the Department of Hematology, Hebrew University Hadassah Medical School, Jerusalem, Israel.)

FIGURE 80-3 Transmission electron micrograph of lymphocyte from normal individual incubated with PHA for 3 days. The transformed cell has a large Golgi zone (G) and many ribosomal aggregates (arrows), and the nucleus is euchromatic. (×7500)

FIGURE 80-4 Transmission electron micrograph of plasmacytoid cell present in culture of lymphocytes from a patient with chronic lymphocytic leukemia incubated with pokeweed mitogen for 7 days. The nucleolus (N) and rough-surfaced endoplasmic reticulum (arrows) are evident. (×9000) (From Cohnen, Douglas, Konig, et al: Pokeweed mitogen response of lymphocytes in chronic lymphocytic leukemia. Blood 42:591, 1973, with permission.)

FIGURE 80-5 This is a diagram of lymphocyte activation that displays the relationship of cell-cycle phases (G0, G1, S, and M) with changes in cell metabolism and cell function. A lymphocyte may become an effector cell or a memory cell in G0 after having traversed the cell cycle. (Klein, Immunology, p 40. Blackwell, Cambridge, MA, 1990, with permission.)

MORPHOLOGIC CHANGES ASSOCIATED WITH ACTIVATION
Lymphocyte stimulation is associated with a complex sequence of morphologic and biochemical events, culminating in the transformation of small lymphocytes into blast or plasmacytoid cells (Fig. 80-3 and Fig. 80-4). Plant lectins, bacterial products, polymeric substances, and enzymes stimulate lymphocyte mitosis. Such agents are called mitogens. Some mitogens are specific for either B cells or T lymphocytes, whereas others stimulate both. The responses of specific lymphocyte subpopulations to various mitogens are complex.33 Nucleolar changes become evident as early as 4 h after exposure to phytomitogens (e.g., phytohemagglutinin, which stimulates T cells). These morphologic changes consist of increases in nucleolar size and in the number and concentration of granules in the granular zone. This is followed by an increase in fibrillar zones and increased intranucleolar chromatin. Nucleolar chromatin becomes more electron-lucent or dispersed. Electron microscopic autoradiography demonstrates that tritiated thymidine, incorporated into newly synthesized DNA, is spread throughout the nucleoplasm but is most concentrated at the nuclear membrane. From 48 to 72 h following the addition of phytohemagglutinin there is an increase in size of the cytoplasm. In addition, the cytoplasm contains an increase in the number of ribosomal clusters and more rough-surfaced endoplasmic reticulum. The transformed cell (lymphoblast) has increased numbers of lysosomes and a larger Golgi complex with more components.19,20,21 and 22 Under some circumstances (e.g., cultures of human lymphocytes stimulated for 7 to 10 days with pokeweed mitogen), some cells may form well-developed Golgi and plasmacytoid features.34 Similar plasmacytoid cells are observed in antigen-stimulated lymph nodes, during graft rejection in vivo, and in some in vitro systems, including the mixed lymphocyte culture.
Following stimulation with antigen or mitogens the lymphocyte enters the cell cycle. The cell-cycle phases and accompanying genetic or morphologic changes are summarized in Fig. 80-5. These parallel genetic and morphologic alterations are necessary correlates of the cell-cycle phases. The fate and function of lymphocytes that traverse the cell cycle may be divided into two pathways. Some lymphocytes may undergo several mitotic cycles and then return to the Go phase, indistinguishable in morphology from the original nonactivated cells. A separate subset of lymphocytes may become memory cells, programmed to remember the stimulating antigen and thus more rapidly respond to reexposure to the original antigen. Finally a small number of lymphocytes are destined to become terminally differentiated lymphocytes, such as plasma cells or cytotoxic T cells.
MICROSCOPY AND HISTOCHEMISTRY OF PLASMA CELLS
MORPHOLOGIC STUDIES
Plasma cells derive from small B lymphocytes after antigenic stimulation and T-cell help. Several sequential mitotic divisions occur during cellular differentiation from the resting lymphocyte to the plasmablast to the immature plasma cell. Immature plasma cells also can undergo successive waves of mitosis in the medullary cords of lymph nodes in response to antigen.37 Cell transfer experiments demonstrated that these transformed cells later mature into antibody-producing plasma cells.38
Pokeweed mitogen induces B lymphocytes to transform into plasma cells after 7 to 10 days’ culture.39 These plasma cells infrequently contain large electron-dense inclusions, which may measure 2 to 3 µm in diameter (Russell bodies) (Fig. 80-6).40 Russell bodies, cytoplasmic immunoglobulin in the endoplasmic reticulum, sometimes are dissolved during the Giemsa staining procedure. They usually occur in pathologic states but may be found in normal lymph nodes or marrow. When cytoplasmic immunoglobulin becomes detectable, the same immunoglobulin isotype is present in the cytoplasm as on the cell membrane.

