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Establishment of a human plasma cell line in vitro.

A human plasma cell line designated ARH-77 has been established and propagated in culture for the past 2 years. The cells exhibited morphological characteristics of plasma cells under light and electron microscopic examination. An average of 40% cells are positive for immunoglobulin G by direct immunofluorescence, while an immunoglobulin G-specific radioimmunoassay reveals the production of 1.21 X 10(4) ng/10(6) plasma cells. The karyotype is aneuploid with a a modal chromosome number of 45 to 46 and no marker chromosome. Growth kinetics characteristics are: doubling time, 110.4 hr; generation time, 56.4 hr; G1 + G2-phase transit time, 45.5 hr; S-phase transit time, 10.9 hr; growth fraction, 74%; mitotic index, 1.5%; labeling index, 14.3%; and cell loss, 31.0%. Some of the growth kinetics characteristics were markedly similar to the properties displayed in vivo by plasma cells of patients with multiple myeloma and suggest that the cell line might be a useful in vitro model for the study of human myeloma.

Aneuploidy

A human tubular array plasma cell.

A human plasma cell is described which is distinct with both the light and electron microscope. With indirect immunofluorescence its cytoplasm is immunoglobulin-associated and the ultrastructure is characterized by tubular arrays of endoplasmic reticulum similar to those described in a number of diseases. Hypotheses regarding these tubular structures are reviewed.

Aged

Virus-bearing plasma cells in peripheral blood of a patient with 'hairy cell' leukemia.

Electron-microscopic examination of peripheral blood from a patient with 'hairy cell' leukemia revealed classical 'hairy cells', atypical lymphoid cells and numerous pathological plasma cells. Osmiophilic granular material coated the cell surface of 'hairy cells' and lymphoid cells but not the plasmalemma of the plasma cells. The most important features of the plasma cells were cytoplasmic protrusions and masses of oncogenic virus A particles in their endoplasmic reticular cisternae.

Cytoplasm

Distribution of IgA 1 and IgA 2 plasma cells in various normal human tissues and in the jejunum of plasma IgA-deficient patients.

The distribution of IgA 1 and IgA 2 plasma cells was studied in normal human tissues. IgA 2 is a minor constituent in peripheral lymph nodes as well as in serum and in bone marrow plasma cells. An increased proportion of IgA 2 plasma cells was observed in gastric and intestinal mucosa, as well as in bronchial mucosa and salivary and mammary glands. Tonsils and mesenteric lymph nodes exhibit values intermediate between those of central and peripheral lymphoid systems. In patients with plasma IgA-deficiency, IgA 2 is the predominant intestinal IgA plasma cells. This may explain the frequent association of an asymptomatic condition and plasma IgA deficiency.

Adolescent

The bone marrow in multiple myeloma: correlation of plasma cell ultrastructure and clinical state.

The bone marrow plasma cells of 52 patients with various kinds of monoclonal gammopathies were studied by electron microscopy, and compared to the bone marrow plasma cells of 22 patients with reactive plasmacytosis. Virtually every marrow from patients with myeloma and macroglobulinemia contained plasma cells with disparity between the nuclear maturation and cytoplasmic differentiation. This asynchronous development was not present in plasma cells of reactive marrows nor in plasma cells from patients with megaloblastic anemias. The degree of asynchrony observed in myeloma and macroglobulinemia was proportional to the extent of disease as judged by clinical criteria. For the most part plasma cells of patients with non-myelomatous monoclonal gammopathy failed to exhibit significant asynchrony. These observations are consistent with the view that multiple myeloma is a neoplastic disorder with a definably malignant-appearing cellular proliferation.

Adult

Growth kinetics of plasma cell myeloma.

