Michael C. Rosner

Phagocytic capacity of hairy cells from seventeen patients

SummaryHairy cells from 17 patients diagnosed as having hairy cell leukemia were evaluated with both light and transmission electron microscopy for their capacity to phagocytose zymosan, latex,Staphylococcus aureus, andPseudomonas aeruginosa. The hairy cells of fifteen of the sixteen patients which were tested with latex showed significant phagocytosis. Cells of thirteen of the seventeen patients showed phagocytosis of staphylococcus; six of the seventeen, phagocytosis of zymosan; and four of seventeen, phagocytosis of pseudomonas. Of the four substances tested, staphylococcus and latex yielded consistently higher numbers of ingested particles per hairy cell than did zymosan or pseudomonas. Observation of serial sections by transmission electron microscopy revealed that each type of particle is fully enclosed within the cytoplasm of the hairy cell.A Lysostaphin assay performed on the hairy cells of two patients showed that lysis of the phagocytosed particles by the hairy cell differs significantly from that by normal mononuclear cells. The results support our earlier study which suggested that hairy cells have phagocytic capabilities which differ not only among patients, but also within a single patient over time.

Surface immunoglobulin, lectin-induced cap formation and phagocytic function in five patients with the leukemic phase of hairy cell leukemia

Hairy cells from 5 patients with greater than 25% hairy cells in the peripheral blood (4 had greater than 45% hairy cells and white blood cell counts (WBC) greater than 10,000/mm3) were studied for surface immunoglobulin (SIg) presence and distribution by two methods, for lectin‐induced cap formation, and for phagocytosis of zymosan. Hairy cells from all 5 cases were found to have distinct monoclonal patterns, 2 Gk 1 MDk, 1 Dk, and 1 GMDk, as well as cap formation with SIg. All 5 cases showed distinct lectininduced cap formation in a percentage of cells similar to the percentage of hairy cells, and 2 of the 5 patients had hairy cells which phagocytosed zymosan. These findings contrasted with the malignant cells from 4 patients with CLL, which had monoclonal SIg but no SIg cap formation and no significant percentage of lectin‐induced cap formation. Cells from 2 cases of T cell lymphomas had no SIg and no lectininduced cap formation as did cells from 2 cases of non‐lymphomas leukemia. Hairy cells not only appear to have SIg cap formation similar to some B‐lymphocytes, which in some patients also phagocytose zymosan, but also demonstrate strong lectin‐induced cap formation. Cancer 46:50–55, 1980.

In vitro response of cells from three patients with hairy cell leukemia to a recombinant leukocyte interferon

SummaryMononuclear cells from the peripheral blood of two patients and from the spleen of one patient, all of whom had hairy cell leukemia, were cultured with a recombinant human leukocyte interferon (RDα2-IFN). The IFN was added at concentrations of 10, 100, 1,000, and 10,000 IU/ml, and the cells were cultured for 1, 3, and 7 days. A cytocidal effect of IFN was observed only on cultured cells from the spleen at day 7. Electron-microscopic observations demonstrated that RDα2-IFN induced the formation of tubuloreticular structures (TRSs) and annulate lamellae (ALs) in hairy cells, as well as in co-isolated non-leukemic cells, from all three patients. Ultrastructural examination revealed a close proximity between ALs and TRSs in co-isolated non-leukemic cells. A variability with respect to the induction of TRSs in hairy cells was observed among the three patients. In two of the three patients, the percentage of hairy cells with TRSs increased with the duration of incubation and with the dose of IFN. In the third patient, few hairy cells showed TRSs after 7 days of incubation with IFN. Our findings indicate that leukemic hairy cells may be heterogenous in their response to IFN.

Hairy cell leukemia: differences in phagocytic capacity of cells in vitro.

SummaryHairy cells from eight patients with hairy cell leukemia were evaluated with both light and transmission electron microscopy for their capacity to phagocytose zymosan, latex, staphylococcus aureus, and pseudomonas aeruginosa. In two patients, there was no phagocytosis of any of these substances; cells from three patients phagocytosed only latex; two, all except pseudomonas ; and one, all 4 substances. Hairy cells became relatively smooth while in culture with staphylococcus, but no surface changes were noted during incubation with the other substances. Of the eight patients studied, one died of pseudomonas pneumonia and sepsis; pseudomonas was the only substance which her hairy cells did not phagocytose. The one patient whose hairy cells phagocytosed all 4 test substances developed a disseminated Mycobacterium intracellulare infection; culture of his hairy cells with this atypical mycobacterium showed no phagocytosis. Hairy cells have different phagocytic capabilities from patient to patient, and the evaluation of these capabilities in vitro might provide early identification of potential infectious complications.

Ultrastructure of Four Human Germ Cell Tumor-Derived Cell Lines: Effect of 12-0-Tetradecanoyl Phorbol-13-Acetate

Cells derived from human germ cell tumors (HGCT) are potential models for the study of human embryonic differentiation. Differentiation in four HGCT-derived lines (1618K, 833KE, 1777Ndif, 2806B) was examined by light and electron microscopy, after exposure of cells to dimethyl sulfoxide (DMSO) and 12-O-tetradecanoyl phorbol-13-acetate (TPA). In line 1618K, the effect of retinoic acid was also examined. After exposure to TPA, the cells of line 1618K had a loss of microvilli, an increased amount of nuclear heterochromatin, and an increase in the number of cytoplasmic vacuoles. The cells of line 833KE exposed to TPA showed only an increase in cytoplasmic vacuoles. No changes were observed in any cell line after exposure to DMSO. Similarly no changes were observed in 1618K after exposure to retinoic acid. These findings indicate that DMSO and retinoic acid are not effective inducers of differentiation in these HGCT-derived cell lines. The phorbol ester, TPA, appears to induce differentiation of some HGCT-derived cell lines.

Ultrastructural Characteristics of the Spleen in Hairy Cell Leukemia

Eighteen spleens derived from patients with hairy cell leukemia (HCL) were analyzed by correlative scanning and transmission electron microscopy. In 15 of the cases, the white pulp areas were markedly decreased or absent when compared to normal spleens, although few hairy cells were observed within this region. In only one case did the white pulp appear normal. In all HCL cases, hairy cells were observed within normal, dilated, and abnormal sinuses. The abnormal sinuses contained hairy cells of typical morphology attached to other hairy cells, to endothelial lining, and to erythrocytes. The degree of sinus filling by hairy cells varied from loosely- to tightly-packed. Endothelial cells exhibiting degenerative changes, such as swelling with smooth surfaces and dilated intercellular spaces, were frequently seen. These results indicate that in addition to the previously described overcrowding of the spleen by hairy cells, the splenic tissue itself is considerably altered and sometimes severely damaged in patients with HCL.