FIGURE 80-6 Intranuclear electron-dense bodies (Russell bodies) (RB) in plasma cell from the marrow of a patient with multiple myeloma. (×7000)

LIGHT MICROSCOPY, HISTOCHEMISTRY, AND ELECTRON MICROSCOPY
When treated with a polychrome stain, the mature plasma cell has a characteristic basophilic cytoplasm and an eccentric nucleus. The nuclear polarity is attributable to a large paranuclear zone, which corresponds to the Golgi apparatus. The typical mature plasma cell spread on a slide is usually round or oval and has a diameter from 9 to 20 µm, with a mean cell diameter of 14.4 µm and a mean nuclear diameter of 8.5 µm27 (see Plate XXI-1). The nuclear heterochromatin is coarse and distributed in a pattern that, in paraffin sections, sometimes resembles the spokes of a wheel (cartwheel nucleus). Plasma cells with two or more nuclei occasionally may be seen in the marrow of normal individuals. The nucleus stains blue-green with methyl green. The cytoplasm is characterized by its intense affinity for cationic dyes. The cytoplasm, basophilic due to ribonucleoprotein, stains selectively with methyl green-pyronine; it stains red with pyronine due to its high content of ribonucleoprotein (pyroninophilia). The cytoplasm also stains with several basic dyes, including toluidine blue and azures.
Plasma cells that are found in patients with certain diseases may have different histochemical properties. These cells may have a larger size and contain cytoplasmic inclusions that may be observed with PAS stains.28 In hemochromatosis and hemosiderosis, plasma cells may contain hemosiderin when examined by electron microscopy.29 Other cytochemical features include absence of peroxidase and nonspecific esterase. Plasma cells are strongly positive for b-glucuronidase and for mitochondrial enzyme markers.30 Plasma cell size and morphology may be altered substantially in multiple myeloma and macroglobulinemia (see Plate XXI-4, Plate XXI-5, Plate XXI-6, Plate XXI-7, Plate XXI-8 and Plate XXI-9). Cells with 2 or 3 nuclei may be seen, even in adults without plasma cell dyscrasias. Under some circumstances amyloid inclusions have been detected by electron microscopy in plasma cells.31
By electron microscopy the plasma cell is packed with a rough-surfaced endoplasmic reticulum that has numerous attached ribosomes. There is a large circumscribed Golgi zone that forms a paranuclear halo when observed by light microscopy. The nucleus has dense areas of heterochromatin. The Golgi zone contains lamellae, vesicles, vacuoles, and a number of granules. Between the strands of endoplasmic reticulum there are mitochondria (Fig. 80-7).32

FIGURE 80-7 (A) Electron micrograph of mature plasma cells in normal human lymph node. (×9000) (B) Diagrammatic representation of normal plasma cell labeling the organelles.

ANTIGENS OF HUMAN LYMPHOCYTES
Human blood lymphocytes possess an array of different membrane antigens. Standardization of monoclonal antibody reagents by identifying clusters of differentiation (CD) and the advent of flow cytometry have facilitated detection of lymphocyte subsets (see Chap. 13). The following sections list those surface antigens and morphologic features that help define the major human lymphocyte subsets.
B-LYMPHOCYTE ANTIGENS
Table 80-1 summarizes the expression of CD antigens on cells of the B-lymphocyte lineage, including committed progenitor B cells and pre-B cells. These cells, and the maturation stages that they represent, are discussed in Chap. 82. Also presented in Table 80-1 are antigens that are expressed or are increased upon B-cell activation. The physiology, structure, and distribution of each of the CD antigens listed in Table 80-1 are presented in Chap. 13 (see Table 13-1).

