Apheresis of Patients With Immunodeficiency

BACKGROUND:

Primary immunodeficiency diseases (PID) represent candidate genetic disorders for new
therapeutic approaches. Our laboratory is developing gene therapy for patients with PID using
autologous CD34+ hematopoietic stem cells (HSC). Gene therapy may circumvent problems with
allogeneic HSC transplantation, especially graft rejection and graft-versus-host-disease. We
are particularly interested in three PID: Dedicator of CytoKinesis-8 (DOCK8) deficiency,
Leukocyte Adhesion Deficiency type 1 (LAD-1), and GATA2 Deficiency. For DOCK8 deficiency, and
LAD-1, the disease gene has been cloned. The genetic basis for MonoMAC has now been
determined to be due to mutations in GATA2.. Testing new vector constructs and transduction
conditions for gene therapy would be considerably enhanced by the acquisition of peripheral
blood CD34+ cells from patients with these immunodeficiency diseases.

OBJECTIVES:

To provide a source of granulocyte colony stimulating factor (G-CSF) mobilized peripheral
blood CD34+ hematopoietic stem cells (HSC) for laboratory research studies including
optimization of vector and transduction conditions for gene therapy for DOCK8 deficiency,
LAD-1, and MonoMAC.

ELIGIBILITY:

Patients 15-40 years old with DOCK8 deficiency, LAD-1, and MonoMAC who meet the eligibility
requirements will be considered for this protocol.

DESIGN:

Patients 15-40 years old with DOCK8 deficiency, LAD-1, and MonoMAC will receive five days of
G-CSF followed by a single apheresis. CD34+ cell will be selected and frozen in aliquots by
the Cell Processing Section of the Department of Transfusion Medicine. No treatments, or
investigational therapy will be administered on this protocol.

- INCLUSION CRITERIA:

1. Patient age of 15-40 years.

2. Diagnosis of DOCK8 deficiency, LAD-1, or MonoMAC:

DOCK8 Deficiency

Homozygous or compound heterozygous mutations in the DOCK8 gene.

LAD-1

Less than 10% CD18 expression on the neutrophil surface.

GATA2 Deficiency

- Onset of immunodeficiency disease beyond infancy.

- Clinical history of at least two episodes of life-threatening infection with
opportunistic organisms.

- Mutation of GATA2 Gene

- Serum creatinine <1.5 mg/dL.

- Total Bilirubin < 3mg/dl, ALT and AST < 5 times upper limit of normal.

- Ability to give informed consent.

- Adequate venous access for peripheral apheresis, or consent to use a temporary central
venous catheter for apheresis.

- Female patients of childbearing age must have a negative urine pregnancy test within
one week of beginning G-CSF administration.

- A patient who is lactating must be willing and able to interrupt breast-feeding or
substitute formula feeding for her infant during the period of filgrastim
administration and for two days following the final dose. Filgrastim may be secreted
in human milk, although its bioavailability from this source is not known. Limited
clinical data suggest that short-term administration of filgrastim or sargramostim to
neonates is not associated with adverse outcomes

EXCLUSION CRITERIA:

- HIV infection.

- Chronic hepatitis B or hepatitis C virus infection.

- History of psychiatric disorder which may compromise compliance with protocol, or
which does not allow for appropriate informed consent.

- Active infection that is not responding to antimicrobial therapy.

- Pregnant. The effects on breast-milk are also unknown and may be harmful to the
infant; therefore, women should not breast feed during the interval from study entry
to collection.

- Sexually active individuals capable of becoming pregnant who are unable or unwilling
to use effective form(s) of contraception during time enrolled on study. Effective
forms of contraception include one or more of the following: intrauterine device
(IUD), hormonal (birth control pills, injections, or implants), tubal
ligation/hysterectomy, partner s vasectomy, barrier methods, (condom, diaphragm, or
cervical cap), or abstinence. Males on the protocol must use an effective form of
contraception at study entry.

- Presence of active malignancy in another organ system other than the hematopoietic
system.

- History of hypertension that is not controlled by medication, stroke, or severe heart
disease. Individuals with symptomatic angina will be considered to have severe heart
disease and will not be eligible.

- Other medical contraindications to stem cell donation (i.e. severe atherosclerosis,
autoimmune disease, iritis or episcleritis, deep venous thrombosis, cerebrovascular
accident)

- Thrombocytopenia (platelets less than 50,000 per microL) at baseline evaluation.

- Patients receiving experimental therapy or investigational agents.

- Sensitivity to filgrastim or to E. coli-derived recombinant protein products.

- Patients must test negative for transfusion-transmissible infectious agents, including
hepatitis B (HBsAg), hepatitis C (anti-HCV), HIV (anti-HIV-1/2).