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Biomedical subjects

M J Cline

Publications and source records attributed to M J Cline.

At least 91 records · Page 5Linked to original sources

Insertion of a new gene of viral origin into bone marrow cells of mice.

DNA containing the herpes simplex virus thymidine kinase (HSVtk) gene was used to transform wild-type tk+ mouse L cells to a tk++ status in vitro using methotrexate as a selective agent. HSVtk DNA was also used to transform mouse bone marrow cells in vitro. Transformed marrow cells injected into irradiated and methotrexate-treated recipient mice gave rise to proliferating cells which in some cases dominated the marrow population and which contained HSVtk gene sequences.

Animals↗

Gene transfer in intact animals.

Resistance to methotrexate was induced in bone marrow cells of mice by transformation in vitro with DNA from a drug-resistant cell line. Transformed cells were injected in vivo and haematopoietic cells expressing resistance were selected by drug treatment of recipients. Transformed cells had elevated levels of dihydrofolate reductase and demonstrated a proliferative advantage over untransformed cells, indicating successful gene transfer.

Animals↗

Amphotericin B plus combination chemotherapy for extensive non-small cell bronchogenic carcinoma.

Sixteen patients with extensive non-small carcinoma of the lung were treated with methotrexate, vincristine, cyclophosphamide, and adriamycin. Cyclophosphamide and adriamycin were administered after pretreatment with amphotericin B. Amphotericin B-related toxicity was mild; cytotoxic chemotherapy-related toxicity was tolerable. The partial response rate was 12.5% and median survival was between 19 and 20 weeks. Response rate and survival were not superior to those of a similar drug combination lacking amphotericin B.

Amphotericin B↗

Serum inhibitors of myelopoiesis.

A modification of the microcytotoxicity assay of Terasaki & McClelland was used to screen for serum inhibitors of myeloid progenitor cells (CFU-C). Sera from 104 patients with neutropenia or bone marrow failure and from 104 controls, including 30 normal subjects and 74 patients with various disorders or multiple transfusions, were studied. Inhibitors of CFU-C were found in 19 of the 104 patients with neutropenia or marrow failure: three with acquited neutropenia and 16 with aplastic anaemia. In 17 of the 19 patients the inhibitor was either complement dependent or chavincingly associated with clinical remission of disease in one patient. In another patient, the inhibitor was an IgG antibody directed against non-HL1 antigens present on CFU-C of all 50 normal target marrows tested; however, it was not clearly autoreactive. Inhibitors were also found in one of nine patients with lupus erythematosus, one with breast carcinoma, one patient with renal transplant rejection, and six of 46 multiply transfused patients without marrow failure. Our findings indicate that most serum inhibitors are associated with multiple transfusion: convincing evidence of pathogenetically important autoreactive antibody was present in only one of 19 patients with inhibitors.

Adult↗

Differentiation of subpopulations of human and murine hemopoietic stem cells by hypotonic lysis.

Both human and mouse bone marrow contain subpopulations of hemopoietic stem cells that greatly vary in their resistance to water exposure: The cells forming erythroid colonies or bursts in methyl cellulose in vitro are most sensitive to hypotonic conditions and are destroyed within 60 s in the hypotonic milieu. The murine pluripotent stem cells assayed by the spleen colony technique, as well as both murine and human myeloid stem cells assayed by the plasma clot diffusion chamber technique, displayed intermediate sensitivity and were nearly completely eliminated by 120 s of exposure to water. Both human and mouse bone marrow stem cells producing myeloid colonies in agar are most resistant to hypotonic conditions. The addition of monocyte-macrophages and lymphoid cells to water-exposed mouse bone marrow cell populations to compensate for losses did not restore either erythroid or myeloid colony formation.

Animals↗

Cytotoxicity of anti-beta 2-microglobulin xenoantisera to human and murine myeloid progenitor cells and lack of cross-reactivity with colony-stimulating activity.

A previous report has suggested an antigenic relationship between beta 2-microglobulin (beta 2 mu) and granulocyte colony-stimulating activity (CSA). Since human myeloid progenitor cells (CFU-C) express HLA antigens and beta 2 mu is a known molecular component of HLA antigens, we wondered whether the reported effect of anti-beta 2 mu heteroantisera on in vitro granulopoiesis might result from cytotoxicity to CFU-C rather than from cross-reactivity with CSA. To test this, we used rabbit antibody reactive with human and murine beta 2 mu (anti-beta 2 mu). Treatment of human and murine bone marrow cells with anti-beta 2 mu and complement resulted in 95+% inhibition of CFU-C colony formation compared to controls. To test for an effect on CSA, anti-beta 2 mu was incubated with human and murine sources of CSA. After addition of goat anti-rabbit Ig antiserum to precipitate immune complexes and unbound anti-beta 2 mu, the supernatant fluid retained CSA but was no longer cytotoxic to CFU-C. These results indicate that human and murine CFU-C express membrane beta 2 mu and that anti-beta 2 mu antibody does not cross-react with human or murine CSA.

Animals↗

Monoclonal antibodies to HLA-A,B, and Ia-like antigens inhibit colony formation by human myeloid progenitor cells.

Hybridomas derived from the fusion of murine myeloma cells with splenocytes from mice immunized with human cultured lymphoid cells secreted monoclonal antibodies to human cell surface antigens. Serologic and immunochemical assays showed that 4 monoclonal antibodies (Ab Q2/47, Q2/61, Q2/70, Q2/80) recognize framework determinants of Ia-like antigens and 1 monoclonal antibody (Ab Q1/28) reacts with determinants expressed on the heavy chain of HLA-A,B antigens. Both anti-HLA-A,B and anti-Ia-like antigen monoclonal antibodies caused complement-dependent inhibition of granulocyte-macrophage colony formation by human bone marrow grown in soft agar. Mixing experiments excluded the possibility of an indirect effect on progenitor cells by lysis of auxiliary cells. These results indicate that human myeloid progenitor cells express HLA-A,B and Ia-like antigens.

