Sharing knowledge. Sharing hope.
Chimerism Reporting
October 12, 2012
Janet Brunner, PA-C
CIBMTR- Milwaukee
CENTER FOR INTERNATIONAL BLOOD & MARROW RESEARCH
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New10_1.ppt
Disclosures
I have no relevant conflicts of interest to
disclose.
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New10_2.ppt
Objective
Participants will be able to:
Report chimerism results using cell
counts, percentages or checking a
non-quantitative box based on the method
used for single donors or for multiple
donors
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Patient Scenario #1
Donor was
her 28 year
old brother
Recipient is a 30
year old female,
with
h/o AML (M5) in
CR2
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Patient Scenario #1
A FISH assay for
the XX/XY
chromosome can
be used to
determine
chimerism due to
sex mismatch.
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Patient Scenario #1
Samples were
sent at Day +30
& Day +90 to
evaluate donor
engraftment.
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FISH Assay Results
Day +30 (March 1)
500 PBMC cells were analyzed
400 cells were XY & 100 cells were XX
Day +90 (May 1)
550 PBMC cells were analyzed
500 cells were XY & 50 cells were XX
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Methods
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Cell Types
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Reporting FISH Results
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Reporting FISH Results
Q81 Date:
03-01-2011
Q82 Method:
02 (FISH)
Q83 Cell Type:
02 (PBMC)
Q84 Total Cells:
500
Q85 # Donor Cells:
400
Q86 # Host Cells:
100
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Reporting FISH results
Do not report the percentage of donor
and recipient if you’re reporting actual
cell numbers.
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Patient Scenario #2
Recipient is a 45 year old male s/p a nonmyeloablative allogeneic HCT for DLBCL
(in 1st relapse- sensitive) from an
unrelated male donor.
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Patient Scenario #2
Samples were
sent for CD3+
chimerism at Day
+30, +60 & Day
+90 to evaluate
donor engraftment
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Example of STR report
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Form 2100
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Reporting STR Results
Q81 Date:
Q82 Method:
Q83 Cell Type:
Q87 % Donor:
Q88 % Host:
06-29-2012
05 (STR)
03 (T-cell)
67%*
33%*
* Based on the DNA marker D3S1358
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Reporting STR/VNTR Results
Results should be
reported as
percentages
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Reporting STR/VNTR Results
If cell sorting was
done to isolate a
particular cell line
(e.g., CD3+ cells)
prior to performing a
DNA based assay
such as STR, do not
report the # of cells
sorted.
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Reporting STR/VNTR Results
150,000 CD3+ cells are isolated from blood
via cell sorting. The results of the STR
assay indicated a mixed chimera-
75% donor & 25% host
Report the percentage of donor & host.
Do not report the number of sorted cells.
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Reporting Chimerism for Multiple Donors
Testing requires DNA markers (loci)
that are informative for both the
recipient & each donor.
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Short Tandem Repeat (STR)
polymorphisms are very useful for this
purpose, since each loci typically have
more than two alleles.
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What does polymorphism mean?
• It’s a variation in the DNA that is too
common to be due merely to a new
mutation.
• A polymorphism must have a
frequency of at least 1% in the
population.
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Chimerism Reporting for Multiple Donors
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Recipient before HCT
Recipient
Genetic Marker
D3S1358
TH01
D21S11
D18S51
Penta E
Identified Alleles
16
6
30,33
14,19
12,14
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Donor 1
Genetic Marker
D3S1358
TH01
D21S11
D18S51
Penta E
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Identified Alleles
14,16
9,10
29,31
14,15
11
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Donor 2
Genetic Marker
D3S1358
TH01
D21S11
D18S51
Penta E
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Identified Alleles
15
8,10
30,33
15,18
5,14
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Informative Alleles
Recipient
An informative
allele is one the
recipient has, but
the donor(s) does
not.
Donor
An informative
allele is one the
donor has, but
the recipient
does not.
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Non-informative Alleles
Both the recipient & donor share the
same allele
R
D
In the case of multiple donors, both
donors share the same allele
D
D
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Recipient Informative Alleles
Genetic Marker
TH01
D18S51
Penta E
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Allele
6
19
12
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10
Donor 1 Informative Alleles
Genetic Marker
D3S1358
TH01
D21S11
D18S51
Penta E
Allele
14
9,10
29,31
15
11
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Donor 2 Informative Alleles
Genetic Marker
D3S1358
TH01
D21S11
D18S51
Penta E
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Allele
15
8,10
---15,18
5
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Donor 1 & 2
• Non-informative donor alleles include
TH01
allele 10
D18S51 allele 15
• Since both donors possess these two
alleles, they cannot be used to
determine the percentage of each
donor post HCT.
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Recipient after HCT
Genetic Marker
Identified Alleles
D3S1358
14,15,16*
TH01
8,9,10
D21S11
29,30*,31,33*
D18S51
14*,15,18
Penta E
5,11,14*
*An allele shared by one of the donors &
the recipient (i.e., not informative)
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Questions
• Is there evidence of one or two donors
post-HCT?
• Is there any evidence of recipient
(host) post-HCT?
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Reporting Chimerism for Multiple Donors
When reporting chimerism results for a
recipient of a double cord HCT, the
combined results must equal 100%.
STR results showed:
Donor X 3%
Donor Y 95%
Host
2%
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Reporting Chimerism for Multiple Donors
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Reporting Chimerism for Multiple Donors
If you get the following STR result:
Donor X = 0%
Donor Y = 100%
Host = 0%
You still must report both donors:
Donor X = 0% & Host = 0%
Donor Y = 100% & Host = 0%
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General Guidelines
• No need to report every chimerism
result.
• Results should be reported near or
around Day +28, Day +100, 6 months
and annually.
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General Guidelines
If a recipient receives an intervention
such as a DCI, report:
• The last chimerism results just prior
to the intervention
• The first results after the intervention
Report the first time any test result shows 100%
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Questions
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