HCT Component Summary and Frequently Asked Questions


1. Which clinical samples should be saved?
Collect all donor-recipient pairs that your laboratory/center is involved in the transplant. All patients with all diagnoses, all ages are to be included.

2. How much of each sample should I save?
  • A minimum of 2 mg of genomic DNA.

  • A minimum of 2 vials of peripheral blood lymphocytes at 5 x 106 PBL each.


    • 3. What should I do with B-LCL?
  • Transform all donors and recipients.
  • If your laboratory does not have the capability to transform, contact Effie Petersdorf immediately.


    • 4. Which samples should I start testing?
  • We will prioritize patients with chronic myeloid leukemia (CML) for the first clinical outcome analysis.
  • If you cannot test all of your samples, prioritize patients with CML and their donors.
  • If you can test more than your CML group, then prioritize acute leukemia, myelodysplastic syndrome, and aplastic anemia.
  • There is no exclusion criteria for the degree of known HLA mismatch at the time of transplant.
  • There is no exclusion criteria for the stage of the disease at the time of transplant.
  • Remember that you should be testing donor-recipient pairs for whom you have plenty of DNA/LCL on both the donor and the recipient.


    • 5. Which loci should I type?
  • HLA-A,B,C,DRB1,DRB3,DRB4,DRB5,DQB1,DPB1 are first priority
  • If you can, type HLA-DQA1 and DPA1


    • 6. What other genes should I type?
  • Our priority after HLA-A,B,C,DRB,DQB1,DPB1 is microsatellite markers in class I, II and III regions.
  • If you can type for microsatellites, or are interested but have not yet typed for these markers, please contact Effie Petersdorf (epetersd@fhcrc.org) immediately.
  • Please use the recommended microsatellite protocol Microsoft Word download or PDF download by Dr. Mary Carrington.


    • 7. What resolution is necessary?
  • Allele-level typing is mandatory.
  • All samples must be allele-level typed at all loci specified above (to the degree of the most recent WHO Nomenclature Committee Report).
  • Samples which are only partially typed to the allele level for some but not all loci will not be informative for clinical outcome, and will not be accepted.
  • Ambigious allele combinations will not be accepted; allele-specific amplification and typing methods must be used to delineate the two alleles at each locus.


    • 8. Which typing techniques should I use?
    SSOP and SBT protocols developed by the Core Components are strongly encouraged. Follow this algorithm:
    1. Select the method(s) by going to the Typing Technology page (see SSOP or SBT). SSOP: refer to website SBT: fill out questionnaire on-line 2. Contact Eric Mickelson (emickels@fhcrc.org) in order to receive the panel of control cells so that you can verify primers/probes are working. Please refer to "Participation in Components Requiring HLA Testing". 3. Contact Jennifer Ng (jenng@erols.com) to receive Quality Control Test Panel. PLEASE READ INSTRUCTIONS on how to sign-up for this panel. 4. Perform the blinded test panel 5. Submit Results SSOP: Submit results as +/- hybridizations. REFER TO INSTRUCTIONS "How to submit QC results" SBT: Submit chromatograms and allele assignments to Marcel Tilanus (M.Tilanus@lab.azu.nl -- for more information, mail Marcel directly). 6. Qualification Data will be accepted into the IHWG database when a laboratory completes QC testing.

    9. How do I report my data to the workshop?
  • Data submission forms will be posted on the website in November.


    • 10. Any other questions?
  • Contact epetersd@fhcrc.org (epetersd@fhcrc.org).


    • 11. What is the schedule of events for the HCT Component meetings in year 2001?
    We are considering meetings at EFI, 2001 and EBMT 2001, however, these have not been formally scheduled as of October 20, 2000. Please re-check this site for updated meeting information.

    		 


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