Animals↗

Enhancement of murine erythroid colony formation in the presence of activated T lymphocytes.

Erythroid colony formation was enhanced by treatment of mouse bone marrow cells with PHA prior to culture in diffusion chambers implanted into intact mice. Addition of PHA-pretreated thymocytes, lymph node lymphocytes, and spleen cells to untreated bone marrow cells also resulted in increased numbers of erythroid colonies in diffusion chambers but had no effect on myeloid colonies. PHA did not augment erythroid colony formation when bone marrow was depleted of Thy 1-bearing cells, although cells capable of erythroid differentiation were still present as judged by the response to erythropoietin. Furthermore, mixing of bone marrow depleted of Thy 1 + cells with PHA-treated thymocytes restored augmented erythroid colony formation. Lack of responsiveness of bone marrow cells to concanavalin A and pokeweed mitogen may indicate that a specific T cell subpopulation reacting with PHA is able to stimulate erythropoiesis. Note that addition of untreated thymocytes, lymph node lymphocytes, and spleen cells into cultures had no effect.

Animals↗

Immunologic classification of acute lymphoblastic leukemia. Implications for normal lymphoid differentiation.

Acute lymphoblastic leukemia (ALL) is a heterogeneous disease as defined by clinical characteristics and immunologic techniques. The standard cell surface markers are sheep erythrocyte receptors for T lymphocytes and surface membrane immunoglobulin for B lymphocytes. Utilizing these markers, three subtypes of ALL designated T-ALL, B-ALL and non-B, non-T or null ALL have been defined. We have studied 70 patients with ALL utilizing these standard cell surface markers. In addition, we have further subclassified these patients by testing each cell for an ALL-associated antigen, the la-like antigen, and thymocyte antigen(s) all defined by well-characterized antisera. We can define 12 subgroups of ALL by their surface antigenic characteristics. These subgroups may have relevance to the clinical expression of disease and may define identifiable stages of normal lymphocyte development.

Animals↗

The effect of granulopoietic stress in mice with "latent" bone marrow failure.

We treated mice with high-dose busulfan to produce a residual or "latent" hematopoietic injury characterized by persistent normal or near-normal peripheral blood counts and marrow cellularity but moderate to marked reduction in hematopoietic stem cells. Control and "latent" mice were then challenged with glucan, a granulopoietic stimulant. We found that glucan administration induced a comparable increase in granulocytic differentiation in control and "latent" mice. This differentiational expansion was not accompanied by a further depletion of pluripotent hematopoietic stem cells. Our results fail to demonstrate competition between self-renewal and differentiation at the level of the CFUS, at least in this model. Stimuli for granulopoiesis act primarily on more distal, committed populations which are apparently unaffected by prior exposure to busulfan.

Anemia, Aplastic↗

[Evidence for CFU-C self-replication (author's transl)].

To explore the proliferative potential of committed myeloid progenitor cells (CFU-C), we placed suspensions of mouse spleen cells in short-term liquid culture with colony stimulating activity and subcultured the cells in agar at intervals to assay for CFU-C. Prior to liquid culture, aliquots of spleen cells were exposed briefly to hypotonic conditions to lyse CFU-S but not CFU-C. A 50-fold increase in CFU-C was observed in control cultures. The pattern of CFU-C proliferation was not altered significantly by hypotonic lysis despite the fact that this treatment effectively removed CFU-S. These results support the hypothesis that CFU-C are capable of self-replication and permit a model of hematopoiesis in which expansion of the granulopoietic pool is controlled, in part, at the level of the CFU-C.

Animals↗

Cure of aplastic anaemia in paroxysmal nocturnal haemoglobinuria by marrow transfusion from identical twin: Failure of peripheral-leucocyte transfusion to correct marrow aplasia.

The ability of syngeneic peripheral leucocytes to cure marrow aplasia was tested in a patient with paroxysmal nocturnal haemoglobinuria (P.N.H.). Transfusion of 7.1X10(10) white cells obtained by leucopheresis from an identical-twin donor, providing 3.4X10(4) myeloid progenitors (C.F.U.-C)/kg, failed to improve marrow function within two months. In contrast, transfusion of 1.3X10(10) nucleated bone-marrow cells, representing 6.4X10(4) C.F.U.-C/kg, from the same donor resulted in prompt bone-marrow recovery. These observations support the hypothesis that aplastic anaemia in P.N.H. is a stem-cell defect that may be corrected by the simple infusion of relatively small numbers of normal bone-marrow cells. They also seem to indicate a distinct advantage of marrow cells over peripheral-blood mononuclear cells in their ability to correct marrow aplasia.

Adult↗

Cellular interactions in haematopoiesis.

In vitro culture of haematopoietic cells has provided some surprising insights into critical interactions of blood-forming cells. Subpopulations of lymphoid cells have been shown to produce colony-stimulating activity, to interact with macrophages, and to have important effects on the very early stages of erythropoiesis. Macrophages have multiple influences on the proliferation and differentiation of other haematopoietic cells.

Animals↗

Hemopoietic precursor cells in human peripheral blood.

Human peripheral blood contains two types of stem cells that differentiate along the granulocytic pathway. They are separable by their ability to form colonies in agar in vitro (CFU-C) and in plasma clots in diffusion chambers in vivo (CFU-DG). Kinetic studies suggest that CFU-DG represents an intermediate between the still hypothetical human pluripotent stem cell and CFU-C.

Animals↗