Compatible Red Blood Cells Research Articles

Innovative Two-Step Negative Selection of Granulocyte Colony-Stimulating Factor–Mobilized Circulating Progenitor Cells: Adequacy for Autologous and Allogeneic Transplantation

A major obstacle in purifying either autologous or allogeneic hematopoietic stem cells from granulocyte colony-stimulating factor (G-CSF) mobilized circulating progenitor cells (CPC) is represented by the huge cellularity present in each apheretic product. To obtain a significant debulking of unwanted cells from the leukapheresis, we developed a modified protocol of immune rosetting whereby human ABO-Rh- compatible red blood cells (RBCs) are treated with chromium chloride and then coated with murine monoclonal antibodies (MoAbs) against leukocyte antigens. When experiments were performed with leukaphereses obtained from normal donors or from T-cell acute lymphoblastic leukemia (T-ALL) patients, RBCs were coated with murine MoAbs against human mature myeloid cells (CD11b) and T cells (CD6); whereas, in the case of patients with B-precursor ALL, B-cell non-Hodgkin's lymphoma (B-NHL), or multiple myeloma (MM), RBCs were coated with anti-CD11b only. After incubation with CPC, rosetting cells (myeloid precursor cells, granulocytes, monocytes, and T cells) were removed by Ficoll-Hypaque density gradient centrifugation with a blood cell processor apparatus, COBE (Lakewood, CO) 2991. After this step, a significant reduction of the initial cellularity was consistently obtained (range, 72% to 97%), whereas the median absolute recovery of the CD34+ cells was above 85% (range, 64 to 100), with a 10-fold relative enrichment ranging from 3% to 41%. In a second step, CPC can be further purged of contaminating T or B cells by incubation with lymphoid-specific magnetic microbeads (anti-CD2 and -CD7 to remove T cells; anti-CD19 to remove B cells) and elution through a type-D depletion column (composed of ferromagnetic fiber) inserted within a SuperMACS separator device (Miltenyi Biotech, Bergisch-Gladbach, Germany). By this approach, a highly effective (three to four logs) T-cell depletion was achieved in all experiments performed with normal donors or T-ALL patients (median loss of CD3+ cells: 99.8% [range 99.2 to 100]) and an equally efficient B-cell depletion was obtained from B-precursor ALL, B-NHL, or MM patients. At the end of the procedure the T- or B-cell depleted fraction retained a high proportion of the initial hematopoietic CD34+ stem cells, with a median recovery above 70% (range 48% to 100%) and an unmodified clonogenic potential. In five patients (two follicular NHL and three ALL) the purified fraction of stem cells was found disease free at the molecular level as assessed by polymerase chain reaction (PCR) analysis of the t(14;18) chromosome translocation or clono-specific DNA sequences of IgH or T-cell receptor gamma and delta chain genes. Purified autologous and allogeneic CPCs were transplanted in three and six patients, respectively, who showed a prompt and sustained hematologic engraftment. In conclusion, this method represents a simple and reproducible two-step procedure to obtain a highly efficient purging of T or B cells from G-CSF expanded and mobilized CPCs. This approach might lead to the eradication of the neoplastic clone in the autologous stem cell inoculum as well as for T-cell depletion during allogeneic transplantation.

Innovative Two-Step Negative Selection of Granulocyte Colony-Stimulating Factor–Mobilized Circulating Progenitor Cells: Adequacy for Autologous and Allogeneic Transplantation

