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Test ID LPA3P Lymphocyte Proliferation to Anti-CD3/Anti-CD28 and Anti-CD3/Interleukin-2 (IL-2) by Flow Cytometry, Blood

Useful For

A second-level test after lymphocyte proliferation to mitogens (specifically phytohemagglutinin) has been assessed. The anti-CD3 proliferation panel is not a first-level test for assessing lymphocyte (T-cell) function


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

Method Name

Flow Cytometry

Reporting Name

Lymphocyte Proliferation, aCD3

Specimen Type

WB Sodium Heparin

Shipping Instructions

Specimens are required to be received in the laboratory weekdays and by 4 p.m. on Friday. Draw 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 draw.


Samples arriving on the weekend and observed holidays may be canceled.

Necessary Information

Date and time of draw and ordering physician's name and phone number are required.

Specimen Required

For serial monitoring, we recommend that specimen draws be performed at the same time of day.


Supplies: Ambient Shipping Box-Critical Specimens Only (T668)

Container/Tube: Green top (sodium heparin)

Specimen Volume:

<3 months: 1 mL

3 months-5 years: 3 mL

6-18 years: 5 mL

>18 years: 20 mL

Collection Instructions: Send specimen is original tube. Do not aliquot.


Blood Volume Recommendations Based on Absolute Lymphocyte Count (ALC)



Blood Volume for Minimum aCD28 Only

Blood Volume for Minimum of aCD3, aCD28, and IL-2

Blood Volume for Full Assay


>15 cc

>28 cc

>50 cc


15 cc

28 cc

50 cc


6.5 cc

12 cc

24 cc


4.5 cc

8.5 cc

16 cc


3.5 cc

6.5 cc

12 cc


2.5 cc

4.5 cc

8 cc


1.8 cc

3.5 cc

6 cc


1.5 cc

2.5 cc

5 cc

Specimen Minimum Volume

1 mL

Specimen Stability Information

Specimen Type Temperature Time Special Container
WB Sodium Heparin Ambient 48 hours GREEN TOP/HEP

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%

Day(s) and Time(s) Performed

Monday through Friday

Do not send specimen after Thursday.

Performing Laboratory

Mayo Clinic Laboratories in Rochester

CPT Code Information

86353 x 1-Anti-CD3 + anti-CD28 stimulation

86353 x 1-Anti-CD3 + IL2 stimulation

86353 x 1-Anti-CD3 stimulation (as indicated)

LOINC Code Information

Test ID Test Order Name Order LOINC Value
LPA3P Lymphocyte Proliferation, aCD3 In Process


Result ID Test Result Name Result LOINC Value
35203 Viab of Lymphs at Day 0 33193-4
35171 Max Prolif, soluble aCD3 as % CD45 81760-1
35172 Max Prolif, soluble aCD3 as % CD3 81756-9
35173 Max Prolif, soluble aCD28 as % CD45 81759-3
35174 Max Prolif, soluble aCD28 as % CD3 81758-5
35176 Max Prolif, soluble IL2 as % CD45 81755-1
35177 Max Prolif, soluble IL2 as % CD3 81757-7
35205 Interpretation 69965-2
35204 aCD3 Comment 48767-8

Test Classification

This test was developed using an analyte specific reagent. Its performance characteristics were determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the U.S. Food and Drug Administration.

Reflex Tests

Test ID Reporting Name Available Separately Always Performed
ADSTM Additional Flow Stimulant No, (Bill Only) No

Testing Algorithm

Testing with 2 stimulants will always be performed. When adequate specimen is available, a third stimulant will be performed and billed separately. If isolated patient’s peripheral blood mononuclear cells (PBMCs) have a low WBC, selected dilutions or stimulants may not be used at the discretion of the laboratory to ensure the most reliable results.

NY State Approved