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HelpOrder Code IABCS B-Cell Phenotyping Profile for Immunodeficiency and Immune Competence Assessment, Blood
Reporting Name
Immune Assessment B Cell Subsets, BUseful For
Screening for common variable immunodeficiency and hyper-IgM syndromes
Assessing B-cell subset reconstitution after stem cell or bone marrow transplant
Assessing response to B-cell-depleting immunotherapy
This test is not indicated for the evaluation of lymphoproliferative disorders (eg, leukemia, lymphoma, multiple myeloma).
Profile Information
| Test ID | Reporting Name | Available Separately | Always Performed |
|---|---|---|---|
| TBBS | QN Lymphocyte Subsets: T, B, and NK | Yes | Yes |
| IABC | Immune Assessment B Cell Subsets, B | No | Yes |
Testing Algorithm
When multiple specimen types are required to perform a panel of tests, the laboratory will perform the tests for which the appropriate specimen type was received. The laboratory will cancel those for which the appropriate specimen was not received. Be advised that this may change the degree of interpretation received with the report.
Performing Laboratory
Mayo Clinic Laboratories in Rochester
Specimen Type
Whole Blood EDTAOrdering Guidance
The IABCS is a panel test. This test requires two separate whole blood EDTA specimens at two different transport temperatures: one ambient and one refrigerate.
The TBBS / Quantitative Lymphocyte Subsets: T, B, and Natural Killer (NK) Cells, Blood is automatically performed, a separate order is not required.
If only an ambient EDTA specimen is received, only the TBBS / Quantitative Lymphocyte Subsets: T, B, and Natural Killer (NK) Cells, Blood will be performed. If only a refrigerate EDTA sample is received, this test will be canceled and converted to RBCS / Relative B-Cell Subset Analysis Percentage, Blood, which provides the relative B-cell subset values without quantitation.
This test is a screening test and further analyses will be required to complete a diagnostic workup for hyper-IgM (XHIM / X-Linked Hyper IgM Syndrome, Blood and CD40 / B-Cell CD40 Expression by Flow Cytometry, Blood).
Shipping Instructions
Testing performed Monday through-Friday. Specimens not received by 4pm (CST) on Friday may be canceled.
Samples arriving on the weekend and observed holidays may be canceled.
Collect and package specimens as close to shipping time as possible.
It is recommended that specimens arrive within 24 hours of collection.
Necessary Information
1. Date of collection is required.
2. Ordering healthcare professional name and phone number are required.
Specimen Required
Two separate EDTA whole blood specimens are required: 1 refrigerate and 1 ambient transport temperature.
For serial monitoring, it is recommended that specimens are collected at the same time of day.
Specimen Type: Whole blood for TBBS / Quantitative Lymphocyte Subsets: T, B, and Natural Killer (NK) Cells, Blood
Container/Tube: 4 mL Lavender top (EDTA)
Specimen Volume: 3 mL
Collection Instructions:
1. Send whole blood specimen in original tube. Do not aliquot.
2. Label specimen as TBBS.
3. Ship ambient.
Specimen Stability Information: Ambient <52 hours
Specimen Type: Whole blood for IABC / B-Cell Phenotyping Screen for Immunodeficiency and Immune Competence Assessment, Blood
Container/Tube: Lavender top (EDTA)
Specimen Volume:
≤14 years: 4 mL
>14 years: 10 mL
Collection Instructions:
1. Send whole blood specimen in original tube. Do not aliquot.
2. Label specimen IABC.
3. Ship refrigerate.
Specimen Stability Information: Refrigerated <48 hours
Specimen Minimum Volume
TBBS: 1 mL; IABC: > 14 years: 5 mL; ≤ 14 years: 3 mL
Specimen Stability Information
| Specimen Type | Temperature | Time | Special Container |
|---|---|---|---|
| Whole Blood EDTA | Varies | 48 hours | PURPLE OR PINK TOP/EDTA |
Reference Values
The appropriate age-related reference values will be provided on the report.
Day(s) Performed
Monday through Friday
CPT Code Information
86355-B cells, total count
86357-Natural killer (NK) cells, total count
86359-T cells, total count
86360-Absolute CD4/CD8 count with ratio
86356 x7 - Mononuclear cell antigen, quantitative
Method Description
Quantitative Lymphocyte Subsets: T, B, and Natural Killer:
The T, B, and natural killer (NK)-cell surface marker assay uses monoclonal antibodies to identify the various membrane antigens and flow cytometry to enumerate the number of cells expressing these differentiation antigens. CD14 is used to exclude monocytes, thereby improving accuracy and enhancing the purity of the lymphocyte population. The results are reported as the percent of lymphocytes that are total T cells (CD3+), CD3+CD4+ T cells, CD3+CD8+ T cells, NK (CD16+56+, CD3-), and B-lymphocytes (CD19+), and the absolute number of each cell type per mL of blood. The assay is a 7-color no-wash procedure, and the absolute counts are calculated from internal bead standards. In addition, the total lymphocyte count and the CD4:CD8 ratio are reported.(Hoffman RA, Kung PC, Hansen WP, Goldstien G. Simple and rapid measurement of human T lymphocytes and their subclasses in peripheral blood. Proc Natl Acad Sci USA. 1980:77[8]:4914-4917; Mandy FF, Nicholson JK, McDougal JS. Guidelines for performing single-platform absolute CD4+T-cell determinations with CD45 gating for persons infected with human immunodeficiency virus. Center for Disease Control and Prevention. MMWR Morb Mortal Wkly Rep. 2003;52[RR-2]:1-13)
Immune Assessment B Cell Subsets:
Peripheral blood mononuclear cells are isolated from whole blood using a Ficoll gradient and used in the staining protocol. The assay involves a multicolor 5-tube panel for the following antibodies: CD45, CD19, CD20, CD27, IgD, IgM, CD38, and CD21. After the staining with specific antibody, the cells are washed and fixed with paraformaldehyde and then analyzed by flow cytometry on a Becton Dickinson FACS Canto instrument. The cell-surface expression is denoted as the percent of CD19+ B cells expressing each of the specific markers. CD19+ and CD20+ B cells are expressed as a percent of the total lymphocytes (CD45+). The absolute counts for the B-cell subsets are derived from flow cytometry analysis of whole blood using monoclonal antibodies to identify CD45, CD3, CD4, CD8, CD19, and CD16+CD56+. CD14 is used to exclude monocytes, thereby improving accuracy and enhancing the purity of the lymphocyte population. The assay is a 7-color, lyse-no wash procedure and the absolute counts are calculated from internal bead standards. The absolute lymphocyte count per microliter is used to calculate the absolute counts of the various B-cell subsets in this assay.(Unpublished Mayo method)
Reject Due To
| Gross hemolysis | Reject |
| Gross lipemia | Reject |
Method Name
Flow Cytometry