Determination of homozygote vs heterozygote of Rh blood group antigens via rosette assays

Loren, Alan; Matsuo, Yoshinobu; Charman, D; Yokoyama, Mitsuo

doi:10.1046/j.1537-2995.1982.22382224939.x

Cited by 7 publications

(3 citation statements)

References 12 publications

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“…Table 1 summarizes a number of leukocyte-rosetting approaches. Protein A-bearing staphylococci, latex particles coated with IgG, and B lymphocytes have been used to characterize Rh(D)-positive erythrocytes sensitized with IgG class anti-Rh(D) Abs (EA; see Table 1) (Loren et al, 1982).…”

Section: Agglutination Involving Two Cell Typesmentioning

confidence: 99%

Precipitation and Agglutination

2000

Journal of Immunoassay

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Section: Agglutination Involving Two Cell Typesmentioning

confidence: 99%

Precipitation and Agglutination

2000

Journal of Immunoassay

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“…Not all reported D variants have been examined and a number of different techniques have been employed. The quantitative techniques employed include aggregation [10][11][12], antibody coated rosette formation [13], radioisotopic labelling [14][15][16][17], ELISA [18] and flow cytometry [19][20][21][22]. The number of D sites per cell (SPC) has been found to vary with different MAb-D for normal and weak D [17] and normal and D variant red cells [23].…”

Section: Introductionmentioning

confidence: 99%

Quantitation of Rh D Antigen Sites on Weak D and D Variant Red Cells by Flow Cytometry

Jones¹,

Lloyd-Evans²,

Kumpel³

1996

Vox Sanguinis

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Information on the number of D sites on weak D (Du) and D variant cells is limited and incomplete. The aim of this study was to use a simple non-isotopic technique utilising a combination of flow cytometry and ELISA to quantitate the number of D sites on an extensive range of these cells. Five human monoclonal IgG anti-D (BRAD-7 (JAC10), BRAD-5, 2B6, BRAD-3, H27) and one affinity-purified polyclonal IgG anti-D were each used at a saturating concentration of 20 micrograms/ml. In general, BRAD-3, BRAD-5 and 2B6 gave the highest number of D sites per cell (SPC), H27 and the polyclonal anti-D were slightly lower, while BRAD-7 gave the lowest SPC with all D-positive cells tested except DFR. Interestingly, BRAD-7 gave the highest SPC with DFR cells. Rh D antigen density for R2R2 cells was approximately double that seen with either R2r or Rzero (presumed R0r) cells. R1R1 cells gave only moderately higher SPC than R1r cells.Higher SPC were obtained with the R1 haplotype if the CW antigen was present. Weak D, Va and VI cells of the R1 haplotype had higher SPC than those of the Rzero or R2 haplotypes. The majority of D variant cells were found to have lower SPC than normal cells, and for polyclonal anti-D, which was the only anti-D to react with all D variants, SPC decreased in the order IIIc > IIIa > HMii > IVa > Va > DFR > DBT > IVb > VII > II > HMi > VI. The number of molecules of IgG anti-D bound to D variant cells varied by up to 10 times with reactive monoclonal antibodies. The highest SPC on weak D and D variant cells were obtained with BRAD-7 (DFR, 9,500), BRAD-3 (Va, 12,500), BRAD-5 (II, 5,500; VII, 5,300; weak D, 1,300), H27 (III, 24,000; VI, 2,900; HMi, 3,000; HMii, 16,800) and polyclonal anti-D (IVa, 9,300; IVb, 4,000; DBT, 4,300).

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“…Not all reported D variants have been ex amined and a number of different techniques have been em ployed. The quantitative techniques employed include ag gregation [10][11][12], antibody coated rosette formation [13], radioisotopic labelling [14][15][16][17], ELISA [18] and flow cy tometry [19][20][21][22]. The number of D sites per cell (SPC) has been found to vary with different MAb-D for normal and weak D [17] and normal and D variant red cells [23], Cells belonging to the same D category also had different numbers of D SPC when quantified using the same anti-D [17,24].…”

mentioning

confidence: 99%

Quantitation of Rh D Antigen Sites on Weak D and D Variant Red Cells by Flow Cytometry

Jones¹,

Loyd-Evans²,

Kumpel³

1996

Vox Sanguinis

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Information on the number of D sites on weak D (Du) and D variant cells is limited and incomplete. The aim of this study was to use a simple non-isotopic technique utilising a combination of flow cytometry and ELISA to quantitate the number of D sites on an extensive range of these cells. Five human monoclonal IgG anti-D (BRAD-7 (JACK)), BRAD-5, 2B6, BRAD-3, H27) and one affinitypurified polyclonal IgG anti-D were each used at a saturating concentration of 20 pg/ml. In general, BRAD-3, BRAD-5 and 2B6 gave the highest number of D sites per cell (SPC), H27 and the polyclonal anti-D were slightly lower, while BRAD-7 gave the lowest SPC with all D-positive cells tested except DFR. Interestingly, BRAD-7 gave the highest SPC with DFR cells. Rh D antigen density for R(2)R(2) cells was approximately double that seen with either R(2)r or R(0) (presumed R(0)r) cells. R(1)R(1) cells gave only moderately higher SPC than R(1)r cells. Higher SPC were obtained with the R, haplotype if the C^w antigen was present. Weak D, Va and VI cells of the R(1) haplotype had higher SPC than those of the R(0) or R(2) haplotypes. The majority of D variant cells were found to have lower SPC than normal cells, and for polyclonal anti-D, which was the only anti-D to react with all D variants, SPC decreased in the order IIIc>IIIa>HMii>IVa> Va>DFR> DBT>IVb>VII>II>HMi>VI. The number of molecules of IgG anti-D bound to D variant cells varied by up to 10 times with reactive monoclonal antibodies. The highest SPC on weak D and D variant cells were obtained with BRAD-7 (DFR, 9,500), BRAD-3 (Va, 12,500), BRAD-5 (II, 5,500; VII, 5,300; weak D, 1,300), H27 (III, 24,000; VI, 2,900; HMi, 3,000; HMii, 16,800) and polyclonal anti-D (IVa, 9,300; IVb, 4,000; DBT, 4,300).

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Determination of homozygote vs heterozygote of Rh blood group antigens via rosette assays

Cited by 7 publications

References 12 publications

Precipitation and Agglutination

Precipitation and Agglutination

Quantitation of Rh D Antigen Sites on Weak D and D Variant Red Cells by Flow Cytometry

Quantitation of Rh D Antigen Sites on Weak D and D Variant Red Cells by Flow Cytometry

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