Biomarkers in Transplant Recipients

All transplant recipients receive periodic monitoring of drug levels and laboratory tests to
assess adequacy of immunosuppression and allograft function. These are performed when the
recipient is admitted to the hospital after transplantation or for a complication such as
acute rejection or toxicity, or when the recipient is an outpatient.

1. Blood samples: Participants may be asked to provide research blood specimens during
regular clinical tests, and may collect up to 15 milliliters of blood as many as 7 times
within the first year of transplant, and then less often thereafter. The total volume
collected will take in account the patient's height, weight and age at the time of the
collection. However, if for any reason participant is unable to provide a sample during
regular clinical test it may be collected at another time. Participants will be asked to
provide these samples indefinitely. This will allow longitudinal assessment of the
stability of biomarker expression as a reflection of clinical drug concentrations in
repeated measurements.

2. Saliva collection: Up to 5 ml of the subject's saliva will be collected no more than
four times, if the previous sample does not provide adequate information. Samples will
be collected in self-collection container at the time of consent or as early as possible
after consents are obtained, and will be stored at room temperature in the Pediatric
Transplantation Laboratory, 3344 Forbes Ave. In recipients where both are available, the
genotyping results as DNA from saliva will be compared between paired blood samples.
Henceforth, saliva collection will only be offered to participants who cannot donate
blood specimens for genotyping. Salivary sampling is considered an acceptable
alternative standard for whole blood genotyping. A saliva sample will be collected only
if the patient or the patient's parent or guardian prefers this option over blood
sampling.

3. Collection of urine, feces, and bile: five mls of any body fluid will be collected in
sterile urine cups for application of proteomics technologies. Collections may be
repeated up to four times, if the first specimen provides suboptimal information.

4. Collection of remaining allograft standard of care biopsy specimens, and tissue from
explants: Any piece of allograft biopsy deemed residual by the pathologist will be
subjected to gene array testing. This will occur when participant is scheduled for their
standard of care biopsy, or while in surgery. Genetic material extracted from the
smallest tissue can be amplified using several approaches.

5. Measurements: Biomarker expression will be evaluated after mitogen and antigen
stimulation of peripheral blood mononuclear cells. (1-3). Briefly, peripheral blood
mononuclear cells (PBMC) are extracted from whole blood by Ficoll gradient separation,
Thereafter, either mitogens such as phytohemaglutinin, pokeweed mitogen, or
phorbol-myristic acid-ionomycin, or viral and major histocompatibility complex (MHC)
peptide antigens, or intact alloantigenic cells will be used to stimulate recipient
PBMC. Cellular responses that can be measured include but are not limited to expressed
pro-inflammatory or anti-inflammatory markers, cytokines, proliferation, cytotoxicity,
and apoptosis.

Inclusion Criteria:

- Recipients of abdominal, thoracic and bone marrow allografts that are receiving
inpatient and outpatient follow-up with routine laboratory tests at the University of
Pittsburgh Medical Center.

- All Ages

- Subject or parents are able to read and understand the informed consent

Exclusion Criteria:

- Subjects and/or their parents who are unable to read and understand informed consent.