Australia. Dr. Frank Christiansen reported for the five centers in Australia (Perth, Adelaide, Melbourne, Sydney, Brisbane). Dr. Jim McCluskey retains all of the cells and EBV cell lines.
DNA = 120 pairs (available)
Over 240 cell lines are available. Typing is being performed by the five centers using workshop techniques. They anticipate discussing results at the November, 2000 ASEATTA meeting in Perth. The current algorithm being followed by the Australian laboratories is commendatory and the Australian data is expected to be complete and definitive. It was confirmed that the workshop would accept unambiguous alleles (any ambiguities should be resolved using allele-specific reagents). Dr. Tilanus confirmed that the protocols being used by the Australian laboratories are workshop protocols because the primers differ in most instances by no more than one nucleotide at 5' or 3' endsfrom IHWG accepted protocols.
Capture of clinical data is an ongoing effort.
Germany. Dr. Shraga Goldmann reported for Ulm and Essen.
800 pairs are available for all diseases
700 URD have A,B serology; High resolution for DRB1 and DQB1
100 related pairs
497 URD pairs and 100 related are 1 antigen mismatched
72 pairs have been typed for HLA-A,B,C using SBT
SSOP is the main typing method for DRB1 and DQB1
DPB has not been performed
Available material on the majority of pairs is in the form of DNA.
Ongoing efforts are needed to assure compliance with human subjects protection.
Norway. Dr. Petersdorf presented Dr. Thorstein Egeland's summary for Oslo: 131 pairs complete
total URD pairs contributed to workshop: 131
Minimal material available: DNA on all samples
Availability of cells: pending
HLA high resolution: pending
Switzerland. Dr. Petersdorf presented Dr. Jean-Marie Tiercy's summary for Geneva
1990 to October 2000:
42 CML
31 AML
21 ALL
13 MDS
6 other
Total = 113 patients
+10-20 BMTs scheduled Nov, 2000-Dec, 2001
· Of these 40-50% patient/donor pairs should be available as frozen cells for EBV transformation.
· For about 30% of the pairs 10-30 ug DNA available (no PBLs available anymore).
Level of resolution of HLA typing for patient and donors:
· ABCw allelic, except for conserved antigens like A1, B8, etc. but ALL pairs tested by CTLpf analysis.
· (CTLpf-negative pairs not tested for all class I allelic subtypes)
· High resolution DRB1, DRB3, DRB5, DQB1
· DPB1 partially done, can be completed in Geneva or through the workshop analysis.
Matching status:
· all pairs AB-serologically, DR-generic matched
· 55% of the pairs A, B, C, DR, DQ-matched, CTLpf-neg
· 5% A or B allele mismatch
· 20% of the pairs with a single Cw allele mismatch
· 15% DRB1 or DRB3 or DQB allele mismatch
· 5% of the pairs with 2 MM (e.g. Cw+DQ)
Clinicians responsible for the clinical data:
Prof. B. Chapuis, Geneva
Prof. A. Gratwohl, Basel
Dr. U. Schanz, Zurich
Prof. R. Seger, Zurich
Seattle. Dr. Petersdorf presented the FHCRC data.
CML study summary
1st priority -myeloablative BM
2nd priority -non-myeloablative PBSC
Number of myeloablative BMT pairs: 507
Number with LCL: 479
|
Number of pairs completed for: | |
| HLA-A | 464 |
| HLA-B | 460 |
| HLA-C | 455 |
| DRB1 | 507 |
| DQB1 | 507 |
| DPB1 | 467 |
Typing still needed
(numbers are in pairs):
| 1 | A only |
| 3 | B only |
| 7 | C only |
| 2 | DP only |
| 1 | A,B |
| 1 | A, C |
| 1 | B,C |
| 1 | B,DP |
| 1 | A,DP |
| 1 | C,DP |
| 1 | A,C,DP |
| 7 | A,B,C |
| 3 | B,C,DP |
| 3 | A,B,DP |
| 28 | A,B,C,DP |
Technical problems in HLA-A,B,C typing;
· PCR failures due to bad DNA prep
· Sequencing failures due to weak PCR amplification and poor template purification · primer-dimer in PCR
~30% of the samples have ambiguous typing combinations, which need to be solved with allele specific primers
Microsatellites
Dr. Mary Carrington from the NCI presented the microsatellite protocol to be used for the HCT Component. A copy of the microsatellite protocol can be found on the web site as a MS Word download or PDF download.
Dr. Petersdorf's lab has tested this protocol and it is technically robust At this time Drs. Carrington and Petersdorf request laboratories to review the protocol and to decide whether microsatellite testing will be performed in local laboratories or whether core laboratories should take on the microsatellite testing for the entire component. Drs. Carrington and Petersdorf recommend that the data analysis (Genotyper software) be conducted by core laboratories who are proficient in interpreting this data. Those labs participating in microsatellite testing will be asked to test a QC panel for microsatellites. This component leaves open the possibility of using 1 common microsatellite protocol for the workshop (CNG protocol and Carrington protocol). The first priority group of transplant pairs for microsatellite testing will be HLA-A, B, C, DRB, DQB1 and DPB1 allele matched pairs.