The plasma cell labeling index(ex) (LI) of 128 patients with multiple myeloma at various stages of disease was compared as a function of tumor mass load and length of treatment. No significant differences in LI were noted for patients with various degrees of tumor reduction. Patients observed during the first 3 months of treatment had significantly higher LI than did untreated patients and those studied after longer intervals. In vivo cell-cycle analysis of myeloma cells by the halving time of the grain-count method failed to establish the length of the cell cycle but defined the length of S+G2 phases as being longer than 60 hours. Similar studies conducted in vitro defined a shorter combined length of S+G2 phase. In 2 patients in whom continuous infusion with [3H] thymidine was performed, the generation time was established as 8 days. Both patients had similar initial LI (5.7 and 6.6) but different growth fractions (19 and 47%). The measured tumor mass doubling time was much longer than the calculated one, suggesting the presence of considerable intrinsic cell loss (83 and 47%). These observations were summarized by a hypothetical two-compartment model for the growth kinetics of multiple myeloma.

Bone Marrow

Cytoplasmic fibrils in plasma cell leukemia.

In a patient with plasma cell leukemia, associated with pleural effusion, ultrastructural studies of the peripheral blood plasma cells showed an abundance of cytoplasmic fibrils. The nature of fibrils was not clarified, but they were not amyloid fibrils. This finding, coupled with a liaterature review, suggests that the cytoplasmic fibrils in plasma cells may be an additional cytological feature of this rare form of leukemia.

Cytoplasm

Ultrastructural aspects of crystal-like inclusions in a case of IgA plasma cell proliferation.

A case of apparently benign plasma cell proliferation is described. Needle-like paracrystalline inclusions were observed in numerous plasma cells outside the cisternae of the rough endoplasmic reticulum. They had a dense protein core surrounded by a layer of carbohydrate. They were found only in plasma cells secreting IgA-kappa immunoglobulin, but did not react with labelled specific antisera. A small amount of paraprotein IgA-kappa was found in the serum but no Bence-Jones protein could be detected in the urine. These observations suggest that these plasma cells synthesize normal IgA immunoglobulin and accumulate abnormal protein in the form of paracrystalline inclusions.

Aged

A method for the separation of lymphocytes and plasma cells from the human palatine tonsil using sedimentation in an isokinetic gradient of Ficoll in tissue culture medium.

Several methods for the dissociation of human tonsils into cell suspensions were compared. Dissociation of tonsils using 0-25 per cent trypsin gave both the largest number of total cells and the largest number of plasma cells per gram of tonsil. Lymphocytes and plasma cells were separated in a previously described isokinetic gradient of Ficoll in tissue culture medium. In the purest gradient fractions, lymphocytes were 97-2 plus or minus 1-9 per cent of nucleated cells. The purest gradient fractions contained 43-1 plus or minus 5-9 per cent plasma cells. More than 95 per cent of purified lymphocytes and plasma cells excluded Trypan Blue.

Cell Separation

Bone marrow kinetic studies on three patients with myelomatosis. Indications for malignant proliferation within both the plasma cell and lymphoid cell compartments.

The proliferative activity has been studied in lymphocyte and plasma cell populations of purified bone marrow from three myeloma patients. In one of them, proliferation was also recorded in blood lymphocytes. Immunofluorescence studies, using idiotypic antisera against the M-component, were performed to identify lymphocytes and plasma cells belonging to the malignant cell clone. Pulse incubations with 3H-thymidine (3H-TdR) were made in vitro. Autoradiographic analyses revealed a higher overall 3H-TdR labelling index in the myeloma cell populations than in normal control cell populations. A few labelled plasma cells were observed in the myeloma cases (2.5-5%) but the major fraction (11.5-14%) and intensity of labelled cells were found among the lymphoid cells. This indicates that the proliferation of malignant cells in myeloma occurs not only within the plasma cell but also in the lymphoid cell population. The results provide additional support for the assumption that B-lymphocytes are part of the malignant cell clone in myelomatosis and most likely precursors to the myeloma plasma cells.

Autoradiography

[Leukemia of plasma cells].

A case of plasma cell leukemia is reported, together with a review of the literature, including a summary of the signs, symptoms and laboratory findings usually present. The relationship of this leukemic state to ordinary multiple myeloma is discussed.

Autopsy

Human B cell differentiation. II. Suppression by T cells of T-dependent and T-independent plasma cell maturation.

