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The common lymphoid progenitor (CLP) responsible for the formation of T, B and NK cells is derived from a hematopoietic stem cell that is first identified in the embryonic aorto-gonad- mesonephros, a descendent of the mesoderm. The signals to initiate and regulate development are due to the control imposed by a variety of marrow stromal cells, transcription factors, and coordinated regulation by the nervous system, extracellular matrix, cytokines and adipocytes found in the bone marrow microenvironment. The general consensus of the ontological steps leading to production of naïve B-cells is summarized as follows; the earliest identifiable committed B-cells derived from the CLP are called progenitor (Pro) B-cells. Pro B-cells arise after obligate stimulation by the transcription factor PAX-5, which engenders CD19 production. These CD34+ CD19+ CD10+ CD38+ TdT+ expressing cells lack the pre-B-cell receptor or surface immunoglobulin (Ig) and initiate VDJ heavy chain rearrangements independent of any antigenic exposure. Pro B cells differentiate into CD34- CD19+ CD10+ CD38+ TdT- precursor (Pre) B-cells that acquire cytoplasmic and then surface mu heavy chain with a transient surrogate immunoglobulin light chain. Next, a CD19+ CD10-CD38- immature B-cell expresses surface IgM+ and physiologic light chain. Ultimately, CD19+ CD20+ B-cells co-expressing IgM and IgD heavy chains exit the bone marrow as transitional B-cells and home to secondary lymphoid organs as naive B-cells.

We are interested in the use of probability state modeling to quantify the locations of antigen modulations during the ontological development of human B-cells to determine the discrete progenitor and B-cell stages that occur during normal maturation. We will use this information to study and predict minimal residual disease in patients with B-lymphoblastic lymphoma.

The MGH Flow Cytometry research laboratories are located on the MGH campus in Simches 3.434 and CNY-5226. These hospital core resources will entertain research collaborations from throughout the pathology laboratories and greater hospital, university & surrounding biotech commumity. The CNY flow laboratory, overseen by Dr. R. Mylvaganam, H. Cahill, M. Drummond and J. Fung, contains a FACSAria II sorter, Fortessa X20, Fortessa, Aurora analyzer, Helios mass cytometer and FACSFusion sorter for BSL2+ operations. The Simches flow and imaging laboratory contains a DiVa cell   sorter and LSR-2 operated by D.  Dombkowski. A FACSFusion sorter permits BSL2+ sorting in   that facility, as well. This laboratory also contains an Amnis ISX mkII imaging flow cytometer which permits bright-field and fluorescent visual analysis of immunophenotyped cells, run by S. Mordecai. The clinical flow cytometry laboratory is located on Warren 5 on the MGH campus in Boston, supervised by M. DeLelys with two FACSCanto-IIs available at that site.

Selected Publications

Maryamchik E, Gallagher K, Preffer F, Kadauke S, Maus M. New Directions in CAR T Cell Therapy and CAR T Flow Cytometry. Cytometry B Clin Cytom 2020; In Press

Human B-cell and progenitor stages as determined by probability state modeling of multidimensional cytometry data.
Bagwell CB, Hill BL, Wood BL, Wallace PK, Alrazzak M, Kelliher AS, Preffer FI. Cytometry B Clin Cytom. 2015 Jul-Aug;88(4):214-26.

Probability state modeling theory. Bagwell CB, Hunsberger BC, Herbert DJ, Munson ME, Hill BL, Bray CM, Preffer FI. Cytometry A. 2015 Jul;87(7):646-60.

Long-term results in recipients of combined HLA-mismatched kidney and bone marrow transplantation without maintenance immunosuppression. Kawai T, Sachs DH, Sprangers B, Spitzer TR, Saidman SL, Zorn E, Tolkoff-Rubin N, Preffer FI, Crisalli K, Gao B, Wong W, Morris H, LoCascio SA, Sayre P, Shonts B, Williams WW Jr, Smith RN, Colvin RB, Sykes M, Cosimi AB. Am J Transplant. 2014 Jul;14(7):1599-611.

Physics of a rapid CD4 lymphocyte count with colloidal gold. Hansen P, Barry D, Restell A, Sylvia D, Magnin O, Dombkowski D, Preffer FI. Cytometry A. 2012 Mar;81(3):222-31.

The role of CD19 and CD27 in the diagnosis of multiple myeloma by flow cytometry: a new statistical model. Cannizzo E, Carulli G, Del Vecchio L, Ottaviano V, Bellio E, Zenari E, Azzarà A, Petrini M, Preffer FI. Am J Clin Pathol. 2012 Mar;137(3):377-86.

Advances in complex multiparameter flow cytometry technology: Applications in stem cell research.Preffer FI, Dombkowski D. Cytometry B Clin Cytom. 2009 Sep;76(5):295-314.