New Research Projects
Dr. Jon van Rood presented a new proposal to study the clinical impact of NIMA in haploidentical related transplants Dr. von Rood presented the results of a study done with Dr. Mary Horowitz and the IBMTR on the effects of NIMA on GVHD. Transplants included 1 or 2 locus mismatches at HLA A, B (serology) or DRB1.
| Patient at risk | Sibling parent RR | p-value | |
| Acute grade 2-4 GVHD | |||
| NIMA sibling | 58 | 1.0 | |
| NIPA sibling | 67 | 1.86 | .02 |
| Mother | 74 | 1.88 | .01 |
| Father | 50 | 2.20 | .003 |
| Chronic GVHD | |||
| NIMA sibling | 62 | 1.0 | |
| NIPA sibling | 69 | 1.76 | .10 |
| Mother | 79 | 2.32 | .01 |
| Father | 57 | 4.11 | .0004 |
| Graft Failure | |||
| NIMA sibling | 63 | 1.0 | |
| NIPA sibling | 67 | .61 | .3 |
| Mother | 79 | .86 | .70 |
| Father | 55 | 1.12 | .78 |
| TRM | |||
| NIMA sibling | 63 | 1.0 | |
| NIPA sibling | 69 | 1.3 | .38 |
| Mother | 79 | 2.02 | .009 |
| Father | 57 | 1.92 | .03 |
The multivariable analyses accounted for all known confounding risk factors.
The following are minimal criteria necessary for a workshop study on NIMA:
1. Clinical Data
2. HLA typing (retrospective)
Propositus
Sibling
Mother
Father
3. Resolution
Low resolution A,B,DR is minimal (this allows compatibility with data above)
4. Demographics of Transplant
T cell replete
Dr. van Rood's preference is to conduct the NIMA analysis as a retrospective collection of families. Dr. Ekkehard Albert also placed family studies as high priority in any workshop. Dr. Petersdorf proposed that we include NIMA analysis as part of the HCT component and to begin with the retrospective study. Those laboratories and centers having samples meeting above eligibility criteria should contact either Drs. Petersdorf or van Rood directly. vanrood@europdonor.nl (email). This proposal was accepted.
Cytokine Gene Polymorphism
Dr. Joannis Mytilineos presented the Cytokine Gene Polymorphism (CGP) study.
It was recommended that CGP analysis be prioritized for donor-recipient pairs having LCL due to the need for additional genomic DNA. Dr. Mytilineos estimates that 15 mg will be required to test all polymorphisms.
Discussion centered on the need to confirm appropriate consenting for human subjects. Dr. Petersdorf requested regional coordinators forward their IRB approved consent forms to the IHWG.
NK - KIR
Dr. Bo Dupont presented the aims of the NK component. A collaborative study of NK - KIR matching in clinical outcome with the Transplantation component represents an important workshop endeavor.
Collaborative Study of Project 5 and Project 9
Role of Donor-Recipient NK-KIR Genotype Matching in
Clinical Outcome after Allogeneic Stem Cell Transplantation
Hypotheses
1. The donor NK-KIR genotype confers cytolytic effector activity against host leukemic cells.
a. The risk of post-transplant relapse is increased in the presence of donor-recipient KIR identity.
b. The pre-transplant donor-recipient genotype can be used to predict patients who may have higher risk of post-transplant relapse.
2. The lack of NK-KIR inhibition by host NK cells increases the risk of graft failure.
a. Donor-recipient NK-KIR genotypic non-identity predicts for increased risk of graft failure.
b. The pre-transplant donor-recipient NK-KIR genotype can be used to predict patients who are at higher risk for graft failure.
Study Design
1. Study population for hypothesis #1: CML, AML, ALL
a. Unrelated donor transplants identical for MHC class I genes.
b. Haploidentical related transplants
c. HLA genotypically-identical sibling transplants having available family studies
2. Study population for hypothesis #2 (no restriction to diagnosis)
a. Unrelated
b. Haploidentical related
c. HLA genotypically identical sibling
3. Clinical Data Requirements
a. definition of engraftment and relapse (parameters)
b. transplant regimens
c. post-grafting immunosuppression regimens
d. treatment regimens for leukemic relapse (identify use of DLI; identify withdrawal of immunosuppression)
e. treatment regimens for graft failure (autologous re-infusion; second transplantation)
4. Methodology
a. Collection of genomic DNA from eligible donor-recipient transplant pairs (study those having complete clinical data and complete class I and II genotyping)
b. Genotyping of donor-recipient pairs and all available family members for NK-KIR
c. Definition of match status for KIR
d. Definition of clinical endpoints for relapse, disease-free survival, and graft failure
e. Statistical methods for cumulative incidence of relpase, Kaplan-Meier probability of disease-free survival, survival; rates of graft failure. Multivariable regresion models to account for known factors affecting relapse (including but not restricted to disease and stage; transplant regimens) and graft failure (including but not restricted to degree of MHC compatibility; stem cell dose; transplant regimens)
Communications
Dr. Petersdorf proposes that in addition to one-on-one communication via e-mail, the HCT component has reached a level of progress requiring group communication. She proposes a quarterly HCT e-mail newsletter to update regional activity, technical issues, etc. These quarterly updates will also be useful in preparing the annual summary reports required by the NIH.
We will post all notices for upcoming committee meetings on the website.