TABLE 80-1 B-LYMPHOCYTE ANTIGENS USED IN CLINICAL MEDICINE

Of the B cell-associated antigens listed in the first column of Table 80-1, only a few are restricted to cells of the B lineage (see Chap. 13). Of these, only CD20, CD79a, and CD79b are not found on other cell types. The latter two antigens associate with Ig to facilitate surface Ig expression and surface-Ig–mediated signal transduction and are expressed only by cells that produce Ig, an exclusive function of B lymphocytes and plasma cells (see Chap. 15 and Chap. 83). CD19 is restricted mostly to B cells but may be expressed weakly by follicular dendritic cells. This antigen, however, is expressed by B cells at all stages of maturation, including the committed B-cell progenitor. As such, it is the best-defined pan-B cell surface antigen. Cytoplasmic CD22 is perhaps the broadest mature B-cell marker.
In addition to the CD antigens, B cells express the three major histocompatibility complex (MHC) class II antigens (DR, DP, DQ). These antigens are heterodimers of a heavy chains and b light chains that are encoded by genes within the D complex of the HLA complex (see Chap. 138).
B-1 B CELLS
A subset of normal B cells express CD5,45 a 67-kDa transmembrane glycoprotein46 that is expressed by T cells (see Chap. 13). These cells are designated CD5 B cells, or, more recently, B-1 B cells.47 B-1 B cells do not express other T-cell markers but do express all other pan-B-cell surface antigens.48 B-cell expression of CD5 can be modulated by various agents.49,50 B-1 B cells are found in umbilical cord blood,51 adult blood, the pleura and peritoneum, and all major secondary lymphoid organs but are rare in the marrow.52 These cells apparently are enriched for cells that spontaneously produce polyreactive autoantibodies.53,54 and 55 They are the cells that are clonally expanded in chronic lymphocytic leukemia (see Chap. 98).
PLASMA CELLS
Most B-cell differentiation antigens are not expressed by the mature plasma cell, including surface immunoglobulin and HLA class II antigens56 (see Table 80-1). Of the cells of the B lineage, plasma cells are distinctive in that they express CD28 and PCA-1. CD28 is found on marrow plasma cells and myeloma cell lines.57 PCA-1 also is found on human plasma cells58 and may function as a threonine-specific protein kinase.59 This latter function may assist in phosphorylation of secretory proteins. PCA-1 also is present at low density on granulocytes and monocytes. In contrast to mature B lymphocytes, plasma cells do not bear surface Ig but express CD38 and very high levels of CD43 and CD85 (see Table 80-1).
T-LYMPHOCYTE ANTIGENS
CD1
CD1 is a family of three membrane glycoproteins, CD1a, CD1b, and CD1c, of 49, 45, and 43 kDa, respectively, that is found on all cortical thymocytes (see Chap. 13). CD1 also can be expressed on monocytes following activation and on Langerhans cells.60 CD1 has a structural relationship to HLA class I and class II proteins. This structural association suggests that CD1 is involved in T-cell interaction with accessory/antigen-presenting cells, perhaps presenting hydrophobic antigens such as lipids.
CD2
This T-cell antigen is expressed early in T-cell development. It is a 50-kDa surface glycoprotein that facilitates T-lymphocyte target cell interactions and T-lymphocyte activation.61 Lymphocyte-function-associated antigen-3 (LFA-3) (CD58) is a ligand for CD2 (see Chap. 13 and Chap. 84). Cross-linking CD2 may activate T cells through a pathway that is distinct from that used by the T-cell receptor for antigen. This may result in augmentation of the immune response in the absence of additional antigenic stimulation. Since anti-CD2 monoclonal antibodies activate early T-lymphocyte progenitors, CD2 antigen probably is a receptor used in the activation of thymocytes prior to appearance of a functional T-cell receptor.62
CD3
CD3 is expressed by early thymocytes and mature T cells.63 It is tightly linked to the T-cell receptor (see Chap. 84). The CD3 molecule serves as a signal transduction unit after T-cell receptor activation. In addition, this antigen frequently is expressed on T-cell acute lymphoblastic leukemia.
CD5
CD5 is found on all T cells. It appears early in T-lymphocyte ontogeny.64 CD5 also may be detected on a subset of blood and cord B cells.
CD7
CD7 is a 40-kDa glycoprotein that is expressed very early in T-cell ontogeny.65 The antigen is lost during the terminal stage of T-cell maturation. The function of CD7 is not known. CD7 also is expressed on monocytes or natural killer cells.66
CD4 AND CD8
CD4 is first expressed on thymocytes along with CD8 (see Chap. 82 and Chap. 84). CD4, a member of the immunoglobulin supergene family, is a single-chain transmembrane glycoprotein.67 CD8 is a homodimeric transmembrane glycoprotein of 34 kDa.68 Each molecule also is associated with the T-cell–specific tyrosine kinase p56.69 CD4 and CD8 act as coreceptors during T-cell activation by antigen (see Chap. 84). CD4 also is a coreceptor for the human immunodeficiency virus70 (see Chap. 89), along with CCR5 or CXCR4.
CD25
