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dothandar last won the day on June 11

dothandar had the most liked content!

About dothandar

  • Rank
    Junior Member
  • Birthday 07/23/1982

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  • Gender
    Not Telling
  • Interests
    I am obsessed with actions going on red cell surface
  • Location
    Seattle, WA
  • Occupation
    IRL tech

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  1. dothandar


    Hello. sorry about the delayed reply. We worked further on this case and turned out the pooled AB plasma that we have frozen did not react with neuraminidase-treated cells (another aliquot of NeuNac treated cells) but fresh AB plasma did. We did test this patient's cells with fresh AB plasma and got a positive reaction. We have to try not to used frozen/thawed AB pooled plasma for this test in the future
  2. dothandar

    wAIHA with IgM and C3c/C3d coating

    I read in Dr.Garratty's Immune Hemolytic Anemias textbook that Spontaneous agglutination is often seen with warm IgM cases. Would it be helpful to spin this patient's cell suspension with either saline or 6-10% albumin if the warm IgM AIHA is in question? And also preform titer and thermal amplitude test to see if antibody titer increases or decreases with higher temperature?
  3. dothandar

    WAA Formation

    Are your cells that is non reactive with eluate either e- or f-? did you have antigen typing on this patient? Was the patient recently transfused?
  4. I recently used this procedure to differentiate anti-Pr from anti-En(a), which is the only time I have used it. Since our lab refer possible Asub for genotyping lab, we have no chance of trying this out to catch weak subgroup of A.
  5. dothandar

    AntiD +Anti G

    Hello, I am wondering what kind of method do you use to quantify anti-D in europe. here, we titer the maternal plasma with D+ cells and freeze an aliquot. Next visit, we would titer the fresh plasma parallel to thawed plasma, see if the titer has gone up compared to previous titer. Seems like having an assay to quantify the amount of anti-D and having a clinical cut off correlation is a better way to approach this.
  6. dothandar

    RESt and DARA

    Anti-CD38 did not react with In(Lu) cells and showed very low signals when anti-CD38 was tested with In(Lu) cells in a flow cytometry. Before we knew the patient was on anti-CD38 and In(Lu) cells has depressed CD38 marker, we have mistaken it being an antibody to high incidence antigen in Lutheran blood group system, based on the reactivity with In(Lu) cells.
  7. dothandar

    5 months with all positive tests!

    Looking at this kid's clinical information may be a good start. What is the diagnostics? is the kid actively hemolyzed? What is the medication history? did you try washing this patient's cells with warm saline and try ABO/D typing with warm-washed cells? That is feasible to do in hospital lab. I would even try using DTT if you have some in-house.
  8. dothandar

    wAIHA with IgM and C3c/C3d coating

    Hello, I am learning so much from you guys here as usual. I also realized this english flag along with the 2+ IgM on the DAT result. I started wondering what kind of direct antiglobulin test is performed to detect anti-IgM in the UK. Do you have liquid antisera (monoclonal, animal antisera etc) or anti-IgM suspended in gel?
  9. dothandar

    C3d positive on cord sample

    Beautiful pictures. Thank you for sharing the cases. I have been on this site for almost a decade and never stopped learning from you guys! I wish we have these gel cards here in the States. There were sometimes we suspected problem due to IgA in DAT negative patients and those cards will be great to investigate such cases. I tend to attempt to "reinvent the wheel" due to my lack of historical knowledge and experience. I wonder if the infants cells from moms with Kidd blood group antibodies screened with anti-C3 in the past to see how many of them have turned up positive. It will be an interesting data to see if that has not already been done.
  10. dothandar

    C3d positive on cord sample

    I am wondering why the cord cells were tested with anti-C3? I believe it is not a common practice to test the cord with anti-C3 I just saw one too a few months ago, mine is a peripheral draw on a new born rather than cord, anti-IgG and -C3 pos. I looked it up on Issitt's textbook. His references indicates that HDN due to complement activaton is an argument that has been challenged, as "C3 coating can be found on normail cord samples". I think if Hb and bili are normal C3 coating probably is not significant??
  11. Very interesting case! Here are my questions related to this case. 1) If it is a case of auto-anti-LW, should we expect the autoantibody to have a "relative specificity" to D antigen, especially with a 3+ reaction by ortho gel. Would a transient suppression of LW antigen effect D antigen typing by a monoclonal reagent? 2) Antigen loss during AIHA is shown in Kell blood group system. Is it possible in Rh blood group system as well? Zimring JC, et al. Antigen loss from antibody-coated red blood cells. Transfus Med Rev. 2009 Jul;23(3):189-204. PMID 19539874
  12. here is a fun case along with a question that I cannot wrap my head around it. Mother: Anti-D and anti-G identified (both strongly reactive by PeG and saline-IAT), Anti-C ruled out by differential adsorption and elution with R2R2 cells and r'r cells. All other common alloantibodies ruled out. She was not previously transfused or pregnant. TITER with R2R2 cells = 1024, titer with r'r cells - 128 (I know this titer is so unhelpful since expression on each of our indicator cells are different) Baby: (here is the fun part.. ready?) - on the birthday D typing negative with MoAb anti-D reagent at room temperature with untreated cells, EGA treated cells, twice EGA treated cells. DAT 4+ with untreated cells, EGA treated cells and twice EGA treated cells. Baby is fine (dont need transfusion, not hemolysing). Plasma and eluate reactive with D+ and C+ cells. insufficient sample to perform adsorption/Elution to differential anti-D from anti-G. eluate- reactive with D+ cells and C+ cells/ we assume it was anti-D and anti-G in there. Genotyping result. RHD*DAU0. Homozygote D deletion detected. So baby is heterogygote DAU-0 without a normal D gene. 1 month later, baby came back hemolysing. DAT 4+ with untreated cells. Anti-D typing 4+ with EGA treated and untreated cells. DAT on EGA treated cells is negative. plasma and eluate reactive with C+ and D+ cells. all other alloabs ruled out. Not enough sample to perform adsorption/Elution My question is.. The baby's red cells were clearly coated with anti-G and/or anti-D why did it not hemolyse at birth but 1 month after. I have heard of blocking mechanism serologically as in this case Does the heavy coating of antibodies makes Fc receptor inaccessible for macrophages for hemolysis? Could it be anti-G alone (not anti-D) coating on the red cells protecting the red cells from hemolysis by anti-D?
  13. dothandar

    Cord cells vs. DTT

    Ours is DTT. We tried cords (alot of different sources) Not every cord cells gave is a negative reaction with plasma from patients on DARA. half of them still reacted microscopically.
  14. dothandar

    SCARF cells

    Thank you very much! This is very helpful to see that cells that are previously frozen in liquid nitrogen can be transferred to -80C and it would recover well. I should have been more clear in my question (#2) above. What I envisioned was making droplets in liquid nitrogen and storing the vials with droplets (instead of an aliquot) to avoid having to thaw and refreeze the same vial every time we use the cells. The plan was to take one frozen droplet out of the vial and use it instead of thawing the entire vial to take out a drop. I assume Alan's statement about previously frozen in liquid nitrogen and recovering well in -80C storage implied that we can make droplet in liquid nitrogen using glycigel and store them in -80C long term?
  15. I am reading the same book right now and am very happy to learn about a history of how this field is eevolved and the key players in the field. Also, realized I was born a little too late. It would have been nice to live in the era of exciting discoveries.

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