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Test Code CD40 B-Cell CD40 Expression by Flow Cytometry, Blood

Performing Laboratory

Mayo Medical Laboratories in Rochester

Reporting Name

CD40 by Flow, QL, B

Specimen Type

Whole Blood EDTA


Specimen Required


Specimens are required to be received in the laboratory weekdays and by 4 p.m. on Friday. It is recommended that specimens arrive within 24 hours of draw. Samples arriving on the weekend may be canceled. Draw and package specimen as close to shipping time as possible.

 

Container/Tube: Lavender top (EDTA)

Specimen Volume: 3 mL

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

Additional Information:

1. Ordering physician name and phone number are required.

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


Reject Due To

Hemolysis

Mild OK; Gross reject

Lipemia

Mild OK; Gross reject

Icterus

NA

Other

NA

Specimen Stability Information

Specimen Type Temperature Time
Whole Blood EDTA Ambient 72 hours

Specimen Minimum Volume

1 mL

Day(s) and Time(s) Performed

Monday through Friday

Do not send specimen after Thursday. Specimen must be received by 10 a.m. on Friday.

Specimen Retention Time

4 days

Analytic Time

3 days

Reference Values

Present (normal)

Useful For

Evaluating patients for hyper-IgM type 3 (HIGM3) syndrome due to defects in CD40, typically seen in patients <10 years of age

 

Assessing B-cell immune competence in other clinical contexts, including autoimmunity, malignancy and transplantation

Method Name

Flow Cytometry

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.

CPT Code Information

88184

LOINC Code Information

Test ID Test Order Name Order LOINC Value
CD40 CD40 by Flow, QL, B In Process

 

Result ID Test Result Name Result LOINC Value
89009 CD40 by Flow, QL, B In Process

Clinical Information

The adaptive immune response includes both cell-mediated (mediated by T cells and natural killer [NK] cells) and humoral (mediated by B cells) immunity. After antigen recognition and maturation in secondary lymphoid organs, some antigen-specific B cells terminally differentiate into antibody-secreting plasma cells. Decreased numbers or aberrant function of B cells result in humoral immune deficiency states with increased susceptibility to infections, and these may be either primary (genetic) or secondary immunodeficiencies. Secondary causes include medications, malignancies, infections, and autoimmune disorders (this does not cause immunodeficiency with increased infection).

 

CD40 is a member of the tumor necrosis factor receptor superfamily, expressed on a wide range of cell types including B cells, macrophages, and dendritic cells.(1) CD40 is the receptor for CD40 ligand (CD40LG), a molecule predominantly expressed by activated CD4+ T cells. CD40/CD40LG interaction is involved in the formation of memory B lymphocytes and promotes immunoglobulin (Ig) isotype switching.(1) CD40LG expression in T cells requires cellular activation, while CD40 is constitutively expressed on the surface of B cells and other antigen-presenting cells.

 

Hyperimmunoglobulin M (hyper-IgM or HIGM) syndrome is a rare primary immunodeficiency characterized by increased or normal levels of IgM with low IgG and/or IgA.(2) Patients with hyper-IgM syndromes may have genetic defects or mutations in 1 of several known genes. Some of these genes are CD40LG, CD40, AICDA (activation-induced cytidine deaminase), UNG (uracil DNA glycosylase), and IKBKG (inhibitor of kappa light polypeptide gene enhancer in B cells, kinase gamma; also known as NEMO).(2) Not all cases of hyper-IgM syndrome fit into these known genetic defects. Mutations in CD40LG and IKBKG are inherited in an X-linked fashion, while mutations in the other 3 genes are autosomal recessive. Elevated IgM is only one of the features of NEMO deficiency and therefore, it is no longer classified exclusively with the hyper-IgM syndromes.

 

Distinguishing between the different forms of hyper-IgM syndrome is very important because of differing prognoses. CD40 and CD40LG deficiency are among the more severe forms, which typically manifest in infancy or early childhood, and are characterized by an increased susceptibility to opportunistic pathogens (eg, Pneumocystis carinii, Cryptosporidium, and Toxoplasma gondii).(3)

 

CD40 deficiency, also known as hyper-IgM type 3 (HIGM3), accounts for <1% of hyper-IgM syndromes. Flow cytometry analysis shows complete lack of CD40 expression on the B cells of these patients.(4) Intravenous injection with IgG is the treatment of choice along with immune reconstitution with hematopoietic cell transplantation. To date, all documented CD40-deficient patients have been diagnosed before age 1. Consequently, when used in the context of HIGM3, this test is only indicated in children (for diagnosis). In the case of CD40L deficiency, this test can be used for male patients or in females of child-bearing age (to identify carriers). A larger age spectrum has been reported with CD40L deficiency, ranging from infancy to early adulthood.

 

CD40 expression on B cells is also an indicator of immune status (eg, after the use of biological immunomodulatory therapy for autoimmune disease, cancer and transplantation).

Interpretation

This assay is qualitative; CD40 expression is reported as present (normal) or absent (abnormal). Normal B cells express surface CD40 on the majority of cells.

 

Hyper-IgM (HIGM3) syndrome patients typically do not express CD40 on the surface of B cells. Genotyping of CD40 is required for a definite diagnosis of HIGM3. Contact Mayo Medical Laboratories for ordering assistance.

Cautions

For questions about appropriate test selection, contact Mayo Medical Laboratories.

 

This test is not used to detect in CD40L expression (CD154), which is responsible for X-linked hyper-IgM syndrome (HIGM1); see XHIM / X-Linked Hyper IgM Syndrome, Blood.

Clinical Reference

Bishop GA, Hostager BS: The CD40-CD154 interaction in B cell-T cell liaisons. Cytokine Growth Factor Rev 2003;14:297-309

2. Lee WI, Torgerson TR, Schumacher MJ, et al: Molecular analysis of a large cohort of patients with hyper immunoglobulin M (IgM) syndrome. Blood 2005;105:1881-1890

3. Kutukculer N, Moratto D, Aydinok Y, et al: Disseminated cryptosporidium infection in an infant with hyper-IgM syndrome caused by CD40 deficiency. J Pediatr 2003;142:194-196

4. Ferrari S, Giliani S, Insalaco A, et al: Mutations of CD40 gene cause an autosomal recessive form of immunodeficiency with hyper IgM. Proc Natl Acad Sci USA 2001;98:12614-12619

Method Description

The assay involves a multicolor panel of antibodies for the following markers: CD45, CD19, and CD40. After the staining, RBCs are lysed. The remaining cells are washed and analyzed by flow cytometry on a BD FACSCanto instrument. The cell surface expression of CD40 in B cells (CD19+) is determined and expressed as being present or absent.(Unpublished Mayo method)