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Pivot Concepts:
Gene/Protein
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Target Concepts:
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Query: EC:3.4.22.6 (
chymopapain
)
407
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Selection of CD34+ hematopoietic progenitor cells from autografts may be performed in multiple myeloma (MM) to minimize contamination with tumor cells. This approach is based on the assumption that the malignant cells do not express the CD34 antigen. Therefore, we first compared the CD34+/CD10+ and CD34+/CD19+ subpopulations from bone marrow (BM) and peripheral blood (PB) of fourteen MM patients and five normal controls. No difference between the respective early B cell subsets of both groups could be observed. Using tricolor flow cytometry, the CD19 expression on CD34+/CD10+ cells in BM was found to increase continuously from CD19- to CD19dim. In contrast, circulating CD34+/CD10+ cells did not coexpress the CD19 antigen. This population may contain myeloid progenitor cells or bipotential progenitor cells of the myeloid and lymphoid lineage as suggested by data obtained with fetal liver cells. Further functional studies are required. Enrichment of CD34+ cells with immunomagnetic beads was performed from BM of three MM patients and four normal donors. The CD34+ cells were selected with the
HPCA
-1 antibody and detached from the beads with
chymopapain
. Compared with the starting cell preparation, a 3.97 +/- 0.48 log (mean +/- SE) reduction of plasma cells could be achieved after CD34 selection. On morphological examination, 84% +/- 4% of the cells in the CD34+ fraction (MM) were immature blasts. The plating efficiency for hematopoietic colony forming cells was 9.7% +/- 2.8% in the CD34 selected fraction of the MM group, reflecting a 51-fold increase as compared with the starting population.(ABSTRACT TRUNCATED AT 250 WORDS)
...
PMID:CD34 selection for purging in multiple myeloma and analysis of CD34+ B cell precursors. 751 57
The hematopoietic system has been one of the major targets for designing human gene therapy protocols. In the present system, we have transduced LNL6, one of the most commonly used retroviral vectors in gene therapy, into purified CD34+ cells from peripheral blood of patients primed with chemotherapy and granulocyte-macrophage colony-stimulating factor (GM-CSF). Purification of CD34+ cells was achieved by incubation with a murine anti-CD34 monoclonal antibody (9C5), and subsequently with paramagnetic microspheres (Dynal) coated with sheep anti-mouse IgG1 (Fc). The CD34+ cells were released from the beads by treatment with
chymopapain
. Flow cytometry analysis using the anti-CD34 antibody
HPCA
-2-FITC targeted at another epitope of CD34 showed that 78-97.5% of the cells thus purified were CD34+. After retroviral-mediated gene transfer, polymerase chain reaction (PCR) analysis revealed that 67-100% of the hematopoietic colonies contained the marker gene neo, indicating that CD34+ cells purified by immunomagnetic microsphere method from peripheral mononuclear cells primed with hematopoietic growth factors are highly susceptible to retroviral-mediated gene transfer. The expression of neo as determined by reverse transcription (RT)-PCR appeared to be unstable and not persistent. Taken together, our data suggest that LNL6 is a suitable vector for gene marking of hematopoietic progenitors but not for gene therapy protocols based on persistent gene expression.
...
PMID:High efficiency retroviral-mediated gene transduction into CD34+ cells purified from peripheral blood of breast cancer patients primed with chemotherapy and granulocyte-macrophage colony-stimulating factor. 751 49