In vitro human plasma cell generation induced by both T-dependent (PWM) and T-independent (NWSM) mitogens was found to be suppressed by peripheral blood lymphocytes preincubated with human aggregated IgG. T cells, but not B lymphocytes, were able to mediate the suppressive activity; since aggregated (Fab)'2 fragments were found unable to generate suppressor cells, it was concluded that the suppressor cell was a T lymphocyte bearing Fcgamma receptors. These cells appeared to be largely radiosensitive. In most cases the proliferative responses remained unchanged. Since NWSM-induced activation is not dependent on the presence of T cells, these results show that, at least in this case, T cells exert their suppressor function directly on B lymphocytes. Whether PWM-induced B cell differentiation is suppressed by the same mechanism or/and by inactivation of T helper lymphocytes remains under investigation.

B-Lymphocytes

Ultrastructural study of pulmonary plasma cell granuloma--report of a case.

Electron microscopy of a tumour diagnosed as a pulmonary plasma cell granuloma showed that the tumour was composed of plasma cells, fibroblasts, histiocytes and other inflammatory cells. The plasma cells were mature, with abundant endoplasmic reticulum. Fibroblasts contained multiple lipid droplets and intracytoplasmic inclusions. The relationship between plasma cell granuloma and lesions believed to be related is discussed.

Adult

Endobronchial plasma cell granuloma (xanthomatous pseudotumor); a light and electron microscopic study.

Plasma cell granuloma (xanthomatous pseudotumor) is polymorphic at light as well as electron microscopic examination. At light microscopy the endobronchial variant of this entity was rich in plasma cells and interwoven, whorl-like, or concentrically arranged spindle cells. Foamy histiocytes and macrophages usually abundant in the intrapulmonary variant were rare. At electron microscopy particles 20 to 50 nm. in size were found at the bronchial mucosal surface but not elsewhere in the lesion. Plasma cells near the bronchial surface contained cytoplasmic fibrils, mitochondria with concentric cristae, and inclusions that bore a close resemblance to adjacent extracellular crystallized hemoglobin. Those in the center of the lesion ordinary round inclusions and none of the other changes. Spindle cells in the interlaced areas were mostly fibroblasts or myofibroblasts, whereas those whorled around capillaries resembled pericytes with basement membranes and nuxes-like intercellular junctions. Year rings like multilayered basal laminae were frequently present between the pericyte-like cells and the endothelial cells of the capillaries. The ultrastructure of plasma cell granuloma, like the histologic and clinical aspects, differs from that of sclerosing hemangioma, pseudolymphoma, and malignant plasma cell tumor affecting the lung.

Adult

Intranuclear inclusions in Bence Jones lambda plasma cell myeloma.

A patient with plasma cell myeloma producing only Bence Jones lambda protein was found to have pale intranuclear inclusions in the majority of the bone marrow plasma cells. These inclusions, previously undescribed in myeloma patients producing only Bence Jones protein, contained Bence Jones lambda protein, were non-electron dense, bound by a single membrane, and contained no cytoplasmic structures. Intracytoplasmic inclusions were not present, and the perinuclear cistern was not dilated. Thus, the inclusions may represent intranuclear protein synthesis with anomalous release in the abnormal cells.

Bence Jones Protein

Heterogeneity of the B cell subpopulation operationally defined by (a) differentiation antigen(s) common to MOPC 104E and mature IgM plasma cells.

An antiserum raised in rabbit against MOPC 104E myeloma cells was extensively absorbed by murine IgM, thymocytes and spleen cells. Using indirect immunofluorescence, the distribution of the corresponding mouse plasma cell antigen(s) (MPCA) was determined among immunized spleen cells. Only 1-3% of the cells were stained but this MPCA-bearing subpopulation included all IgM plasma cells, a sizable proportion of IgG plasma cells and about one third of the antigen binding cells identified following deliberate immunization. It is therefore proposed that MPCA is transiently expressed during the antigen-induced differentiation of virgin B lymphocytes into memory cells as it does during the maturation process into Ig-producing cells and thus reflects the ontogenic relationship of these two differentiation pathways.

Antibody-Producing Cells