Question 35: Were tests performed to detect persistence of the cellular product since the date of last report?

Methods such as PCR assays, flow cytometry (immunophenotyping) or immunohistochemistry can be used to detect persistence of the cellular product in the recipient.

If tests were performed to detect persistence of the cellular product in the current reporting period, select Yes.

If tests were not performed to detect persistence of the cellular therapy product in the current reporting period, select No and continue with question 60.

Question 36: Was persistence evaluated by molecular assay (PCR)?

Molecular assessment involves testing blood, bone marrow, tumor or other source for the presence of known molecular markers. Molecular assessments are the most sensitive test and involve amplifying regions of cellular DNA by polymerase chain reaction (PCR), typically using RNA to generate complementary DNA through reverse transcription (RT-PCR). The amplified DNA fragments are compared to a control, providing a method of quantifying log increase of genetic mutation transcripts. Each log increase is a 10-fold increase of gene transcript compared to control.

Indicate Yes or No whether molecular assay testing was performed to detect the persistence of the genetically modified cellular therapy product within the reporting period. If persistence was not evaluated by molecular assay, report No and continue with question 41.

Question 37: Date Sample collected:

Report the date (YYYY-MM-DD) the sample was collected for molecular assay. If multiple tests were performed in the reporting period and

  • all tests were negative, report the date of the first negative test result
  • there were positive and negative results, report the date of the last positive test (do not report negative results)

If the exact date is unknown, please view General Instructions, General Guidelines for Completing Forms for more information on reporting partial and unknown dates.

Questions 38-39: Specify the cell source: (check all that apply)

Specify the cell source of the sample collected for evaluation by molecular assay. Select all that apply. If multiple cell sources were used and persistence was detected in some but not all the samples, report ONLY the cell sources that were positive. If Other source is selected, specify the source in question 39.

Question 40: Were the infused cells detected?

Indicate Yes or No if the infused cells were detected by molecular assay.

Question 41: Was persistence evaluated by flow cytometry testing (immunophenotyping)?

Flow cytometry is a technique that can be performed on blood, bone marrow, or tissue preparations where cell surface markers can be quantified on cellular material. The nature of flow cytometry is to detect cells based on a specific probe. To report flow cytometry results, the test must have been performed to specifically detect the genetically modified cellular therapy product.

Indicate Yes or No if flow cytometry testing was performed to detect the persistence of the genetically modified cellular therapy product within the reporting period. If flow cytometry was not performed, select No and continue with question 49.

Question 42: Date sample collected:

Report the date (YYYY-MM-DD) the sample was collected for flow cytometry testing (immunophenotyping). If multiple tests were performed in the reporting period and

  • all tests were negative, report the date of the first negative test result
  • there were positive and negative results, report the date of the last positive test (do not report negative results)

If the exact date is unknown, please view General Instructions, General Guidelines for Completing Forms for more information on reporting partial and unknown dates.

Question 43-44: Specify the cell source (check all that apply)

Specify the cell source of the sample collected for evaluation by flow cytometry. Select all that apply. If multiple cell sources were used and persistence was detected in some but not all the samples, report ONLY the cell sources that were positive. If Other source is selected, specify the source in question 44.

Question 45: Were the infused cells detected?

Indicate Yes or No if the infused cells were detected by flow cytometry testing (immunophenotyping).

Question 46: Were B-cell counts monitored after infusion?

CAR-T cells that target antigens (CD19, CD20, CD22, BCMA) on B-cells do not distinguish between cancerous and normal B-cells. As result, the recipient can develop B-cell aplasia (low number or absence of B-cells). B-cell aplasia can be used as a surrogate to track persistence of the product. If the recipient has B-cell aplasia, then the product may still be present.

Indicate Yes or No if B-cell counts were monitored during the current reporting period. If B-cell counts were not monitored, select No and continue with question 49.

Question 47: Was there B-cell recovery?

A guideline for B-cell aplasia is a B-cell count of <50 cells>50 cells/µL), select Yes. If B-cells never recovered, report No and continue with question 49.

B-cell counts in the blood do vary with age, and children have much higher counts than adults. The younger the child, the higher is the concentration.

Question 48: Date of B-cell recovery

Report the date (YYYY-MM-DD) the flow cytometry report showed B-cell recovery.

Question 49: Was persistence evaluated by immunohistochemistry?

Immunohistochemistry is a process that uses antibodies to test for certain antigens (markers) in a sample. When the antibodies bind to the antigen in the tissue sample, the enzyme or dye is activated, and the antigen can then be seen under a microscope.

Indicate Yes or No if immunohistochemistry testing was performed to detect the persistence of the genetically modified cellular product within the reporting period. If immunohistochemistry testing was not performed in the current reporting period, report No and continue with question 54.

Question 50: Date sample collected:

Report the date (YYYY-MM-DD) the sample was collected for immunohistochemistry studies. If multiple tests were performed in the reporting period and

  • all tests were negative, report the date of the first negative test result
  • there were positive and negative results, report the date of the last positive test (do not report negative results)

If the exact date is unknown, please view General Instructions, General Guidelines for Completing Forms for more information on reporting partial and unknown dates.

Question 51-52: Specify the cell source:

Specify the cell source of the sample collected for evaluation by immunohistochemistry. Select all that apply. If multiple cell sources were used and persistence was detected in some but not all the samples, report ONLY the cell sources that were positive. If Other source is selected, specify the source in question 52.

Question 53: Were the infused cells detected?

Indicate Yes or No if the infused cells were detected by immunohistochemistry testing.

Questions 54-55: Was persistence evaluated by other method?

Indicate Yes or No if persistence of cells was tested by a method not listed above. If Yes, specify the other method used to evaluate persistence of cells in question 55. If persistence of cells was not tested by another method, select No and continue with question 60.

Question 56: Date sample collected:

Report the date (YYYY-MM-DD) the sample was collected for the other method. If multiple tests were
performed in the reporting period and

  • all tests were negative, report the date of the first negative test result
  • there were positive and negative results, report the date of the last positive test (do not report negative results)

If the exact date is unknown, please view General Instructions, General Guidelines for Completing Forms for more information on reporting partial and unknown dates.

Question 57-58: Specify the cell source:

Specify the cell source of the sample collected for evaluation by other method. Select all that apply. If multiple cell sources were used and persistence was detected in some but not all the samples, report ONLY the cell sources that were positive. If Other source is selected, specify the source in question 58.

Question 59: Were the infused cells detected?

Indicate Yes or No if the infused cells were detected by other method.

Section Updates:

Question Number Date of Change Add/Remove/Modify Description Reasoning (If applicable)
. . . . .
Last modified: Jan 22, 2021

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