AbstractA major obstacle in purifying either autologous or allogeneic hematopoietic stem cells from granulocyte colony-stimulating factor (G-CSF) mobilized circulating progenitor cells (CPC) is represented by the huge cellularity present in each apheretic product. To obtain a significant debulking of unwanted cells from the leukapheresis, we developed a modified protocol of immune rosetting whereby human ABO-Rh– compatible red blood cells (RBCs) are treated with chromium chloride and then coated with murine monoclonal antibodies (MoAbs) against leukocyte antigens. When experiments were performed with leukaphereses obtained from normal donors or from T-cell acute lymphoblastic leukemia (T-ALL) patients, RBCs were coated with murine MoAbs against human mature myeloid cells (CD11b) and T cells (CD6); whereas, in the case of patients with B-precursor ALL, B-cell non-Hodgkin's lymphoma (B-NHL), or multiple myeloma (MM), RBCs were coated with anti-CD11b only. After incubation with CPC, rosetting cells (myeloid precursor cells, granulocytes, monocytes, and T cells) were removed by Ficoll-Hypaque density gradient centrifugation with a blood cell processor apparatus, COBE (Lakewood, CO) 2991. After this step, a significant reduction of the initial cellularity was consistently obtained (range, 72% to 97%), whereas the median absolute recovery of the CD34+ cells was above 85% (range, 64 to 100), with a 10-fold relative enrichment ranging from 3% to 41%. In a second step, CPC can be further purged of contaminating T or B cells by incubation with lymphoid-specific magnetic microbeads (anti-CD2 and -CD7 to remove T cells; anti-CD19 to remove B cells) and elution through a type-D depletion column (composed of ferromagnetic fiber) inserted within a SuperMACS separator device (Miltenyi Biotech, Bergisch-Gladbach, Germany). By this approach, a highly effective (three to four logs) T-cell depletion was achieved in all experiments performed with normal donors or T-ALL patients (median loss of CD3+cells: 99.8% [range 99.2 to 100]) and an equally efficient B-cell depletion was obtained from B-precursor ALL, B-NHL, or MM patients. At the end of the procedure the T- or B-cell depleted fraction retained a high proportion of the initial hematopoietic CD34+ stem cells, with a median recovery above 70% (range 48% to 100%) and an unmodified clonogenic potential. In five patients (two follicular NHL and three ALL) the purified fraction of stem cells was found disease free at the molecular level as assessed by polymerase chain reaction (PCR) analysis of the t(14;18) chromosome translocation or clono-specific DNA sequences of IgH or T-cell receptor γ and δ chain genes. Purified autologous and allogeneic CPCs were transplanted in three and six patients, respectively, who showed a prompt and sustained hematologic engraftment. In conclusion, this method represents a simple and reproducible two-step procedure to obtain a highly efficient purging of T or B cells from G-CSF expanded and mobilized CPCs. This approach might lead to the eradication of the neoplastic clone in the autologous stem cell inoculum as well as for T-cell depletion during allogeneic transplantation.

The clinical significance of anti-Bga-51 chromium survival studies.

An attempt to determine the clinical significance of anti-Bga was performed by transfusing 51Cr-labelled red blood cells (RBC) from HLA-B7 positive donors to four patients who had histories of anti-Bga. Pre- and post-transfusion samples showed no change in antibody titer scores, haptoglobin or complement levels. Three of the patients were then transfused with 51Cr-labelled RBC from HLA-B7 negative donors and the half-life (T50Cr) of both were compared. Although low, the T50Cr of HLA-B7 incompatible RBC appeared to be similar to the T50Cr of HLA-B7 compatible RBC.

赤血球抗原刺激によらない抗Ｋ（Ｋ１）の１例

An example of anti-K1 found in a Japanese patient with occipitalcraniomenigiomata, aged 19 years, is reported. His red cells had not K1 antigen. The patient had no history of prior blood transfusion, but he received twelve units of K1 negative and compatible red blood cells at the time of surgery in July 1982. No clinical and haematological signs of red cell destruction were noted. Unexpected antibodies were not detectable by routine pretransfusion screening.However, in September 1982, his serum was found to contain an IgG antibody with anti-K (K1) specificity; the titer of which was 16 to 32 with K:1 and 8 to 16 with K:1, 2 cell samples by the indirect antiglobulin test using anti-IgG.His red cells gave a doubtful direct antiglobulin test using polyspecific antiglobulin and anti-C3, but not anti-IgG. Anti-K1 specificity was demonstrated in an eluate from his red blood cells.The antibody stimulation of the anti-K1 remains unknown.

Transient LW‐Negative Red Blood Cells and Anti‐LW in a Patient with Hodgkin's Disease1

LW-negative (LW4) red blood cells and anti-LW antibody were discovered in the blood of a 50-year-old white male with Hodgkin's disease and blood type O. Compatible red blood cells for transfusion could not be obtained because available LW-negative cells were either type A, LW4, or type O, LW3. Following cytotoxic therapy for his basic disease, the anti-LW disappeared and his red blood cells developed normal reactions with anti-LW sera.

Hemolytic Anemia Secondary to Stibophen Therapy

STIBOPHEN (Fuadin) (sodium antimony III biscatechol-2-4 disulfonate heptahydrate), is an anthelmintic which since 1929 has been used extensively in the treatment of schistosomiasis. The manufacturers claim that toxicity is low and that severe side effects have not been observed. Transient nausea, vomiting, headache, diarrhea, and muscular and joint pains have been the most frequently observed side effects. Thrombocytopenia has been the most common hematological complication.1 In 1956 Harris2,3described the occurrence of hemolytic anemia in a patient who was receiving stibophen therapy. Furthermore, he found that when stibophen was added to the patient's serum, antibodies active against cells from the patient and compatible red blood cells could be demonstrated. The patient had a positive direct and indirect Coombs' test. In 1960, a similar case occurred but the antibodies appeared to be of the saline agglutinating type. During 1961, two cases of stibophen-induced hemolytic anemia came to our attention.