CD25 originally was defined as a component of the interleukin-2 receptor (IL-2R).71 This antigen is expressed on activated T and B cells. CD25 also is found on thymocytes and monocytes. The IL-2 receptor is made of three distinct proteins [a chain (CD25), b chain (CD122), and g chain, also a component of the receptors for IL-4, 7, and 9]. When bound to the a chain alone, IL-2 binds with higher affinity to the IL-2R b chain. This then results in formation of a high-affinity complex of IL-2 bound to both IL-2R a and b chains.73 The g chain is involved in signaling to the cell after IL-2 binding. IL-2 binding to the high-affinity receptor signals T cells to proliferate and differentiate into specific effector T cells (e.g., helper T or cytotoxic T cells).
CD45
CD45 is tyrosine-specific phosphatase.74 It is expressed by virtually all hematopoietic cells. However, different isoforms exist due to alternative splicing of the CD45 transcript and differential glycosylation.75 These different isoforms of CD45 are expressed differentially by different cell subsets (see Chap. 84). CD4+ helper T cells can be divided into functionally disparate subsets based on their expression of the different CD45 isoforms and CD29 (the VLA b chain). Naive CD4+ helper T cells are CD45RA+ and express low levels of CD29. These CD4+ T cells can induce CD8+ T cells to downregulate B-cell IgG synthesis.76 After activation, T-cell expression of CD45RA is diminished, but that of CD29 and CD45RO (a lower-molecular-weight isoform of CD45) is enhanced.77 CD4+ CD45RO+ CD29 high-affinity T cells are called memory-helper T cells, in that they apparently facilitate induction of B-cell IgG synthesis in response to secondary challenge with antigen. In addition, CD45 is necessary for activation of either CD4+ T cells or CD8+ T cells. T cells that lack expression of CD45 do not respond well to various activation stimuli.77
CD28
CD28 is a 44-kDa homodimer that is expressed on resting T cells. CD28 is a surface receptor for a cyclosporine-resistant T-cell signal-transduction pathway.78 CD28 binds the CD80 (B7/BB1) antigen expressed by activated B cells and professional antigen-presenting cells79 (see Chap. 84). This interaction provides a second costimulatory signal to T cells that are activated by the cross-linking of their T-cell antigen receptors in response to specific interactions with antigenic peptide bound to the major histocompatibility antigens expressed by antigen-presenting cells.80,81 Without this second costimulatory signal, the T cell may be induced into anergy.82 In this state the T cells fail to respond to the antigen(s) presented by the antigen-presenting cell because of a block in IL-2 gene expression.83
T-CELL RECEPTOR
The T-cell antigen receptor (TCR) is present on all mature T cells and on developing immature thymocytes (see Chap. 84). The TCR may have unique determinants that are found on all T cells within a given T-cell clone. It is expressed by most T-cell lymphomas.
NATURAL KILLER CELLS
The NK cell is defined as an effector cell that is not MHC-restricted and has the capacity for spontaneous cytotoxicity toward various target cells (see Chap. 85). The large granular lymphocyte was identified as being the blood cell responsible for NK-cell function because these cells, when enriched by sedimentation, accounted for almost all the blood NK-cell activity.84 However, not all NK cells have large granular lymphocyte morphology.
Large granular lymphocytes have a unique morphology (see Plate XX-3). These cells typically have round or indented nuclei and abundant pale cytoplasm containing a few coarse pink granules (1.0 to 2.0 µm in diameter).85 Large granular lymphocytes have membrane-bound granules that stain for acid hydrolases, including acid phosphatase, a napthyl acetate esterase, and b glucuronidase. These granules may be related to the cytolytic capacity of these cells. These cells do not express surface immunoglobulin and lack adherent or phagocytic properties. They may form rosettes with sheep erythrocytes and express immunoglobulin Fc receptors.25,86 Despite their relative morphologic homogeneity, they comprise several subpopulations with distinct phenotypes. NK cells express class I but not class II antigens of the MHC. Human NK cells characteristically express CD16 (FcgRIII) and CD56, but not CD3.87,88 CD16 (FcgRIII) is a low-affinity receptor that binds to IgG that is bound specifically to antigens present on cells targeted for destruction in antibody-dependent cell-mediated cytotoxicity (ADCC). CD16 is expressed on all NK cells, neutrophils, and tissue macrophages. CD56 is the neural adhesion protein (N-CAM) and is seen on most NK cells, albeit at low density.89 This 200-kDa protein is expressed at higher levels following activation.
NK cells have some T-cell antigens on their cell membrane, including CD8, found on approximately 30 to 50 percent of NK cells; CD2, present on about half of all NK cells; and CD38, present at low density on most NK cells. However, NK cells do not express CD4.90 Upon activation, NK cells express increased levels of CD25, CD56, and class II antigens of the MHC. Three cytokines that can activate NK cells are IL-2, interferon-alpha, and IL-12. IL-2 also can induce NK cells and some T cells to differentiate into lymphokine-activated killer cells in vitro (see Chap. 85).
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Books@Ovid
Copyright © 2001 McGraw-Hill
Ernest Beutler, Marshall A. Lichtman, Barry S. Coller, Thomas J. Kipps, and Uri Seligsohn
Williams Hematology

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