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Test Code LAB3960 Lymphocyte Proliferation to Mitogens

Important Note

Note: Limited specimen stability 
Specimens must be received within 24 hours of collection on SCH Seattle Main Campus and by noon Monday-Friday. Testing is not performed on weekends or holidays; adjust collections accordingly.
Cell Markers Lab can only set this test up between 8 am-12 pm on weekdays, due to the nature of this test. Consistent timing of setup and subsequent test steps is imperative to quality patient results.

Clinical System Name

Lymphocyte Proliferation to Mitogens

Synonyms

Anti CD3 Stimulant
Lymphocyte Function Analysis
Mitogen Stimulation Study
PHA (Phytohemaglutinin) Stimulant
T cell proliferation to mitogens

Lymphocyte Mitogen Stimulation

MSS

Lymphocyte proliferation

Lymphocyte stimulation

PHA stimulation

Anti-CD3 stimulation

Description

The Lymphocyte Proliferation to Mitogens assay is used to evaluate cellular immune system functionality and is useful for diagnosing disorders of T-cell immunity.  This assay measures the in vitro proliferation of lymphocytes after exposure to molecules that non-specifically stimulate cells to undergo mitosis, known as mitogens. The mitogens utilized include phytohemagglutinin (PHA) and anti-CD3, both of which stimulate T-cells to proliferate, independent of prior antigen exposure. The magnitude of proliferation, quantified as the percentage of EdU-positive cells within specified populations, reflects global T-cell proliferative capacity and functional cellular immune competence. Please note, abnormal results are not specific for a particular disease state, and the magnitude of the abnormality detected may not be directly related to the degree of immunodeficiency.

The Click-iT EdU assay (EdU assay) is a flow cytometry-based proliferation assay that enables simultaneous measurement of DNA synthesis and immunophenotypic identification of T-cells. Cell proliferation is quantified as the percentage of EdU-positive cells within defined populations, including live singlet CD45+ cells (CD45+EdU+%) and live singlet CD3+CD45+ T-cells (CD3+EdU+%). 

Sample Requirements

Specimen:  Whole Blood

Container(s):  Dark Green/Sodium Heparin (no serum separator)

Preferred Vol:  10 mL

Minimum Vol:  10 mL (if unable to collect 10 mL, please contact lab at 206-987-2560)

Note:  Specimens stable for only 24 hours at room temperature.

Call the Cell Markers Lab (206) 987-2560 with any questions regarding specimen collection or handling. If unable to collect minimum volume, please collect as close to 10 mL as possible and notify Cell Markers at 206-987-2560 (leave voicemail after hours).

Draw an additional 10 mL Na-Hep if an antigen stimulation studies (Candida, Tetanus) are also ordered.

Processing Instructions

Rejection criteria: specimens greater than 24 hours old. If specimen is near the 24 hour stability limit at receipt, notify Cell Markers at x72560 to determine ability to perform test.

Requisition Entry: If no stimulants are indicated on the requisition, please select both PHA & CD3. 

Spin: N

Aliquot: N

Temp: RT

Storage  

Days: Transport specimen, copy of Cell Markers Peripheral Blood Requisition (if applicable), and labels to the Cell Markers lab (station #181). Notify Cell Markers at x72560 right away if specimen is near 24 hour specimen stability limit - billing/ordering problems should not delay delivery to lab.

Eves/Nights: Store specimen, copy of Cell Markers Peripheral Blood Requisition (if applicable), and labels in the Cell Markers RT box in CPA.

Off-site collection:  A normal control should accompany the specimen. It is critical samples be kept at room temperature; use extra packing to maintain temperature. If specimen will not arrive at Children's before noon for same day testing, blood should be drawn as late in the day as possible, maintained at room temperature, and sent overnight to allow test set-up the following morning. Testing must be set up within 24 hours of collection. Dark Blue Na Heparin okay, but dark green tops preferred.Transport all tubes at RT to Seattle Children's Hospital address on requisition: Laboratory FB.2.441, 4800 Sand Point Way NE, Seattle, WA  98105. 

Stability

Temperature Time
Room temp 24 hours
Refrigerated N
Frozen N

 

Availability

STAT Performed TAT
N

Monday - Friday; must be in lab before noon

 7 - 10 days

Performing Laboratory

Seattle Children's Laboratory    

Department

Department:  Cell Markers

Phone Number: 206-987-2560

 

 

Methodology

Method:  Lymphocyte proliferation in response to mitogen stimulation with enumeration using the Click-iT EdU flow cytometry-based assay

Analytical Volume: dependent on patient lymphocyte count

Limitations:  None specified

Reference Range

Reference values accompany patient report.

CPT Codes

  CPT CODE
Set up and Initial Mitogen Stimulant

86353
Each additional Mitogen Stimulant 86353

 

Requisition

Cell Marker Requisition