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HelpOrder Code LPA3P Lymphocyte Proliferation to Anti-CD3/Anti-CD28 and Anti-CD3/Interleukin-2 (IL-2), Flow Cytometry, Blood
Useful For
A second-level test after lymphocyte proliferation to mitogens (specifically phytohemagglutinin) has been assessed
Evaluating patients suspected of having impairment in cellular immunity
Evaluation of T-cell function in patients with primary immunodeficiencies, either cellular (DiGeorge syndrome, T-negative severe combined immunodeficiency [SCID], etc) or combined T- and B-cell immunodeficiencies (T- and B-negative SCID, Wiskott-Aldrich syndrome, ataxia telangiectasia, common variable immunodeficiency, among others) where T-cell function may be impaired
Evaluation of T-cell function in patients with secondary immunodeficiency, either disease related or iatrogenic
Evaluation of recovery of T-cell function and competence following bone marrow transplantation or hematopoietic stem cell transplantation
Evaluation of T-cell function in patients receiving immunosuppressive or immunomodulatory therapy
Evaluation of T-cell function in the context of identifying neutralizing antibodies in patients receiving therapeutic anti-CD3 antibody immunosuppression for solid organ transplantation or autoimmune diseases, such as type 1 diabetes
This panel is not useful as a first-level test for assessing lymphocyte (T-cell) function.
Reflex Tests
| Test ID | Reporting Name | Available Separately | Always Performed |
|---|---|---|---|
| ADSTM | Additional Flow Stimulant | No, (Bill Only) | No |
Testing Algorithm
To ensure the most reliable results, if insufficient peripheral blood mononuclear cells are isolated from the patient's specimen due to low white blood cell counts or specimen volume received, selected dilutions or stimulants may not be tested at the discretion of the laboratory.
Testing performed with at least one stimulant will be reported. When adequate specimen is available, the second and third stimulants will be evaluated, each at an additional charge.
Reporting Name
Lymphocyte Proliferation, aCD3Specimen Type
WB Sodium HeparinShipping Instructions
Testing performed Monday through Friday. Specimens not received by 4 p.m. Central time on Friday may be canceled.
Specimens arriving on the weekend and observed holidays may be canceled.
Collect and package specimen as close to shipping time as possible. Ship specimen overnight in an Ambient Shipping Box-Critical Specimens Only (T668) following the instructions in the box. It is recommended that specimens arrive within 24 hours of collection.
Necessary Information
1. Date and time of collection are required.
2. The ordering healthcare professional's name and phone number are required.
Specimen Required
Supplies: Ambient Shipping Box-Critical Specimens Only (T668)
Container/Tube: Green top (sodium heparin)
Specimen Volume: 20 mL
See table for information on recommended volume based on absolute lymphocyte count
Pediatric Volume:
<3 months: 1 mL
3-24 months: 3 mL
25 months-18 years: 5 mL
Collection Instructions: Send whole blood specimen in original tube. Do not aliquot.
Additional Information: For serial monitoring, it is recommended that specimen collection be performed at the same time of day.
Table. Blood Volume Recommendations Based on Absolute Lymphocyte Count (ALC)
|
ALC |
Blood volume for minimum aCD28 only |
Blood volume for minimum of aCD3, aCD28, and IL-2 |
Blood volume for full assay |
|
<0.5 |
>15 mL |
>28 mL |
>50 mL |
|
0.5-1.0 |
15 mL |
28 mL |
50 mL |
|
1.1-1.5 |
6.5 mL |
12 mL |
24 mL |
|
1.6-2.0 |
4.5 mL |
8.5 mL |
16 mL |
|
2.1-3.0 |
3.5 mL |
6.5 mL |
12 mL |
|
3.1-4.0 |
2.5 mL |
4.5 mL |
8 mL |
|
4.1-5.0 |
1.8 mL |
3.5 mL |
6 mL |
|
>5.0 |
1.5 mL |
2.5 mL |
5 mL |
Specimen Minimum Volume
See Specimen Required
Specimen Stability Information
| Specimen Type | Temperature | Time | Special Container |
|---|---|---|---|
| WB Sodium Heparin | Ambient | 48 hours | GREEN TOP/HEP |
Reject Due To
| Gross hemolysis | Reject |
| Gross lipemia | OK |
Reference Values
Viability of lymphocytes at day 0: ≥75.0%
Maximum proliferation of anti-CD3 as % CD45: ≥19.4%
Maximum proliferation of anti-CD3 as % CD3: ≥20.3%
Maximum proliferation of anti-CD3 + anti-CD28 as % CD45: ≥37.5%
Maximum proliferation of anti-CD3 + anti-CD28 as % CD3: ≥44.6%
Maximum proliferation of anti-CD3 + IL-2 as % CD45: ≥41.7%
Maximum proliferation of anti-CD3 + IL-2 as % CD3: ≥46.2%
Method Description
Peripheral blood mononuclear cells in RPMI 1640 medium supplemented with L-glutamine and 5% human AB serum are incubated unstimulated or stimulated with varying concentrations of anti-CD3, anti-CD3+anti-CD28, or anti-CD3+IL-2. A daily experimental normal control is included with each batch of patient samples to serve as an internal control.
Following incubation, the cells are assessed for proliferation and stained with the following markers: CD45+ lymphocytes, CD3+ T cells, and CD69+ activated cells. Results are reported for the percent viable cells on day 0, as well as the percentage of proliferating cells of total lymphocytes and T cells.(Unpublished Mayo method)
Day(s) Performed
Monday through Friday
Performing Laboratory
Mayo Clinic Laboratories in Rochester
CPT Code Information
86353 x 2
86353 (as appropriate)
Method Name
Flow Cytometry