Human B-lymphocyte antigens expressed by lymphocytic and myelocytic leukemia cells. I. Detection by rabbit antisera.

Billing, Ronald; B, Rafizadeh; Drew, Ian; Hartman, Gary A.; Gale, Robert Peter; Terasaki, Paul I.

doi:10.1084/jem.144.1.167

Cited by 170 publications

(40 citation statements)

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“…It has previously been reported that rabbit antisera prepared with papain digests of human malignant spleen cell membranes recognize antigens expressed by normal and neoplastic human B lymphocytes (1)(2)(3). The antigens recognized by these antisera appear to be similar to, and may be identical with, the p23, 30 antigen identified by Humphreys and colleagues and by Schlossman and colleagues on normal B cells, 15-20% of null cells, and the majority of acute leukemia cells (4,5).…”

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Antigens expressed by human B lymphocytes and myeloid stem cells.

Cline¹,

Billing²

1977

The Journal of Experimental Medicine

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It has previously been reported that rabbit antisera prepared with papain digests of human malignant spleen cell membranes recognize antigens expressed by normal and neoplastic human B lymphocytes (1-3). The antigens recognized by these antisera appear to be similar to, and may be identical with, the p23, 30 antigen identified by Humphreys and colleagues and by Schlossman and colleagues on normal B cells, 15-20% of null cells, and the majority of acute leukemia cells (4,5). A surprising and unexplained observation was the frequent detection of similar antigens on acute and chronic myeloid (granulocytic) leukemia cells. It was not clear why neoplastic myeloid cells should express an antigen found on mature B cells but not mature granulocytes. To answer this question, we examined the effects of similar antisera on normal human myeloid stem cells. These cells, colony-forming unit-culture (CFU-C), were assayed for their ability to form granulecyte and macrophage colonies in agar, in vitro (6, 7) in the presence or absence of antiserum and complement (C). It is now clear that the antigens recognized by such antisera also occur on normal myeloid progenitors. Materials and MethodsAntisera. As previously described, antisera were raised in rabbits immunized with papain digests of cell membranes from the spleen of patients with a variety of hematologic neoplasms (1).Cells. Heparinized aspirates of bone marrow were obtained from appropriately informed normal adult volunteers, and single cell suspensions were prepared as previously described (8).Heparinized peripheral blood of patients with chronic lymphocytic leukemia (CLL) was used to prepare lymphocyte populations of >98% purity for absorption of antisera. Permanent tissue culture lines of B cells (pooled lines B44, 45, and 47) and T cells (MOLT-4) were also used for absorption (1).Cytotoxicity Testing. Normal bone marrow cells, 2 × 106/ml, in 15% heat-inactivated fetal calf serum in McCoy's 5A medium were incubated with 1/10 volume of heated rabbit antiserum or heated normal rabbit serum at room temperature for 30 min. Normal rabbit serum, 1/10 volume, as a source of C was added for an additional 60 min. Cells were then plated in 0.3% agar above a feeder layer of 1 × 10 ~ peripheral blood leukocytes for assay of CFU-C as previously described (8). After 10 days incubation the number of colonies with >40 cells were determined. Results were expressed as the percentage inhibition (mean _+ SD) of colony formation in experimental assays relative to appropriate controls. The concentration of fresh rabbit serum used to provide C had no inhibitory effect on human marrow colony formation. ResultsAntisera to papain-digested spleen membranes inhibited normal marrow

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confidence: 73%

“…Permanent tissue culture lines of B cells (pooled lines B44, 45, and 47) and T cells (MOLT-4) were also used for absorption (1).…”

Section: Methodsmentioning

confidence: 99%

Antigens expressed by human B lymphocytes and myeloid stem cells.

Cline¹,

Billing²

1977

The Journal of Experimental Medicine

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“…Later, they were detected by hetero-antisera raised against the isolated protein from cell membranes (2)(3)(4). In addition to their presence on B cells and monocytes, they have been detected on the leukemic blast in eases of acute lymphocytic leukemia, acute myelogenous leukemia, and chronic myelogenous leukemia in blastic crisis (4)(5)(6).…”

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confidence: 98%

Ia-bearing T lymphocytes in man. Their identification and role in the generation of allogeneic helper activity.

Fu¹,

Chiorazzi²,

Wang³

et al. 1978

The Journal of Experimental Medicine

203

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The human Ia-like antigens were recognized initially as a series of HLA-linked alloantigens primarily represented on B lymphocytes with multiple pregnancy sera (I). Later, they were detected by hetero-antisera raised against the isolated protein from cell membranes (2-4). In addition to their presence on B cells and monocytes, they have been detected on the leukemic blast in eases of acute lymphocytic leukemia, acute myelogenous leukemia, and chronic myelogenous leukemia in blastic crisis (4-6). Recently, these Ia antigens were also shown to be present on the surface of precursor cells responsible for colony formation of both the myeloid monocytic and erythroid series (7-9) and on the surface of nonblood cells such as epidermal Langerhans cells (10).The presence of certain types of Ia antigens on T lymphoeytes has been demonstrated in murine systems, although the exact molecular character remains unclear (11). The presence of Ia antigens on human T cells has not been well documented. The present studies demonstrate the presence of Ia antigens on a small population of normal circulating human T lymphocytes, on T lymphocytes grown in long-term cultures, and on certain leukemic T cells. Evidence also is presented showing that cells responsible for the generation of helper activity during a mixed lymphocyte reaction are contained in this Ia-bearing T-cell population. Materials and MethodsIsolation of Lymphocytes. Mononuclear cells from the peripheral blood of normal individuals and patients with leukemia and various lymphoproliferative states and tonsillar mononuclear cells were isolated as described previously (12). Spontaneous rosette formation between human lymphocytes and sheep erythrocytes (SRBC) were performed with neuraminidase-treated SRBC (E). The rosette-forming cells (E-RFC) were separated from the nonrosette-forming cells by Ficoll-Hypaque (Pharmacia Fine Chemicals, Div. of Pharmacia Inc., Piscataway, N. J.) gradient centrifugation. The RFC-enriched fraction was purified further by repeated gradients until no significant numbers of cells were observed at the interphase. The E-RFCs were recovered after lysis of SRBC by a Tris-buffered ammonium chloride solution. Tonsillar B cells were obtained by the depletion of E-RFCs from the mononuclear cell preparations.

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“…Anti-Ta-like serum was obtained from Alpha Gamma Labs (Sierra Madre, California, U.S.A). The characteristics of the antiserum have been described in detail elsewhere (Billing et al, 1976 Sternberger et al (1970) as modified by Taylor (1974). The procedure has been described in detail by Papadimitriou et al (1978).…”

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confidence: 99%